Relationship Between Disease Resistive Materials With Apple Ringspot Morbidity And Identification Of AFLP Markers

Apple is the most important fruit which popular in Chinese consumers．And it isone of the most important processing and marketing fruits in the world. However,Apple ring rot disease, the most serious and prevalent apple disease in china, whichleads to numerous rotten fruits and weaken tree vigorous, affects apple production inChina,，generates．it becomes an outstanding problem in apple production.We areplanning to study interaction of Host-Pathogen at biochemical level.We observed thevariation regularity of resistive materials such as polyphenols,phenylanineammonialyase(PAL), peroxidase(POD)and polyphenoloxidase(PPO) among appletrees with different level of resistance under natural condition from Apirl to August,and the changes of these resistive material after inoculated with the Botryosphaeriaberengriana f．piricolca, and deduced the effective period of each resistive materialaccording to the variation rule of resistive materials.thereby screened specificresistive material in specific periods as resistance index This research in combinationwith the study of inhibitive effect of polyphenols on Botryosphaeria berengriana invitro will laid the theoretical foundation,to identify resistant apple stocks,and toelucidate the mechanisms of how resistive material confer their resistance to applering rot. On the other hand, F1hybrids segregation population of seedling plants from“wumei×M9”were researched searching for the molecular marker linked to apple ringrot resistance gene locus by using AFLP to be used in marker assisted breeding forhigh resistant apple varieties in the future.1Under natural condition,the variation regularity of different resistivematerials (total leaves polyphenol, activities of protective enzymes) in different appletrees with different rot disease resistance were determined. The results showed thatfrom April to July, the content of polyphenols and enzymatic activities from1eaves ofhighly resistance trees presented a gradually increasing trend, and showed higher concentration or activities than those of moderate resistance trees and suspitiblematerials in about May to June according to the kinds of resistive materials,then theseresistive materials reached the maximum in July, which is the main infection period.while the maximum resistive materials of moderate resistance materials andsuspitible materials occurred in May or June,then appear downtrend. The maximum ofeach resistive materials showed that highly resistance trees>moderate resistancetrees> suspitible trees2The different trees with different level of resistance to apple ring rot diseasewere studied by inoculating the Botryosphaeria berengriana.f sp.piricola on theirstems. The results showed the inoculation can arouse the change of each resistivematerials obviously. The average and the maximum of polyphenols content and theactivities of enzymes showed following rules: high resistant trees> moderateresistance trees> suspitible trees. And high resistance trees maintained longer periodof high level of resistive materials than that of other resistant grades trees.3To a certain extent, the resistive materials were correlated with plantsusceptible level. the correlate level of total phenol content with susceptible level isremarkable. In some stage and period, the activities of defendant enzymes with thesusceptible level also had remarkable correlation, for example, PAL activity of highresistant trees had significance level（P<0.05） with moderate resistance trees in the7days to21days after inovulation and had high significance level（P<0.01） withsuspitible trees; PPO activity of high resistant trees had significance level（P<0.05）with moderate resistance trees and suspitible trees in the14days to21days; PODactivity of high resistant trees had significance level（P<0.05） with moderateresistance trees and suspitible trees in the21days to28days. The data showed thatPAL was the earliest activated resistive substance and maintains for the longesttime. The peak of POD activity and Phenols content appeared later.4The inhibitive effects of different concentrations of tea-polyphenol and applepolyphenol on Botryosphaeria berengriana.f sp.piricola were tested, The resultsshowed that tea-polyphenol and apple polyphenols inhibited pathogen growthsignificantly(p<0.05). The higher the concentration was, the higher the inhibitive ratewas．Among the series concentrations，the100and50mg/mL had the highestinhibitive rate.5We constructed three different susceptible level segregation populations usingthe thirteen-years old F1seedling of “wumei×M9” based on the natural incidence inthe field of apple branches, and screened for molecular markers closely linked withresistance genes using AFLP.Among all the64primer pair we used,primer pairE4/M4generated bands showing the lowest recombination rate(10.7%). An460bpband, named R4-4,amplified in E4/M4primer combination was found to be linkedto the resistance population...