工业大麻抗旱基因的挖掘及功能分析

Drought is one of the main factors which severely decrease the production of hemp. However, studies on drought-resistance of hemp are quite few at home and abroad let alone the molecular mechanism of drought-resistant studies. The tremendous progress on Arabidopsis drought mechanism studies has laid the theoretical foundation for hemp. The completion of the hemp genome sequencing work and the development of expression profiling technology has provided technical support to carry out the study on the drought resistance mechanism of hemp. The team of applicant has proved the BM001 is excellent material for drought and salt tolerance in preliminary work. So in this project, BM001 will be used as research object with DGE technology, comparative genomics and bio-informatics methods to obtain the first drought resistance expression profiling of hemp, analyze the different of functional genes and the metabolic pathways. RACE technology also will be used to clone the full length of candidate genes. qRT-PCR technology can determine the gene expression characteristics and the signal response. For the first time from the level of transcriptome analysis and drought response mechanism of hemp, this study will lay solid foundation for the further work on drought molecular mechanism of hemp. It is also valuable to find drought resistant gene which is applicable with independent intellectual property rights and provide gene source for cultivation of hemp with resistance.

干旱影响植株正常生长，导致纤维产量降低、品质下降，是限制工业大麻生产的主要环境因素之一，但目前国内外对工业大麻抗旱的研究较少，且均没有开展抗旱分子机理方面的研究。拟南芥等植物在抗旱研究中取得的巨大进步为工业大麻开展抗旱研究奠定了较好基础；工业大麻全基因组测序完成和表达谱技术的发展为开展大麻抗旱机理研究提供了技术保障。申请者团队前期研究证明BM001是优异的抗旱、耐盐材料，本项目拟以BM001为研究对象，利用DGE技术、比较基因组学和生物信息学方法获得首份工业大麻干旱胁迫表达谱，分析差异表达基因及其相关代谢途径；RACE技术克隆候选基因全长并进行亚细胞定位；qRT-PCR确定基因的时空表达特性和信号响应情况。本研究将首次从转录组水平分析工业大麻的抗旱应答机制，为深入解析工业大麻的抗旱分子机制奠定基础；挖掘具有自主知识产权、有应用价值的工业大麻抗旱基因，为培育抗逆工业大麻新种质提供基因资源。

干旱影响植株正常生长，导致纤维产量降低、品质下降，是限制工业大麻生产的主要环境因素之一，但目前国内外对工业大麻抗旱的研究较少，且均没有开展抗旱分子机理方面的研究。本研究首次从转录组水平分析工业大麻的抗旱应答机制，共得到1292个差异表达基因，其中上升表达883个，下降表达409个。共影响代谢路径11条，94个基因。其中激素信号转导通路为受影响最大的代谢路径，共有15个基因受到显著影响，而7个基因与ABA代谢路径密切相关，说明ABA在工业大麻干旱响应中起到了关键作用。以此为基础对40个大麻WRKY转录因子进行了功能鉴定，对干旱、盐和镉有显著性反应的转录因子分别有23个，15个和14个， 6个基因同时响应3种逆境反应。同时利用大麻转录组开发了3442对SSR引物，选取115份国内外大麻种质资源进行了种质资源鉴定，根据亲缘关系将种质资源分为4个区域，其中中国划分为3个区域，欧洲材料与中国北方地区的种质资源具有更近的亲缘关系，已顺利完成预期工作任务，并为大麻分子育种提供了良好基础。研究结果整理研究论文3篇，其中两篇SCI已接收，一篇正在修改，超额完成了项目设定的各项指标。