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Study on tobacco disease resistances mediated by the elicitor gene cryptogein from Phytophthora cryptogea

Published online by Cambridge University Press:  12 February 2007

Jiang Donghua ,
Guo Zejian ,
Chen Xujun ,
Cheng Zhiqiang  and
Zheng Zhong
Jiang Donghua
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China
Guo Zejian*
Affiliation:
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100094, China
Chen Xujun
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
Cheng Zhiqiang
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
Zheng Zhong
Affiliation:
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China
*
*Corresponding author: Email: [email protected]
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Abstract

The cryptogein (Crypt) gene was obtained by PCR amplification of genomic DNA from Phytophthora cryptogea and confirmed by DNA sequencing. A promoter of the rice phenylalanine ammonia-lyase (PAL) gene was used to regulate the expression of Crypt, because this promoter has a low level of constitutive expression, is strongly induced by pathogen infection and is expressed in epidermal tissues. These promoter characteristics may be suitable for potential inhibition of pathogen attack on epidermal tissues. For functional interaction of Crypt with its outer plasma membrane binding sites, Crypt was led by a signal sequence of the extracellular pathogenesis-related protein (PR1b) of tobacco (Nicotiana tabacum). The fused gene was constructed into a binary vector and the final plasmid (CHF-PAL::Crypt) was transformed into tobacco using the Agrobacterium-mediated transformation method. Twenty-two lines of transformants were obtained from selection medium containing 100 mg/l of kanamycin. Results from PCR amplification and Southern blot analysis demonstrated that Crypt was integrated into the tobacco genome. In the assay of pathogen challenge, nearly two-thirds (68.2%) of the transgenic plants showed significantly enhanced resistance against black shank fungal (Phytophthora parasitica var. nicotianae), brown spot fungal (Alternaria alternata) and wild fire bacterial (Pseudomonas syringae pv. tabaci) diseases. This observation indicates that low-level constitutive expression of Crypt gene in tobacco could have potential use in generating broad-spectrum disease-resistant plants.

Keywords

elicitorcryptogeinconstruction of vectortransformationdisease resistance
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 1 , Issue 2 , August 2004 , pp. 125 - 131
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

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