The RAPD Maker Of The Resistant Gene For Late Blight In Tomato

Tomato late blight is a major disease on tomato of northem facility and southern open ground,resistance breeding is the effective way to resolve the serious harm of tomato late blight.The use of modern biotechnology to find the molecular markers for the resistance gene of tomato late blight,for the theoretical basis of efficient breeding for disease resistance and the cultivation of improved technologies.In this paper CLN2037(resistant parent),T2-03(susceptible parent) and the F₁ hybrid generation,F₂ generation,through the artificial inoculation with disease resistance identification,the purpose is use BSA,to find the RAPD markers closely linked with tomato late blight resistance gene Ph-3.For late blight resistance breeding MAS has laid a good foundation.The results are as Follow:1 Through the artificial inoculation with disease resistance identification,the Disease class index of CLN2037 is 0.1,the type of anti-influenza is R,the Disease class index of T2-03 is 6.0,the type of anti-influenza is S,the Disease class index of F₁ is 2.1,the type of anti-influenza is R,the F₂-generation plant identification results:43 individual high resistance,high sense of the individual or the death of 49 individuals,F2 generation of 220 plants,disease-resistant strain 154,a sense of disease for 66,X²=2.38＜X²_0.05=3.84, Therefore,the late blight of tomato genetic traits should be controlled by the quality characters.2 The random amplified polymorphic DNA(RAPD)system for tomato was optimized. The results as follows:In the 25μL RAPD system of tomato,10×Buffer was 2.5μL,Mg²⁺ was 1.5 mmol·L^-1,dNTPs was 0.15 mmol·L-1,Random primer was 5pmol·L-1,Taq DNA polymerase was 1.25U and DNA was 25ng.the most suitable program was one cycle at 94℃/5 min,45cycles at 94℃/1min,32℃/1min,72℃/1.5min and one cycle at 72℃/10min.3 The objective of this study was to identify RAPD maker for resistance to late blight in tomato.BSA(Bulked Segregant Analysis) was used to search for RAPD markers linked to late blight resistant gene Ph-3,using F₂ segregating population(147 individuals)derived from a cross of CLN2037(resistant) and T2-03(susceptible).230 decamer primers with arbitrary sequences are chosen for polymerase chain reaction amplication,one RAPD marker CCPB272-03₇₄₀(the sequence of the primer:GGTCGATCTG) was tightly linked to the resistant gene Ph-3 and it was 5.8cM from the resistant gene.