Sequence Characterization and Expression Analysis of OsRhoGAP1 Gene in Rice

doi:10.13560/j.cnki.biotech.bull.1985.2017.05.014

Abstract

Abstract: A function-unknown gene was isolated from a cDNA library of young spike of rice and designated as OsRhoGAP1. Based on the bioinformatics and modern molecular biology,we systematically surveyed its expression patterns at different development stages and under different hormones treatments. Bioinformatics analysis showed that OsRhoGAP1 had typical structural feature of plant RhoGAP protein,including CRIB motif and RhoGAP domain. qRT-PCR analysis revealed that OsRhoGAP1 gene presented specifically spatio-temporal characteristics,i.e.,relative high expression levels in root and leave at the seedling stage,in young panicle and leave at the panicle stage,and in root at the mature stage. Furthermore,the transcription of OsRhoGAP1 was strongly induced by ABA,SA,GA,and MeJA,but suppressed by IAA and 6-BA. It was speculated that the expression of OsRhoGAP1 was regulated by hormones.

Key words: rice, OsRhoGAP1gene, sequence analysis, expression analysis, plant hormones

YAN Xin-tian HUANG Jun-jun LOU Chen GE Hui-wen LIANG Wei-hong. Sequence Characterization and Expression Analysis of OsRhoGAP1 Gene in Rice[J]. Biotechnology Bulletin, 2017, 33(5): 94-101.

References

[1] Nagawa S, Xu T, Yang ZB. RHO GTPase in plants：conservation and invention of regulators and effectors[J]. Small GTPases, 2010, 1：78-88.
[2] Amin E, Jaiswal M, Derewenda U, et al. Deciphering the molecular and functional basis of RhoGAP family proteins：a systematic approach towards selective inactivation of Rho family proteins[J]. The Journal of Biological Chemistry, 2016, 291：20353-20371.
[3] Hwang JU, Vernoud V, Szumlanski A, et al. A tip-localized RhoGAP controls cell polarity by globally inhibiting Rho GTPase at the cell apex[J]. Current Biology Cb, 2008, 18：1907-1916.
[4] Klahre U, Kost B. Tobacco RhoGTPase activating protein1 spatially restricts signaling of RAC/Rop to the apex of pollen tubes[J]. Plant Cell, 2006, 18：3033-3046.
[5] Hoefle C, Huesmann R. A barley ROP GTPase activating protein associates with micro-tubules and regulates entry of the barley powdery mildew fungus into leaf epidermal cells[J]. Plant Cell, 2011, 23：2422-2439.
[6] Baxter-Burrell A, Yang Z, Springer PS, et al. RopGAP4-dependent Rop GTPase rheostat control of Arabidopsis oxygen deprivation tolerance[J]. Science, 2002, 296：2026-2028.
[7] 米志勇, 王树生, 吴乃虎. 水稻低分子量G T P结合蛋白基因OsRacD的分离[J]. 科学通报, 2000, 45（19）：2047-2055.
[8] Ye JR, Huang MJ, Wu NH. Fertility analysis of the Arabidopsis transformed with antisense rice osRACD gene[J]. Progress in Natural Science, 2003, 13：424-428.
[9] 叶建荣, 黄美娟, 赵淑慧, 等. osRACD 基因表达与光敏核不育水稻光周期育性转换的相关性[J]. 自然科学进展, 2004, 14（2）：166-172.
[10] Liang WH, Tang CR, et al. Cloning and characterization of a new actin gene from Oryza sativa L.[J]. Progress In Natural Science, 2004, 14：867-874.
[11] 梁卫红, 唐朝荣, 吴乃虎. 两种水稻GDP解离抑制蛋白基因的分离及特征分析[J]. 中国生物化学与分子生物学报, 2004, 20（6）：785 -791.
[12] Yoshida S, Forno DA, Cock JH. Laboratory manual for physiological studies of rice[M]. International Rice Research Institute, 1976.
[13] Mcelroy D, Blowers AD, Jenes B, et al. Construction of expression vectors based on the rice Actin1（Act1）5' region for use in monocot transformation[J]. Molecular and General Genetics, 1991, 231：150-160.
[14] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2（-Delta DeltaC（T））method[J]. Methods, 2001, 25：402-408.
[15] Wu G, Li H, Yang Z. Arabidopsis RopGAPs are a novel family of rho GTPase-activating proteins that require the Cdc42/Rac-interadtive binding motif for rop-specific GTPase stimulation[J]. Plant Physiology, 2000, 124：1625-1636.
[16] 张之为, 赵君, 樊明寿, 等. 植物小G蛋白的研究进展[J]. 西北植物学报, 2009, 29（3）：622-628.
[17] Nibau C, Wu HM, Cheung AY. RAC/ROP GTPases：‘hubs’ for signal integration and diversification in plants[J]. Trends Plant Sci, 2006, 11：309-315.
[18] Yang Z, Fu Y. ROP/RAC GTPase signaling[J]. Curr Opin Plant Biol, 2007, 10：490-494.
[19] Schaefer A, Miertzschke M, Berken A, et al. Dimeric plant RhoGAPs are regulated by its CRIB effector motif to stimulate a sequential GTP hydrolysis[J]. Journal of Molecular Biology, 2011, 411：808-822.
[20] Hoefle C, Huesmann C, Schultheiss H, et al . A barley rOP GTPase activating protein associates with microtubules and regulates entry of the barley powdery mildew fungus into leaf epidermal cells[J]. Plant Cell, 2011, 23：2422-2439.
[21] Ying G, Yang Z. ROP GTPase regulation of pollen tube growth through the dynamics of tip-localized F-actin[J]. Journal of Experimental Botany, 2003, 54：822-824.
[22] Yamauchi Y, Ogawa M, Kuwahara A, et al. Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds[J]. Plant Cell, 2004, 16：367-378.
[23] Itoh H, Ueguchi-Tanka M, Sato Y, et al. The gibberellins signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei[J]. Plant Cell, 2002, 14：57-70.
[24] Bright J, Desikan R, Hancock JT, et al. ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H 2 O 2 synthesis[J]. Plant Journal, 2006, 45：113-122.
[25] Du H, Liu H, Xiong L. Endogenous auxin and jasmonic acid levels are differentially modulated by abiotic stresses in rice[J]. Frontiers in Plant Science, 2013, 4：397-397.
[26] 林群婷, 梁卫红, 李辉, 等. 水稻Rop基因OsRac5的表达特性[J]. 植物生理学报, 2013, 49（12）：1400-1406.