百合抗逆基因遗传转化及转化后植株的抗逆性鉴定

Genetic transformation of lily stress resistance gene and identification of stress resistance of transformed plants

【目的】获得抗逆性强的东方百合新材料。【方法】从卷丹(Lilium lancifolium)中克隆得到的抗逆相关基因LlZFHD 4和LlMYB3同源转化至东方百合‘索邦’和‘西伯利亚’。利用普通PCR检验2种基因是否成功整合到受体植物基因组。利用qPCR筛选受体植物基因组中过表达2种基因的植株。通过测定寒冷和干旱胁迫后转基因植株的可溶性糖含量、相对电导率以及失水率来验证2种基因的功能。【结果】PCR结果表明,成功在受体植物基因组内检测到目的基因。qPCR结果表明,有4株植株的目的基因表达量高于对照组。可溶性糖含量和相对电导率结果表明,LlZFHD 4和LlMYB3基因可提高2种百合的抗寒性。失水率结果表明,LlZFHD4和LlMYB3基因可提高2种百合的抗旱性。【结论】得到4株转基因植株,检测结果表明,LlZFHD4和LlMYB3基因可提高2种百合的抗寒性和抗旱性。

【Objective】In order to obtain new materials of oriental lily with strong stress resistance.【Method】The stress-resistant genes LlZFHD 4 and LlMYB 3 cloned from Lilium lancifolium were found to be homologous to transform Oriental lilies‘Sorbonne’and‘Siberia’.The successful integration of two genes into the recipient plant genome was tested by using standard PCR.Plants that overexpressed both genes in the recipient plant genome were identified using qPCR.The two genes’functions were confirmed by evaluating the soluble sugar level,relative conductivity,and water loss rate of transgenic plants under cold and drought stress.【Result】The target gene was successfully found in the recipient plant genome according to the PCR data.qPCR results revealed that the expression of the target gene in four plants was higher than that of control group.The soluble sugar content and relative conductivity data indicated that the LlZFHD 4 and LlMYB 3 genes could increase the cold tolerance of the two lilies.The water loss rate data revealed that the LlZFHD 4 and LlMYB 3 genes could improve drought resistance in the two lilies.【Conclusion】Four transgenic plants were obtained,and the results revealed that the LlZFHD 4 and LlMYB 3 genes may improve the two lilies’cold and drought resilience.