| The interaction between tomatoes and C.fulvum abides "gene for gene"hypothesis and this pathosystem is one of the model systems to study plant-pathogen interaction.By this model system,a number of resistance(R)genes,corresponding avirulence(Avr)genes and the key components in the signal transduction pathway have been identified.Cf-4E/Avr4E is a pair of complementary genes.Compared with other gene pairs such as Cf-4/Avr4,Cf-9/Avr9 and Cf-2/Avr2,the mechanism of interaction and signaling pathway determined by Cf-4E/Avr4E are less known.Cf-4E driven by inducible promoter transforming Nb and tomatoes was carried out in order to study mechanism of interaction between Cf-4E and Avr4E.(1)Because many transgenic plant lines needed to be screened,a high-through method for selecting transgenic progenies was required,in the thesis,an improving selection method was established.The unsterilized T1 generation transgenic seeds were sowed on filtering paper which contained both 1/2 MS and corresponding antibiotic content.When the difference occurred among T1 seedlings,those with keeping green leaves,which were positive,were transferred onto the soil or other culture medium.Compared to traditional ways,its advantage was to both efficiently identify positive seedlings and avoid injury of roots.More importantly,it simplified process and saved the time.(2)Avr4E and Cf-4E was inserted into the pER8 vector,which is induced by estradiol.This constructed vector was respectively transformed into Nb expressing Avr4E and tomatoes expressing Cf-4E.28 T1 lines,whose background is Avr4E 12-3-1-T3,out of 101 transgenic Nb were screed by estradiol and 10 independent lines 1,2,5,6,7,11,14,16,32 and 34,appeared hypersensitive response under the injecting estradiol.In addition,all the transgenic lines were screed for copy numbers by the method improved.The two lines which contained single copy Cf-4E were obtained and confirmed by chi-square value.Currently,the work of genetic transformation of tomato is carrying out. |
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