猕猴桃应答果实软腐病菌侵染的转录组研究及关键抗性基因挖掘

Fruit rot is the most serious fungal disease in kiwifruit storage period, which caused great losses to post-harvest storage and transportation. Clarify the molecular resistance mechanism of kiwifruit response to fruit rot pathogens is critical important to the disease resistance breeding. On the basis of pathogens identification and resistance evaluation of 32 main kiwifruit cultivars, the project plan to employ suspectible cultivar ‘Hongyang’ and resistant variety ‘H-2' as research materials to invesitige the molecular response of kiwifruit to main pathogen Diaporthe. actinidiae. The study includes the application of transcriptomic analysis to investigate the expression profile of kiwifruit in response to D. actinidiae and the ultilization of cluster analysis to discover defense-related genes; as well as function verification of the crucial resistance genes via Agrobacterium tumefaciens-mediated RNA interference, gene overexpression and transient expression technology. The result of this project will expound the molecular resistance mechanism of kiwifruit cultivars response to fruit rot pathogen from transcript level, mining and verify the function of 3 to 5 crucial resistance genes, provide a theoretical basis for the molecular breeding of kiwifruit.

果实软腐病是猕猴桃贮藏期最严重的真菌性病害，给采后储运造成了巨大的损失。明确猕猴桃果实采后应答软腐病菌的分子抗性机理，对猕猴桃的抗病育种至关重要。在前期已鉴定中国猕猴桃果实软腐病菌种类并获得32份猕猴桃主栽品种抗性评价结果的基础上，本项目拟以遗传背景清晰的高感品种‘红阳’和高抗品系‘H-2’为研究材料，开展软腐病致病菌D. actinidiae侵染实验，运用转录组测序技术分析软腐病菌侵染抗/感猕猴桃材料的差异基因表达，聚类关联分析并挖掘抗性相关基因；最后，利用农杆菌介导的RNAi、过表达及瞬时表达技术进行关键抗性基因的功能验证。本项目的开展将初步揭示猕猴桃应答果实软腐病菌侵染的分子抗性机理，挖掘并验证3-5个关键抗性基因，为猕猴桃的分子抗性育种提供理论依据。

果实软腐病是猕猴桃贮藏、转运及销售期间最严重的真菌性病害，给世界猕猴桃产业造成了重大经济损失。明确猕猴桃采后应答软腐病菌的分子抗性机理，挖掘关键抗性基因，对猕猴桃抗病育种至关重要。在前期研究基础上，本项目对中国猕猴桃果实软腐病的致病菌种类及主栽品种抗性进行了更深入的鉴定评价；此外，以4对高感及高抗品种、检出率最高且致病力最强的D.actinidiae菌株为研究材料，运用链特异性转录组测序技术分析了DA菌株侵染抗/感猕猴桃材料的差异基因表达，聚类关联分析并挖掘抗性相关基因，筛选得到10个在高抗品种中均显著上调表达的基因；随后，利用农杆菌介导的RNAi、过表达技术，结合瞬时转化后组织的抗病表型，对10个上调基因的功能进行验证，最终筛选得到了4个关键抗病基因。本项目的开展为猕猴桃软腐病及其它真菌病害的分子抗性机理分析、抗性基因挖掘验证及抗性种质的创制奠定了理论及技术基础。