基于CRISPR/Cas9系统探究乙酰甲胺磷影响番茄光合作用机制

Acephate is an important organophosphorus pesticide, which has been proved with an ability to affect the photosynthesis in tomato (Solanum lycopersicum) leaves. Low concentration of the pesticide would improve the photosynthesis, whereas high concentration of that would inhibit the bioprocess. With tomato as the test organism and CRISPR/Cas9 as main method, this project aims to 1): Construct lhcB1、LHCP mutant and determine effects of acephate against the photosynthesis factors in mutants and wild tomatoes; 2) Construct the complementation of the mutants with transgene method, and detect the effects of acephate against the photosynthesis factors in wild tomatoes and mutants as well as its complementation; 3) Analyze functions of lhcB1、LHCP playing in influence in tomato leaves by acephate. Therefore, this project would be meaningful to: 1)confirm the mechanism of acephate against photosynthesis within tomato leaves; 2) provide clues to identify the modulation mechanism against acephate stress; 3)provide theoretical basis for efficient and reasonable application of organophosphorus pesticide, as well as to study the phytotoxicity by insecticides.

乙酰甲胺磷是重要的有机磷农药，前期研究表明该杀虫剂可对番茄叶片光合作用有影响。其中低浓度处理对光合作用有提升效果，而高浓度却能抑制光合作用。本项目拟以番茄为供试生物，采用CRISPR/Cas9构建lhcB1、LHCP基因突变体，并检测乙酰甲胺磷对突变体与野生型叶片光合效率以及相关参数的差异；通过转基因技术进行突变体的互补研究，探究互补结果与突变体以及野生型叶片光合作用受乙酰甲胺磷影响的差异，分析、推测lhcB1、LHCP基因在乙酰甲胺磷影响光合作用中的功能。研究结果可以为明确有机磷杀虫剂乙酰甲胺磷对番茄叶片光合作用的影响机制以及番茄自身对此胁迫的调节机制提供线索，可以为高效、合理利用有机磷农药提供参考，也可以为研究杀虫剂药害的产生提供理论依据。