芒果果实抗炭疽病相关基因的克隆及功能分析

炭疽病菌是引起贮运期芒果果实腐烂的主要诱因，国内外报道不同品种的芒果对炭疽病的感病性存在较大差异，表明有抗病基因可以利用。但迄今为止，芒果抗炭疽病的分子机理还不清楚。前期工作我们已完成接种胶孢炭疽病菌后抗病品种'Zill'与感病品种'台农'的果实转录组测序。本项目仍以上述抗/感品种为材料，对接种后不同时间点的抗/感果实进行RNA-seq测序，通过横向比较抗病品种与感病品种基因表达谱、以及纵向比较相同品种接种前后不同时间点的基因表达谱，筛选差异表达的基因, 并对差异表达基因进行后续的代谢通路分析、与小RNA的联合分析、以及与其它物种已知的抗病基因比较等，寻找芒果抗炭疽病相关基因；并通过果实接种后实时荧光定量PCR检测候选基因的表达水平，和构建植物表达载体，对拟南芥同源基因进行抑制或过表达来研究候选基因功能。研究结果有利于揭示芒果抗炭疽病的分子机理，可为培育抗病新品种提供基因资源和奠定基础。

The plant fungal pathogen Colletotrichum gloeosporioides (Penz) is the causal agent of mango fruit anthracnose during storage and transportation. Domestic and overseas reports have evidenced that there were great differences in disease resistance among different variety of mangos and suggested that disease resistance genes can be isolated from mango fruit. But till now, the molecular mechanism of mango fruit with resistance to anthracnose is still unclear. In our early study, we have accomplished mango fruit transcriptome sequencing of resistant varieties (Zill) and susceptible varieties (Tai Nong 1) when inoculated with C. gloeosporioides conidia. In this study, we will study fruit transcriptomes through RNA sequencing(RNA-seq) and compare the differential genes expression profiling between resistant and susceptible varieties at different time points after inoculation. To identify the differentially expressed genes with potential relevance in disease resistance, subsequent metabolic pathway analysis of the differential expressed genes, combinational analysis of small RNAs and the comparison with known resistance gene of other species are needed. Moreover, to analyses the candidate genes function, the candidate genes functional analysis will be conducted by qRT-PCR technology, or by over expression and gene silencing technology in Arabidopsis. The research result will be advantageous in uncovering mango fruit disease resistant molecular mechanism; It is hopeful to provide disease resistance genes resource and help establish stable basis in mango disease resistance breeding.