Cloning and functional analysis of the genes involved in signal transduction in tomato <i>Cf</i>-4-<i>Avr</i>4 pathosystem

doi:10.1007/s11703-007-0009-8

Front. Agric. China

2007, Vol. 1

Issue (1) : 52-57 https://doi.org/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:

https://academic.hep.com.cn/fag/EN/10.1007/s11703-007-0009-8
https://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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