忍冬属植物抗旱性综合评价及其相关MYB转录因子的克隆与鉴定

The plants of Flos Lonicerae are not only the resource plant of medical herb jinyinhua, but also are adaptable plant to Karst environment; MYB factors are important in regulatory networks controlling development, metabolism and responding to biotic and abiotic stresses. The cloning and functional verification of MYB transcription factors related to drought is important to explain the adaptation mechanism of plant to Karst environment and control rocky desertification.. In this study, the comprehensive drought resistance ability of different Flos Loniceraes. is evaluated according to their physiological indexes under drought through principal component analysis and subordinate function method. Then the specie with the highest resistance to drought is used as the material from which the gene of MYB factors is selected through digital gene expression profile of the samples under different degree of drought. The candidate genes selected are cloned and functional verification by the transgenic Arabidopsis and Flos Lonicerae.. This study can provide the resistant genes for the economic plant, and also be useful to explain the relation of medical quality and environment of Lonicerae genus and the mechanism of adaptation to Karst.

忍冬属植物是药典中金银花和山银花的来源植物，同时也是岩溶区的适生植物；MYB转录因子家族成员广泛参与植物次生代谢调控和对激素及环境因子应答；忍冬属植物抗旱综合评价及其抗旱相关的MYB转录因子克隆与功能验证对石漠化治理的引种工作具有实践意义，对于阐明植物适应岩溶环境机制具有理论意义。.本项目首先用主成分分析和隶属函数法对几种忍冬属植物进行抗旱综合评价；以干旱胁迫第一主成分因子的生理指标为依据，对其中抗性最好的种通过数字基因差异表达谱分析,结合q-PCR的方法进行响应干旱的相关MYB转录因子筛选；然后将侯选基因以农杆菌介导的方式转化到拟南芥和忍冬属植物中进行功能验证，期望筛到重要的与抗逆相关的MYB转录因子。.本项目能够为有经济价值的植物提供优良的抗逆基因，在为阐明忍冬属植物药用品质与环境之间的关系提供线索的同时也能为阐明植物对岩溶环境的适应机制提供数。

忍冬属植物是药典中金银花和山银花的来源植物，同时也是岩溶区的适生植物。MYB转录因子家族成员广泛参与植物次生代谢调控和环境因子应答。本课题对种质资源创新和广西石漠化治理的引种工作具有重要意义。.对红色金银花、四季花、华南忍冬、水忍冬和红腺忍冬这5份种质进行盆栽控水实验，以SOD酶活性、POD酶活性、APX酶活性、CAT酶活性、可溶性蛋白质含量、叶绿素含量、丙二醛含量、脯氨酸含量、光合速率、水份利用率等10种生理指标为评价指标评价抗旱性能，结果表明红腺忍冬其中抗性最好的种质。用PEG6000处理红腺忍冬种子苗，SPSS分析叶片抗旱生理指标，结果表明蛋白质含量、APX酶活性和叶绿素含量是第一主成分指标。依据这三个指标确定0, 10%和15%的PEG6000处理样品为测序样品进行转录组测序。依测序结果，共克隆了8个全长MYB转录因子基因，确定其中6个为候选基因参加功能验证。共构建过表达和干扰表达载体12个，分别转化红腺忍冬。总计DNA阳性苗共162株。Cluster-1328.31475和C39455.graph-c0的转化苗占了绝大多数。对阳性苗进行PEG6000胁迫处理，测定生理指标的同时进行相关基因的QPCR检测。结果表明，基因Cluster-1328.31475相对表达量与CAT酶活和SOD酶性相关，相关系数分别为-0.76和0.61； 与TRINITY_DN21544_c0_g2（SOD）、TRINITY_DN24342_c2_g1(APX)和TRINITY_DN29963_c1_g1(CHLH)相对表达量相关系数分别为-0.731、0.600和-0.511。以上初步说明此MYB转录因子参与了植物对干旱的响应过程。进一步功能验证正在进行中。