Os03g0411500 - Revision history

192.168.72.52: /* Structured Information */

2015-06-12T05:54:33Z

‎Structured Information

Xiaoyanjie2012: /* Expression */

2014-06-08T08:39:00Z

‎Expression

Xiaoyanjie2012: /* 6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT */

2014-06-08T08:38:46Z

‎6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT

Xiaoyanjie2012: /* 6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT */

2014-06-08T08:38:17Z

‎6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT

Xiaoyanjie2012: /* 6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT */

2014-06-08T08:37:56Z

‎6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT

Xiaoyanjie2012: /* 6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT */

2014-06-08T08:30:28Z

‎6.Interactions between VYL, OsClpP3, OsClpP4, OsClpP5, and OsClpT

Xiaoyanjie2012: /* 5.The vyl Locus Maps to a Putative Gene Encoding OsClpP6 */

2014-06-08T08:27:45Z

‎5.The vyl Locus Maps to a Putative Gene Encoding OsClpP6

Xiaoyanjie2012: /* Expression */

2014-06-08T08:24:09Z

‎Expression

Xiaoyanjie2012: /* 5.The vyl Locus Maps to a Putative Gene Encoding OsClpP6 */

2014-06-08T08:19:55Z

‎5.The vyl Locus Maps to a Putative Gene Encoding OsClpP6

Xiaoyanjie2012: /* 4.Altered Expression of Genes Associated with Chloroplast Biogenesis and Photosynthesis in vyl Mutants */

2014-06-08T08:18:20Z

‎4.Altered Expression of Genes Associated with Chloroplast Biogenesis and Photosynthesis in vyl Mutants

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 </references>
 ==Structured Information==
 [[Category:Genes]]
 [[Category:Japonica mRNA]]

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 To investigate the expression patterns of VYL during chloroplast and leaf development, researchers analyzed VYL expression in different sections of leaves at various leaf developmental stages by qRT-PCR (Fig. 2A). They found that in wild-type plants, VYL was most highly expressed in the L4 section at the early chloroplast and leaf development stage (Fig. 6A). Furthermore, qRT-PCR analysis and histochemical staining of the pVYL::GUS reporter gene transgenic plants showed that VYL was constitutively expressed in young buds, young roots, stems, leaves, leaf shoots, and panicles (Fig. 6, B and C). To test whether VYL expression is regulated by light, They analyzed VYL expression during greening of etiolated seedlings. Wild-type rice plants were grown in continuous darkness for 10 d and subsequently exposed to light for 3, 6, 9, 12, 15, 18, 21, or 24 h. The expression of VYL was highly induced after 3 h of illumination and peaked after 6 h of illumination, then its expression gradually decreased over time, and by 15 h after illumination, its expression returned to the preillumination basal level (Fig. 6D). These observations suggested that VYL likely plays a role in the light regulation of chloroplast development. We next conducted qRT-PCR analysis to examine a possible effect of the vyl mutation on the expression of other genes encoding various components of the rice Clp. We
 found that OsClpPs, OsClpT, and OsClpR4 all had similar expression patterns to VYL, with a peak accumulation at the early stage of chloroplast development, but the expression levels of these genes were higher in the vyl mutants compared with the wild type (Fig. 7). This observation suggested that there may be a compensatory mechanism to increase the expression of OsClpPs, OsClpT, and OsClpR4 in vyl mutants.
 ==Evolution==

← Older revision		Revision as of 08:38, 8 June 2014
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−	[[File:Figure 5 .png]][[File:Figure 6 ~~.png]][[File:Figure 7 .png]][[File:Figure 8~~ .png]]	+	[[File:Figure 5 .png]][[File:Figure 6 .png]]

	===Expression===		===Expression===

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 Fig. 8C). Furthermore, we found that the yeast strain (Gold Saccharomyces cerevisiae) carrying BD-VYL+AD-OsClpP4 grew and developed the blue color faster than the yeast strain (Gold Saccharomyces cerevisiae) carrying BD-DVYL+AD-OsClpP4 or BD-VYL+AD-OsClpP3 on yeast growth medium containing 5-Bromo-4-chloro-3-
 indoylla-galactoside (X-a-Gal). Furthermore, the yeast strain(GoldSaccharomycescerevisiae)carryingBD-DVYL+AD-OsClpP3 did not grow even after incubation for 5 d in a growth chamber (Fig. 8D). This result suggests that the C-terminal polypeptide-binding site of VYL likely plays a role in mediating the interaction between VYL with OsClpP3 and OsClpP4. In addition, they also found that OsClpP3 interacted with OsClpT, OsClpP4 interacted with OsClpP5 and OsClpT, and that both OsClpP4 and OsClpT interacted with themselves but OsClpP5 and OsClpP3 did not (Fig. 8E). Due to the self-activation activity of VYL-AD, we were not able to test whether VYL can homodimerize.
 [[File:Figure 5 .png]][[File:Figure 6 .png]][[File:Figure 7 .png]][[File:Figure 8 .png]]

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 Fig. 8C). Furthermore, we found that the yeast strain (Gold Saccharomyces cerevisiae) carrying BD-VYL+AD-OsClpP4 grew and developed the blue color faster than the yeast strain (Gold Saccharomyces cerevisiae) carrying BD-DVYL+AD-OsClpP4 or BD-VYL+AD-OsClpP3 on yeast growth medium containing 5-Bromo-4-chloro-3-
 indoylla-galactoside (X-a-Gal). Furthermore, the yeast strain(GoldSaccharomycescerevisiae)carryingBD-DVYL+AD-OsClpP3 did not grow even after incubation for 5 d in a growth chamber (Fig. 8D). This result suggests that the C-terminal polypeptide-binding site of VYL likely plays a role in mediating the interaction between VYL with OsClpP3 and OsClpP4. In addition, they also found that OsClpP3 interacted with OsClpT, OsClpP4 interacted with OsClpP5 and OsClpT, and that both OsClpP4 and OsClpT interacted with themselves but OsClpP5 and OsClpP3 did not (Fig. 8E). Due to the self-activation activity of VYL-AD, we were not able to test whether VYL can homodimerize.
 ===Expression===

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 which the C terminus of VYL was fused with a Hisbiotin tag (Tagwerker et al., 2006), and the fusion gene was driven by the Ubiquitin promoter. The VYL-HBH
 fusion construct was introduced into the vyl mutant via Agrobacterium tumefaciens-mediated transformation. The VYL-HBH fusion protein transgene recovered the chlorotic phenotype of the vyl mutant, indicating that the fusion protein is biologically functiona SDS-PAGE and immunoblot analyses showed that the VYL-HBH fusion protein and several additional proteins could be effectively purified from the pUbi:: VYL-HBH transgenic plants using Ni 2+ -Sepharose and streptavidin beads (Fig. 8A). The copurified protein bands were excised from the SDS-PAGE gel and analyzed by matrix-assisted laser-desorption ionization time of flight (MALDI-TOF). Three bands were identified with confidence (P , 0.05), including two putative components of the Clp complex (LOC_Os03g29810/ VYL/OsClpP6 and LOC_Os10g43050/OsClpP4) and a vacuolar ATP synthase subunit E (Loc_Oso1g46980/ v-ATP-E; Fig. 8B; Table II). This result suggested that VYL is a bona fide component of the OsClp complex in vivo.
 ===Expression===

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 lines. qRT-PCR analysis revealed that the expression of VYL was reduced in three tested transgenic lines compared with the wild-type plants. These VYL knockdown transgenic plants showed reduced accumulation of chlorophyll a and chlorophyll b. Together, these results confirmed that LOC_Os03g29810 indeed corresponds to the VYL gene.
 [[File:Figure 4 .png]]
 ===Expression===

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 Chloroplast biogenesis and physiological changes are tightly regulated by the coordinated expression of plastid and nuclear genes during leaf development and facilitated by protein quality control (Kusumi et al., 2010; Clarke, 2012). To examine whether the expression of genes associated with chloroplast biogenesis and photosynthesis was altered in vyl mutants, quantitative real-time reverse transcription (qRT)-PCR was performed on total RNAs extracted from the L4 and
 L3U leaf sections of wild-type and vyl plants. Compared with the wild type, the transcript levels of Virescent1 (V1), V2, and V3 genes and some other genes encoding components of the plastid and nuclear transcription apparatus Sigma factor 2A (OsSig2A), Ribosomal Protein S15 (rps15) and the subunit RNA polymerase (RpoA and RpoTp) that are highly expressed in early stages of chloroplast development were significantly increased in vyl mutants. In contrast, several genes encoding components of the photosynthesis apparatus RuBisCO large subunit (RbcL), the subunit of photosystem I (PsaA), a core component of Photosystem II (PsbA), light-harvesting complex protein (Lhcp2) and Chlorophyll A/B binding protein1 (Cab1) that are highly expressed in later stages of chloroplast development were significantly reduced in vyl mutants (Fig. 3). These results suggested that VYL plays an important role in regulating chloroplast biogenesis.
 ====5.The vyl Locus Maps to a Putative Gene Encoding OsClpP6====
 Genetic analysis showed that the virescent yellow leaf phenotype in vyl mutants is controlled by a single recessive nuclear locus, VYL. Using a BC 1 F 2 mapping population of the vyl mutant and 93-11 (an indica variety), we mapped the vyl gene to a 137-kb genomic region on chromosome 3 between the insertion/deletion polymorphism (Indel) markers F17 and I53. Within this region, 17 open reading frames (ORFs) were predicted from published data (http://