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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front. Agric. China    2007, Vol. 1 Issue (1) : 52-57     DOI: 10.1007/s11703-007-0009-8
Research article |
Cloning and functional analysis of the genes involved in signal transduction in tomato Cf-4-Avr4 pathosystem
Qing LIU,Dongxin FENG,Xiaowu WANG,Yongchen DU()
Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
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Abstract

Hypersensitive response (HR) is one of the most efficient and common resistance mechanisms in plants. Cloning signaling genes are very important to elucidate the resistance mechanisms. A gene in tomato homologous to several resistance proteins in plant was involved in HR and named as RGL (Resistance Gene Like). RGL protein was used as a bait to screen interacting protein(s) from tomato cDNA library through the yeast two-hybrid system. Two interacting proteins were found, which were called as RGLIP-1 and RGLIP2 (RGL Interacting Protein), respectively. RGLIP-1 is a protein of 291 amino acids with significant homology with thylakoid lumen protein. RGLIP-2 is a protein of 248 amino acids with significant homology with transducin protein. Virus-Induced Gene Silencing (VIGS) of the two genes results in a partial and complete suppression of Avr4induced HR, which indicates that both genes are involved in hypersensitive response.

Keywords tomato      Cladosporium fulvum      hypersensitive response      yeast two-hybrid system      virus-induced gene silencing     
Issue Date: 22 February 2016
 Cite this article:   
Qing LIU,Dongxin FENG,Xiaowu WANG, et al. Cloning and functional analysis of the genes involved in signal transduction in tomato Cf-4-Avr4 pathosystem[J]. Front. Agric. China, 2007, 1(1): 52-57.
 URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-007-0009-8
http://academic.hep.com.cn/fag/EN/Y2007/V1/I1/52
Fig. 1  Sequence and putative conserved domain of RGL
Plate I  Screening result of yeast two-hybrid

A: Screen on Try-/Leu-/Ade-/His- medium (arrows indicate candidate clones)

B: Auto activation test (the upper three clones are the yeast with empty BD plasmid and AD plasmid; the lower three clones are the yeast with bait BD plasmid and corresponding AD plasmid)

Plate II  Functional analysis of RGLIP genes using VIGS in Cf-4 transgenic Nicotiana benthamiana Plants

C: Negative control; D: Positive control; E and F: RGLIP-1 and RGLIP-2

Fig. 2  Nucleotide sequence and deduced amino-acid sequence of RGLIP-1 isolated from tomato cDNA library through yeast two-hybrid system
Fig. 3  Nucleotide sequence and deduced amino acid sequence of RGLIP-2 isolated from tomato cDNA library through yeast two-hybrid system
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