Os06g0181700

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Annotated Information

Function

ERF proteins, Arabidopsis WIN1/SHN1 and Medicago WXP1/2, have been shown to induce the production of epidermal waxes when overexpressed in plants.

The WIN1-like OsWR1 controls wax formation by direct or collaborative activation of wax synthesis genes, and this regulation, distinct from its homology regulator of WIN1/SHN1 in cutin synthesis, subsequently results in reduced water loss and enhanced drought tolerance.

To investigate the functional role of this region,Youhua Wang fused the coding regions of OsWR1 and its deletion of the predicted acidic activation region to the DNA-binding-domain expres- sion vector of GAL4 and examined the behavior of GAL4– OsWR1 and GAL4–OsWR1–DAD constructs as potential transcriptional activators in yeast. In the presence of the activation domain, the OsWR1 protein fused to the DNA- binding domain activated transcription of the lacZ reporter gene, whereas GAL4–OsWR1–DAD could not, indicating that the OsWR1 protein may act as a transcriptional activator.

Datas reveal that OsWR1 is responsive to drought, salt and ABA in rice.

OsWR1 is a positive regulator for drought response through affecting water loss.

OsWR1 might participate in the regulation of oxidative stress response and membrane stability, and this regulation might be a compensation for the modulation of OsWR1 in drought response.

OsWR1 that at least partially directly activates the expression of wax synthesis related genes to enhance wax production, through the regulation of the components of wax biosyn- thesis. This indicate that OsWR1 is a positive regulator of wax synthesis in rice.

OsWR1 modulates the expression of wax biosynthesis related genes in rice and OsWR1 physically interacts with the GCC box and DRE in the wax synthesis related gene promoters.

Expression

To test nuclear localization, an in vivo targeting experiment was performed by fusing the coding region of OsWR1 to GFP, and the resulting constructs (WR1–GFP and GFP only) were introduced into rice calli by agroinfiltration. These fusion proteins, under the control of the CaMV 35S promoter, were expressed in rice callus cells,and the localization of the fusion protein was determined by visualization with a fluorescence microscope. The result indicates that WR1–GFP was localized primarily in the nucleus, while GFP-only was observed in cytosol.

Real-time PCR (Q-PCR) amplifications showed that the expression of OsWR1 was quickly responded at 1 h, and then gradually increased to peak at 2.5 h after drought treatment. The transcripts of OsWR1 induced by ABA treatment were clearly observed at 6 h, and reached max- imum induction at 9 h and then decreased, whereas the induction of OsWR1 by salt treatment was dramatically increased at 3 h and reached a maximum at 6 h, and sustained high induction thereafter.

Evolution

One of the rice WIN1-like genes, OsWR1(Os02g0202000), encodes a protein of 205 amino acids with a predicted protein molecular weight of 22.45 kDa. The amino acid of full length OsWR1 shows a high similarity to WIN1/SHN1 (68.8%), SHN2 (58%), SHN3(57.8%) and OsWR2 (Os06g0604000, 81%), but low similarity to OsWR3 (Os02g0797100, 41.4%).these all contain the conserved ERF domain , suggesting that OsWR1 is a homology of WIN1/SHN1.

References

Youhua Wang;Liyun Wan;Lixia Zhang;Zhijin Zhang;Haiwen Zhang;Ruidang Quan;Shirong Zhou;Rongfeng Huang An ethylene response factor OsWR1 responsive to drought stress transcriptionally activates wax synthesis related genes and increases wax production in rice Plant Molecular Biology, 2012, 78(3): 275-288

Aharoni A, Dixit S, Jetter R, Thoenes E, van Arkel G, Pereira A (2004) The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis. Plant Cell 16:2463–2480

Broun P, Poindexter P, Osborne E, Jiang CZ, Riechmann JL (2004) WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis. Proc Natl Acad Sci USA 101:4706–4711

Jung KH, Han MJ, Lee DY, Lee YS, Schreiber L, Franke R, Faust A, Yephremov A, Saedler H, Kim YW, Hwang I, An G (2006) Wax-deficient anther1 is involved in cuticle and wax production in rice anther walls and is required for pollen development. Plant Cell 18:3015–3032

Rowland O, Zheng H, Hepworth SR, Lam P, Jetter R, Kunst L (2006) CER4 encodes an alcohol-forming fatty acyl-coenzyme a reductase involved in cuticular wax production in Arabidopsis. Plant Physiol 142:866–877

Labs working on this gene

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

National Key Facility of Crop Gene Resources and Genetic Improvement, Beijing 100081, China

National Center for Plant Gene Research (Beijing), Beijing 100081, China