Screening The Mutants Of Late Blight Resistance Signal Transduction In Tomato

Tomato late blight caused serious loss.Chemical control of late blightnot only affect the environment but it will cause resistance of pathogenicbacteria.Therefor it,s important to explore the mechanism of anti-disease gene and understandthe transmission channel of disease resistance signal,so controling late blight will stepin a new field if the key gene of signaling pathway be founded.Using a novel transgenic tomato plant conferred potato late blight resistancegene R3a and glucocorticoid-inducible avirulence gene Avr3a was constructed：MM-R3a-Avr3a.，Screening the survival mutants from the MM-R3a-Avr3a EMSMutants by spray inducer. Molecular identification and genetic analysis of themutants which HR were interrupted, and provide research foundation for cloning thekey gene of resistance signal transduction。The experiment efficiency will beimproved while identified the mutant by molecular using alkaline lysis method extracttomato DNA.The main results of our research are showed as follows:1.Using tomato leaves, cotyledons as as the materials, with this methodextracting DNA, the DNA mass concentrations and purity are relatively highdetermined by UV spectrophotometer,30times dilution DNA amplified by PCR isrelatively stable.2.After three generations of testing and screening, the final six major families begot, and identified the mutant characteristics of the six pedigrees.130,208,1926aredominant mutation, and130are single-gene dominant mutation;208,821,1057arerecessive mutations.3.hypersensitive response of the hybrids were analyzed and determine the1057 and821two-families as the key gene mutation;1601as R3a, Avr3a+GVG mutation.4.Mutant emerged in a special field mutant traits: potato leaf, limited growth type,duplex inflorescence, dwarf, leaves drooping, leaf. It maybe chain with mutant geneand provide reference information for the succeeding gene location.