蛋白激发子基因pemG1转化三生烟中及其对TMV抗性的提高

doi:10.3724/SP.J.1006.2008.02070

摘要/Abstract

摘要：

通过根癌农杆菌(Agrobactrium tumefaciens)介导转化法，将含有稻瘟菌蛋白激发子基因pemG1的植物表达载体pCAMBIA2300-Ubi-pemG1-Ocs转化三生烟(Nicotiana tobacum cv. Samsun NN)，获得了转基因植株。用PCR检测抗卡那霉素烟苗确认阳性转化株，用Southern、Northern和Western杂交进一步证实了pemG1基因的整合、转录和表达。对T2代转基因阳性株进行烟草花叶病毒(Tobacco Mosaic Virus)接种试验。接种5 d后发现，与非转基因对照相比，表达pemG1的烟草叶片枯斑数量减少，表明稻瘟菌蛋白激发子基因pemG1的表达提高了转基因烟草对TMV的抗性。

关键词: 蛋白激发子, pemG1, 烟草, 表达, 烟草花叶病毒

Abstract:

Protein elicitors are important signal molecules that trigger plants disease resistance. Defence responses will be induced once the elicitors are recognized by acceptors in plants. It is revealed that elicitor protein PemG1 from Magnaporthe grisea is able to increase hydrogen peroxide content of tobacco suspension cells. To study PemG1’s functions in plants, pemG1 gene was transferred into tobacco in the study. For this, plant expression vector pCAMBIA2300-Ubi-pemG1-Oc harboring elicitor-encoding gene pemG1 from Magnaporthe grisea was constructed. The maize ubiquitin promoter/octopine synthase terminator system and kanamycin-resistant gene npt II (neomycin phosphotransfers II) were used for constitutive expression systems. The vector was then introduced into Agrobacterium tumefaciens (strain AGL-1) by freeze-thaw method. Tobacco (Nicotiana tobacum cv. Samsun NN) primary transformants were produced by leaf disc transformation. The kanamycin-resistant regenerated plants were confirmed to be electropositive by PCR. Integration and expression of the pemG1 gene were further confirmed by Southern blotting and Western blotting, respectively. Then, transgenic tobacco plants of T₂ generation were inoculated with Tobacco Mosaic Virus (TMV) at two different virus concentration. In comparison with TMV-infected wild-type SNN plants, PemG1-expressed plants displayed reduced hypersensitive-response lesions in both treatments. Furthermore, accumulation level of pemG1 steady-state transcripts was examined at 24 h after inoculation. The results indicated that the reduction of lesions corresponded to the accumulation of pemG1 steady-state transcripts as monitored by Northern analysis. All these indicated that the expression of pemG1 in tobacco plants improved the resistance to TMV.

Key words: Protein elicitor, pemG1, Tobacco, Expression, TMV

毛建军;邱德文;杨秀芬;曾洪梅;袁京京. 蛋白激发子基因pemG1转化三生烟中及其对TMV抗性的提高[J]. 作物学报, 2008, 34(12): 2070-2076.

MAO Jian-Jun;QIU De-Wen;YANG Xiu-Feng;ZENG Hong-Mei;YUAN Jing-Jing. Expression of Protein Elicitor-Encoding Gene pemG1 in Tobacco (Nicotiana tobacum cv. Samsun NN) Plants and Enhancement of Resistance to TMV[J]. Acta Agron Sin, 2008, 34(12): 2070-2076.

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