Cloning Of The Full-Length CDNA Of Melon Ethylene Receptor Gene Cm-ETR1 And Transformaton Of Its Mutants Into Melon

Ethylene, as a very important phytohormone, regulates a variety of developmental process in plants. Especially for climacteric fruit, ethylene accelerates the ripening process. Over the last decade of genetic studies of model plant Arabidopsis, a largely linear model of plant ethylene signal transduction pathway has been established. Since as a key component of ethylene biosynthsis and ethylene signal transduction pathway, ethylene receptor ETR1 determines the sensitivity of plants to ethylene and controls plant senescence process. In Arabidopsis and tomato, the gain of function mutants of ETR1 displays ethylene insensitive phenotype.The full-length cDNA of Cm-ETRI was amplified via RT-PCR from the pericarp of Cucumis melo L. cv Hetao. We designed five pairs of primers according to amino acid sequences of the ETR1 mutants of gain of function from Arabidopsis and tomato. Then the five mutants of Cm- ETR1 were amplified through gene splicing by overlap extension PCR (SOE-PCR) and were designated Cm-ETR1A31V, Cm-ETR1P36L, Cm-ETR1C65Y, Cm-ETR1H69A and Cm-ETR1A102T respectively. We have constructed the five plant expression vectors of these five mutants genes, then introduced them into Cucumis melo L. cv Hetao by pollen tube pathway transformation method and obtained transgenic Cucumis melo L. cv Hetao, respectively.