小麦Ta4CL1基因的克隆及其在促进转基因拟南芥生长和木质素沉积中的功能  

Cloning of Ta4CL1 and its function in promoting plant growth and lignin deposition in transgenic Arabidopsis plants

作　　者：孟颖 邢蕾蕾 曹晓红 郭光艳[1] 柴建芳[2] 秘彩莉[1] MENG Ying;XING Lei-Lei;CAO Xiao-Hong;GUO Guang-Yan;CHAI Jian-Fang;BEI Cai-Li(Life Science College,Hebei Normal University,Shijiazhuang 050024,Hebei,China;Institute of Genetics and Physiology,Hebei Academy of Agriculture and Forest Sciences,Plant Genetic Engineering Center of Hebei Province,Shijiazhuang 050051,Hebei,China)

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024 [2]河北省农林科学院遗传生理研究所/河北省植物转基因中心,河北石家庄050051

出　　处：《作物学报》2022年第1期63-75,共13页Acta Agronomica Sinica

基　　金：河北省自然科学基金项目(C2019205102),河北省重点研发计划项目(20326348D),河北师范大学重点基金项目(L2019Z05)和河北师范大学博士基金项目(L2018B14)资助。

摘　　要：4-香豆酸辅酶A连接酶(4CL;EC 6.2.1.12)位于苯丙烷途径分支点的上游,是苯丙烷代谢途径的核心酶,可产生木质素、黄酮类等化合物,这些化合物对植物的生长发育及环境适应性均具有重要作用。在双子叶植物中,有关4CL的研究较多,而在单子叶植物尤其是作物中的研究相对较少。本研究利用RACE技术从普通小麦中克隆了一个4CL基因Ta4CL1。系统发育分析表明,Ta4CL1与水稻、玉米和高粱等植物中在木质素合成中具有重要作用的4CLs聚成一类;利用Ta4CL1过表达、拟南芥4CLs突变体at4cl1、at4cl3和at4cl14cl3及其功能回复株系进行的分析表明,Ta4CL1与At4CL1功能相似,在植物木质素合成中具有重要作用,但未参与黄酮类化合物生物合成的调控过程;Ta4CL1是转基因拟南芥中4CL酶活性的主要贡献者。过表达Ta4CL1的转基因拟南芥叶片增大、茎更粗;Ta4CL1的表达还受茉莉酸甲酯(Methyl jasmonic acid,MeJA)、赤霉素(Gibberellin,GA)和生长素(Indoleacetic acid,IAA)等激素处理的影响。本研究为利用基因工程将Ta4CL1应用于改善小麦秸秆的利用效率提供了理论依据。4-Coumarate:coenzyme A ligase(4CL;EC 6.2.1.12)acts upstream of the branch point of phenylpropanoid,which is a key enzyme in the phenylpropanoid metabolic pathways for monolignol and flavonoid biosynthesis,and these compounds play important roles in plant growth and development as well as stress adaptability.Many 4CLs had been extensively studied in di-cotyledons,but their function in monocotyledons,especially in crops,was relatively poorly understood.In this study,Ta4CL1,which encoded 4-Coumarate:coenzyme A ligase,was isolated from common wheat by RACE technique.Polygenetic analysis revealed that Ta4CL1 could be clustered to the same group of 4CLs from rice,maize,and sorghum,which was mainly involved in lignin biosynthesis.Ta4CL1-overexpressed Arabidopsis lines,at4cl1,at4cl3,and at4cl14cl3 as well as their corresponding func-tional recovery lines were used to elucidate the function of Ta4CL1 in the phenylpropanoid metabolic pathway.The results sug-gested that Ta4CL1 had similar function with At4CL1 in regulating lignin biosynthesis but it had no effect on flavonoid biosyn-thesis.Ta4CL was the major contributor of 4CL enzyme activity in transgenic Arabidopsis plants.Overexpression of Ta4CL led to enlarged leaves and thickened stems in transgenic Arabidopsis seedlings,the expression of Ta4CL was also affected by MeJA,GA,and IAA treatments.These results provide the theoretical basis for improving the utilizing efficiency of plant straws using Ta4CL1 by genetic engineering.

关 键 词：4-香豆酸辅酶A连接酶 Ta4CL1 木质素沉积 促进生长 小麦 拟南芥 

分 类 号：S512.1[农业科学—作物学]