百合LiP5CS、LiGDH和LiOAT基因克隆及表达分析  

Cloning and Expression Analysis of LiP5CS,LiGDH and LiOAT Genes in Lilium

作　　者：阎俊卉 刘立 谢天 赵子贤 赵家惠 文锦芬 Yan Junhui;Liu Li;Xie Tian;Zhao Zixian;Zhao Jiahui;Wen Jinfen(College of Architecture and Urban Planning,Kunming University of Science and Technology,Kunming,650500)

机构地区：[1]昆明理工大学建筑与城市规划学院,昆明650500

出　　处：《分子植物育种》2022年第6期1776-1787,共12页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31760588,32160721)资助。

摘　　要：为探究百合脯氨酸生物合成途径中部分相关基因的功能及其对非生物胁迫的响应,本研究以百合‘西伯利亚’作为试验材料,克隆了3个脯氨酸生物合成相关基因:Δ^(1)-吡咯啉-5-羧酸合成酶基因(LiP5CS)、谷氨酸脱氢酶基因(LiGDH)和鸟氨酸氨基转移酶基因(LiOAT),分析其蛋白特性,并测定3个基因在百合不同组织、花不同发育时期以及失水和低温胁迫下的相对表达量和酶活性。结果表明,LiP5CS长度为1 068 bp,编码355个氨基酸残基;LiGDH长度为429 bp,编码142个氨基酸残基;LiOAT长度为870 bp,编码289个氨基酸残基。LiP5CS、LiGDH和LiOAT都含有特有保守motif,皆为脂溶性、亲水性的稳定蛋白,都具有磷酸化位点,3个蛋白在进化上具有一定保守性。表达分析发现,3个基因在百合鳞茎中的表达量最低,LiP5CS与LiOAT在根中的表达量最高,LiGDH在花中的表达量最高;在花蕾的不同发育时期中LiP5CS基因表达随发育呈下降趋势,而LiGDH与LiOAT表达呈升高趋势。在失水和低温胁迫下,LiP5CS、LiGDH和LiOAT的表达均被诱导升高,其中LiP5CS的表达量峰值出现较早,3个酶活性变化趋势与各自基因表达变化相似。本研究结果暗示百合切花中的谷氨酸途径是响应低温和失水胁迫的主途径,而鸟氨酸途径为辅助途径。In order to explore the function of some rel ated genes in proline biosynthesis pathway and their response to abiotic stress in lily,three proline biosynthesis related genes were cloned from lily ’Siberia’ as experimental material:Δ^(1)-pyrroline-5-carboxylate synthetase gene(LiP5CS),glutamate dehydrogenase gene(LiGDH) and ornithine amino transferase gene(LiOAT),then,the protein characteristics were analyzed and the relative gene expression levels in different tissues,flower development stages,water loss and low temperature stress were determined.Meanwhile the enzyme activities of the three genes were also checked under water loss and low temperature stress.The results showed that the length of LiP5CS was 1 068 bp,encoding 355 amino acid residues.LiGDH was 429 bp in length,encoding 142 amino acid residues.The length of LiOAT was 870 bp,encoding 289 amino acid residues.LiP5CS,LiGDH and LiOAT proteins were lipid soluble and hydrophilic stable proteins with phosphorylation sites.All the three proteins were conserved in different families.Expression analysis showed that the expression levels of the three genes were the lowest in bulbs,while the expression levels of LiP5CS and LiOAT were the highest in roots,and LiGDH was the highest in flowers.LiP5CS gene expression decreased with the development of bud,while LiGDH and LiOAT expression increased.Under water loss and low temperature stress,LiP5CS,LiGDH and LiOAT expressions were induced to increase,and the peak of LiP5CS expression appeared earlier,and the change trends of the three enzymes activities were similar to the changes of gene expression.The results suggested that glutamate pathway was the main pathway in response to low temperature and water loss stress,ornithine pathway is the auxiliary pathway in lily cut flowers.

关 键 词：百合 Δ^(1)-吡咯啉-5-羧酸合成酶 谷氨酸脱氢酶 鸟氨酸氨基转移酶 非生物胁迫 

分 类 号：S682.29[农业科学—观赏园艺]