Os04g0645100 - Revision history

192.168.72.52: /* Structured Information */

2015-06-12T07:14:23Z

‎Structured Information

Beiyuanyuan at 01:46, 10 June 2014

2014-06-10T01:46:53Z

Beiyuanyuan at 01:43, 10 June 2014

2014-06-10T01:43:01Z

Beiyuanyuan at 01:40, 10 June 2014

2014-06-10T01:40:55Z

Beiyuanyuan at 01:29, 10 June 2014

2014-06-10T01:29:23Z

Beiyuanyuan at 01:22, 10 June 2014

2014-06-10T01:22:52Z

Beiyuanyuan at 01:07, 10 June 2014

2014-06-10T01:07:30Z

Show changes

Beiyuanyuan: /* Function */

2014-06-09T23:33:08Z

‎Function

Beiyuanyuan: /* Expression and Mutation */

2014-06-09T23:23:39Z

‎Expression and Mutation

Beiyuanyuan: /* Annotated Information */

2014-06-09T15:39:59Z

‎Annotated Information

@@ Line 103: / Line 103: @@
 ==Structured Information==
 [[Category:Genes]]
 [[Category:Japonica mRNA]]

@@ Line 10: / Line 10: @@
 The transcript of ''FLO2'' is abundant in immature seeds and mature leaves, but not abundant in panicles before heading, stems, and roots.<ref name="ref1"/>
-The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The ''flo2'' grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of ''flo2'' grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).
+The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The ''flo2'' grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of ''flo2'' grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).<ref name="ref1"/>
-[[File:bf1.jpg|center|thumb|2000px|'''Figure 1.''' ''Phenotype of the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
+[[File:bf1.jpg|center|thumb|1000px|'''Figure 1.''' ''Phenotype of the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 (A) Representative seed of the wild type (Kinmaze), the ''flo2'' mutant (EM37), vector control line (VC), and complemented line (CL). CL and VC are transformants with/without a wild-type ''FLO2'' gene in the ''flo2'' mutant EM37. The top panel shows the grain shapes of the seeds, and the bottom panel shows seeds illuminated with backlight. The ''floury'' phenotype (EM37 and VC) is indicated by a dark image. Wild-type and CL grains were more packed and transmitted light, whereas the ''flo2'' mutant and VC grains were chalky and floury, resulting in dark images when illuminated with backlight. Bar = 1 cm.
 (B) Scanning electron microscopy images of the transverse sections of the wild-type, ''flo2'' mutant, VC, and CL grains. Bars = 50 μm.
 Grain size was significantly smaller than that of the wild type (89% on average, ''n'' > 40, P < 0.01), as were both grain weight and grain length. In the endosperm of the ''flo2'' mutant, the amylose content was lower than that of the wild type (Figure 2A), and structural changes in amylopectin were also noted, with both the short and the long chains consisting of 9 to 21 degrees of polymerization (DP) and ≥38 DP, respectively, decreasing and the middle chains with 22 to 38 DP increasing (Figure 2B).
 [[File:bf2.jpg|center|thumb|1000px|'''Figure 2.''' ''Amylose Content and Amylopectin Composition of the ''flo2'' and Wild-Type Grain.(from reference) <ref name="ref1" />.'']]
 (A) Amylose content of the wild type (Kinmaze), ''flo2'' mutant (EM37), VC, and CL. Representative VC and CL were analyzed. Error bars show sd (''n'' = 6).
@@ Line 22: / Line 23: @@
-Real-time quantitative RT-PCR indicated that expression levels of many genes that participated in starch biosynthesis were significantly decreased, such as genes for BEI, BEIIb, AGPases (AGPL1, AGPL2, AGPL3, AGPL4, AGPS1, AGPS2a, and AGPS2b), soluble SSs (SSI, SSIIa, SSIIc, SSIIIa, and SSIVb), GBSSI, isoamylases (ISA1 and ISA2), and pullulanase (PUL). Expression of genes for SSIIb, SSIIIb, SSIVa, GBSSII, BEIIa, and ISA3 was little decreased (Figure 3). Gene expression for sucrose synthase (Susy1 and Susy2), PPDKB, glucose-6-phosphate isomerases (PGIa and PGIb), and α-amylase (Amy3C, Amy3D, and Amy3E) was also reduced, while expression of the genes for Susy3 and Amy3B was little decreased (Figure 3). The expression of storage protein genes encoding glutelins (GluA1, GluA2, GluA3, and GluB1), globulins (Globulin1, Globulin2, 11S-globulin, and 19-kD globulin), prolamins (10 kD, 13 kD, and 17 kD), major allergenic protein genes (14 to 16 kD; ''RA16, RA17, RAG5B, RAG2'', and ''RG21''), and the alanine aminotransferase gene was also significantly reduced, whereas expression of GluB4 and protein disulfate isomerase was little decreased (Figure 3). In short, the ''flo2'' mutant showed reduced expression of a large portion of the genes participating in storage starch and storage protein biosynthesis in developing rice seeds.
+Real-time quantitative RT-PCR indicated that expression levels of many genes that participated in starch biosynthesis were significantly decreased, such as genes for BEI, BEIIb, AGPases (AGPL1, AGPL2, AGPL3, AGPL4, AGPS1, AGPS2a, and AGPS2b), soluble SSs (SSI, SSIIa, SSIIc, SSIIIa, and SSIVb), GBSSI, isoamylases (ISA1 and ISA2), and pullulanase (PUL). Expression of genes for SSIIb, SSIIIb, SSIVa, GBSSII, BEIIa, and ISA3 was little decreased (Figure 3). Gene expression for sucrose synthase (Susy1 and Susy2), PPDKB, glucose-6-phosphate isomerases (PGIa and PGIb), and α-amylase (Amy3C, Amy3D, and Amy3E) was also reduced, while expression of the genes for Susy3 and Amy3B was little decreased (Figure 3). The expression of storage protein genes encoding glutelins (GluA1, GluA2, GluA3, and GluB1), globulins (Globulin1, Globulin2, 11S-globulin, and 19-kD globulin), prolamins (10 kD, 13 kD, and 17 kD), major allergenic protein genes (14 to 16 kD; ''RA16, RA17, RAG5B, RAG2'', and ''RG21''), and the alanine aminotransferase gene was also significantly reduced, whereas expression of GluB4 and protein disulfate isomerase was little decreased (Figure 3). In short, the ''flo2'' mutant showed reduced expression of a large portion of the genes participating in storage starch and storage protein biosynthesis in developing rice seeds.<ref name="ref1"/>
-[[File:bf3.jpg|center|thumb|1500px|'''Figure 3.''' ''Expression Levels of the Genes Involved in Production of Storage Starch and Protein in the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
+[[File:bf3.jpg|center|thumb|1000px|'''Figure 3.''' ''Expression Levels of the Genes Involved in Production of Storage Starch and Protein in the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 Total RNA extracted from 10 DAF developing seeds was used for real-time RT-PCR analysis. Expression of representative genes involved in storage starch production, storage proteins, and carbon metabolism in the ''flo2'' mutant (EM37) are shown relative to the wild type (Kinmaze), which is set as 1. Each gene name is indicated by a simplified representation with the accession number of the corresponding full-length cDNA. BE1, BEIIa, and BEIIb, branching enzyme I, IIa, and IIb, respectively; AGPL (AGPL1, AGPL2, AGPL3, and AGPL4) and AGPS (AGPS1, AGPS2a, and AGPS2b), ADP glucose pyrophosphorylase large subunit and small subunit, respectively; SSI, SSIIa, SSIIb, SSIIc, SSIIIa, SSIIIb, SSIIIc, SSIVa, and SSIVb, soluble starch synthase I, IIa, IIb, IIc, IIIa, IIIb, IIIc, IVa, and IVb, respectively; GBSSI and GBSSII, granule-bound starch synthase I and II; ISA1, ISA2, and ISA3, corresponding isoamylase isozymes; PUL, pullulanase; Susy1, Susy2, and Susy3, sucrose synthase 1, 2, and 3, respectively; PPDKB, pyruvate phosphate dikinase B; PGI-a and PGI-b, glucose-6-phosphate isomerase a and b; Amy3B, Amy3C, Amy3D, and Amy3E, α-amylase 3B, 3C, 3D, and 3E, respectively; GluA1, GluA2, GluA3, GluB1, and GluB4, glutelin A1, A2, A3, B1, and B4, respectively; Globulin1, Globulin2, 11S-globulin, and 19-kD globulin, corresponding globulin species; 10-, 13-, and 17-kD prolamin, prolamins with each size; RA16, RA17, RAG5B, RAG2, and RG21, species of rice allergenic proteins; AlaAT, alanine aminotransferase; PDI, protein disulfate isomerase. Asterisks show subunits of the plastidic AGPase. Error bar shows sd (''n'' = 3).
-In the ''flo2'' grains, reduced accumulation of glutelin, a representative storage protein, was observed (Figure 4A), suggesting decreased production of storage proteins. Protein gel blot analyses also detected an extreme reduction in the amount of BEI and of major allergenic proteins, as well as of RA16 and homologs, in the grain of the ''flo2'' mutant (Figures 4B and 4C). These results suggest that the ''flo2'' mutation reduced the production of storage substances in total in the rice endosperm.
+In the ''flo2'' grains, reduced accumulation of glutelin, a representative storage protein, was observed (Figure 4A), suggesting decreased production of storage proteins. Protein gel blot analyses also detected an extreme reduction in the amount of BEI and of major allergenic proteins, as well as of RA16 and homologs, in the grain of the ''flo2'' mutant (Figures 4B and 4C). These results suggest that the ''flo2'' mutation reduced the production of storage substances in total in the rice endosperm.<ref name="ref1"/>
 [[File:bf4.jpg|center|thumb|1000px|'''Figure 4.''' ''Accumulation of Glutelins, BEI, and RA16 in Developing Seeds..(from reference) <ref name="ref1" />.'']]
 (A) Accumulation of glutelins in developing seeds of the wild type (Kinmaze), ''flo2'' mutant (EM37), VC, and CL. Each lane contains proteins from one-hundredth of the total glutelin fraction, which was extracted from the mixture of 10 rice grains of each line. Proteins are shown by Coomassie blue staining after separation on SDS-PAGE. Arrows indicate 51-kD glutelin precursor, 30- to 36-kD α-subunits of glutelins, and 19- to 22-kD β-subunits of glutelins, respectively. Size markers are shown on the left.
@@ Line 64: / Line 65: @@
 It has been reported that high-temperature stress during seed development apparently reduces the accumulation of storage starch in rice grains. In the case of Nipponbare, which is a rice cultivar sensitive to high-temperature stress, high temperature during seed development resulted in significant reduction of ''FLO2'' expression, while ''FLO2'' was expressed most abundantly under normal ripening conditions. The timing of elevation of FLO2 expression seemed to be delayed for several days in response to high-temperature stress (Figure 7B). This result suggests that ''FLO2'' expression was apparently repressed by high-temperature stress in this cultivar. On the other hand, in Kinmaze, which is relatively tolerant to high-temperature stress, it seemed that ''FLO2'' expression was upregulated by high-temperature stress (Figure 7A). These facts support the idea that ''FLO2'' plays an important role in the trait for tolerance to high-temperature stress.<ref name="ref1"/>
-''FLO2'' was abundantly expressed in immature seeds (Figures 8 and and 7 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
+''FLO2'' was abundantly expressed in immature seeds (Figures 7 and 8 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
-[[File:bf9.jpg|center|thumb|2000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
+[[File:bf9.jpg|center|thumb|1000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
 (A) Expression of ''FLO2'' in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).
 (B) and (C) Expression level of ''FLL1'' (B) and ''FLL2'' (C) in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Closed box, wild type (Kinmaze); open box, ''flo2'' mutant (EM37). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).
@@ Line 76: / Line 77: @@
 ===Evolution===
-In the rice genome ''FLO2'' have two homologous genes, designated ''FLO2-LIKE1'' (''FLL1'') (Os02g0255700) and ''FLL2'' (Os07g0422000). The predicted proteins encoded by ''FLL1'' and ''FLL2'' showed 59.8 and 52.7% similarity in amino acid sequence, respectively, to FLO2. Orthologous genes to ''FLO2'' and its homologs were also found in other plant genomes, such as in wheat, sorghum (''Sorghum bicolor''), poplar (''Populus spp''), grape (''Vitis vinifera''), castor bean (''Ricinus communis''), ''Arabidopsis thaliana'', and moss (''Physcomitrella patens'') (Figure 10), but not in animals or yeast. These homologs in higher plants were classified into three groups, which contained ''FLO2'', ''FLL1'', and ''FLL2'', respectively, whereas homologs of moss diverged from this group. These results suggest that ''FLO2'' constitutes a novel conserved gene family in plants.
+In the rice genome ''FLO2'' have two homologous genes, designated ''FLO2-LIKE1'' (''FLL1'') (Os02g0255700) and ''FLL2'' (Os07g0422000). The predicted proteins encoded by ''FLL1'' and ''FLL2'' showed 59.8 and 52.7% similarity in amino acid sequence, respectively, to FLO2. Orthologous genes to ''FLO2'' and its homologs were also found in other plant genomes, such as in wheat, sorghum (''Sorghum bicolor''), poplar (''Populus spp''), grape (''Vitis vinifera''), castor bean (''Ricinus communis''), ''Arabidopsis thaliana'', and moss (''Physcomitrella patens'') (Figure 10), but not in animals or yeast. These homologs in higher plants were classified into three groups, which contained ''FLO2'', ''FLL1'', and ''FLL2'', respectively, whereas homologs of moss diverged from this group. These results suggest that ''FLO2'' constitutes a novel conserved gene family in plants.<ref name="ref1"/>
 [[File:bf6.jpg|center|thumb|1000px|'''Figure 10.''' ''Phylogenetic Tree of the Representative FLO2 Homologs.(from reference) <ref name="ref1" />.'']]

@@ Line 10: / Line 10: @@
 The transcript of ''FLO2'' is abundant in immature seeds and mature leaves, but not abundant in panicles before heading, stems, and roots.<ref name="ref1"/>
-The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The ''flo2'' grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of ''flo2'' grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).<ref name="ref1"/>
+The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The ''flo2'' grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of ''flo2'' grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).
 [[File:bf1.jpg|center|thumb|2000px|'''Figure 1.''' ''Phenotype of the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 (A) Representative seed of the wild type (Kinmaze), the ''flo2'' mutant (EM37), vector control line (VC), and complemented line (CL). CL and VC are transformants with/without a wild-type ''FLO2'' gene in the ''flo2'' mutant EM37. The top panel shows the grain shapes of the seeds, and the bottom panel shows seeds illuminated with backlight. The ''floury'' phenotype (EM37 and VC) is indicated by a dark image. Wild-type and CL grains were more packed and transmitted light, whereas the ''flo2'' mutant and VC grains were chalky and floury, resulting in dark images when illuminated with backlight. Bar = 1 cm.
@@ Line 16: / Line 16: @@
 Grain size was significantly smaller than that of the wild type (89% on average, ''n'' > 40, P < 0.01), as were both grain weight and grain length. In the endosperm of the ''flo2'' mutant, the amylose content was lower than that of the wild type (Figure 2A), and structural changes in amylopectin were also noted, with both the short and the long chains consisting of 9 to 21 degrees of polymerization (DP) and ≥38 DP, respectively, decreasing and the middle chains with 22 to 38 DP increasing (Figure 2B).
 [[File:bf2.jpg|center|thumb|1000px|'''Figure 2.''' ''Amylose Content and Amylopectin Composition of the ''flo2'' and Wild-Type Grain.(from reference) <ref name="ref1" />.'']]
 (A) Amylose content of the wild type (Kinmaze), ''flo2'' mutant (EM37), VC, and CL. Representative VC and CL were analyzed. Error bars show sd (''n'' = 6).
@@ Line 23: / Line 22: @@
-Real-time quantitative RT-PCR indicated that expression levels of many genes that participated in starch biosynthesis were significantly decreased, such as genes for BEI, BEIIb, AGPases (AGPL1, AGPL2, AGPL3, AGPL4, AGPS1, AGPS2a, and AGPS2b), soluble SSs (SSI, SSIIa, SSIIc, SSIIIa, and SSIVb), GBSSI, isoamylases (ISA1 and ISA2), and pullulanase (PUL). Expression of genes for SSIIb, SSIIIb, SSIVa, GBSSII, BEIIa, and ISA3 was little decreased (Figure 3). Gene expression for sucrose synthase (Susy1 and Susy2), PPDKB, glucose-6-phosphate isomerases (PGIa and PGIb), and α-amylase (Amy3C, Amy3D, and Amy3E) was also reduced, while expression of the genes for Susy3 and Amy3B was little decreased (Figure 3). The expression of storage protein genes encoding glutelins (GluA1, GluA2, GluA3, and GluB1), globulins (Globulin1, Globulin2, 11S-globulin, and 19-kD globulin), prolamins (10 kD, 13 kD, and 17 kD), major allergenic protein genes (14 to 16 kD; ''RA16, RA17, RAG5B, RAG2'', and ''RG21''), and the alanine aminotransferase gene was also significantly reduced, whereas expression of GluB4 and protein disulfate isomerase was little decreased (Figure 3). In short, the ''flo2'' mutant showed reduced expression of a large portion of the genes participating in storage starch and storage protein biosynthesis in developing rice seeds.<ref name="ref1"/>
+Real-time quantitative RT-PCR indicated that expression levels of many genes that participated in starch biosynthesis were significantly decreased, such as genes for BEI, BEIIb, AGPases (AGPL1, AGPL2, AGPL3, AGPL4, AGPS1, AGPS2a, and AGPS2b), soluble SSs (SSI, SSIIa, SSIIc, SSIIIa, and SSIVb), GBSSI, isoamylases (ISA1 and ISA2), and pullulanase (PUL). Expression of genes for SSIIb, SSIIIb, SSIVa, GBSSII, BEIIa, and ISA3 was little decreased (Figure 3). Gene expression for sucrose synthase (Susy1 and Susy2), PPDKB, glucose-6-phosphate isomerases (PGIa and PGIb), and α-amylase (Amy3C, Amy3D, and Amy3E) was also reduced, while expression of the genes for Susy3 and Amy3B was little decreased (Figure 3). The expression of storage protein genes encoding glutelins (GluA1, GluA2, GluA3, and GluB1), globulins (Globulin1, Globulin2, 11S-globulin, and 19-kD globulin), prolamins (10 kD, 13 kD, and 17 kD), major allergenic protein genes (14 to 16 kD; ''RA16, RA17, RAG5B, RAG2'', and ''RG21''), and the alanine aminotransferase gene was also significantly reduced, whereas expression of GluB4 and protein disulfate isomerase was little decreased (Figure 3). In short, the ''flo2'' mutant showed reduced expression of a large portion of the genes participating in storage starch and storage protein biosynthesis in developing rice seeds.
 [[File:bf3.jpg|center|thumb|1500px|'''Figure 3.''' ''Expression Levels of the Genes Involved in Production of Storage Starch and Protein in the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 Total RNA extracted from 10 DAF developing seeds was used for real-time RT-PCR analysis. Expression of representative genes involved in storage starch production, storage proteins, and carbon metabolism in the ''flo2'' mutant (EM37) are shown relative to the wild type (Kinmaze), which is set as 1. Each gene name is indicated by a simplified representation with the accession number of the corresponding full-length cDNA. BE1, BEIIa, and BEIIb, branching enzyme I, IIa, and IIb, respectively; AGPL (AGPL1, AGPL2, AGPL3, and AGPL4) and AGPS (AGPS1, AGPS2a, and AGPS2b), ADP glucose pyrophosphorylase large subunit and small subunit, respectively; SSI, SSIIa, SSIIb, SSIIc, SSIIIa, SSIIIb, SSIIIc, SSIVa, and SSIVb, soluble starch synthase I, IIa, IIb, IIc, IIIa, IIIb, IIIc, IVa, and IVb, respectively; GBSSI and GBSSII, granule-bound starch synthase I and II; ISA1, ISA2, and ISA3, corresponding isoamylase isozymes; PUL, pullulanase; Susy1, Susy2, and Susy3, sucrose synthase 1, 2, and 3, respectively; PPDKB, pyruvate phosphate dikinase B; PGI-a and PGI-b, glucose-6-phosphate isomerase a and b; Amy3B, Amy3C, Amy3D, and Amy3E, α-amylase 3B, 3C, 3D, and 3E, respectively; GluA1, GluA2, GluA3, GluB1, and GluB4, glutelin A1, A2, A3, B1, and B4, respectively; Globulin1, Globulin2, 11S-globulin, and 19-kD globulin, corresponding globulin species; 10-, 13-, and 17-kD prolamin, prolamins with each size; RA16, RA17, RAG5B, RAG2, and RG21, species of rice allergenic proteins; AlaAT, alanine aminotransferase; PDI, protein disulfate isomerase. Asterisks show subunits of the plastidic AGPase. Error bar shows sd (''n'' = 3).
-In the ''flo2'' grains, reduced accumulation of glutelin, a representative storage protein, was observed (Figure 4A), suggesting decreased production of storage proteins. Protein gel blot analyses also detected an extreme reduction in the amount of BEI and of major allergenic proteins, as well as of RA16 and homologs, in the grain of the ''flo2'' mutant (Figures 4B and 4C). These results suggest that the ''flo2'' mutation reduced the production of storage substances in total in the rice endosperm.<ref name="ref1"/>
+In the ''flo2'' grains, reduced accumulation of glutelin, a representative storage protein, was observed (Figure 4A), suggesting decreased production of storage proteins. Protein gel blot analyses also detected an extreme reduction in the amount of BEI and of major allergenic proteins, as well as of RA16 and homologs, in the grain of the ''flo2'' mutant (Figures 4B and 4C). These results suggest that the ''flo2'' mutation reduced the production of storage substances in total in the rice endosperm.
 [[File:bf4.jpg|center|thumb|1000px|'''Figure 4.''' ''Accumulation of Glutelins, BEI, and RA16 in Developing Seeds..(from reference) <ref name="ref1" />.'']]
 (A) Accumulation of glutelins in developing seeds of the wild type (Kinmaze), ''flo2'' mutant (EM37), VC, and CL. Each lane contains proteins from one-hundredth of the total glutelin fraction, which was extracted from the mixture of 10 rice grains of each line. Proteins are shown by Coomassie blue staining after separation on SDS-PAGE. Arrows indicate 51-kD glutelin precursor, 30- to 36-kD α-subunits of glutelins, and 19- to 22-kD β-subunits of glutelins, respectively. Size markers are shown on the left.
@@ Line 65: / Line 64: @@
 It has been reported that high-temperature stress during seed development apparently reduces the accumulation of storage starch in rice grains. In the case of Nipponbare, which is a rice cultivar sensitive to high-temperature stress, high temperature during seed development resulted in significant reduction of ''FLO2'' expression, while ''FLO2'' was expressed most abundantly under normal ripening conditions. The timing of elevation of FLO2 expression seemed to be delayed for several days in response to high-temperature stress (Figure 7B). This result suggests that ''FLO2'' expression was apparently repressed by high-temperature stress in this cultivar. On the other hand, in Kinmaze, which is relatively tolerant to high-temperature stress, it seemed that ''FLO2'' expression was upregulated by high-temperature stress (Figure 7A). These facts support the idea that ''FLO2'' plays an important role in the trait for tolerance to high-temperature stress.<ref name="ref1"/>
-''FLO2'' was abundantly expressed in immature seeds (Figures 7 and 8 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
+''FLO2'' was abundantly expressed in immature seeds (Figures 8 and and 7 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
 [[File:bf9.jpg|center|thumb|2000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
 (A) Expression of ''FLO2'' in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).
@@ Line 77: / Line 76: @@
 ===Evolution===
-In the rice genome ''FLO2'' have two homologous genes, designated ''FLO2-LIKE1'' (''FLL1'') (Os02g0255700) and ''FLL2'' (Os07g0422000). The predicted proteins encoded by ''FLL1'' and ''FLL2'' showed 59.8 and 52.7% similarity in amino acid sequence, respectively, to FLO2. Orthologous genes to ''FLO2'' and its homologs were also found in other plant genomes, such as in wheat, sorghum (''Sorghum bicolor''), poplar (''Populus spp''), grape (''Vitis vinifera''), castor bean (''Ricinus communis''), ''Arabidopsis thaliana'', and moss (''Physcomitrella patens'') (Figure 10), but not in animals or yeast. These homologs in higher plants were classified into three groups, which contained ''FLO2'', ''FLL1'', and ''FLL2'', respectively, whereas homologs of moss diverged from this group. These results suggest that ''FLO2'' constitutes a novel conserved gene family in plants.<ref name="ref1"/>
+In the rice genome ''FLO2'' have two homologous genes, designated ''FLO2-LIKE1'' (''FLL1'') (Os02g0255700) and ''FLL2'' (Os07g0422000). The predicted proteins encoded by ''FLL1'' and ''FLL2'' showed 59.8 and 52.7% similarity in amino acid sequence, respectively, to FLO2. Orthologous genes to ''FLO2'' and its homologs were also found in other plant genomes, such as in wheat, sorghum (''Sorghum bicolor''), poplar (''Populus spp''), grape (''Vitis vinifera''), castor bean (''Ricinus communis''), ''Arabidopsis thaliana'', and moss (''Physcomitrella patens'') (Figure 10), but not in animals or yeast. These homologs in higher plants were classified into three groups, which contained ''FLO2'', ''FLL1'', and ''FLL2'', respectively, whereas homologs of moss diverged from this group. These results suggest that ''FLO2'' constitutes a novel conserved gene family in plants.
 [[File:bf6.jpg|center|thumb|1000px|'''Figure 10.''' ''Phylogenetic Tree of the Representative FLO2 Homologs.(from reference) <ref name="ref1" />.'']]

@@ Line 11: / Line 11: @@
 The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The ''flo2'' grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of ''flo2'' grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).<ref name="ref1"/>
-[[File:bf1.jpg|center|thumb|1000px|'''Figure 1.''' ''Phenotype of the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
+[[File:bf1.jpg|center|thumb|2000px|'''Figure 1.''' ''Phenotype of the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 (A) Representative seed of the wild type (Kinmaze), the ''flo2'' mutant (EM37), vector control line (VC), and complemented line (CL). CL and VC are transformants with/without a wild-type ''FLO2'' gene in the ''flo2'' mutant EM37. The top panel shows the grain shapes of the seeds, and the bottom panel shows seeds illuminated with backlight. The ''floury'' phenotype (EM37 and VC) is indicated by a dark image. Wild-type and CL grains were more packed and transmitted light, whereas the ''flo2'' mutant and VC grains were chalky and floury, resulting in dark images when illuminated with backlight. Bar = 1 cm.
 (B) Scanning electron microscopy images of the transverse sections of the wild-type, ''flo2'' mutant, VC, and CL grains. Bars = 50 μm.
@@ Line 24: / Line 24: @@
 Real-time quantitative RT-PCR indicated that expression levels of many genes that participated in starch biosynthesis were significantly decreased, such as genes for BEI, BEIIb, AGPases (AGPL1, AGPL2, AGPL3, AGPL4, AGPS1, AGPS2a, and AGPS2b), soluble SSs (SSI, SSIIa, SSIIc, SSIIIa, and SSIVb), GBSSI, isoamylases (ISA1 and ISA2), and pullulanase (PUL). Expression of genes for SSIIb, SSIIIb, SSIVa, GBSSII, BEIIa, and ISA3 was little decreased (Figure 3). Gene expression for sucrose synthase (Susy1 and Susy2), PPDKB, glucose-6-phosphate isomerases (PGIa and PGIb), and α-amylase (Amy3C, Amy3D, and Amy3E) was also reduced, while expression of the genes for Susy3 and Amy3B was little decreased (Figure 3). The expression of storage protein genes encoding glutelins (GluA1, GluA2, GluA3, and GluB1), globulins (Globulin1, Globulin2, 11S-globulin, and 19-kD globulin), prolamins (10 kD, 13 kD, and 17 kD), major allergenic protein genes (14 to 16 kD; ''RA16, RA17, RAG5B, RAG2'', and ''RG21''), and the alanine aminotransferase gene was also significantly reduced, whereas expression of GluB4 and protein disulfate isomerase was little decreased (Figure 3). In short, the ''flo2'' mutant showed reduced expression of a large portion of the genes participating in storage starch and storage protein biosynthesis in developing rice seeds.<ref name="ref1"/>
-[[File:bf3.jpg|center|thumb|1000px|'''Figure 3.''' ''Expression Levels of the Genes Involved in Production of Storage Starch and Protein in the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
+[[File:bf3.jpg|center|thumb|1500px|'''Figure 3.''' ''Expression Levels of the Genes Involved in Production of Storage Starch and Protein in the ''flo2'' Mutant.(from reference) <ref name="ref1" />.'']]
 Total RNA extracted from 10 DAF developing seeds was used for real-time RT-PCR analysis. Expression of representative genes involved in storage starch production, storage proteins, and carbon metabolism in the ''flo2'' mutant (EM37) are shown relative to the wild type (Kinmaze), which is set as 1. Each gene name is indicated by a simplified representation with the accession number of the corresponding full-length cDNA. BE1, BEIIa, and BEIIb, branching enzyme I, IIa, and IIb, respectively; AGPL (AGPL1, AGPL2, AGPL3, and AGPL4) and AGPS (AGPS1, AGPS2a, and AGPS2b), ADP glucose pyrophosphorylase large subunit and small subunit, respectively; SSI, SSIIa, SSIIb, SSIIc, SSIIIa, SSIIIb, SSIIIc, SSIVa, and SSIVb, soluble starch synthase I, IIa, IIb, IIc, IIIa, IIIb, IIIc, IVa, and IVb, respectively; GBSSI and GBSSII, granule-bound starch synthase I and II; ISA1, ISA2, and ISA3, corresponding isoamylase isozymes; PUL, pullulanase; Susy1, Susy2, and Susy3, sucrose synthase 1, 2, and 3, respectively; PPDKB, pyruvate phosphate dikinase B; PGI-a and PGI-b, glucose-6-phosphate isomerase a and b; Amy3B, Amy3C, Amy3D, and Amy3E, α-amylase 3B, 3C, 3D, and 3E, respectively; GluA1, GluA2, GluA3, GluB1, and GluB4, glutelin A1, A2, A3, B1, and B4, respectively; Globulin1, Globulin2, 11S-globulin, and 19-kD globulin, corresponding globulin species; 10-, 13-, and 17-kD prolamin, prolamins with each size; RA16, RA17, RAG5B, RAG2, and RG21, species of rice allergenic proteins; AlaAT, alanine aminotransferase; PDI, protein disulfate isomerase. Asterisks show subunits of the plastidic AGPase. Error bar shows sd (''n'' = 3).
@@ Line 66: / Line 66: @@
 ''FLO2'' was abundantly expressed in immature seeds (Figures 7 and 8 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
-[[File:bf9.jpg|center|thumb|1000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
+[[File:bf9.jpg|center|thumb|2000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
 (A) Expression of ''FLO2'' in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).
 (B) and (C) Expression level of ''FLL1'' (B) and ''FLL2'' (C) in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Closed box, wild type (Kinmaze); open box, ''flo2'' mutant (EM37). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).

@@ Line 65: / Line 65: @@
 It has been reported that high-temperature stress during seed development apparently reduces the accumulation of storage starch in rice grains. In the case of Nipponbare, which is a rice cultivar sensitive to high-temperature stress, high temperature during seed development resulted in significant reduction of ''FLO2'' expression, while ''FLO2'' was expressed most abundantly under normal ripening conditions. The timing of elevation of FLO2 expression seemed to be delayed for several days in response to high-temperature stress (Figure 7B). This result suggests that ''FLO2'' expression was apparently repressed by high-temperature stress in this cultivar. On the other hand, in Kinmaze, which is relatively tolerant to high-temperature stress, it seemed that ''FLO2'' expression was upregulated by high-temperature stress (Figure 7A). These facts support the idea that ''FLO2'' plays an important role in the trait for tolerance to high-temperature stress.<ref name="ref1"/>
-''FLO2'' was abundantly expressed in immature seeds (Figures 8 and and 7 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
+''FLO2'' was abundantly expressed in immature seeds (Figures 7 and 8 ), and its expression pattern was similar to that of genes involved in the production of storage starch and storage proteins. This temporal expression may account for the appearance of the loss-of-function phenotype observed in the endosperm of the ''flo2'' mutant seeds, such as reduced production of these storage substances. While the ''flo2'' mutant produced smaller grains, the OX lines had larger seeds than those of the wild type (Figures 2 and 9), exhibiting the gain-of-function phenotype of the ''FLO2'' gene. The increase in gene expression for enzymes involved in starch biosynthesis, however, is not directly linked to the level of ''FLO2'' expression. As a matter of fact, the increases in some of the parameters of grain size measured are barely significant (Figure 9). This could be attributed to the fact that expression of the analyzed genes was measured at a single time point and that the impact of ''FLO2'' may not coincide with its expression level at that time. This work suggests that ''FLO2'' has a pivotal role in the accumulation of storage substances in the endosperm and sets the foundation for the understanding of grain size and quality regulation in rice.<ref name="ref1"/>
 [[File:bf9.jpg|center|thumb|1000px|'''Figure 8.''' ''Expression Pattern of ''FLO2, FLL1, and FLL2''.(from reference) <ref name="ref1" />.'']]
 (A) Expression of ''FLO2'' in the stem (S), leaf blade (LB), flag leaf (FL), panicles before heading (P), 10 DAF developing seed (DS), and root (R). Amounts of transcripts are shown as the relative values to those of ''Actin I''. Error bar shows sd (''n'' = 3).

@@ Line 5: / Line 5: @@
 ==Annotated Information==
 ===Function===
-Rice (Oryza sativa) endosperm accumulates a massive amount of storage starch and storage proteins during seed development. However, little is known about the regulatory system involved in the production of storage substances. The rice flo2 mutation resulted in reduced grain size and starch quality. Map-based cloning identified FLOURY ENDOSPERM2 (FLO2), a member of a novel gene family conserved in plants, as the gene responsible for the rice flo2 mutation. FLO2 harbors a tetratricopeptide repeat motif, considered to mediate a protein-protein interactions. FLO2 was abundantly expressed in developing seeds coincident with production of storage starch and protein, as well as in leaves, while abundant expression of its homologs was observed only in leaves. The flo2 mutation decreased expression of genes involved in production of storage starch and storage proteins in the endosperm. Differences between cultivars in their responsiveness of FLO2 expression during high-temperature stress indicated that FLO2 may be involved in heat tolerance during seed development. Overexpression of FLO2 enlarged the size of grains significantly. These results suggest that FLO2 plays a pivotal regulatory role in rice grain size and starch quality by affecting storage substance accumulation in the endosperm.<ref name="ref1"/>
+Rice (''Oryza sativa'') endosperm accumulates a massive amount of storage starch and storage proteins during seed development. However, little is known about the regulatory system involved in the production of storage substances. The rice ''flo2'' mutation resulted in reduced grain size and starch quality. Map-based cloning identified ''FLOURY ENDOSPERM2'' (''FLO2''), a member of a novel gene family conserved in plants, as the gene responsible for the rice ''flo2'' mutation. FLO2 harbors a tetratricopeptide repeat motif, considered to mediate a protein-protein interactions. ''FLO2'' was abundantly expressed in developing seeds coincident with production of storage starch and protein, as well as in leaves, while abundant expression of its homologs was observed only in leaves. The ''flo2'' mutation decreased expression of genes involved in production of storage starch and storage proteins in the endosperm. Differences between cultivars in their responsiveness of ''FLO2'' expression during high-temperature stress indicated that ''FLO2'' may be involved in heat tolerance during seed development. Overexpression of ''FLO2'' enlarged the size of grains significantly. These results suggest that ''FLO2'' plays a pivotal regulatory role in rice grain size and starch quality by affecting storage substance accumulation in the endosperm.<ref name="ref1"/>
 ===Expression and Mutation===

@@ Line 10: / Line 10: @@
 The transcript of FLO2 is abundant in immature seeds and mature leaves, but not abundant in panicles before heading, stems, and roots.<ref name="ref1"/>
-The flo2 mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The flo2 grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of flo2 grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).
+The ''flo2'' mutants showed no apparent differences in visible features during the vegetative stage; plant height, the number of leaves, the number of tillers, and the number of panicles were similar to those of wild-type plants. No significant difference was found in the timing of flowering. The number of panicles per plant, the number of spikelets per panicle, and the ratio of ripened grains did not significantly differ from those of the wild-type plant. The flo2 grains had white and floury endosperm (Figure 1A), and scanning electron microscopy images of transverse sections of flo2 grains indicated that this endosperm was filled with loosely packed, small, and spherical starch granules with large air spaces, while the wild-type endosperm consisted of densely packed, large, and irregularly polyhedral starch granules (Figure 1B).
 [[File:bf1.jpg|center|thumb|1000px|'''Figure 1.''' ''Phenotype of the flo2 Mutant.(from reference) <ref name="ref1" />.'']]
 (A) Representative seed of the wild type (Kinmaze), the flo2 mutant (EM37), vector control line (VC), and complemented line (CL). CL and VC are transformants with/without a wild-type FLO2 gene in the flo2 mutant EM37. The top panel shows the grain shapes of the seeds, and the bottom panel shows seeds illuminated with backlight. The floury phenotype (EM37 and VC) is indicated by a dark image. Wild-type and CL grains were more packed and transmitted light, whereas the flo2 mutant and VC grains were chalky and floury, resulting in dark images when illuminated with backlight. Bar = 1 cm.

@@ Line 88: / Line 88: @@
 The FLO2 gene belongs to a gene family that consists of FLO2, FLL1, and FLL2 in the rice genome. Orthologs of these genes widely exist in plant species, including mosses, but none was found in yeast or animals. The phylogenetic tree indicates that there are three major clades for higher plants, each of which consists of the orthologs of FLO2, FLL1, and FLL2 (Figure 6). The interspecies similarity between members is larger than the similarities among FLO2, FLL1, and FLL2 within a species. These facts suggest that this gene family was acquired in an ancestor plant and diverged within the plant kingdom. The FLO2 homologs in moss (P. patens) form an independent branch in the phylogenetic tree, suggesting that the moss FLO2 family members may have evolved independently.<ref name="ref1" />
-FLL1 and FLL2 have similar protein structures to FLO2, and presumably have similar functions, but have somewhat different expression patterns. FLO2 was expressed in organs such as leaves and developing seeds (Figure 6A) and is therefore assumed to function in these organs. However, the flo2 mutant showed no apparent phenotype other than in seeds. In both the wild type and the flo2 mutant, a normal level of expression of both FLL1 and FLL2 was observed in leaves, whereas they were poorly expressed in developing seeds. It is presumed that the function of FLO2 is specific to developing seeds.<ref name="ref1"/>
+FLL1 and FLL2 have similar protein structures to FLO2, and presumably have similar functions, but have somewhat different expression patterns. FLO2 was expressed in organs such as leaves and developing seeds (Figure 6A) and is therefore assumed to function in these organs. However, the flo2 mutant showed no apparent phenotype other than in seeds. In both the wild type and the flo2 mutant, a normal level of expression of both FLL1 and FLL2 was observed in leaves, whereas they were poorly expressed in developing seeds. It is presumed that the function of FLO2 is specific to developing seeds.<ref name="ref1" />
 ==Labs working on this gene==