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利用荞麦种子蛋白酶抑制剂基因提高烟草和马铃薯植株对生物胁迫的抗性。

Use of buckwheat seed protease inhibitor gene for improvement of tobacco and potato plant resistance to biotic stress.

作者信息

Khadeeva N V, Kochieva E Z, Tcherednitchenko M Yu, Yakovleva E Yu, Sydoruk K V, Bogush V G, Dunaevsky Y E, Belozersky M A

机构信息

Vavilov Institute of General Genetics, Russian Academy of Sciences, 119991 Moscow, Russia.

出版信息

Biochemistry (Mosc). 2009 Mar;74(3):260-7. doi: 10.1134/s0006297909030031.

Abstract

The possibility to use agrobacterial transformation of leaf discs to produce resistance to bacterial infections in tobacco and potato plants by introduction of a single gene encoding the serine proteinase inhibitor BWI-1a (ISP) from buckwheat seeds is shown. All studied PCR-positive transgenic plants exhibited antibacterial activity in biotests. It was shown that the presence of just a single gene of serine proteinase inhibitor provides sufficient protection at least against two bacterial phytopathogens, Pseudomonas syringae pv. tomato and Clavibacter michiganensis sbsp. michiganensis. The biotest including tobacco plant infection by the white wings butterfly in the green house has also demonstrated the existence of protective effect in transgenic tobacco plants. Significant genotypic variations in the protection efficiency were found between members of different genera of the same family (potato and tobacco) as well as between different lines of the same species. Northern blot analysis of four transgenic potato lines and three tobacco lines transformed by a vector plasmid containing the ISP gene of serine proteinases BWI-1a from buckwheat seeds has shown the presence of the expected size mRNA transcript.

摘要

研究表明,通过农杆菌介导的叶盘转化法,将来自荞麦种子的编码丝氨酸蛋白酶抑制剂BWI-1a(ISP)的单个基因导入烟草和马铃薯植株中,有可能使其产生对细菌感染的抗性。所有经研究的PCR阳性转基因植株在生物测试中均表现出抗菌活性。结果显示,仅一个丝氨酸蛋白酶抑制剂基因的存在就能为植株提供足够的保护,至少可抵御两种细菌性植物病原体,即丁香假单胞菌番茄致病变种和密执安棒形杆菌密执安亚种。在温室中用白翅蝴蝶感染烟草植株的生物测试也证明了转基因烟草植株具有保护作用。在同一科不同属(马铃薯和烟草)的成员之间以及同一物种的不同品系之间,发现了保护效率上显著的基因型差异。对四个转基因马铃薯品系和三个经含有来自荞麦种子的丝氨酸蛋白酶BWI-1a的ISP基因的载体质粒转化的烟草品系进行Northern印迹分析,结果显示存在预期大小的mRNA转录本。

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