Os01g0262600 - Revision history

192.168.72.52 at 04:39, 14 May 2015

2015-05-14T04:39:33Z

Tracy: /* References */

2014-06-04T06:20:48Z

‎References

Tracy: /* Labs working on this gene */

2014-06-04T05:59:33Z

‎Labs working on this gene

Tracy: /* Labs working on this gene */

2014-06-04T05:57:58Z

‎Labs working on this gene

Tracy: /* Function */

2014-06-04T05:55:01Z

‎Function

Tracy: /* Evolution */

2014-06-04T05:50:51Z

‎Evolution

Tracy: /* Expression */

2014-06-04T05:37:35Z

‎Expression

Tracy: /* Function */

2014-06-04T05:31:41Z

‎Function

Huanghs at 14:38, 2 June 2014

2014-06-02T14:38:06Z

Huanghs: /* Annotated Information */

2014-06-02T14:37:49Z

‎Annotated Information

← Older revision		Revision as of 04:39, 14 May 2015
Line 116:		Line 116:

	==Structured Information==		==Structured Information==
−	~~{{JaponicaGene\|~~	+	[[Category:Genes]][[Category:Oryza Sativa Japonica Group]][[Category:Japonica Chromosome 1]]
−	~~GeneName = Os01g0262600\|~~
−	~~Description = Glycosyl transferase, group 1 domain containing protein\|~~
−	~~Version = NM_001049188.1 GI:115435789 GeneID:4325125\|~~
−	~~Length = 5685 bp\|~~
−	~~Definition = Oryza sativa Japonica Group Os01g0262600, 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 1\|Chromosome 1]]\|~~
−	~~AP = Chromosome 1:8884740..8890424\|~~
−	~~CDS = 8884740..8884997,8886068..8886130,8887345..8887607,8887708..8887862,8888174..8888445<br>,8889573..8889715,8890130..8890424\|~~
−	~~GCID = <gbrowseImage1>~~
−	~~name=NC_008394:8884740..8890424~~
−	~~source=RiceChromosome01~~
−	~~preset=GeneLocation~~
−	~~</gbrowseImage1>\|~~
−	~~GSID = <gbrowseImage2>~~
−	~~name=NC_008394:8884740..8890424~~
−	~~source=RiceChromosome01~~
−	~~preset=GeneLocation~~
−	~~</gbrowseImage2>\|~~
−	CDNA = <cdnaseq>atggcgaagccctcgtcggcggcggcggcgtcgggcggcggccgaggcccggcccaccaccacaggacccgcctcctcctcctgctcctcctcgccgtcgccgcctgcgcatccaccgcgggcttcctcctctgcggcgccatgctggacccctgtgacgtcgacgcgcgcaggggctcgggctcgtcggcggcggccgtcgcgaccacccgcaccggcgccgtcgcagggaaccccctcgagttcatgaggtccaagctcatgctgctcgtctcccacgagctctccctctccgggggaccattgttactgatggagttggcatttcttctgcggcaggttggttcgcaagtggtgtggataacaaaccagagatcggaagaaacaaatgatgttacatatagcttggagcataagatgttgagccatggagtgcaggtattacctgctaggggacatgaggcgattgatactgccctaaaggctgatcttgttatcttgaacactgctgttgctggcaagtggcttgatgctgtcctaaatgaccatgttccccaagttcttcccaagattttgtggtggatccatgaaatgcgagggcattactttaagcttgaatatgtcaaacatcttcctctggttgctggagccatgattgattctcatacaacggctgaatattggaagaccaggactcatgaccgtttgaaaatacagatgccacaaacttatgttgttcacctcgggaatagtaaggagttaatggaagttgctgaagataatgttgcaagaagagtcctacgggagcatatccgtgagttccttggagtacggagtgaagatcttgtgtttgcaataataaacagtgtttcacgtggaaaagggcaggacttatttcttcaagcgttttatcagggtgtccagctcatcgaacagaagaagttaaaagtgcccacaatgcatgctgtagttgtgggaagtgacataaatgctcagaccaagtttgagacacaattacgtgattttgcggtgaagaatgggattcaggaccgggttcactttgtaaacaagacattagcagtggctccttatttggctgcaactgatgtgcttgttcagaactctcaggcccgtggagaatgctttggaaggataacgattgaagcaatggcattcaagttgccagtgttgggcacggcggccggagggaccacggagatcgtcgtggacggctcaaccggcctcctgcaccccgccgggaaggaaggcgtcgcgcccctcgcgaagaacatggtaaggctcgcgagccacgaagaggacagggtctccatgggtaggaaaggctatggcagggtgaaggaaatgttcatggagcaccacatggctggtaggatcgcggccgtgttgcaggaagttctgaagaaatcgcgggaacattctcactcttga</cdnaseq>\|
−	AA = <aaseq>MAKPSSAAAASGGGRGPAHHHRTRLLLLLLLAVAACASTAGFLL CGAMLDPCDVDARRGSGSSAAAVATTRTGAVAGNPLEFMRSKLMLLVSHELSLSGGPL LLMELAFLLRQVGSQVVWITNQRSEETNDVTYSLEHKMLSHGVQVLPARGHEAIDTAL KADLVILNTAVAGKWLDAVLNDHVPQVLPKILWWIHEMRGHYFKLEYVKHLPLVAGAM IDSHTTAEYWKTRTHDRLKIQMPQTYVVHLGNSKELMEVAEDNVARRVLREHIREFLG VRSEDLVFAIINSVSRGKGQDLFLQAFYQGVQLIEQKKLKVPTMHAVVVGSDINAQTK FETQLRDFAVKNGIQDRVHFVNKTLAVAPYLAATDVLVQNSQARGECFGRITIEAMAF KLPVLGTAAGGTTEIVVDGSTGLLHPAGKEGVAPLAKNMVRLASHEEDRVSMGRKGYG RVKEMFMEHHMAGRIAAVLQEVLKKSREHSHS</aaseq>\|
−	DNA = <dnaseqindica>5428..5685#4295..4357#2818..3080#2563..2717#1980..2251#710..852#1..295#atggcgaagccctcgtcggcggcggcggcgtcgggcggcggccgaggcccggcccaccaccacaggacccgcctcctcctcctgctcctcctcgccgtcgccgcctgcgcatccaccgcgggcttcctcctctgcggcgccatgctggacccctgtgacgtcgacgcgcgcaggggctcgggctcgtcggcggcggccgtcgcgaccacccgcaccggcgccgtcgcagggaaccccctcgagttcatgaggtccaagctcatgctgctcgtctcccacgagctctccctctccggtaaggagtagcagtgtgcttcagtatattctgttgaggaatcttgcgatttgttcatgttaccttttgagaatgcgatgcctccaagatcgttatatggattttatttccagtagcaaacagactgataggaaagtgaaatttctactcccagtgctcagcacgaactcatgatccctgaaaaggaaaaaaagaaaaactggaaattcggattatcacagatcacaaataaaattgtcgtatgcgtcgctctgcttgagttgtttataagcgggaaacttagtttggaagttttagcatatctacgcagttctcattgtcacagatctcatagaattgtttgctgttagtgacatctgtaactatgtctgattgtctactggtgttactggaaacacttttgttttgatgcagggggaccattgttactgatggagttggcatttcttctgcggcaggttggttcgcaagtggtgtggataacaaaccagagatcggaagaaacaaatgatgttacatatagcttggagcataagatgttgagccatggagtgcaggtatgttctacgctacttagttctgcaaattacccgtatgtcagcagaatatctaataggagttcacctcggtgaaagccaccgccatgctattcaacactaccttaaaatatggtccaggatatcatttgttcggattggaagttagatatgaaaaataacttttttttctgttcagtttgataattggactttaaagcaaagttcgatgggcaaaaatggaatttcttcaatgaatcatagtgtaaacaaaattgttgaataaatcatcttgtgggaaacaagaaattgaattaccacaatgtaatgttttagttctgaattagaacctatttaaatttttagagaatactagattgattgtttatgttgagtcacacaagaaggcacaatggaataaaatattgcacggtaagggaaaataaaagaagggtggaagttttcatataacacttgaaacttcaaggatcaaacagttcagtataaattggcattaactgtttgatccttgaaacaacatcttgtagcatgctgtgtcatgcttttgctcatgtgcattgttgagtcctaatatagaccaatatatatatatacattcattaagactaacaggatttcttgtggctggtctaatcttatcccatcattcatctgcatgttactatatgcacagaagccataatatttcttgcattgcaaatttataactgattagtatatagtagttatatactacttaagattgttgataaaccatattgatggggtgataaataaatagctacagatgaatttactctgttattattgtcttgtagtcttggagtgttagttttgagttctcttgcttgtttatttgtcagtactgctaattttgatgttctacatactcgttgcagtcctatcttggtctcatgtactatgattaccctcttaaatttgttccaaatggctgttttctcctgagataattacttaaacgtttgaaataattactacgcaatatttgatagaatatgttggagtacaaaagaaaaacaggtccaaatactttcccttcttttggcagacttaacaaacattccatgctctcgtcaattttcacactatcactgcttgcttcataggtattacctgctaggggacatgaggcgattgatactgccctaaaggctgatcttgttatcttgaacactgctgttgctggcaagtggcttgatgctgtcctaaatgaccatgttccccaagttcttcccaagattttgtggtggatccatgaaatgcgagggcattactttaagcttgaatatgtcaaacatcttcctctggttgctggagccatgattgattctcatacaacggctgaatattggaagaccaggactcatgaccgtttgaagtatgtgcaaaattattcacattctttgtaatataaatgctggatcatgtaacatttgtcaccgtttctgcttattggacttgctagtgcttttatattatttactgccctagatgatgataaacttctttggccctttgcttctctgctgaactcattggcaacttttctcttcttcaattagcctatatatgaggtttcactctataaaatgaatcatgatagctatgatcaaatgcttgttcaatgcttttaaatttcctaaatttgtacttcgttgttatataattctgatttgcttccatgtaaagaatacagatgccacaaacttatgttgttcacctcgggaatagtaaggagttaatggaagttgctgaagataatgttgcaagaagagtcctacgggagcatatccgtgagttccttggagtacggagtgaagatcttgtgtttgcaataataaacagtatgcgccatttgcagcttcttatccctgttgttattccaactatatatgtatatattgattataccaattcatttctctgattgatgttaaaatacaggtgtttcacgtggaaaagggcaggacttatttcttcaagcgttttatcagggtgtccagctcatcgaacagaagaagttaaaagtgcccacaatgcatgctgtagttgtgggaagtgacataaatgctcagaccaagtttgagacacaattacgtgattttgcggtgaagaatgggattcaggaccgggttcactttgtaaacaagacattagcagtggctccttatttggctgcaactgatgtgcttgttcagaactctcaggtgagtaataattgtaataacacttccattttctcttgttttcccagttcagtggtgtttcgtgatgcaaactgtatgccttttgtgtatttgcaattttgcatcatagtttgttatttctcttcatattctttgtgtgcgtatgtgcgcgtgcgtggaatcaatcaactgttggttaagtgagtgtgctgctagcagaaagtcaagaatttaatgtgatttacaatttttctgccattttcttttttttttgggagagttttgaagcacatctgattttaagagtaatactaactactattttttttttcatctgggaacagtaagcttgataactcgataggttgaaccactgtattatggagttaattctattattctagactattctgtcagaactattagcctgaatcctaaatagcatccaacttatcttcattgtcacaacatgcttttaagtaacttgacaggagatgttatttctacatagatagtgcagaatgcttcagtatattggcatagaatgttcagcaattgtgtttattctctacgcaatactgacaaaccattttgattactttctctagcttttgattttttacatggtctcatgctcggtcgctcacccatgaggccataaccatggttatatgtattgtttcagtgtggctaggactgcctgtaccaatcacattcatcagtaaaatatctggaactgttggtccattttttattacttgttaattgttatgccgcaacgatcagtaataatgcatatttttgcttttgtatgttccgttatggctaacaggcaataccagtctccaatttttaagaacaagacagaaaaataatattgttttcgtccaaataaaaacatatatacttgtatagatttgttattcctggatttgcctccaatttatctatggaatgtttccgtacaataataatgtctagatcaagtcaatgagcacatagttgaaagcaatgtaaaagtagggtatggtcagtctgcaaatagctgaaagggccttagctccagattggtttatgggtgtgccagttgcaaatggttctgttggcacagagccctgctcctgattaggcacgtggagaatgctgtatgttaggtatcaataagtagagctcttcaaagctagaatatccataagctgatgtgcttttgtttttgctgttcttttcgtacttaggcccgtggagaatgctttggaaggataacgattgaagcaatggcattcaagttgccagtgttggtaaaactccctccctccctctcttgaagtgctcctagttataccccattgatccacaaacacaaatagtttttgactcactagtctcgatgcaagagttgagtacaattcttttttttatcaaagtatattagaaacaaaaatatatcataattttactggggaatgcaaatccaatgacactaatttcacatgatccatttctataacctcacagttcagtggtcatgaatgattaactgcatagatctcaaggtaacacgtacgtatttatcgatggagggagtataattttctacttctattaaccatcaatctgccagctagaaagccttacgcccttaccataatatagtttggaacaaaggaattttataaatactccctccactctaaaatataaaaatataagtatttctaggattcaaagtttgtatcacaatataagcatttctgggtctggggctctatacattttcagaaatacttatatttggaatggagggaactgagggagaagttcatttcccatgggagtcaaaaaaaaaaaaggattcccacgttccaaaggagccctggaatgtgggccagtatgcctctgaatacagagttattattgacaatcatgagccttggactcaactgtggactgtctgactgtccctacaaaccctcggctcttgagcctgccatcctctgtttcagccggcatctcctgcctcatgcttccatatgacagggtgtgtttggttgcaagctacactttgccacagtttgccacgcctaaggttaggcaaatttgacaggtgtttggttgtagccacagttgtggcaagattcccctccaacaaattgggtcccacgtgtcaatggctcaaaaaagtgtggcaagattcctttaggcttagtaagttgtggctaacaatttgatcacctcaccttaggcaaggtgtggcaacttttgttggcaagtaatggtaaagtgtggctgggaaccaaacagccccacaatttcattgacttgggtttaatgtcacactgcaccctcctatatctatctatactatcagggcacggcggccggagggaccacggagatcgtcgtggacggctcaaccggcctcctgcaccccgccgggaaggaaggcgtcgcgcccctcgcgaagaacatggtaaggctcgcgagccacgaagaggacagggtctccatgggtaggaaaggctatggcagggtgaaggaaatgttcatggagcaccacatggctggtaggatcgcggccgtgttgcaggaagttctgaagaaatcgcgggaacattctcactcttga</dnaseqindica>\|
−	~~Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001049188.1 RefSeq:Os01g0262600]\|~~
−	}}
−	[[Category:Genes]]
−	~~[[Category:Japonica mRNA]]~~
−	[[Category:Oryza Sativa Japonica Group]]
−	[[Category:Japonica ~~Genes]]~~
−	~~[[Category:Japonica Chromosome 1]]~~
−	~~[[Category:~~Chromosome 1]]

@@ Line 112: / Line 112: @@
 ==References==
-Please input cited references here.
 ==Structured Information==

@@ Line 108: / Line 108: @@
 ==Labs working on this gene==
 Crop Biotech Institute and Department of Plant Molecular Systems Biotechnology, Kyung Hee University, Yongin 446–701, Korea
 State Key Laboratory of
-Hybrid Rice, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
+State Key Laboratory of Hybrid Rice, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
 ==References==

@@ Line 107: / Line 107: @@
 ==Labs working on this gene==
-Please input related labs here.
+Crop Biotech Institute and Department of Plant Molecular Systems Biotechnology, Kyung Hee University, Yongin 446–701, Korea
 ==References==

← Older revision		Revision as of 05:55, 4 June 2014
Line 55:		Line 55:

	''Figure 4. TEM analyses of developing anthers from wild-type (A–C, G–I, and M–O) and OsGT1/osgt1-1 (D–F, J–L, and P–R) plants at stage 11a (A–F), stage 11b (G–L), and stage 12 (M–R). Ba, Bacula; BP, binuclear pollen; Cy, cytosol; DP, defective microspores; En, endothecium; Ep, epidermis; Ex, exine; In, intine; MP, mature pollen; Ne, nexine; Or, orbicule; SG, starch granule; Te, tectum. Bars = 10 mm (A, D, G, J, M, and P), 2 mm (B, E, H, K, N, and Q), and 0.5 mm (C, F, I, L, O, and R).''		''Figure 4. TEM analyses of developing anthers from wild-type (A–C, G–I, and M–O) and OsGT1/osgt1-1 (D–F, J–L, and P–R) plants at stage 11a (A–F), stage 11b (G–L), and stage 12 (M–R). Ba, Bacula; BP, binuclear pollen; Cy, cytosol; DP, defective microspores; En, endothecium; Ep, epidermis; Ex, exine; In, intine; MP, mature pollen; Ne, nexine; Or, orbicule; SG, starch granule; Te, tectum. Bars = 10 mm (A, D, G, J, M, and P), 2 mm (B, E, H, K, N, and Q), and 0.5 mm (C, F, I, L, O, and R).''
		+
		+
		+	'''OsGT1 Functions at a Late Stage of Pollen Development'''
		+
		+	Recent genetic and biochemical investigations have uncovered several key genes required for pollen wall development in rice. For example, UDP-glucose pyrophosphorylase1 (UGP1) catalyzes the reversible production of Glc-1-P and UTP to UDP-Glc and pyrophosphate. In Ugp1-silenced plants, callose deposition is disrupted during meiosis. Consequently, the pollen mother cells begin to degenerate at the early meiosis stage, eventually
		+	resulting in complete pollen collapse (Chen et al., 2007). This indicates that Ugp1 plays a very early role (stage 6) in anther development (Fig. 8). TAPETUM DEGENERATION RETARDATION (TDR), GAMYB, PERSISTENTTAPETAL CELL1 (PTC1), CYP704B2, DEFECTIVE POLLEN WALL (DPW), POST-MEIOTIC-DEFICIENT ANTHER1 (PDA1), OsC6, and WAX-DEFICIENT ANTHER1 (WDA1) are critical for exine development (Jung et al., 2006; Zhang et al., 2008, 2010; Aya et al., 2009; Hu et al., 2010; Li et al., 2010a, 2011; Shi et al., 2011). In their mutants, microspores display a defective exine layer. TDR and GAMYB are transcription factors that control lipid biosynthesis and metabolism. PTC1 is orthologous to MS1 and regulates timely degradation of tapetal cells as well as pollen exine development. CYP704B2 catalyzes the production of v-hydroxylated fatty acid with 16- and 18-carbon chains. DPW is a fatty acid reductase and produces 1-hexadecanol with higher specificity against the palmiltoyl-acyl carrier protein (Shi et al., 2011). OsC6 is a lipid transfer protein; expression of OsC6 is positively regulated by TDR. WDA1 is involved in the biosynthesis of cuticular waxes. Mutants defective in these genes start to show defects at stages 8 through 10 (Fig. 8). We observed that the defective phenotypes of osgt1 pollen began to appear at stage 11b, which is much later than the stages when the phenotypes became obvious in the exine-defective mutants. Mutationsin RICE IMMATURE POLLEN1 (RIP1), which encodes a WD40 protein, result in defects at late stage 11b (Han et al., 2006). Because the rip1 mutant develops sperm cells normally, and the mutant pollen grains are nearly normal at the mature stage, we conclude that RIP1 functions downstream of OsGT1 (Fig. 8).
		+
		+	[[File:image8.png]]
		+
		+	''Figure 8. Schematic diagram of rice pollen wall development based on stages described by Li et al. (2010). Genes involved inthis process are represented. Arrows indicate times when mutant phenotypes become visible.''

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

@@ Line 70: / Line 70: @@
 ===Evolution===
-Please input evolution information here.
+'''OsGT1 Encodes a Glycosyltransferase'''
 You can also add sub-section(s) at will.

@@ Line 57: / Line 57: @@
 ===Expression===
-Please input expression information here.
+'''Expression of OsGT1 Is High at Later Pollen Developmental Stages'''
 ===Evolution===

@@ Line 4: / Line 4: @@
 '''T-DNA Insertions in OsGT1 Cause a Defect in Male Gametophyte Development'''
 We previously generated transfer DNA (T-DNA)-tagged lines of japonica rice (Jeon et al., 2000; Jeong et al., 2002) and determined flanking sequences for the insertion sites (An et al., 2003; Jeong et al., 2006). To identify genes essential for gametophyte development, we genotyped the T-DNA insertion lines; those with a segregation ratio close to 1:1:0 (wild type:heterozygote: homozygote) were selected. From 541 independentlines, we obtained eight with the segregation distortion phenotype. Here, we report detailed analyses of line 3C-00590, which carries a T-DNA insertion in the fifth intron of LOC_Os01g15780 (Fig. 1A). Because the gene encodes a protein in the glycosyltransferase family, we named it OsGT1. We found that another allele (osgt1-2) also harbors T-DNA in the fourth exon of OsGT1 (Fig.1A). This line also exhibited a 1:1:0 segregation (Table I;Supplemental Fig. S1), confirming that this phenotype is due to the T-DNA insertion to OsGT1.
 These segregation phenotypes suggested that the defect results from a failure in gene transfer through either the male or female gamete. To clarify the function of OsGT1 in the gametophyte, we performed reciprocal crosses between heterozygotes (OsGT1/osgt1-1) and the
@@ Line 9: / Line 10: @@
 '''osgt1 Grains Are Defective at the Mature Pollen Stage'''
 To examine further the morphological defects in male gametes of osgt1-1, we collected pollen grains from fully developed anthers and observed them with a bright field microscope. Pollen from wild-type anthers was normal: only a small fraction (nine out of 1,393) was shrunken at stage 12 (Fig. 1B; Supplemental Fig. S2). However, anthers from the heterozygote produced shrunken grains at a 49.5% frequency (915 of 1,847; Fig. 1C; Supplemental Fig. S2), indicating that OsGT1 is required for male gamete development. To determine whether those shrunken grains were caused by the T-DNA insertion, we used a Suc density gradient to separate them from normal grains (Bedinger and Edgerton, 1990; Fig. 1D). The gradient containedfour discrete bands, two lower bands that contained normal pollen with a globular shape (Fig. 1, E and F) plus two upper bands with defective grains that were shrunken or underdeveloped (Fig. 1, G and H). To see whether this defect was due to the mutation, we prepared DNA from each band and performed PCR with gene-specific primers. This resulted in amplification of the OsGT1 fragment from the two lower bands but not from the upper bands (Fig. 1I). Amplification with a gene-specific primer (R2) and a T-DNA primer (L1) did not generate bands from any of the four samples, indicating that normal pollen in the lower bands did not carry T-DNA and that abnormal pollen in the upper bands lacked genomic DNA because they were dead (Fig. 1J).
 To investigate the exact time when this mutant pollen began to show the phenotypic defect, we compared wild-type and osgt1 pollen at five stages of development (Fig. 2). At the young microspore stage (stage 9), cells from the wild type were released as free microspores from the tetrad (Fig. 2A). These gradually swelled and underwent vacuolation, causing the cytoplasm and nuclei to be distributed in the periphery (Fig. 2B). Afterward, the pollen started to divide mitotically (stage 11a; Fig. 2C; Zhang et al., 2011). Until this stage, all gametophytes in the OsGT1/osgt1-1 anthers appeared normal and were identical to those of the wild type (Fig. 2, F–H). However, at stage 11b, two different types of pollen were observed from the heterozygous anthers: normal grains showing even cytosolic staining and empty pollen with a lumped staining
@@ Line 14: / Line 16: @@
 We tested pollen viability with fluorescein diacetate, which stains only live cells (Heslop-Harrison and Heslop- Harrison, 1970).Whereas globular pollen emitted a bright green signal, no signal was observed from the collapsed grains from mutant plants (Supplemental Fig. S3). These
 findings indicated that osgt1 pollen cannot form normal mature grains.
 '''osgt1 Pollen Is Defective in Intine Formation'''
 To investigate the defects in osgt1 pollen further, we stained the mature grains with auramine O, which binds to exine (Dobritsa et al., 2011). This revealed that the exine was not markedly different between mutant and wild-type pollen (Fig. 3, A–D; Supplemental Fig. S4). We also monitored for the presence of intine via calcofluor white staining.Whereas defective pollen from heterozygous plants exhibited a very weak signal, wildtype pollen emitted bright blue fluorescence, demonstrating that the mutant pollen was defective in intine (Fig. 3, E–H; Supplemental Fig. S4).
 Staining with 496-diamidino-2-phenylindole (DAPI) showed that the defective pollen did not carry any nuclei at stage 12, whereas wild-type grains contained one vegetative nucleus and two generative nuclei (Fig. 3, I–L). Because mitotic division precedes intine construction in rice (Lu et al., 2002; Lin et al., 2009), we performed DAPI staining at stage 11, when the binuclear grains are formed. All pollen from the heterozygous plants contained two nuclei (Fig. 3, N and P), suggesting that osgt1 pollen underwent normal mitosis I. Furthermore, amido black staining showed that osgt1 grains did not accumulate cytosolic contents such as proteins (Supplemental Fig. S5), supporting our belief that the pollen abnormality developed later in the mutant.
 To understand the subcellular changes in osgt1 pollen, we performed transmission electron microscopy (TEM) analysis at stages 11a, 11b, and 12 (Zhang and Wilson, 2009). As revealed via bright-field microscopy, wild-type and osgt1 pollen did not differ significantly at stage 11a (Fig. 4, A–F). At the end of this stage, exine formation is generally considered complete (Lu et al., 2002; Lin et al., 2009). We noted a complete exine structure for both the wild type (Fig. 4, A–C) and osgt1 (Fig. 4, D–F) at stage 11a. However, differences were observed at stage 11b. Whereas wild-type grains had evenly stained cytoplasm (Fig. 4, G and H), mutant pollen showed severely degraded cytoplasm (Fig. 4, J and K). At higher magnification, intine accumulation was observed in wild-type pollen (Fig. 4I), but the intine was defective in osgt1 grains (Fig. 4L). At stage 12, these differences were more apparent. Whereas wildtype grains had accumulated starch granules as well as a thick intine (Fig. 4, M–O), approximately 50% (72 of 129) of the pollen from OSGT1/osgt1 anthers had an altered cytoplasm density and a severe defect in intine accumulation (Fig. 4, P–R). Because intine development precedes the accumulation of starch grains, proteins, and other inclusions in the cytosol (Lu et al., 2002; Lin et al., 2009), we speculated that OsGT1 plays a key role in the former process.
-[[File:rice1]]
 ===Expression===
 Please input expression information here.

@@ Line 1: / Line 1: @@
-Please input one-sentence summary here.
+OsGT1 is a Golgi-localized glycosyltransferase essential for intine construction and pollen maturation, providing new insight into male reproductive development.
 ==Annotated Information==
 ===Function===

← Older revision		Revision as of 14:37, 2 June 2014
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	==Annotated Information==		==Annotated Information==
−	~~OsGT1 is a Golgi-localized glycosyltransferase essential for intine construction and pollen maturation, providing new insight into male reproductive development.~~
	===Function===		===Function===