基于转录组测序的人参自毒胁迫响应分子机制研究

As an intractable problem, continuous cropping influence the yield and quality of Panax ginseng severely. More and more research indicated that autotoxicity is one of the major factors induced continuous cropping problem in Panax ginseng. But unfortunatedly, what changes have happened in gene expression and regulation system under autotoxic atrss was still unknown yet. Based on the work we have done, in the present project, we plan to research the different expression of functional genes after and before induced by autotoxic stress in Pananx ginseng on transcriptome level by high-throughput sequencing. To identify functional genes that tightly linked with autotoxic stress by bioinformatic analysis. To clone the full-length sequence of target gene by RACE, and verify the function of gene by transgenic method. In addition illustrated the molecular mechanism of Panax ginseng answer autotoxic stress, results in the present project will also be helpful to interprect the molecular mechanism of continuous cropping problem in Panax ginseng.

连作障碍是影响人参产量和品质的突出问题。已有研究表明，自毒作用是导致人参连作障碍的主要因素之一。人参在自毒胁迫作用下，基因表达调控系统如何变化从而导致生长异常，至今尚无研究报道。本项目在课题组前期研究基础上，拟借助高通量测序平台，从转录组水平研究自毒胁迫诱导前后人参功能基因的差异表达情况；通过生物信息学分析，明确差异表达基因所在的信号通路及其表达调控模式，根据注释信息找到与自毒胁迫相关的功能基因。用RACE技术克隆目的基因全长序列，并通过转基因手段验证其功能。本项目研究结果不仅可揭示人参响应自毒胁迫的应答分子机制，也有助于全面阐明人参连作障碍的作用机理。

连作障碍是影响人参产量和品质的突出问题，揭示人参基因表达调控系统在自毒胁迫作用下的变化规律，将有助于深入阐述人参响应自毒胁迫的分子机制。本项目在课题组前期研究基础上借助Illumina高通量测序平台，从转录组水平研究了苯甲酸胁迫诱导前后人参根、茎、叶功能基因的差异表达情况，结合实时荧光定量PCR验证，获得了7个与人参生理指标相关基因、7个人参皂苷合成相关基因和13个人参转录因子相关基因；用RT-PCR方法及RACE技术克隆到1条WRKY转录因子编码基因WRKY7，通过农杆菌介导法将其转化到拟南芥，获得了能够稳定遗传的转基因株系，为后续相关基因功能研究奠定了基础。本研究结果有助于揭示人参响应自毒胁迫应答的分子机制以及全面阐明人参连作障碍作用机理。