| Grape has become one of the fruit tree species with the largest cultivation area in my country due to its high economic value and nutritional value.During the entire growth process of grapes,the infection of Botrytis cinerea will affect it to varying degrees,which has seriously affected the quality and yield of grapes and caused huge losses to the grape industry.At present,chemical control is the most commonly used control method to resist the invasion of Botrytis cinerea,but this method will not only increase the economic cost,but also have a certain destructive effect on the environment.Based on this,the breeding work to improve the resistance of grape to Botrytis is also becoming more and more important.In this experiment,the hybrid F1 generation of the female parent ’Red Globe’ and the male parent ’Beta’ was used as the experimental material.First,the SSR molecular marker technology was used to identify the true and false hybrids of the offspring of the two cultivars ’Red Globe’ and ’Beta’.There are 189 hybrids,and the leaves of the 189 hybrids were tested for resistance to Botrytis cinerea,by inoculating leaves in vitro.strains.Finally,RNA-Seq technology was used to compare and analyze the differences in the transcriptional level of the resistant variety ’Beta’ and the susceptible variety ’Red Globe’ ind uced by Botrytis cinerea,so as to discover candidate genes related to botrytis resistance,which could be used for grapes.Breeding for botrytis resistance provides a reference.The main results are as follows:1.The SSR molecular marker technology was used to identify the authenticity of the hybrid progeny plants of the female parent ’Red Globe’ and the male parent ’Beta’,and the suitable SSR primer VRZAG25 was screened out,and the optimal annealing temperature of the primer was determined by gradient PCR method.Finally,189 true hybrids were identified by primer VRZAG25,and the true hybrid rate was94.5%.2.The leaves of the hybrid F1 generation with the susceptible variety ’Red Globe’ as the female parent and the resistant variety ’Beta’ as the male parent were used as the test materials,and the spore suspension was used to infect the isolated leaves to identify the resistance to Botrytis cinerea.Through three repeated experiments,it was found that the incidence of the hybrid offspring population was quite different.The disease resistance of the hybrid offspring of the population showed a continuous distribution,with the characteristics of quantitative trait inheritance.The disease resistance coefficient of variation of the progeny population is relatively large,indicating that the breeding of lines with resistance to gray mold has great potential,and 8 progeny lines(No.19,No.38,No.47,No.137,No.156,No.178,No.179 and 193).3.Transcriptome sequencing of the parental leaves at different stages after Botrytis cinerea infection,a total of 18 candidate genes that may be related to Botrytis cinerea resistance were screened and verified by fluorescence quantitative(qRT-PCR),of which 14 were The candidate genes for botrytis cinerea were completely consistent with the transcriptome sequencing results,and the other four candidate genes against botrytis cinerea were basically consistent with the transcriptome sequencing results.4.The transcript data was analyzed,and four genes associated with Botrytis cinerea resistance were determined: F3 H,WRKY33,TIR2,and EDS1 were determined.The expression patterns of these genes were positively correlated with the resistance to Botrytis cinerea,and they were all up-regulated.It was speculated that these genes were important for the resistance of grapes to Botrytis cinerea infection. |
