Os06g0604000

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Annotated Information

The ethylene response factor (ERF) gene WIN1/SHN1 was shown to transcriptionally activate the expression of wax synthesis genes and wax production, resulting in drought tolerance in Arabidopsis. To further investigate the regulation of ERF proteins in rice, we started with homology search for rice genes in the National Center of Biological Informatics using the Arabidopsis WIN1/SHN1 as a probe and found 4 rice orthologues with high amino acid identities in N-terminal region. These rice WIN1-like genes, named rice wax synthesis regulator (OsWR), all encode ERF proteins that contain a basic N-terminal region that might function as an nuclear localization signal and an acidic C-terminal region that might act as an activation domain for transcription. OsWR2 (Os06g0604000) is one of them.

Function

The Wax Synthesis Regulatory 2 gene (OsWR2) in rice (Oryza sativa L.) is highly expressed in epidermal tissues and contributes to the transcriptional regulation of both cuticular wax and cutin biosynthesis in rice cuticle. Overexpression of OsWR2 in rice increased the total cuticular wax level by 48.6%in leaves and by 72.4% in panicles. Of the major wax classes, aldehydes increased most in leaves, and alkanes increased most in panicles. Total cutin amounts were increased by 48.1 % in leaves and 65.9 % in panicles of rice overexpressing OsWR2, and these increases were due primarily to the increase inω-OH and di-OH acids. Our results showed that 19 genes previously associated with wax and cutin biosynthesis were up-regulated in OsWR2 overexpressors. Overexpression of OsWR2 also altered cuticular wax crystallization and cuticlemembrane ultrastructure. Furthermore, OsWR2 overexpression in rice decreased leaf chlorophyll leaching rate, reduced water loss rate, and enhanced tolerance to water-limited conditions. We demonstrate in this report that OsWR2 regulates wax and cutin biosynthesis differently than does the OsWR1 homologue, and plays a major role in controlling cuticle permeability. The increased resistance to water deficit conditions by OsWR2 overexpression in rice elucidates a potential new strategy for genetic improvement of plant drought tolerance.

Expression

Fig-1 OsWR2 expression profile in different organs of wild type (WT) Nipponbare and under different stresses. a: Semiquantitative RT-PCR analysis of OsWR2 expression profile in different organs of Nipponbare. R Root, C culm, L leaf, Sh sheath, P panicle, S seed, Sl seedling. OsActin1 gene was used as a control. b: Histochemical staining of GUS activity in pOsWR2::GUS transgenic rice plants. Bars: Root 80 μm; sheath, leaf blade 10 μm; culm, stamen 0.5 mm; anther 80 μm; glume, seed 2mm; lemma 1 μm. c: Semiquantitative RT-PCR analysis of OsWR2 expression profile in different stress treated Leaves. NT, Untreated Nipponbare. OsActin1 gene was used as a control

RT-PCRanalysis of the OsWR2 expression in different organs of WT Nipponbare showed that OsWR2 expression was high in culm, leaf blade, leaf sheath, panicle and seedling, but low in root and very low in seed (Fig. 1a). Further OsWR2 expression analysis using the pOsWR2:GUS reporter construction revealed ubiquitous expression of OsWR2 in the epidermis of aerial parts of transgenic rice (Fig. 1b). In roots, GUS activity was detectedmainly in the exodermal and endodermal cells, whereas no expression was detected in cortical cell and vascular cylinder. In leaf sheath and blade, OsWR2 was expressed through the entire leaf epidermis, most strongly in trichomes and guard cells of the stomatal apparatus. In the cross section of culm,GUS staining was observed specifically in endodermal cells, with no blue staining in epidermal or vascular bundle cells. In glumous flower, GUS staining was restricted to the anther of the stamens, the paleas and lemmas, and especially in the trichomes of paleas and lemmas. However, no expressionwas found in the filaments of stamens and any part of the pistil including stigmas, styles and ovaries. When anthers were observed by phase-contrast microscopy, only the microspores showed GUS staining. In addition, intense GUS staining was observed in the seed embryo (scutellum, epiblast, plumule and rachilla) with only faint staining in starchy endosperm (Fig. 1b). The ubiquitous expression pattern of OsWR2 suggests that OsWR2 is expressed in a variety of tissues, intimating that cuticle plays a role in diverse developmental processes in rice. For analyzing the responses of OsWR2 to abiotic stresses, 3-week-old rice seedlings were treated with water deficit in the potting medium for 5 days or with 150mMNaCl for 8 h. Semiquantitative RT-PCR analysis showed that both water deficit and NaCl treatments enhanced the expression of OsWR2 (Fig. 1c), suggesting that OsWR2 mayplayan important role in plant response to stresses. OsWR2 expression was increased more under dehydration treatment than NaCl treatment.

Evolution

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Labs working on this gene

Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Hunan Agricultural University

References

Overexpression of Transcription Factor OsWR2 RegulatesWax and Cutin Biosynthesis in Rice and Enhances its Tolerance toWater Deficit

Structured Information

Gene Name	Os06g0604000
Description	Similar to Ethylene response factor 1
Version	NM_001064549.2 GI:297606135 GeneID:4341466
Length	1563 bp
Definition	Oryza sativa Japonica Group Os06g0604000, 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 6
Location	Chromosome 6:24775260..24776822
Sequence Coding Region	24775899..24775916,24776044..24776275,24776508..24776590
Expression	GEO Profiles:Os06g0604000
Genome Context	<gbrowseImage1> name=NC_008399:24775260..24776822 source=RiceChromosome06 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008399:24775260..24776822 source=RiceChromosome06 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atgggacagtcgaagaagaagttccgcggagtcaggcagcgccactggggctcctgggtctccgagatcaggcaccctctccttaagaggagggtgtggctgggtacctttgagacggcggaggaggcggcgcgggcgtacgacgaggccgccatcctgatgagcggccgcaacgccaagaccaacttcccagtcgcgaggaacgccacgggggagctcacaccggcggctgcggtggcagggcgggatggccgtgtcggcggcggcagcggcagctcgtcctcaatgacggccaacggcggcgggaacagcctgactccagctgggtcatga</cdnaseq>
Protein Sequence	<aaseq>MGQSKKKFRGVRQRHWGSWVSEIRHPLLKRRVWLGTFETAEEAA RAYDEAAILMSGRNAKTNFPVARNATGELTPAAAVAGRDGRVGGGSGSSSSMTANGGG NSLTPAGS</aaseq>
Gene Sequence	<dnaseqindica>907..924#548..779#233..315#attgaacgcacgctacactgcaagaaaggcaagtataggggtggcaggtagtttatattcactggaaaccgagaacttcactcctcccagcactccacactaggaggatgctcactgtctgactgactgactggctgctgctgctgctcattccttctcctactctgcctgcctcctctgctgtgtccttgggagaattcttcagggacagtatccctgcagaggtgagatcatgggacagtcgaagaagaagttccgcggagtcaggcagcgccactggggctcctgggtctccgagatcaggcaccctctcctgtaagtctctatctcactagtgatgatcactagctagctatgttgctctgatgtgttatgccactgtcgtgatagatacattttgatggatagctagtggttctggtttgttgttgctttctttctttttttctttctctttcatggtatgttagtagctaactcaaggtgcttgggtgcgtgcatgcatgaatgatggatgatgttttttttgctacgtgcgttgtggcagtaagaggagggtgtggctgggtacctttgagacggcggaggaggcggcgcgggcgtacgacgaggccgccatcctgatgagcggccgcaacgccaagaccaacttcccagtcgcgaggaacgccacgggggagctcacaccggcggctgcggtggcagggcgggatggccgtgtcggcggcggcagcggcagctcgtcctcaatgacggccaacggcggcgggaacagcctgtctcagatcctcagcgccaagctccgcaagtgctgcaagacgccgtcgccgtcgctcacctgcctccgccttgacccggagaagtcccacattggcgtctggcagaagcgcgccggcgcacgcgctgactccagctgggtcatgaccgtcgagctcaacaaggacacggccgtgtcgtcggctgcgacggtggcagcagcaacagcagtgtcgtccagcgaccagccgactccgagtgacagcacagtcacaacgacgtccacgtccaccacgggctcgccgtcgccaccacctccggcaatggacgacgaggagaggatcgcgctgcagatgatcgaggagctgctgggcaggagcggcccgggctcgccgtcacatgggctgctgcacggtggtgaaggtagcctcgtcatctgaattccggaagaacaggaaagaaattgaaaatgcaaggttaaaacagcatgatcaggtcaccatctaagatcaaggatctggtagggtggttggtgcacaggcagttaagattgctacatatgataggtatatctctattactactacatatccagcttaattaggaaacaattataagatcgattactactgtgtgaagtgaagctgtgtatttattaaagacttacttgtatgtacaagacgtccccgtcattatagtcatactggtgaaagctctgctatgtatcaacgtcatcagagatcagactacatatatataagtggtgtcaacttctaataaaagtaaataaaaacattgtaattgtgt</dnaseqindica>
External Link(s)	NCBI Gene:Os06g0604000, RefSeq:Os06g0604000