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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (21): 207-214.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

苦荞中查尔酮合成酶基因（CHS）的克隆

高帆 张宗文 李艳琴 吴斌 宋韡   

  • 收稿日期:2011-04-12 修回日期:2011-05-30 出版日期:2011-08-25 发布日期:2011-08-25
  • 基金资助:

    作物种质资源保护专项

Cloning of Chalcone Synthase Gene in Tartary Buckwheat

  • Received:2011-04-12 Revised:2011-05-30 Online:2011-08-25 Published:2011-08-25

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摘要：

获得完整的苦荞查尔酮合成酶基因（CHS）信息并评价其进化地位，对分子辅助选育高黄酮含量的苦荞品种具有重要的指导意义。用RACE法克隆苦荞CHS基因，用生物信息学手段分析预测苦荞CHS基本理化性质和同源性，用临接法构建了该酶的系统发生树。获得1250 bp的CHS cDNA全长，含241 bp的3’UTR和185 bp的5’UTR；等电点（pI）和分子量（Mr）分别为5.33和35340.75 Da；克隆获得975 bp的开放性阅读框（ORF）。预测该基因编码含325个氨基酸残基的蛋白，氨基酸同源比对结果表明，苦荞CHS与蓼科的虎杖相似性很高；临接法构建的系统发生树结果表明，苦荞CHS与其他双子叶植物有共同的起源，与蓼科的金荞麦、甜荞、虎杖及石竹科的满天星亲缘关系较近。成功获得苦荞CHS基因的cDNA全长，克隆出完整的ORF，并确定了苦荞中该酶的进化地位和方向。

关键词: GIS, GIS, MapX, 木薯, 现代产业体系, 管理系统

Abstract:

The key chalcone synthase (CHS) gene was cloned in tartary buckwheat, and its evolution status was evaluated, which would play an important role for molecular assistant breeding for the excellent Fagopyrum tataricum accessions with high flavonoids content. The gene was cloned by rapid amplification of cDNA Ends (RACE) and analyzed the basic physicochemical properties and homology by bioinformatics methods. Moreover, the phylogenetic tree of chalcone synthase family was constructed by neighbor-joining. Full length of CHS cDNA was 1250 bp containing 241 bp 3'UTR and 185 bp 5'UTR. Isoelectric point (pI) and molecular weight (Mr) were 5.33 and 35340.75 Da, respectively. Meanwhile, 975 bp open reading frame (ORF) was cloned. We predicted the gene encoding 325 amino acid residues. Amino acid sequences alignment results showed that chalcone synthase in tartary buckwheat had higher similarity with Polygonum cuspidatum. The phylogenetic tree showed that chalcone synthase of tartary buckwheat had one same origin with other dicots. The genetic relationships were closed among Gypsophila paniculata, Polygonum cuspidatum, F. dibotrys, F. esculentum, F. tataricum. We obtained the cDNA of CHS in tartary buckwheat, cloned the whole ORF, determine the status and direction of the enzyme in tartary buckwheat.