国家重点基础研究发展计划(973计划);国家自然科学基金项目(面上项目,重点项目,重大项目)
The National Basic Research Program of China (973 Program);The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)
Rop家族蛋白是植物特有的一类小GTP结合蛋白,其属于Rho家族,在调控植物生长发育、抗逆和抗病过程中发挥重要的作用。本研究利用同源克隆法,筛选非亲和条锈菌小种CYR32侵染诱导的以感病品种Taichung29为遗传背景的抗条锈病基因Yr5近等基因系cDNA文库,分离获得1个Rop家族基因的全长cDNA序列,并分别命名为TaRop2。TaRop2包含1个591 bp开放阅读框,编码含197个氨基酸残基的蛋白质,分子量为21.52 kDa,理论等电点为9.49。通过转化烟草表皮细胞发现TaRop2定位于细胞核和细胞膜。RT-PCR分析结果表明,在高盐处理、非亲和条锈菌小种CYR32和亲和混合白粉菌菌株侵染时TaRop2基因的表达水平升高,但被干旱、高温、低温和ABA处理抑制。说明TaRop2可能参与小麦的生长发育、抗病防卫和抗逆反应过程。
The members of Rho family of small GTP-binding proteins isolated from plant species form a plant-specific one designated Rop (Rho-related proteins of plant)family. They play important roles in regulating plant development, stress tolerance and defense responses. In this study, a full-length cDNA encoding a Rop protein was isolated from the cDNA library of the near-isogenic line (Taichung29*6/Yr5) of wheat stripe rust resistance gene Yr5 using homologous cloning method, and it was designated TaRop2(Triticum aestivum Rop2). It contained a 591-bp open reading frame encoding a 197-amino-acid protein, whose molecular weight is about 21.52 kDa with isoeletric point (pI) 9.49. It was confirmed that TaRop2 were mainly distributed in the nuclei and on the membrane in tobacco epidermal cells through subcellular localization experiment. By RT-PCR expression analyses, TaRop2 expression level was increased after salt treatment or inoculation with incompatible race of stripe rust pathogen and compatible mixed powdery mildew strains, but was suppressed by treatments of drought, heat, cold and ABA. Based on these results, we proposed that TaRop2 may be involved in regulation of wheat development, defence and stress responses.
李媛媛,安艳秋,王凤涛,等.小麦中小GTP结合蛋白基因TaRop2克隆及其表达分析[J].植物遗传资源学报,2015,16(2):315-322.
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