Os10g0471100

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Annotated Information

Function

Wax-deficient anther1 (Wda1), a rice homolog of Arabidopsis CER1, is a male-sterile mutant of rice, that was generated through T-DNA insertional mutation, and is preferentially expressed in epidermis of various floral organs, including Anthers. This mutant shows morphological changes in its pollen and anther walls as well as alterations in its wax composition. Wda1 mutant shows defects in the biosynthesis of very-long-chain fatty acids in both layers of anthers. Scanning electron microscopy analysis shows that epicuticular wax crystals were absent in the outer layer of the anther and that microspore development was severely retarded and finally disrupted as a result of defective pollen exine formation in the mature anthers. Biochemical analysis of the wda1 mutant anthers showed that the levels of very-long-chain alkanes, alkens, fatty acids, and primary alchols are severely reduced.

Expression

Wda1 is strongly expressed in the epidermal cells of rice anthers. Wda1 transcript is abundant during flowering stages.Because the GUS gene located in T-DNA was fused to Wda1, the level and pattern of expression of the fusion gene could be assayed for GUS activity. Experiments have showed temporal and spatial expression patterns of the Wda1-GUS fusion product in heterozygous spikelets at various flowering stages(Fig.1). High levels of GUS activity were observed in spikelets from the premeiosis stage to the heading stage (Fig. 1A and 1K ). GUS activity was also prominent in the outer and inner epidermis (Fig. 1H) and trichomes (Fig. 1G) of the palea/lemma, lodicules (Fig. 1C, 1E, 1F, and 1I), stigmas (Fig. 1C), and anthers (Fig. 1C, 1D, 1I, and 1J). Cross sections of heterozygotic anthers at the vacuolated pollen stage revealed that GUS was expressed strongly in the epidermis but only weakly in the tapetum, endothecium, and connective tissue (Fig. 1J). In vegetative tissue, GUS activity was mostly absent except at collar regions (between the leaf sheath and the leaf blade) and the base of shoots. Semiquantitative RT-PCR analyses have been performed to examine whether GUS activity truly represented Wda1 expression patterns. Wda1 was highly expressed in panicles at various developmental stages (Fig. 1K) but only very weakly in the roots and shoots of 7-d-old seedlings. Only low transcript levels were detected in the developing seeds and flag leaves. In developing spikelets, transcript was detected in all floral organs (Fig. 1K and 1L), but it was barely detectable in the wda1 anthers (Fig. 1L). LOC_Os02g40780, which is the most similar rice gene to Wda1, was also expressed throughout panicle development. However, its expression was much higher in the shoots and flag leaves. In mature spikelets, transcript levels were low in the anthers and palea/lemma and barely detectable in the ovaries and lodicules (Fig. 1L). These results indicate that Wda1 functions primarily in the panicles, especially in the reproductive organs, whereas LOC_Os02g40780 may play roles in both vegetative and reproductive organs.

Figure 1. Expression Profiles of Wda1 by GUS Assay and RT-PCR.

Evolution

The Wda1 Gene derived from a T-DNA Insertional Mutation. The transcript of the wda1 gene is 2337 bp long, including an 1866-bp coding sequence. The locus name of this gene has been registered as OSJNBa0079L16.17 in GenBank (http://www.ncbi.nlm.nih.gov) and as LOC_Os10g33250 in The Institute for Genomic Research database (http://tigrblast.tigr.org/euk-blast/). The primary structure of the gene comprises 10 exons and 9 introns (Fig. 2A ). The T-DNA insertion was located at 311 bp downstream from the ATG start codon, in the first intron of the gene. Multiple alignments of plant proteins showing high similarity with the WDA1 protein(Fig. 2B). WDA1 (top line) was aligned with three rice proteins (LOC_Os02g56920, LOC_Os02g40780, and Os Wax2-like [LOC_Os09g25850]) and four Arabidopsis proteins (CER1, CER1-like, At2g37700, and WAX2). Black background indicates amino acid residues that are >70% conserved, and gray background indicates amino acids that are >30% conserved. Asterisks indicate perfectly matched amino acids, colons indicate highly conserved amino acids, and dots indicate low-matched amino acids among these eight proteins. Phylogenic analysis of proteins similar to WDA1 suggests that WDA1 is likely involved in VLCFA metabolism.

Figure 2. Scheme of the Wda1 Gene, Multiple Alignments, and Phylogenic Analysis of WDA1-Related Proteins.

Labs working on this gene

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References

<references> <ref name="ref1">Ki-Hong Jung,Min-Jung Han,Dong-yeun Lee,et al.Wax-deficient anther1 Is Involved in Cuticle and Wax Production in Rice Anther Walls and Is Required for Pollen Development.The Plant Cell,2006,18(11):3015-3032.

Structured Information

Gene Name	Os10g0471100
Description	Sterol desaturase family protein
Version	NM_001071361.1 GI:115482465 GeneID:4348869
Length	5861 bp
Definition	Oryza sativa Japonica Group Os10g0471100, 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	Chromosome 10
Location	Chromosome 10:17906207..17912067
Sequence Coding Region	17906494..17906655,17906746..17906922,17907019..17907117,17907202..17907402,17907731..17907838 ,17907926..17908307,17908870..17909089,17910772..17911004,17911137..17911360 ,17911825..17911884
Expression	GEO Profiles:Os10g0471100
Genome Context	<gbrowseImage1> name=NC_008403:17906207..17912067 source=RiceChromosome10 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008403:17906207..17912067 source=RiceChromosome10 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atggccacaaacccaggactcttcacggagtggccatggaagaagcttggcagcttcaagtacgtgttgctggcgccgtgggtagcgcacgggtggtatgaggtggcgaccaaggggtggcgggaggtggatctcgggtacatcgccatcctcccgtcgttgctgctgcggatgctgcacaaccaagcctggatcaccatctcccgcctccaaaacgcccgtggccgccgccagattgttcgccgcggcatcgagttcgaccaggttgaccgcgagaggaactgggacgaccagatcatcctgagcggtatcctactatacctcggcgcactgtacgtaccgggcgggcaacacttaccgctgtggaggacggatggcgcggggctgatcgccttgctgcatgcggggccagtggagttcctctactactggttccatcgtgcgttgcaccaccactttctctacacccactaccactcgcaccaccactcttcaatcgtcactgagcccatcacatccgtcatccatccgtttgctgagctcgttgcctacgaattgctcttctcgatcccactgatcgcctgtgccttgactggaactgcttccataattgcattcgagatgtatttgatttacattgatttcatgaacaacatggggcactgcaacttcgagttggtacccagctggctcttcacatggttccctcctctcaagtatctcatgtacacgccatcgtttcattctctgcaccacactcagtttcggacaaactactcccttttcatgccgttctacgattatatctacaacaccatggacaagtcatcagacactctgtatgagaactcactgaaaaacaatgaggaagaagaagcagtggatgtggttcaccttacacacctgaccaccctgcattccatctaccacatgaggcccggtttcgccgaatttgcgtccaggccctacgtttccaggtggtacatgaggatgatgtggcctctgtcatggctctccatggtgctgacatggacgtacggttcttcattcactgttgagaggaatgtcatgaagaagatcaggatgcagtcatgggccataccaagatacagtttccattatggattggattgggagaaggaagctatcaatgatcttatcgaaaaggcggtatgtgaagctgacaagaatggggctaaagttgtaagccttggactcctgaatcaggcacataccctgaataaaagtggagaacaatatctgctgaaatacccaaagttgggagcaagaattgtcgatggaaccagcttagctgctgcagtggttgtcaacagtatcccccaaggcacagaccaagtaattcttgcaggaaatgtttccaaggtggcgcgtgctgtagcgcaagcattatgcaagaaaaatatcaaagtcacgatgacaaacaagcaagattatcatttactcaagccagaaataccagaaactgtagctgacaatctttcgttttcgaaaacaggcaccgcaaaggtttggttaattggtgatggtctagattctgctgaacaattcagagcacagaaaggaactctgtttatcccatactcacaatttcctccaaagatggtacgcaaggacagctgcagctactcgacaactcctgcaatggctgtaccaaaaacgctgcagaatgtgcattcatgcgagaattggctgccaaggagggttatgagcgcatggcgaatcgcgggaattcttcatgcgttggagggatggaatgagcatgaatgtggtgacaaggtgcttgatatggacaaagtttggtctgctgcaattatgcatgggttctgcccagttgctcaaggttga</cdnaseq>
Protein Sequence	<aaseq>MATNPGLFTEWPWKKLGSFKYVLLAPWVAHGWYEVATKGWREVD LGYIAILPSLLLRMLHNQAWITISRLQNARGRRQIVRRGIEFDQVDRERNWDDQIILS GILLYLGALYVPGGQHLPLWRTDGAGLIALLHAGPVEFLYYWFHRALHHHFLYTHYHS HHHSSIVTEPITSVIHPFAELVAYELLFSIPLIACALTGTASIIAFEMYLIYIDFMNN MGHCNFELVPSWLFTWFPPLKYLMYTPSFHSLHHTQFRTNYSLFMPFYDYIYNTMDKS SDTLYENSLKNNEEEEAVDVVHLTHLTTLHSIYHMRPGFAEFASRPYVSRWYMRMMWP LSWLSMVLTWTYGSSFTVERNVMKKIRMQSWAIPRYSFHYGLDWEKEAINDLIEKAVC EADKNGAKVVSLGLLNQAHTLNKSGEQYLLKYPKLGARIVDGTSLAAAVVVNSIPQGT DQVILAGNVSKVARAVAQALCKKNIKVTMTNKQDYHLLKPEIPETVADNLSFSKTGTA KVWLIGDGLDSAEQFRAQKGTLFIPYSQFPPKMVRKDSCSYSTTPAMAVPKTLQNVHS CENWLPRRVMSAWRIAGILHALEGWNEHECGDKVLDMDKVWSAAIMHGFCPVAQG</aaseq>
Gene Sequence	<dnaseqindica>5413..5574#5146..5322#4951..5049#4666..4866#4230..4337#3761..4142#2979..3198#1064..1296#708..931#184..243#atatgagactctgacacgtacgcacagctagatcgaagccctggtcgagtttataaattgagatatatccgtcctccattctcaggccagttcttgcaagactcttgccgtgttgagactgcagaaacgagagctagctagccagaggctgcaagaagattctgcacccatagatcgtctaccatggccacaaacccaggactcttcacggagtggccatggaagaagcttggcagcttcaaggtaatcaaaattaaccttccatcttcttctccttaattcagttacttcactgtcttctggcagctcgatcgagcaaataaacgcacagcttctagttccatccatggcagcactgcaccatgaccattatgttcttgtttgagttcgctagcttctgtttgatatcaaagcatacattgccgcatgtgcagctagcagtgcatgagattaattctcttgcatctgctgcagatcagtaatgcaggggcatgaactttcctgcagcacctctgtgcagcaaataaactgttaaagagtacatggttaaataagcttttgccacatcgtctttgattaggcaaaaacattcagtatacacaacttccgacctttacaaattacaacagtaacttcagctataaattatagcacatgtgagttaattcgatggatggattcatgcatgtatatgtgtgcatatgcagtacgtgttgctggcgccgtgggtagcgcacgggtggtatgaggtggcgaccaaggggtggcgggaggtggatctcgggtacatcgccatcctcccgtcgttgctgctgcggatgctgcacaaccaagcctggatcaccatctcccgcctccaaaacgcccgtggccgccgccagattgttcgccgcggcatcgagttcgaccaggttgaccgcgagaggaactggtatgccaaacaccataaaattacgtacaacccacgcggtgtgttcgtggcggctagcttcaccttcgagattaattaattcaacctcattatagttgatgtgttgttacgtgtatggtggtgtgcgtgcagggacgaccagatcatcctgagcggtatcctactatacctcggcgcactgtacgtaccgggcgggcaacacttaccgctgtggaggacggatggcgcggggctgatcgccttgctgcatgcggggccagtggagttcctctactactggttccatcgtgcgttgcaccaccactttctctacacccactaccactcgcaccaccactcttcaatcgtcactgagcccatcacatgtacgtaattaattgccactagagacggtttatcctcgaggctccaaacattacattctcttttaggcaataacctttgagcttctatattagtatcgaattgtgttgcgtcataaatatttttacaaaatttttattcatcatctaaagttatcaatgatcaaagttcaatcaatactaatagttacaagtaatttgaaatagaggaagtaagttgtactacctagttgtaccatcattgctccaaacaacctattacatgatattactttcgtcccaaacctctaaatgaatattaagattttggtaatacaaattcttaaaattttgagacagaaatattactagacctgtaaccttgggataaattaaaccacggtaatcttccaatagaacaaagtaaaaaacaagtcttttccataaggacacaaaacaggaggtgtacttcgtcagatgcaagtgtcgcgcgtgtccgttcctcaggaaacacagagtgttcctgtggccagtcgatcgctacttcaattccctcctccttcaatgcatgcctgaacaataattgtaggcctagagagtgtgtgagttaaggaatgcaaaggacacacgatatttcatggaccattgagatcgaagtaagccgaccgatcgatcgcgagctaaatttatgtagcccatcgaatccgttgctgaatgcctgaccaaacggagaagcaaacgtggtccatgaacaacgtatcagagacttttagacaagagagaaatcacattctaaggggttttttcccccttttttttgagtggaagagttttttttattgagaggtatcgagaagtactatatttttcgtgtaagctagccaaatatcagtgcgttgcaacttgcaacggattattaaaaatagcaaaattaagtccttgttaaattttttttaaaaaaaaatggttgaaacacaacttttcatcttaaagaaacaaacaaaaatgccatgcttagaaaattaaagagctaaagtatagtacagtatatatagtacaggcgactctcccaaaccttaataccatggcccatatatcagtgacccaggccctgggcaaactggtccacggcccagggcctgagatgaaaggcccgatgagaaatatgcatttttactaggaataagtctatttttcctcctttcaaccatgtaactttgtgttgggccaaaatttccatggggcggcctaggccttggcctatttctgggtccgccccttatcagtgattgtggtgatggcagattcactaccaccactactatcttttaatagtagtaaagattttggtgtcatcttgtaattttaagatgtacaagacggtaccataaattgactttaaagtgatttgaatggatgtgtaatgggagaaatcgcttttcaaatgtaaaataatgcactacttgtattagtactaaattttacattagagaacacgatatattcagtgattcttgataaccataaaatccctttaagattatggtgttttctttaagtactgttaaaaaaaaaccaatagtgtctactggttacactgcagaagaagagaaccctgcagactgcagtatttgtaattgagcaagcagatacatatacctaaatgcatatatgttcattacctacaattcctaacgttgttgcagccgtcatccatccgtttgctgagctcgttgcctacgaattgctcttctcgatcccactgatcgcctgtgccttgactggaactgcttccataattgcattcgagatgtatttgatttacattgatttcatgaacaacatggggcactgcaacttcgagttggtacccagctggctcttcacatggttccctcctctcaagtatctcatgtacacgccatcgtaagtacttcatcaggctcttaataatacttccttcttccttaaatgtttgacgccgttgacttttttaaacatgtttgaccgttcgtcttattcaaaaacttttgtgaaatgtgtaaaactatatgtatacataaaagtatatttaacaataaatcaaatgatagaaaaagaattaataattatttaaatttttgaataagacgaacggtcaaatatttttaaaaaagtcaatggcatcaaatattttgggatggagggagtatgtaaaagagtaactttacagtttttaagaaagcataggggaagtaccatattttatattggagaaattttacggttttcagtatttcttttaagataaaggaggctttattgatcttggatacaatatactctaaaaccactcctgatatctgcatagctaggatgcacatggtctatagttattgaaaaagccaaataaaagaagatgatgtgcatgtcactctgcaaaataaataacttgatcatacatgctgttttcactgttataataatctgaacttatgtgatatattaggtttcattctctgcaccacactcagtttcggacaaactactcccttttcatgccgttctacgattatatctacaacaccatggacaagtcatcagacactctgtatgagaactcactgaaaaacaatgaggaagaagaagcagtggatgtggttcaccttacacacctgaccaccctgcattccatctaccacatgaggcccggtttcgccgaatttgcgtccaggccctacgtttccaggtggtacatgaggatgatgtggcctctgtcatggctctccatggtgctgacatggacgtacggttcttcattcactgttgagaggaatgtcatgaagaagatcaggatgcagtcatgggccataccaagatacagtttccatgtaagagtaaccatgttcagtgcatgattcacagatggtgtgcatgataaattgataatcaggttagaaacattttgttttgtgcagtatggattggattgggagaaggaagctatcaatgatcttatcgaaaaggcggtatgtgaagctgacaagaatggggctaaagttgtaagccttggactcctgaatcaggtagctcatttcctaatgaattcagagcaagttgaaacattatcatccatacttcagaattcagatgttagctatatgtccacatgtactgccaaattttagtcaagataaaacagttagtgcttaataattgtttatcttcagggaaaaggtttagtcataaaggtaggaaaataaaaaaattaagtagcaaagagtccgttgacagtagtccaacagaactaataaattctgtacatttcatgagaatacaatatgtagaaattagggaagtatgtactcatgcatggttgaactggtaattgacatacaatatcttcaatataaggcacataccctgaataaaagtggagaacaatatctgctgaaatacccaaagttgggagcaagaattgtcgatggaaccagcttagctgctgcagtggttgtcaacagtatcccccaaggcacagaccaagtaattcttgcaggaaatgtttccaaggtggcgcgtgctgtagcgcaagcattatgcaagaaaaatatcaaagtaattaatccactactcacctctgatttattttatagcatgcaaaaggataccatgttgtaacaaaccactattcttgtacaggtcacgatgacaaacaagcaagattatcatttactcaagccagaaataccagaaactgtagctgacaatctttcgttttcgaaaacaggcaccgcaaaggtaagaatagatttcctcatttttcctacattctctggacaaatgaatagaatgtataagagaaatatcattgcctaataaaccaaaaattttcaggtttggttaattggtgatggtctagattctgctgaacaattcagagcacagaaaggaactctgtttatcccatactcacaatttcctccaaagatggtacgcaaggacagctgcagctactcgacaactcctgcaatggctgtaccaaaaacgctgcagaatgtgcattcatgcgaggtagtagtaataacctgttgaaataatccagagattcagacgaacttcacatgatctaatcacaaaatttttgaactttttttttgacagaattggctgccaaggagggttatgagcgcatggcgaatcgcgggaattcttcatgcgttggagggatggaatgagcatgaatgtggtgacaaggtgcttgatatggacaaagtttggtctgctgcaattatgcatgggttctgcccagttgctcaaggttgatgattcagttgcaaagtatcagaataagttttagctggattatgctatgtggtcaccgtatcagaaagagagatgttctgttggcaagaagagatatagtatctgagtaatttcatctgccatggtttttaaaacattgttcatcatctcctttgatcaaagactggccatggctactttctgtataagtatctgtcctttttagtccatagtgtaattcagcaacaaaacactcttggcattgttatgcagtctcatgaatgtttttccagcatacggaattacgg</dnaseqindica>
External Link(s)	NCBI Gene:Os10g0471100, RefSeq:Os10g0471100