Os03g0654600 - Revision history

Liman at 16:05, 7 August 2016

2016-08-07T16:05:20Z

Liman at 16:02, 7 August 2016

2016-08-07T16:02:21Z

Liman: /* Expression */

2016-08-07T15:58:29Z

‎Expression

Liman: /* Function */

2016-08-07T15:52:11Z

‎Function

Liman at 15:47, 7 August 2016

2016-08-07T15:47:32Z

Liman: /* Function */

2016-08-07T15:46:44Z

‎Function

Liman at 15:38, 7 August 2016

2016-08-07T15:38:27Z

Liman: /* Function */

2016-08-07T14:13:21Z

‎Function

Liman at 14:10, 7 August 2016

2016-08-07T14:10:42Z

Liman: /* Mutation */

2016-08-07T14:07:37Z

‎Mutation

@@ Line 22: / Line 22: @@
 [[File:Os03g0654600-2.png|center|thumb|400px|'''Figure 2. Physiological characterization of nol-1 during senescence. (a) Change in the chlorophyll content during dark incubation. Chlorophyll was extracted with 80% acetone and its content was measured spectrophotometrically. Solid line, nol-1; dotted line, wild-type; closed circles, Chl a; open circles, Chl b. Bars indicate standard errors (n = 3). (b) Change in Fv/Fm values during dark incubation. Solid line, nol-1; dotted line, wild-type. Bars indicate standard errors (n = 3). (c) Change in membrane ion leakage during dark incubation. Solid line, nol-1; dotted line, wild-type. Bars indicate standard errors (n = 3).''' '' <ref name="ref1" />.'']]
-* To examine the behavior of the chlorophyll–protein complexes in nol mutants, we performed a ‘green-gel’ analysis (Figure 3a). The amounts of chlorophyll–protein complexes, namely the PSI reaction center, LHCII trimer, CP43/47, LHCI dimer and LHCII monomer, were similar between the wildtype and nol-1 before dark treatment. During dark-induced senescence, all chlorophyll–protein complexes were degraded in the wild-type, and the LHCII monomer and LHCII trimer were selectively retained in nol-1 (Figure 3a). Immunoblot analysis revealed that all LHCI proteins
+* To examine the behavior of the chlorophyll–protein complexes in nol mutants, we performed a ‘green-gel’ \ analysis (Figure 3a). The amounts of chlorophyll–protein complexes, namely the PSI reaction center, LHCII trimer, CP43/47, LHCI dimer and LHCII monomer, were similar between the wildtype and nol-1 before dark treatment. During dark-induced senescence, all chlorophyll–protein complexes were degraded in the wild-type, and the LHCII monomer and LHCII trimer were selectively retained in nol-1 (Figure 3a). Immunoblot analysis revealed that all LHCI proteins
 examined, namely Lhca1, Lhca3 and Lhca4, and PsaF, a component of the PSI core complex, were degraded at the late stage of senescence in both wild-type and nol-1 (Figure 3b)<ref name="ref1" />.
@@ Line 65: / Line 65: @@
 Kusaba, M., Ito, H., Morita, R. et al. (2007) Rice NON-YELLOW COLORING1 is involved in light-harvesting complex II and grana degradation during leaf senescence. Plant Cell, 19, 1362–1375.
 </ref>
 </references>
 ==Structured Information==
       [[Category:Genes]][[Category:Oryza Sativa Japonica Group]][[Category:Japonica Chromosome 3]]

@@ Line 1: / Line 1: @@
-Please input one-sentence summary here.
+NONYELLOW COLORING1 (NYC1) and NYC1-LIKE (NOL) are thought to encode Chl b reductases, which catalyze the conversion of Chl b to 7-hydroxymethyl Chl a (HMChl)<ref name="ref2" />.
 ==Annotated Information==
@@ Line 5: / Line 5: @@
 * The researchers therefore predicted that NYC1 and NOL are Chl b reductases with distinct roles in Chl b degradation<ref name="ref2" />.
-* To examine the hypothesis thatt NYC1 and NOL directly interact with each other, the researchers performed immunoprecipitation analysis. NOL and NYC1 proteins were produced in insect cells infected with baculovirus harboring NOL–HA, NYC1–FLAG or both. The produced proteins were detected by immunoblot analysis using anti-FLAG and anti-HA antibodies (Figure 8a). NYC1– FLAG was precipitated using anti-HA only in the presence of NOL–HA, indicating that NOL and NYC1 directly interact with each other in vitro (Figure 8b).
+* To examine the hypothesis thatt NYC1 and NOL directly interact with each other, the researchers performed immunoprecipitation analysis. NOL and NYC1 proteins were produced in insect cells infected with baculovirus harboring NOL–HA, NYC1–FLAG or both. The produced proteins were detected by immunoblot analysis using anti-FLAG and anti-HA antibodies (Figure 8a). NYC1– FLAG was precipitated using anti-HA only in the presence of NOL–HA, indicating that NOL and NYC1 directly interact with each other in vitro (Figure 8b)<ref name="ref1" />.
 [[File:Os03g0654600-7.png|center|thumb|400px|'''Figure 8. Physical interaction between NOL and NYC1 in vitro. (a) Expression of NOL–HA and NYC1–FLAG proteins in insect cells infected with recombinant baculoviruses. (b) Immunoprecipitation analysis of NOL–HA and NYC1–FLAG proteins. Proteins extracted from the insect cells were immunoprecipitated using anti-HA Sepharose conjugate.''' '' <ref name="ref1" />.'']]
-* Because NYC1 is localized in plastids and has transmembrane domains, it is likely thatNOL and NYC1 are co-localized in thylakoid membranes facing the stromal side where they form a complex that functions as a Chl b reductase (Figure 10).
+* Because NYC1 is localized in plastids and has transmembrane domains, it is likely thatNOL and NYC1 are co-localized in thylakoid membranes facing the stromal side where they form a complex that functions as a Chl b reductase (Figure 10)<ref name="ref1" />.
 [[File:Os03g0654600-8.png|center|thumb|400px|'''Figure 10. A model for the regulation of chlorophyll and LHCII degradation during leaf senescence in rice.''' '' <ref name="ref1" />.'']]
 ===Mutation===
-* As seen in the screening of stay-green mutants, nol-1, nol-2 and nol-3 showed the stay-green phenotype 7 days after dark incubation (DAD), in comparison to the wild-type, which showed full yellowing at this stage (Figure 1b). The leaves of nol-1 retained more greenness relative to the wildtype and did not turn yellow even at the very late stage of natural senescence (Figure 1c).
+* As seen in the screening of stay-green mutants, nol-1, nol-2 and nol-3 showed the stay-green phenotype 7 days after dark incubation (DAD), in comparison to the wild-type, which showed full yellowing at this stage (Figure 1b). The leaves of nol-1 retained more greenness relative to the wildtype and did not turn yellow even at the very late stage of natural senescence (Figure 1c)<ref name="ref1" />.
 [[File:Os03g0654600-1.png|center|thumb|400px|'''Figure 1. Mutation positions and the stay-green phenotype of nol mutants.''' '' <ref name="ref1" />.'']]
-* The change in chlorophyll content during dark-induced senescence was analyzed in nol-1, which showed prominent inhibition of Chl b degradation until 8 DAD (Figure 2a). Inhibition of Chl a degradation in nol-1 was not obvious until 4 DAD, but became significant at 6 DAD and thereafter. The ratio of Chl a to Chl b was close to 1 at 8 DAD. These characteristics were also observed in nol-2 and nol-3. As other parameters of leaf senescence, we measured the change in the Fv/Fm value (the ratio of variable to maximum fluorescence; a parameter of PSII activity) and membrane ion leakage during dark-induced senescence. The Fv/Fm value for nol-1 during dark-induced senescence decreased in a similar manner to the wild-type (Figure 2b), and membrane ion leakage also increased similarly to the wild-type (Figure 2c). These results indicate that nol shows the stay-green phenotype due to the retention of chlorophyll, but leaf functionality decreases during senescence.
+* The change in chlorophyll content during dark-induced senescence was analyzed in nol-1, which showed prominent inhibition of Chl b degradation until 8 DAD (Figure 2a). Inhibition of Chl a degradation in nol-1 was not obvious until 4 DAD, but became significant at 6 DAD and thereafter. The ratio of Chl a to Chl b was close to 1 at 8 DAD. These characteristics were also observed in nol-2 and nol-3. As other parameters of leaf senescence, we measured the change in the Fv/Fm value (the ratio of variable to maximum fluorescence; a parameter of PSII activity) and membrane ion leakage during dark-induced senescence. The Fv/Fm value for nol-1 during dark-induced senescence decreased in a similar manner to the wild-type (Figure 2b), and membrane ion leakage also increased similarly to the wild-type (Figure 2c). These results indicate that nol shows the stay-green phenotype due to the retention of chlorophyll, but leaf functionality decreases during senescence<ref name="ref1" />.
 [[File:Os03g0654600-2.png|center|thumb|400px|'''Figure 2. Physiological characterization of nol-1 during senescence. (a) Change in the chlorophyll content during dark incubation. Chlorophyll was extracted with 80% acetone and its content was measured spectrophotometrically. Solid line, nol-1; dotted line, wild-type; closed circles, Chl a; open circles, Chl b. Bars indicate standard errors (n = 3). (b) Change in Fv/Fm values during dark incubation. Solid line, nol-1; dotted line, wild-type. Bars indicate standard errors (n = 3). (c) Change in membrane ion leakage during dark incubation. Solid line, nol-1; dotted line, wild-type. Bars indicate standard errors (n = 3).''' '' <ref name="ref1" />.'']]
 * To examine the behavior of the chlorophyll–protein complexes in nol mutants, we performed a ‘green-gel’ analysis (Figure 3a). The amounts of chlorophyll–protein complexes, namely the PSI reaction center, LHCII trimer, CP43/47, LHCI dimer and LHCII monomer, were similar between the wildtype and nol-1 before dark treatment. During dark-induced senescence, all chlorophyll–protein complexes were degraded in the wild-type, and the LHCII monomer and LHCII trimer were selectively retained in nol-1 (Figure 3a). Immunoblot analysis revealed that all LHCI proteins
-examined, namely Lhca1, Lhca3 and Lhca4, and PsaF, a component of the PSI core complex, were degraded at the late stage of senescence in both wild-type and nol-1 (Figure 3b).
+examined, namely Lhca1, Lhca3 and Lhca4, and PsaF, a component of the PSI core complex, were degraded at the late stage of senescence in both wild-type and nol-1 (Figure 3b)<ref name="ref1" />.
 [[File:Os03g0654600-3.png|center|thumb|400px|'''Figure 3. Protein degradation during senescence in nol-1, nyc1-2 and nol-1 nyc1-2.''' '' <ref name="ref1" />.'']]
-* To examine the behavior of photosynthetic pigments during senescence, the researchers performed HPLC analysis of extracts from detached leaves of nol-1 and the wild-type (Figure 4). The chromatograms were very similar before dark incubation, but at 7 DAD nol-1 showed apparent retention of not only Chl a and Chl b but also neoxanthin and lutein, all of which are LHCII-bound pigments. The stability of violaxanthin, a weakly LHCII-bound pigment, was less affected in nol-1. This result suggests that the inhibition of photosynthetic pigment breakdown in nol is due to the stability of LHCII during senescence, as is the case for nyc1.
+* To examine the behavior of photosynthetic pigments during senescence, the researchers performed HPLC analysis of extracts from detached leaves of nol-1 and the wild-type (Figure 4). The chromatograms were very similar before dark incubation, but at 7 DAD nol-1 showed apparent retention of not only Chl a and Chl b but also neoxanthin and lutein, all of which are LHCII-bound pigments. The stability of violaxanthin, a weakly LHCII-bound pigment, was less affected in nol-1. This result suggests that the inhibition of photosynthetic pigment breakdown in nol is due to the stability of LHCII during senescence, as is the case for nyc1<ref name="ref1" />.
 [[File:Os03g0654600-4.png|center|thumb|400px|'''Figure 4. HPLC elution profiles of photosynthetic pigments extracted from non-senescent (0 DAD) and fully senescent (7 DAD) leaves of the wild-type and nol-1. Absorption was measured at 440 nm. The same weight of leaves was extracted with the same volume of 80% acetone. N, neoxanthin; V, violaxanthin; L, lutein; La, lutein 3-acetate; b, Chl b; a, Chl a; C, b-carotene.''' '' <ref name="ref1" />.'']]
-* The researchers therefore examined the ultrastructure of chloroplasts during senescence in the wild-type and nol-1 (Figure 5). There was no obvious difference at 0 DAD, but at 4 DAD chloroplasts became round in shape and plastoglobules became obvious in both the wild-type and nol-1, and initial degradation of grana stacks was observed in the wild-type. At 7 DAD, only a small volume of thylakoid membranes remained in the wildtype, but very thick and wide grana stacks were observed in nol-1. These observations indicate that grana stacks are not properly degenerated during senescence in nol.
+* The researchers therefore examined the ultrastructure of chloroplasts during senescence in the wild-type and nol-1 (Figure 5). There was no obvious difference at 0 DAD, but at 4 DAD chloroplasts became round in shape and plastoglobules became obvious in both the wild-type and nol-1, and initial degradation of grana stacks was observed in the wild-type. At 7 DAD, only a small volume of thylakoid membranes remained in the wildtype, but very thick and wide grana stacks were observed in nol-1. These observations indicate that grana stacks are not properly degenerated during senescence in nol<ref name="ref1" />.
 [[File:Os03g0654600-5.png|center|thumb|400px|'''Figure 5. Ultrastructures of chloroplasts during leaf senescence in the wild-type and nol-1. (a)–(c) wild-type; (d)–(f) nol-1. (a,d), 0 DAD; (b,e), 4 DAD; (c,f), 7 DAD. g, grana; p, plastoglobule. The arrowhead indicates residual grana stacks. Scale bars = 500 nm.''' '' <ref name="ref1" />.'']]
-* The observations described above indicate that nol and nyc1 show a very similar phenotype, suggesting that they act in the same process in chlorophyll degradation. To examine the genetic interaction between NOL and NYC1, the researchers raised a double mutant of the probable null mutations nol-1 and nyc1-2. In both the nol-1 and nyc1-2 mutants, Chl b was slightly degraded at the late stage of senescence. The chlorophyll contents at 0 and 6 DAD were also measured in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 (Figure 6). The degree of inhibition of chlorophyll degradation was broadly similar among the nol-1, nyc1-2 and nol-1 nyc1-2 mutants, with a slight enhancement of Chl b retention in nol-1 nyc1-2.
+* The observations described above indicate that nol and nyc1 show a very similar phenotype, suggesting that they act in the same process in chlorophyll degradation. To examine the genetic interaction between NOL and NYC1, the researchers raised a double mutant of the probable null mutations nol-1 and nyc1-2. In both the nol-1 and nyc1-2 mutants, Chl b was slightly degraded at the late stage of senescence. The chlorophyll contents at 0 and 6 DAD were also measured in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 (Figure 6). The degree of inhibition of chlorophyll degradation was broadly similar among the nol-1, nyc1-2 and nol-1 nyc1-2 mutants, with a slight enhancement of Chl b retention in nol-1 nyc1-2<ref name="ref1" />.
 [[File:Os03g0654600-6.png|center|thumb|400px|'''Figure 6. Chlorophyll degradation during leaf senescence in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2. Chl a and Chl b contents at 0 and 6 DAD in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 were measured spectrophotometrically. Bars indicate standard errors (n = 6).''' '' <ref name="ref1" />.'']]
 ===Subcellular localization===
-* To examine the intracellular localization of NOL, we fused the region encoding the putative transit peptide of NOL to the green fluorescent protein (GFP) gene and introduced the fusion into the epidermal cells of onion (Allium cepa) by particle bombardment. The GFP signal co-localized with that of red fluorescent protein fused to the transit peptide of plastid-localizing rice S9 ribosomal protein. This result indicates that NOL is localized in the plastids.
+* To examine the intracellular localization of NOL, we fused the region encoding the putative transit peptide of NOL to the green fluorescent protein (GFP) gene and introduced the fusion into the epidermal cells of onion (Allium cepa) by particle bombardment. The GFP signal co-localized with that of red fluorescent protein fused to the transit peptide of plastid-localizing rice S9 ribosomal protein. This result indicates that NOL is localized in the plastids<ref name="ref1" />.
 ===Expression===

@@ Line 44: / Line 44: @@
 ===Expression===
-Please input expression information here.
+* Expression of both NYC1 and NOL is induced during senescence in rice<ref name="ref2" />.
 ===Evolution===

← Older revision		Revision as of 15:52, 7 August 2016
Line 8:		Line 8:

	[[File:Os03g0654600-7.png\|center\|thumb\|400px\|'''Figure 8. Physical interaction between NOL and NYC1 in vitro. (a) Expression of NOL–HA and NYC1–FLAG proteins in insect cells infected with recombinant baculoviruses. (b) Immunoprecipitation analysis of NOL–HA and NYC1–FLAG proteins. Proteins extracted from the insect cells were immunoprecipitated using anti-HA Sepharose conjugate.''' '' <ref name="ref1" />.'']]		[[File:Os03g0654600-7.png\|center\|thumb\|400px\|'''Figure 8. Physical interaction between NOL and NYC1 in vitro. (a) Expression of NOL–HA and NYC1–FLAG proteins in insect cells infected with recombinant baculoviruses. (b) Immunoprecipitation analysis of NOL–HA and NYC1–FLAG proteins. Proteins extracted from the insect cells were immunoprecipitated using anti-HA Sepharose conjugate.''' '' <ref name="ref1" />.'']]
		+
		+	* Because NYC1 is localized in plastids and has transmembrane domains, it is likely thatNOL and NYC1 are co-localized in thylakoid membranes facing the stromal side where they form a complex that functions as a Chl b reductase (Figure 10).
		+
		+	[[File:Os03g0654600-8.png\|center\|thumb\|400px\|'''Figure 10. A model for the regulation of chlorophyll and LHCII degradation during leaf senescence in rice.''' '' <ref name="ref1" />.'']]

	===Mutation===		===Mutation===

@@ Line 59: / Line 59: @@
 </ref>
 * <ref name="ref2">
+Kusaba, M., Ito, H., Morita, R. et al. (2007) Rice NON-YELLOW COLORING1 is involved in light-harvesting complex II and grana degradation during leaf senescence. Plant Cell, 19, 1362–1375.
 </ref>
 * <ref name="ref3">

← Older revision		Revision as of 14:10, 7 August 2016
Line 31:		Line 31:

	[[File:Os03g0654600-6.png\|center\|thumb\|400px\|'''Figure 6. Chlorophyll degradation during leaf senescence in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2. Chl a and Chl b contents at 0 and 6 DAD in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 were measured spectrophotometrically. Bars indicate standard errors (n = 6).''' '' <ref name="ref1" />.'']]		[[File:Os03g0654600-6.png\|center\|thumb\|400px\|'''Figure 6. Chlorophyll degradation during leaf senescence in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2. Chl a and Chl b contents at 0 and 6 DAD in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 were measured spectrophotometrically. Bars indicate standard errors (n = 6).''' '' <ref name="ref1" />.'']]
		+
		+	===Subcellular localization===
		+	* To examine the intracellular localization of NOL, we fused the region encoding the putative transit peptide of NOL to the green fluorescent protein (GFP) gene and introduced the fusion into the epidermal cells of onion (Allium cepa) by particle bombardment. The GFP signal co-localized with that of red fluorescent protein fused to the transit peptide of plastid-localizing rice S9 ribosomal protein. This result indicates that NOL is localized in the plastids.

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

← Older revision		Revision as of 14:07, 7 August 2016
Line 23:		Line 23:

	[[File:Os03g0654600-4.png\|center\|thumb\|400px\|'''Figure 4. HPLC elution profiles of photosynthetic pigments extracted from non-senescent (0 DAD) and fully senescent (7 DAD) leaves of the wild-type and nol-1. Absorption was measured at 440 nm. The same weight of leaves was extracted with the same volume of 80% acetone. N, neoxanthin; V, violaxanthin; L, lutein; La, lutein 3-acetate; b, Chl b; a, Chl a; C, b-carotene.''' '' <ref name="ref1" />.'']]		[[File:Os03g0654600-4.png\|center\|thumb\|400px\|'''Figure 4. HPLC elution profiles of photosynthetic pigments extracted from non-senescent (0 DAD) and fully senescent (7 DAD) leaves of the wild-type and nol-1. Absorption was measured at 440 nm. The same weight of leaves was extracted with the same volume of 80% acetone. N, neoxanthin; V, violaxanthin; L, lutein; La, lutein 3-acetate; b, Chl b; a, Chl a; C, b-carotene.''' '' <ref name="ref1" />.'']]
		+
		+	* The researchers therefore examined the ultrastructure of chloroplasts during senescence in the wild-type and nol-1 (Figure 5). There was no obvious difference at 0 DAD, but at 4 DAD chloroplasts became round in shape and plastoglobules became obvious in both the wild-type and nol-1, and initial degradation of grana stacks was observed in the wild-type. At 7 DAD, only a small volume of thylakoid membranes remained in the wildtype, but very thick and wide grana stacks were observed in nol-1. These observations indicate that grana stacks are not properly degenerated during senescence in nol.
		+
		+	[[File:Os03g0654600-5.png\|center\|thumb\|400px\|'''Figure 5. Ultrastructures of chloroplasts during leaf senescence in the wild-type and nol-1. (a)–(c) wild-type; (d)–(f) nol-1. (a,d), 0 DAD; (b,e), 4 DAD; (c,f), 7 DAD. g, grana; p, plastoglobule. The arrowhead indicates residual grana stacks. Scale bars = 500 nm.''' '' <ref name="ref1" />.'']]

	* The observations described above indicate that nol and nyc1 show a very similar phenotype, suggesting that they act in the same process in chlorophyll degradation. To examine the genetic interaction between NOL and NYC1, the researchers raised a double mutant of the probable null mutations nol-1 and nyc1-2. In both the nol-1 and nyc1-2 mutants, Chl b was slightly degraded at the late stage of senescence. The chlorophyll contents at 0 and 6 DAD were also measured in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 (Figure 6). The degree of inhibition of chlorophyll degradation was broadly similar among the nol-1, nyc1-2 and nol-1 nyc1-2 mutants, with a slight enhancement of Chl b retention in nol-1 nyc1-2.		* The observations described above indicate that nol and nyc1 show a very similar phenotype, suggesting that they act in the same process in chlorophyll degradation. To examine the genetic interaction between NOL and NYC1, the researchers raised a double mutant of the probable null mutations nol-1 and nyc1-2. In both the nol-1 and nyc1-2 mutants, Chl b was slightly degraded at the late stage of senescence. The chlorophyll contents at 0 and 6 DAD were also measured in the wild-type, nol-1, nyc1-2 and nol-1 nyc1-2 (Figure 6). The degree of inhibition of chlorophyll degradation was broadly similar among the nol-1, nyc1-2 and nol-1 nyc1-2 mutants, with a slight enhancement of Chl b retention in nol-1 nyc1-2.