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OsTOGR1 是定位于核仁的 DEAD-Box RNA 解旋酶,其异位表达赋予转基因白菜耐热性。

Ectopic expression of nucleolar DEAD-Box RNA helicase OsTOGR1 confers improved heat stress tolerance in transgenic Chinese cabbage.

机构信息

State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China.

University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

Plant Cell Rep. 2020 Dec;39(12):1803-1814. doi: 10.1007/s00299-020-02608-x. Epub 2020 Sep 29.

Abstract

The DEAD-Box RNA helicase OsTOGR1 positively regulates heat stress tolerance in Chinese cabbage. Non-heading Chinese cabbage (Brassica rapa L. ssp. chinensis) is primarily cultivated vegetable crop in Asian countries. Heat stress is one of the major threats for its growth and yield. Numerous regulatory genes in various crops have shown to contribute thermotolerance. Among them, Thermotolerant growth required 1 (TOGR1) is an important DEAD-box RNA helicase. To examine whether its role is conserved in other crops, we constructed pCAMBIA1300-pHSP:OsTOGR1 expression vector driven by the rice small heat shock protein promoter (pHSP17.9) and successfully produced transgenic non-heading Chinese cabbage plants expressing OsTOGR1 gene via Agrobacterium-mediated vacuum infiltration transformation. In total, we generated three independent transgenic cabbage lines expressing TOGR1 gene. Expression and integration of TOGR1 was confirmed by PCR, RT-PCR and qPCR in T and T generations. The relative leaf electrical conductivity of transgenic seedlings was reduced subjected to high temperature (38 °C) compared to heat shock treatment (46 °C). In addition, hypocotyl length of transgenic seedlings increased compared to wild-type plants under high temperature and heat shock treatment. Furthermore, the transgenic plants exhibited higher chlorophyll content than wild-type plants under high temperature and heat shock treatment. The transgenic seeds displayed better germination under heat shock treatment. Tested heat stress-responsive genes were also up-regulated in the transgenic plants subjected to high temperature or heat shock treatment. To the best of our knowledge, this is the first report on describing the role of DAED-Box RNA helicases in improving heat stress tolerance of transgenic plants.

摘要

拟南芥 DEAD-box RNA 解旋酶 OsTOGR1 正向调控白菜耐热性。不结球白菜( Brassica rapa L. ssp. chinensis )是亚洲国家主要栽培的蔬菜作物。热胁迫是影响其生长和产量的主要威胁之一。众多不同作物中的调控基因已被证明有助于耐热性。其中,耐热生长必需 1(TOGR1)是一种重要的 DEAD-box RNA 解旋酶。为了检验其在其他作物中的作用是否保守,我们构建了由水稻小热激蛋白启动子(pHSP17.9)驱动的 pCAMBIA1300-pHSP:OsTOGR1 表达载体,并通过农杆菌介导的真空渗透转化成功获得了表达 OsTOGR1 基因的非结球白菜转基因植株。总共生成了三个独立的表达 TOGR1 基因的转基因白菜株系。通过 PCR、RT-PCR 和 qPCR 在 T 和 T 代中证实了 TOGR1 的表达和整合。与热激处理(46°C)相比,转 TOGR1 基因的幼苗在高温(38°C)下的相对叶片电导率降低。此外,与野生型植株相比,高温和热激处理下转基因幼苗的下胚轴长度增加。此外,高温和热激处理下,转基因植株的叶绿素含量高于野生型植株。在热激处理下,转基因种子的萌发率更好。在经受高温或热激处理的转基因植株中,测试的热应激响应基因也上调。据我们所知,这是首次描述 DEAD-box RNA 解旋酶在提高转基因植物耐热性方面的作用的报道。

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