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作 者 ：王名雪1,陆 平2，许 菲1，潘琪芳1，唐克轩1，赵静雅1*

期 刊 ：西北植物学报 2012年 12期 页码：2365~2373

关键词：长春花；丙二烯氧化物合酶；茉莉酸生物合成途径；克隆；表达分析；

Keywords:Catharanthus roseus, allene oxide synthase, jasmonate biosynthetic pathway, clone, expression,

摘 要 ：利用RT-PCR和RACE相结合的方法，从长春花中克隆了丙二烯氧化物合酶（AOS）基因。结果显示：长春花AOS基因（CrAOS）cDNA全长为2 118 bp，包括5′和3′非翻译区，polyA尾和一个长1 638 bp的开放阅读框，其基因组中不含内含子；CrAOS基因编码的蛋白含545个氨基酸。多重比对表明CrAOS蛋白与其他的AOS蛋白具有较高的相似性，CrAOS蛋白序列中含有AOS家族应有的保守氨基酸残基。Southern杂交表明：CrAOS基因在长春花中为低拷贝。qRT-PCR结果显示：CrAOS在各个组织均有表达但表达量存在差异，在老叶中最高，在幼花中表达最低。对长春花幼苗进行不同处理，结果表明：伤害、低温、甲基茉莉酸、乙烯利处理等可使CrAOS基因表达量显著提高，水杨酸处理对基因表达影响不大。

Abstract:In this study,a full-length cDNA of allene oxide synthase (AOS) gene (named as CrAOS,JQ364955) was cloned from Catharanthus roseus.The gene was 2 118 bp in size containing an open reading frame (1 638 bp) encoding 545 amino acids.Comparative and bioinformatic analysis revealed that the deduced protein of CrAOS was highly homologous to AOSs from other plant species.Southern blot analysis revealed that it was a low-copy gene.Real-time Quantitative PCR (qRT-PCR) analysis showed that CrAOS mRNA accumulated most abundantly in old leaves and least in young alabastrums.The qRT-PCR analysis revealed that wound,low temperature,methyl jasmonic acid,ethylene treatments significantly enhanced CrAOS transcript expression,and salicylic acid had no influence.

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