江西铅山红芽芋是国家地理标志农产品，由于长期无性繁殖导致其病毒积累、种性退化、产量和品质下降。植物脱毒能恢复种性是学术界一个共识。超低温疗法是超低温保存技术在植物脱毒上的一个新应用，可代替传统茎尖培养，实现种质保存和植物脱毒双重目标。目前对超低温疗法脱毒的研究多集中于操作程序、病毒检测、遗传变异和细胞凋亡等方面，而关于植物对超低温疗法脱毒的响应机制少见报道。本课题以江西铅山红芽芋为研究对象，通过超低温疗法对其进行脱毒，获得脱毒苗，然后采用RNA-Seq技术，对其进行转录组分析，筛选出与超低温疗法脱毒密切相关的候选基因，克隆其全长cDNA序列，并通过遗传转化过量表达研究其确切功能及其精确的亚细胞定位，分析其作用机制模式，再结合光合生理、生理生化指标、DNA甲基化、抗氧化酶基因和激素相关基因表达等分析，初步诠释红芽芋超低温疗法脱毒的响应机制，为植物脱毒恢复种性以及植物超低温保存提供理论依据。

Jiangxi Yanshan red bud taro is “national agricultural product of geographical indication”. The long-term asexual reproduction has caused its virus accumulation, variety degeneration, yields and quality decline. It is a common understanding in the academic field that plant virus-free can recover its variety. Cryotherapy is a new application of cryopreservation technique in plant virus-free, it can replace the traditional stoot-tip culture, and achieve the dual objective of germplasm conservation and plant virus-free. At present, the study on virus-free by cryotherapy is mainly focused on the operation process, virus detection, genetic variation, apoptosis and so on, while the response mechanism of plants to virus-free by cryotherapy is rarely reported. On the base of working foundation obtained in our lab, in this project, using Jiangxi Yanshan red bud taro as experimental material, the virus-free plantlets will be obtained by shoot-tip virus-free by cryotherapy. Then its transcriptome will be analyzed by RNA-Seq technique and the candidate genes closely related to virus-free by cryotherapy will be selected; The full-length cDNA sequence of candidate genes will be cloned and their exact function and precise subcellular localization will be studied through the overexpression of genetic transformation, whose mechanism model will be analyzed. At last combined with photosynthesis, physiological and biochemical analysis, DNA methylation and expression of antioxidant enzyme genes and genes related to hormones, the response mechanism of plants to shoot tip virus-free by cryotherapy will be preliminarily interpretated, which will provide some theoretical foundations for plant virus-free recovering the variety and plant germplasm cryopreservation.