GC-MS内标法定量分析植物组织中的JA、IAA和ABA  

Quantitative analysis of jasmonic acids,indole-3-acetic acid and abscisic acid in plant tissues by GC-MS

作　　者：李金克[1] 陈华君[1] 陈少良[2,3] 

机构地区：[1]北京林业大学实验室与设备管理处 [2]林木育种国家工程实验室,林木花卉遗传育种教育部重点实验室,国家林业局树木花卉育种与生物工程重点开放实验室,北京林业大学生物科学与技术学院 [3]湖北民族学院生物资源保护与利用湖北省重点实验室

出　　处：《北京林业大学学报》2010年第5期143-148,共6页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(30872005;30430430);"863"国家高技术研究发展计划项目(2006AA10Z131);北京市优秀博士学位论文指导教师科技项目(YB20081002201);教育部重点项目(2009-84);全国高校优秀博士学位论文作者专项基金项目(200152);高等学校优秀青年教师教学科研奖励计划(2002-323);霍英东教育基金会高等院校青年教师基金项目(91031);湖北省自然科学基金项目(2007ABB003)

摘　　要：提出了利用气相色谱-质谱联用仪同时定量测定植物组织中茉莉酸(JA)、吲哚-3-乙酸(IAA)和脱落酸(ABA)的方法。实验以欧洲灰杨叶片为材料,在用80%冷甲醇提取激素时,一并加入ABA、IAA和JA的内标(D3-ABA300ng,13C6-IAA60ng和DHJA80ng),激素提取液浓缩至水相,调pH2.5～3.0后经乙酸乙酯萃取、Sep-PakC18小柱纯化,甲酯化后,GC-MS定量测定ABA、JA。IAA经甲酯化后再经TMS化(三甲基硅烷化),用GC-MS定量测定。经检测:盆栽欧洲灰杨叶组织中脱落酸含量为120.170～128.771ng/g(鲜质量),茉莉酸含量为1.697～6.974ng/g(鲜质量),吲哚乙酸含量为4.990～14.776ng/g(鲜质量);脱落酸含量最高,其次是吲哚乙酸,茉莉酸含量最低。GC-MS was used to determine the concentrations of jasmonic acid（JA）,indole-3-acetic acid（IAA） and abscisic acid（ABA） in Populus canescens leaves in this paper.Abundant cold methanol（80%,v/v） was used to extract JA,IAA and ABA,and three internal standards,D3-ABA 300 ng,13C6-IAA 60 ng and DHJA 80 ng were simultaneously introduced into the leaf samples.Crude extract was condensed by vacuum evaporating and hormones were re-extracted by ethyl acetate at pH 2.5-3.0（× 3）.Then condensed JA,IAA and ABA extracts were purified by Sep-Pak C18 columns.Thereafter,partially purified hormones were methylated using trimethylsilyldizomethane and then analyzed by GC-MS（JA and ABA）.IAA was analyzed by GC-MS after an additional TMS reaction.Results showed that concentrations of ABA,JA,and IAA in leaf tissues of P.canescens were 121.064-128.771 ng/g,2.190-6.974 ng/g and 4.990-15.012ng/g Fw,respectively.In concentration in P.canescens leaves,ABA 〉 IAA 〉 JA.

关 键 词：气相色谱-质谱联用仪 内标法 茉莉酸 吲哚乙酸 脱落酸 欧洲灰杨 

分 类 号：Q947.885[生物学—植物学]