RiceWiki - User contributions [en]

File:3.jpg

2014-06-06T08:19:57Z

Weilovefairy: uploaded a new version of "File:3.jpg"

Os05g0465800

2014-06-06T08:19:29Z

Weilovefairy: /* Expression */

File:2.jpg

2014-06-06T08:17:59Z

Weilovefairy: uploaded a new version of "File:2.jpg"

Gross morphology of wildtype (left) and gid1-1 (right) plants. Scale bar, 10 cm. Inset: higher magnification of gid1-1. Scale bar, 1 cm

Os05g0465800

2014-06-06T08:16:44Z

Weilovefairy: /* References */

Os05g0465800

2014-06-06T08:10:57Z

Weilovefairy: /* Evolution */

Os05g0465800

2014-06-06T08:07:40Z

Weilovefairy: /* Annotated Information */

Os05g0465800

2014-06-06T07:42:43Z

Weilovefairy: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination
[[File:1.jpg|right|thumb|275px|'''Figure 1.''' ''Chromosome behavior in Wild type.(from reference) <ref name="ref1" />.'']]

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===

Although many papers reported that the seed of PI3K mutant
showed a reduced germination rate compared with wild type [23],
it has rarely been directly assayed so far. In this study, we
elaborately characterized the effects of PI3K on the rice seed
germination by promoting NADPH oxidase activity. LY294002
and Wortmannin, two sorts of PI3K specific inhibitor, obviously
inhibited the rice seed germination. Our mainly study here focused
on the inhibition of ROS generation (Figure 3) and the
relationship between PI3K and NADPH oxidase in seed
germination.
PI3K activity is closely correlated with rice seed
germination
In plant cells, the level of PI3P, as the product of PI3K, were
relatively low, but turn over rapidly following the alteration of
external environment [14]. This character determined that the
level of PI3P maybe has more sensitive response to the external
stimulation. Thus, we firstly examined the expression of PI3Kin
rice seed germination (Figure 1A). From 0 hour to 12 hours, the
expression of PI3K almost raised thirty times than initial
expression, which obviously indicated the key role of PI3K in
rice seed germination. Interestingly, the exogenous Ca
2+
promoted
the expression of PI3K(Figure 1B). As we know, the level of Ca
2+
rapidly increased following by the uptake of water in the early
germination. Here, PI3K seemed to be one of the important relay
station of Ca
2+
signal. In previous publication, it had been
reported that Wortmannin and LY294002 inhibited Ca
2+
oscillation induced by ABA [37], which supported the role of
PI3P in ABA-induced Ca
2+
oscillation. In contrast with the above
studies, our results showed that Ca
2+
promoted the expression of
PI3K. Meanwhile, it was reported that PI3K could also raise the
levels of intracellular Ca
2+
[37]. Therefore, we speculated that
PI3K and Ca
2+
seemed to have a feedback regulated relationship.
This cycle rapidly altered the total quantities of PI3K and Ca
2+
,
which laid the foundation for fulfilling their functions in the seed
germination.

==Labs working on this gene==

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|Jian Liu;Jun Zhou;Da Xing
Phosphatidylinositol 3-Kinase Plays a Vital Role in Regulation of Rice Seed Vigor via Altering NADPH Oxidase Activity
PLoS ONE, 2012, 7(3): e33817
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0465800

2014-06-06T07:42:17Z

Weilovefairy: /* Function */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination
[[File:Pair3_zhengchang.jpg|right|thumb|275px|'''Figure 1.''' ''Chromosome behavior in Wild type.(from reference) <ref name="ref1" />.'']]

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===

Although many papers reported that the seed of PI3K mutant
showed a reduced germination rate compared with wild type [23],
it has rarely been directly assayed so far. In this study, we
elaborately characterized the effects of PI3K on the rice seed
germination by promoting NADPH oxidase activity. LY294002
and Wortmannin, two sorts of PI3K specific inhibitor, obviously
inhibited the rice seed germination. Our mainly study here focused
on the inhibition of ROS generation (Figure 3) and the
relationship between PI3K and NADPH oxidase in seed
germination.
PI3K activity is closely correlated with rice seed
germination
In plant cells, the level of PI3P, as the product of PI3K, were
relatively low, but turn over rapidly following the alteration of
external environment [14]. This character determined that the
level of PI3P maybe has more sensitive response to the external
stimulation. Thus, we firstly examined the expression of PI3Kin
rice seed germination (Figure 1A). From 0 hour to 12 hours, the
expression of PI3K almost raised thirty times than initial
expression, which obviously indicated the key role of PI3K in
rice seed germination. Interestingly, the exogenous Ca
2+
promoted
the expression of PI3K(Figure 1B). As we know, the level of Ca
2+
rapidly increased following by the uptake of water in the early
germination. Here, PI3K seemed to be one of the important relay
station of Ca
2+
signal. In previous publication, it had been
reported that Wortmannin and LY294002 inhibited Ca
2+
oscillation induced by ABA [37], which supported the role of
PI3P in ABA-induced Ca
2+
oscillation. In contrast with the above
studies, our results showed that Ca
2+
promoted the expression of
PI3K. Meanwhile, it was reported that PI3K could also raise the
levels of intracellular Ca
2+
[37]. Therefore, we speculated that
PI3K and Ca
2+
seemed to have a feedback regulated relationship.
This cycle rapidly altered the total quantities of PI3K and Ca
2+
,
which laid the foundation for fulfilling their functions in the seed
germination.

==Labs working on this gene==

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|Jian Liu;Jun Zhou;Da Xing
Phosphatidylinositol 3-Kinase Plays a Vital Role in Regulation of Rice Seed Vigor via Altering NADPH Oxidase Activity
PLoS ONE, 2012, 7(3): e33817
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

File:1.jpg

2014-06-06T07:40:49Z

Weilovefairy: uploaded a new version of "File:1.jpg"

Gross morphology of GID1-overexpressor and control plants. Scale
bar, 50 cm.

File:1.jpg

2014-06-06T07:30:50Z

Weilovefairy: uploaded a new version of "File:1.jpg": Figure 1. Changes ofPI3Kexpression in rice seed after imbibition.RT-PCR assayed the expression ofPI3Kfrom rice seed, which was imbibed from 0 hour to 12 hours in the presence (A) or absence (C) of 20 m

Gross morphology of GID1-overexpressor and control plants. Scale
bar, 50 cm.

Os05g0465800

2014-06-06T07:26:56Z

Weilovefairy: /* Structured Information */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===

Although many papers reported that the seed of PI3K mutant
showed a reduced germination rate compared with wild type [23],
it has rarely been directly assayed so far. In this study, we
elaborately characterized the effects of PI3K on the rice seed
germination by promoting NADPH oxidase activity. LY294002
and Wortmannin, two sorts of PI3K specific inhibitor, obviously
inhibited the rice seed germination. Our mainly study here focused
on the inhibition of ROS generation (Figure 3) and the
relationship between PI3K and NADPH oxidase in seed
germination.
PI3K activity is closely correlated with rice seed
germination
In plant cells, the level of PI3P, as the product of PI3K, were
relatively low, but turn over rapidly following the alteration of
external environment [14]. This character determined that the
level of PI3P maybe has more sensitive response to the external
stimulation. Thus, we firstly examined the expression of PI3Kin
rice seed germination (Figure 1A). From 0 hour to 12 hours, the
expression of PI3K almost raised thirty times than initial
expression, which obviously indicated the key role of PI3K in
rice seed germination. Interestingly, the exogenous Ca
2+
promoted
the expression of PI3K(Figure 1B). As we know, the level of Ca
2+
rapidly increased following by the uptake of water in the early
germination. Here, PI3K seemed to be one of the important relay
station of Ca
2+
signal. In previous publication, it had been
reported that Wortmannin and LY294002 inhibited Ca
2+
oscillation induced by ABA [37], which supported the role of
PI3P in ABA-induced Ca
2+
oscillation. In contrast with the above
studies, our results showed that Ca
2+
promoted the expression of
PI3K. Meanwhile, it was reported that PI3K could also raise the
levels of intracellular Ca
2+
[37]. Therefore, we speculated that
PI3K and Ca
2+
seemed to have a feedback regulated relationship.
This cycle rapidly altered the total quantities of PI3K and Ca
2+
,
which laid the foundation for fulfilling their functions in the seed
germination.

==Labs working on this gene==

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|Jian Liu;Jun Zhou;Da Xing
Phosphatidylinositol 3-Kinase Plays a Vital Role in Regulation of Rice Seed Vigor via Altering NADPH Oxidase Activity
PLoS ONE, 2012, 7(3): e33817
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0465800

2014-06-06T07:25:42Z

Weilovefairy: /* Labs working on this gene */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===

Although many papers reported that the seed of PI3K mutant
showed a reduced germination rate compared with wild type [23],
it has rarely been directly assayed so far. In this study, we
elaborately characterized the effects of PI3K on the rice seed
germination by promoting NADPH oxidase activity. LY294002
and Wortmannin, two sorts of PI3K specific inhibitor, obviously
inhibited the rice seed germination. Our mainly study here focused
on the inhibition of ROS generation (Figure 3) and the
relationship between PI3K and NADPH oxidase in seed
germination.
PI3K activity is closely correlated with rice seed
germination
In plant cells, the level of PI3P, as the product of PI3K, were
relatively low, but turn over rapidly following the alteration of
external environment [14]. This character determined that the
level of PI3P maybe has more sensitive response to the external
stimulation. Thus, we firstly examined the expression of PI3Kin
rice seed germination (Figure 1A). From 0 hour to 12 hours, the
expression of PI3K almost raised thirty times than initial
expression, which obviously indicated the key role of PI3K in
rice seed germination. Interestingly, the exogenous Ca
2+
promoted
the expression of PI3K(Figure 1B). As we know, the level of Ca
2+
rapidly increased following by the uptake of water in the early
germination. Here, PI3K seemed to be one of the important relay
station of Ca
2+
signal. In previous publication, it had been
reported that Wortmannin and LY294002 inhibited Ca
2+
oscillation induced by ABA [37], which supported the role of
PI3P in ABA-induced Ca
2+
oscillation. In contrast with the above
studies, our results showed that Ca
2+
promoted the expression of
PI3K. Meanwhile, it was reported that PI3K could also raise the
levels of intracellular Ca
2+
[37]. Therefore, we speculated that
PI3K and Ca
2+
seemed to have a feedback regulated relationship.
This cycle rapidly altered the total quantities of PI3K and Ca
2+
,
which laid the foundation for fulfilling their functions in the seed
germination.

==Labs working on this gene==

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0465800

2014-06-06T07:25:02Z

Weilovefairy: /* Evolution */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===

Although many papers reported that the seed of PI3K mutant
showed a reduced germination rate compared with wild type [23],
it has rarely been directly assayed so far. In this study, we
elaborately characterized the effects of PI3K on the rice seed
germination by promoting NADPH oxidase activity. LY294002
and Wortmannin, two sorts of PI3K specific inhibitor, obviously
inhibited the rice seed germination. Our mainly study here focused
on the inhibition of ROS generation (Figure 3) and the
relationship between PI3K and NADPH oxidase in seed
germination.
PI3K activity is closely correlated with rice seed
germination
In plant cells, the level of PI3P, as the product of PI3K, were
relatively low, but turn over rapidly following the alteration of
external environment [14]. This character determined that the
level of PI3P maybe has more sensitive response to the external
stimulation. Thus, we firstly examined the expression of PI3Kin
rice seed germination (Figure 1A). From 0 hour to 12 hours, the
expression of PI3K almost raised thirty times than initial
expression, which obviously indicated the key role of PI3K in
rice seed germination. Interestingly, the exogenous Ca
2+
promoted
the expression of PI3K(Figure 1B). As we know, the level of Ca
2+
rapidly increased following by the uptake of water in the early
germination. Here, PI3K seemed to be one of the important relay
station of Ca
2+
signal. In previous publication, it had been
reported that Wortmannin and LY294002 inhibited Ca
2+
oscillation induced by ABA [37], which supported the role of
PI3P in ABA-induced Ca
2+
oscillation. In contrast with the above
studies, our results showed that Ca
2+
promoted the expression of
PI3K. Meanwhile, it was reported that PI3K could also raise the
levels of intracellular Ca
2+
[37]. Therefore, we speculated that
PI3K and Ca
2+
seemed to have a feedback regulated relationship.
This cycle rapidly altered the total quantities of PI3K and Ca
2+
,
which laid the foundation for fulfilling their functions in the seed
germination.

==Labs working on this gene==
Please input related labs here.

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0465800

2014-06-06T07:20:49Z

Weilovefairy: /* Expression */

Please input one-sentence summary here.

==Annotated Information==
===Function===
Phosphatidylinositol 3-kinase (PI3K) has been reported to be important in normal plant growth and stress responses. In this
study, it was verified that PI3K played a vital role in rice seed germination through regulating NADPH oxidase activity.
Suppression of PI3K activity by inhibitors wortmannin or LY294002 could abate the reactive oxygen species (ROS) formation,
which resulted in disturbance to the seed germination. And then, the signal cascades that PI3K promoted the ROS liberation
was also evaluated. Diphenylene iodonium (DPI), an NADPH oxidase inhibitor, suppressed most of ROS generation in rice
seed germination, which suggested that NADPH oxidase was the main source of ROS in this process. Pharmacological
experiment and RT-PCR demonstrated that PI3K promoted the expression of Osrboh9. Moreover, functional analysis by
native PAGE and the measurement of the 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazo-lium-5- carboxanilide (XTT)
formazan concentration both showed that PI3K promoted the activity of NADPH oxidase. Furthermore, the western blot
analysis of OsRac-1 demonstrated that the translocation of Rac-1 from cytoplasm to plasma membrane, which was known
as a key factor in the assembly of NADPH oxidase, was suppressed by treatment with PI3K inhibitors, resulting in the
decreased activity of NADPH oxidase. Taken together, these data favored the novel conclusion that PI3K regulated NADPH
oxidase activity through modulating the recruitment of Rac-1 to plasma membrane and accelerated the process of rice seed
germination

===Expression===
Changes of PI3K expression in rice seed after imbibition and the expression of PI3K is regulated by Ca2+. To determine the dynamic expression ofPI3Kin rice seed germination, the total RNA was extracted from the rice seed embryo after imbibition. The result of RT-PCR analysis suggested that PI3K might involve in the regulation of seed germination. Meanwhile, it was found that the expression ofPI3K was increased with the prolongation of imbibition time (Figure 1A).This expression peaked at 12 h and reached 30 times as many as seeds without treatment (Figure 1C). Interestingly, low concentration of Ca
2+(below10 mM) could further increase the expression of PI3Kin rice seeds after germinated for 12 h. However, when the concentration of Ca2+ was up to 20 mM, thePI3Kexpression began to
decrease slightly compared with the treatment with 10 mM
CaCl2(Figure 1B, D).
LY294002 and Wortmannin inhibit rice seed germination
Next, Wortmannin and LY294002, two kinds of PI3K
inhibitors with different action mechanisms, were used to test
the role of PI3K in the de-coated (without pericarp) rice seed
germination [21]. Following treatment with 60mM LY294002 at
27uC for four days, the seed germination rate decreased to about
67.3% compared with the control. And it was further decreased to
54.0% when the concentration of LY294002 was increased to
90mM (Figure 2A). The seed germination rate was 55.7% of the
control in the presence of 20mM Wortmannin and it would be
decreased to 31.7% of the control with 30mM Wortmannin
(Figure 2B). Dynamics of seed germination revealed that, relative
to the control, 60 mM LY294002 or 20mM Wortmannin
displayed both significant inhibitory effects on the seed germination at any time point (Figure 2C). From these results, we
reasonably concluded that PI3K played a positive role in rice seed
germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination.
LY294002 and Wortmannin inhibit ROS production
PI3K inhibitors suppressed the ROS production in guard cell,
root hair, and pollen tube [18,19,23]. However, it was uncertain
that PI3K inhibitors restrained the ROS formation in rice seed
germination. In our case, it was found that exogenous hydrogen
peroxide (H2O2) could partly rescue the inhibitory effects of PI3K
inhibitors on rice seed germination (Figure S1A). This finding
allowed us to assume the possible association of PI3K with ROS
production during rice seed germination To further investigate this relationship between PI3K and ROS,
ROS probe H2
DCFDA was used to examine the characteristics of
ROS production under the treatment of pharmacological
inhibitors of PI3K LY294002 (60 mM) or Wortmannin (20mM).
Results demonstrated that, during the imbibition period of 48 h,
treatment with PI3K inhibitors resulted in lower level of ROS
production compared with the control (Figure 3A). Meanwhile,
the change of superoxide anion was also examined through NBT
staining [33] and XTT test [34]. As shown in Figure 3B, 3C, the
alteration of superoxide anion was similar with the change of ROS
level. Interestingly, PI3K inhibitors seemed to suppress ROS not
only by NADPH oxidase but also other sources (Figure S2). The
above findings led us to get the conclusion that PI3K inhibitors
could suppress the formation of ROS in rice seed germination.
NADPH oxidase is an important source of ROS in rice
seed germination
Several studies have showed that ROS plays a key role in seed
germination [7,35,36]. Here, we also found that rice seed
germination could be inhibited by ROS scavengers, 10 mM KI
or 1 Mm ASA (Figure S1B). Subsequently, 100mM diphenylene
iodonium (DPI), one of the highly effective inhibitor of NADPH
oxidase [27], was used to explore whether NADPH oxidase is
involved in ROS production responsible for rice germination. As
shown in Figure S1B, the germination rate in rice seed treated
with DPI was lower than that of control. Interestingly, exogenous
H2O2 (20 mM) could partly rescue the decrease in seed
germination caused not only by KI and ASA but also by DPI
(Figure S1B). These phenomenons indicated that NADPH oxidase
might be the important source of ROS formation in rice seed
embryo under germination.
To prove this hypothesis, the concentration of superoxide anion
was under determination in the absence and presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 4A, NBT
was applied to evaluate the quantity of superoxide anion. DPI
(100 mM) was added before NBT staining. Compared with
control, exogenous DPI could significantly suppress the formation
deposits in the embryos.
To further verify this phenomenon, superoxide anion was
quantified by XTT [34]. As was illustrated in Figure 4B, under the
treatment of PI3K inhibitors LY294002 (60 mM) or Wortmannin
(20 mM), the formation of superoxide anion was obviously
restricted in contrast with control. These observations confirm
the idea that NADPH oxidase is an important source of ROS in
seed germination.
LY294002 and Wortmannin inhibit the expression of
NADPH oxidase
The earlier studies have reported that NADPH oxidasegenerated ROS plays a key role in seed germination [7,35,36].
As mentioned above, PI3K inhibitors can prevent ROS
production during rice seed germination. Therefore, it is worth
assuming that PI3K might control ROS level via regulating
NADPH oxidase. In the previous study, it has been proved that
PI3K is associated with nuclear transcription sites in higher plant
[12]. Thus, we examined whether PI3K regulated the transcription of NADPH oxidase in rice seed germination. Firstly, in order
to investigate the types of NADPH oxidase in the rice seed
germination, the RNA was extracted from wild type rice seed
imbibed for 24 h. RT-PCR experiments revealed that NADPH
oxidase Os rboh2, Os rboh4, Os rboh5 and Os rboh9 played a
primary role in rice seed germination (Figure 5A, B). Besides, the
dynamics of these types of NADPH oxidase expression was also assayed during the initial 24 h of rice seed germination (Figure 5C,
D). To investigate whether PI3K promote the transcription of
NADPH oxidase in the rice seed germination, the expression of
NADPH oxidase was examined in the presence of 60mm
LY294002 or 20mm Wortmannin. As shown in Figure 5E, F,
the PI3K inhibitors notably restrained the expression of NADPH
oxidae Osrboh9 and slightly suppressed the expression of Osrboh4
in comparison with other types of NADPH oxidase. On the basis
of the above results, we drew the conclusion that PI3K could
promote the transcription of Osrboh4and Osrboh9 other than the
rest types of NADPH oxidase.
PI3K is required for subcellular translacaiton of Rac-1
In mammalian cells, it has been proved that PI3K class I can
regulate the translocation of cytosolic factors, such as Rac-1. In fact,
it is necessary for the assembly of the active NADPH oxidase
complex to translocate Rac-1 to the cell membrane in rice cells [31].
However, it has been unclear whether PI3K regulates NADPH
oxidase activity through mediating the translocation of Rac-1. To investigate the possible mechanisms of Rac1 regulated by
PI3K, western blot was used. The experiment was performed with
total or membrane protein isolated from rice seed embryo cells
imbibed for 24 h with or without 60 mM LY294002 or 20mM
Wortmannin. Western blot analysis of membrane protein
indicated that the translocation of Rac-1, following the treatment
of LY294002, was suppressed compared with control, whereas the
quantity of Rac-1 from the total protein was not obviously altered.
In addition, compared with that of LY294002, another PI3K
inhibitor Wortmannin seemed to abate the amount of total Rac1
while it reduced the amount of membrane Rac1 severely
(Figure 7A, 7B). These results (Figure 7C, 7D) confirmed the
speculation that PI3K promoted the translocation of Rac-1 to the
membrane and thus facilitated the activity of NADPH oxidase

===Evolution===
Please input evolution information here.

You can also add sub-section(s) at will.

==Labs working on this gene==
Please input related labs here.

==References==
Please input cited references here.

==Structured Information==
{{JaponicaGene|
GeneName = Os05g0465800|
Description = Similar to RbohAp108|
Version = NM_001062318.1 GI:115464366 GeneID:4339045|
Length = 4894 bp|
Definition = Oryza sativa Japonica Group Os05g0465800, complete gene.|
Source = Oryza sativa Japonica Group

ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
Chromosome = [[:category:Japonica Chromosome 5|Chromosome 5]]|
AP = Chromosome 5:22905974..22910867|
CDS = 22906100..22906400,22907201..22907363,22907440..22907488,22907582..22907695,22907768..22908190 ,22908263..22908649,22908733..22908828,22908934..22909049,22909126..22909228 ,22909348..22909436,22909542..22909692,22909773..22909922,22910013..22910141 ,22910232..22910310,22910391..22910500|
GCID = <gbrowseImage1>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage1>|
GSID = <gbrowseImage2>
name=NC_008398:22905974..22910867
source=RiceChromosome05
preset=GeneLocation
</gbrowseImage2>|
CDNA = <cdnaseq>atggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacgacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttgctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgataatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaa</cdnaseq>|
AA = <aaseq>MAGDYVDVPLGGGGQSTLPPVAPLKKQPSRLASGMKRLASMVPD TMKLKRTHSSAQPALRGLRFLDKTSAGKDGWKNVEKRFDEMSADGRLPQESFAKCIGM ADSKEFASEVFVALARRRSIKPEDGITKEQLKEFWEELTDQNFDSRLRIFFDMCDKNG DGQLTEDEVKEVIVLSAAANKLAKLKSHAATYASLIMEELDPDHRGYIEIWQLETLLR GMVTAQGPPEKVKLASASLARTMVPSSHRSPMQRRFNKTVDFIHENWKRIWVLSLWAI LNIALFMYKFVQYSRRDAFQVMGYCVCIAKGAAETLKLNMAVILLPVCRNTLTRLRST ALSKVVPFDDNINFHKVIALTIAIGAATHTLAHVTCDFPRLVSCPRDKFEATLGPYFN YVQPTYSSLVASTPGWTGILMILIMSFSFTLATHSFRRSVVKLPSPLHHLAGFNAFWY AHHLLVIAYILLVLHSYFIFLTKQWYNRTTWMFLAVPVLFYSCERTIRRVRESSYGVT VIKAAIYPGNVLSIHMNKPSSFKYKSGMYMFVKCPDVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTTELRNLFGKACEAQVSSKKATLARLETTIIADGLKEETCFPKVFIDGPF GAPAQNYKKYDILLLIGLGIGATPFISILKDLLNNIKSNGDVQSTHDAELGCTFKSNG PGRAYFYWVTREQGSFEWFKGVMNDVAESDHDNVIEMHNYLTSVYEEGDARSALIAMV QSLQHAKNGVDIVSGSKIRTHFARPNWRKVFSDLANAHQNSRIGVFYCGSPTLTKMLR DLSLEFSQTTTTRFHFHKENF</aaseq>|
DNA = <dnaseqindica>127..427#1228..1390#1467..1515#1609..1722#1795..2217#2290..2676#2760..2855#2961..3076#3153..3255#3375..3463#3569..3719#3800..3949#4040..4168#4259..4337#4418..4527#tttcttcattggcctggaacgacgacgaccacgtacgccatggcgcccggggccacgctcctcctgcggtgagggcacccaccaccaccaccagcagcagcagcgggctacctagttcggatcaggatggcgggggactacgtggacgtgccgctgggcggcggcgggcagagcacgctgccgccggtggcgccgctgaagaagcagccgtcgcggctggcgtccgggatgaagcggctggcgtccatggtgcccgacacgatgaagctgaagcggacgcactccagcgcgcagccggcgctgcgcggcctgcgcttcctcgacaagacgtcggcggggaaggacgggtggaagaacgtcgagaagcggttcgacgagatgagcgccgacggacgcctgccccaggagagcttcgccaaatgcatcggtgagtgctcctgcacatgcccccatggtgttctccatcctccaaagtgattagtgattcaccacagcatgaaaagagagacggtgaaattgcattgttagcaagaacacatcgaaaattgacaagctttgttttccctgcaaaaaaaaaaggggaaaatgttgtcaaagcttttgtgcatgtgatggaaaaaggggaacagttagtagatgagaaattggtacagatcattcctagctcgtatgtcgatggtgtttgtccaaaatttcagccatttcaactgtttaagcagccagcctcctcctgccatccatgcatacatgtttgcagcacttctttgataaagtatacataatcattcctattcacaattttattttaacaaaaaacggaggaaagaaaattggaaaatttaagaggatataattggtcgaccacgggttgcacaaagttcagactttggcccccccaaatataactcggagacaatacttaccacgtagcaagaataggatatacttcctccgtttcataatgtaagactttcaatcattgcccacattcatatagatgttaatgaatctagacatatatatatgtgtatagattcattaacatctatatgaatatgggcaatgctagaaagtcttacaatatgaaacggagggagtactcgtgcttacgacattttcttttaaaagaagtgtagtcaggtaaataaatgacatcagaatagcaataaatccatcctctgtatataaattgaagctctaagcttagaatgcctctgcaacttaaaggtatggcggattctaaggagtttgcaagcgaggtgtttgtggcattggcgagaaggaggagcatcaaaccagaagatgggataacaaaggaacagctgaaagaattttgggaggagttgactgatcagaactttgattcacgactacgaatattctttgacatgtaaaggtctaccttccttgtcttcaatggaaaaaaccttatgacttgccccttgatgagattgctttgcaatcaggtgtgacaagaatggtgatgggcagcttacagaggacgaggtcaaagaggtcagttcccaatcatctctgaatttaccttttttccacaaagcttgtggaaatgatctcaataagaatttacctttctctttccatatgcaggttattgtgttgagtgccgcagcaaacaaacttgccaagttgaaaagccatgctgcaacctacgcctcactgatcatggaagagctggatcctgatcatcgcggttacattgaggtaccaagacatgaaattatctcccttttttttgacaaacagcagcatacagatatgcttgtctcaatgcagatttggcagctggagacgctactccgtggtatggtgacagcccaagggcctccagagaaggtgaagctggcttcagcaagccttgcaaggacaatggtcccttccagtcaccggagtccaatgcagaggcgtttcaacaagactgttgacttcatccatgagaattggaagaggatatgggtgctctccttgtgggcgatcctcaatattgcattattcatgtacaagtttgtgcagtacagtaggcgggatgcatttcaggtgatgggctactgtgtctgcatagcaaagggtgctgccgaaacactcaagctgaacatggccgttatactcctcccggtgtgccgaaacacactaacaaggctccgatcaacagcactcagcaaagtcgtaccatttgatgataacataaacttccacaaggtacacgtgtggaactctttgccatttccaggattacacagcctgacctctttgttctgatcaccatatcaggttatcgcactgacgattgcaattggagcggctactcatactcttgctcatgtaacctgcgacttcccaagattggtatcatgtccaagggacaagttcgaggccacactggggccttacttcaactatgttcaaccaacatactcatcgctggttgcaagcactccagggtggactggcatcctcatgatcttgataatgtcattctcgttcacacttgcgacacattcgttcaggaggagcgttgtgaagcttccatcaccactgcaccaccttgctggtttcaatgccttttggtacgcccatcacctactggtgattgcatatatcctcttggtgctgcactcctacttcatatttctcaccaagcagtggtacaatcgaacggtatttctctggcatatgtttggcacaactatttcttttgtgaattcagtggttgtgcataatattgacgcttgtgtttacagacgtggatgttcttggcagttccagtcctcttttactcctgcgagagaactatcagaagagttcgtgagagcagttatggggtgaccgtcatcaaggtaagcaacatatttcacatgaattgcacgatgagtattttcaatttattctagcttctaagtgtacaagaataccattaattgactatcaaatgtgccatgcaggcagcaatttaccctggaaatgtgctctctattcacatgaataaaccatcaagtttcaagtacaaaagtgggatgtatatgtttgtaaaatgcccagatgtttcgccttttgaatggtaccatctgatcgtctctgtccaaatgtcagtgtttatccaggctatggattaatagattcttcaatgcttccaggcatcccttctccataacttctgcacctggagatgactacttgagtgttcatatccgtacattaggtgactggacaacagaacttagaaacctatttgggaaggtcagttgagcaaacagaaacaaaaagaaaaggagaatagttcaggcataatgggagcttatgcagtatttatatgctaaattaacaaagttgctaaagggttatgctaattcatgcaggcttgtgaggcacaagtaagttccaagaaggctacacttgcacgacttgaaaccactatcatagcagatggtctgaaagaggagacttggtaattatgccctgtttgtttcatattctacattcatctactttttgaaggaatttctatacaattgtttcactaatcttcctgactttggttctcttcatttagctttcccaaagtctttatagatggtcctttcggcgcaccagctcaaaattacaagaaatatgacattcttttgcttatcgggcttggaattggagcaacgcctttcatcagcatactgaaggatctcctgaacaacataaaatccaatggagtaaactgcttaactatagcaaattctttcttatacagtaaaccaagtgcataatatctaaagtgaattggattcaacaggatgtgcaaagcacgcatgatgctgagttaggctgcacctttaagagcaatgggccaggaagagcttatttctactgggtcaccagagagcaaggttcctttgaatggtttaaaggcgtgatgaatgatgttgctgaaagtgatcatgatgtactactcagtctaactgcttcagtgcaacttccatgttctctccttcagattaataaatagatactaacataatcactcactatacagaatgtaatagagatgcacaattacctgacaagcgtgtatgaagaaggagatgcaagatcagctctgattgccatggtccaatcacttcaacatgccaaaaatggtgtggatatcgtctctggcagcaaggtttttcccacttctatcctcctgcaaaaataaaaataaaaactaaatcacatgtgcaaaatttgattcaatatttctcgctttatgcagatccggacacattttgcaaggccgaactggagaaaggtattctcagatctggccaatgcccaccagaactctcgtataggtcagttctatgtgataaatctcaagtgatggttcctagtttacaggatagcaatgctgagatgtttcataaatctgcaggcgttttctactgtgggtctccaacacttacgaaaatgctgagggatctttcactagaattcagccagacgacaacgactcggttccatttccacaaggagaacttctaagaccagaccaggaaaaaaacaccaaaataatcgcaggcgctcttatatagaacaagaaattttagcatcggttagcaactctttctgtgtaaagttccatcaagaactggaacattggtgtacaaaggtacgtgggcattagttagtgctgctgccgtttatagatggaggaaaaagaaaggcatgtattttctgagcaatttgattgtttcttagaatgtactagaaacaacgtgcggctttgccgcgcccaatttagattaggccaatctattttgtcagtggccttgtttaggatcaatggttttttttcttaacatattgaaccaaaaacctaaagaaataaattaatagacgttgggacttg</dnaseqindica>|
Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001062318.1 RefSeq:Os05g0465800]|
}}
[[Category:Genes]]
[[Category:Japonica mRNA]]
[[Category:Oryza Sativa Japonica Group]]
[[Category:Japonica Genes]]
[[Category:Japonica Chromosome 5]]
[[Category:Chromosome 5]]

Os05g0465800

2014-06-06T07:16:05Z

Weilovefairy: /* Annotated Information */

Os05g0465800

2014-06-06T07:09:03Z

Weilovefairy: /* Annotated Information */