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Acta Agron Sin ›› 2008, Vol. 34 ›› Issue (12): 2070-2076.doi: 10.3724/SP.J.1006.2008.02070

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Expression of Protein Elicitor-Encoding Gene pemG1 in Tobacco (Nicotiana tobacum cv. Samsun NN) Plants and Enhancement of Resistance to TMV

MAO Jian-Jun,QIU De-Wen*,YANG Xiu-Feng,ZENG Hong-Mei,YUAN Jing-Jing   

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2008-04-21 Revised:2008-07-08 Online:2008-12-12 Published:2008-10-10
  • Contact: QIU De-Wen

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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

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