阳春砂茉莉酸生物合成酶基因鉴定与时空表达分析

Identification and Spatiotemporal Expression Analysis of Jasmonic Acid Biosynthetic Enzyme Genes in Amomum villosum

为探究阳春砂(Amomum villosum)花器中茉莉酸(JA)生物合成酶基因表达与雌雄蕊生长的关系,本研究利用cDNA末端快速扩增(RACE)技术克隆阳春砂JA生物合成酶基因的全长序列,并对各基因进行生物信息学分析、表达模式测定及各基因表达与阳春砂雌雄蕊生长的相关性分析,以探究阳春砂花器中JA生物合成酶基因表达与雌雄蕊生长的关系。结果表明,阳春砂中JA生物合成酶基因AvLOX、AvAOS、AvAOC、AvOPR3序列全长分别为2733、1798、930、1400 bp,分别包含2577、1485、744、1179 bp的开放阅读框(ORF),其编码蛋白分别含有858、494、247、392个氨基酸残基。4个JA生物合成酶基因的表达随阳春砂雄蕊的发育进程逐渐增加,但在雌蕊中的表达则呈现多样性。在雌蕊中,AvLOX、AvAOS基因在开花后1天表达最高,AvAOC在标记后第9天表达最高,AvOPR3基因在标记后第6天表达最高。除AvAOS基因在小花在标记后第3天以及AvOPR3基因在小花标记后第6天外,4个JA生物合成酶基因在雄蕊中的表达在各发育阶段显著高于同时期在雌蕊中的表达。与对照组相比,在喷施茉莉酸甲酯(MeJA)后,4个JA生物合成酶基因在雄蕊中的表达模式,以及AvLOX和AvOPR3基因在雌蕊中的表达模式没有改变,但AvAOS和AvAOC基因在雌蕊中的表达模式有变化。在对照组和MeJA处理组中,4个JA生物合成酶基因的表达与阳春砂小花花丝和花柱的长度有极显著的相关性。本研究结果为进一步探究JA与阳春砂花器官生长发育的关系奠定了基础。

To explore the relationship between the expression of jasmonic acid(JA)biosynthetic genes and the growth of stamen and pistil of A.villosum.The full-length JA biosynthase genes were cloned using RACE technology,and their sequences were analyzed using the bioinformatics method.The gene expressions and their correlation with stamen and pistil growth were then examined.The results showed that in A.villosum,the full lengths of the JA biosynthase genes AvLOX,AvAOS,AvAOC,and AvOPR3 were 2733 bp,1798 bp,930 bp,and 1400 bp respectively,with ORFs of 2577 bp,1485 bp,744 bp,and 1179 bp,encoding 858,494,247,and 392 amino acids.The expressions of these four JA biosynthetic enzyme genes increased as the stamen developed in A.villosum,however,there was diverse expressions in the pistil.In pistil,the expressions of AvLOX and AvAOS peaked on the first day after flowering.The expression peak of AvAOC occurred on the 9th day after marking,and the expression peak of AvOPR3 occurred on the 6th day after marking.Futhermore,the expressions of these four JA biosynthase genes in stamen were significantly higher than those in pistil at the same developmental stage,except for AvAOS on the 3rd day and AvOPR3 on the 6th day in floweret.Compared to the control,the expression patterns for four JA biosynthase genes in stamen,as well as AvLOX and AvOPR3 in pistil,remained unchanged after spraying MeJA(methyl jasmonate).However,there were differences observed for both AvAOS and AvAOC in the pistil.Under both the control and MeJA treatments,the expressions of four JA synthase genes showed significant correlations with the length of the filament and style within A.villosum floweret.This study lays a foundation for further research on the regulation of JA on the growth and development of the floral organ of A.villosum.