苹果内根-贝壳杉烯合成酶基因的克隆及表达分析

doi:10.3969/j.issn.1000-7091.2013.02.009

华北农学报 ›› 2013, Vol. 28 ›› Issue (2): 46-51. doi: 10.3969/j.issn.1000-7091.2013.02.009

所属专题： 苹果；生物技术；

苹果内根-贝壳杉烯合成酶基因的克隆及表达分析

邓晓云, 戴洪义, 梁美霞

青岛农业大学园林园艺学院, 山东青岛 266109

收稿日期:2012-12-02 出版日期:2013-04-28
通讯作者: 梁美霞(1970－)，女，山东莱阳人，副教授人，硕士生导师，主要从事果树生物技术研究。
作者简介:邓晓云(1986－)，女，山东潍坊人，在读硕士，主要从事果树遗传育种。
基金资助:
国家现代农业(苹果)产业技术体系项目(CARS-28-01-07);山东良种产业化工程项目;山东省教育厅项目(J10LC12);青岛市科技局项目(10-3-4-5-2-jch)

Cloning and Expression Analysis of Ent-Kaurene Synthasegene MdKS in Apple(Malus domestica Borkh)

DENG Xiao-yun, DAI Hong-yi, LIANG Mei-xia

College of Landscape and Horticulture, Qingdao Agricultural University, Qingdao 266109, China

Received:2012-12-02 Published:2013-04-28

摘要/Abstract

摘要： 以柱型苹果茎尖为试材,采用同源克隆的方法克隆得到赤霉素合成代谢关键酶内根-贝壳杉烯合成酶的cDNA全长序列,命名为MdKS。MdKS的开放阅读框(ORF)全长2 214 bp,编码737个氨基酸。KS属于萜类合成酶亚家族,具有DDXXD保守结构域。氨基酸聚类分析表明,MdKS与梨同源性最高,为98%,其次是板栗,为73%|实时荧光定量PCR分析表明,在苹果的生长季节,MdKS基因在柱型和普通型苹果中均能表达。在苹果的生长初期,普通型苹果MdKS基因的表达量均明显高于柱型苹果,而在6月初,其表达量却略低于柱型苹果。

关键词: 苹果, 内根-贝壳杉烯合成酶, 矮化, 实时荧光定量PCR

Abstract: Ent-kaurene synthase(KS) is a critical enzyme in the pathway ofgibberellins biosynthesis. In this research,ent-kaurene synthasegene in columnar apple,designated as MdKS,was isolated from apple(Malus domes-tica Borkh)stem apical tissue by homologous cloning technology. The full-length cDNA consisted of 2 214 nucleo-tides encoding a putative protein of 737 amino acids. MdKS contained a core functional domain DDXXD of plant class terpene synthase. Amino acids cluster analysis showed that KS from apple was highly homologous with PpKS firstly,had 98% similarity,and followed by that from Castanea sativa,had 73% similarity. The Real-time quantita-tive PCR analysis showed that MdKSgene were always expressed in columnar and standard apple trees during thegrowing season. In the earlygrowing season,the MdKS of standard apples were expressed at higher level than that of columnar apples,but on early June,it was opposite.

Key words: Apple, Ent-kaurene synthase, Dwarf, Real-time PCR

中图分类号:

S661. 1
Q78

邓晓云, 戴洪义, 梁美霞. 苹果内根-贝壳杉烯合成酶基因的克隆及表达分析[J]. 华北农学报, 2013, 28(2): 46-51. doi: 10.3969/j.issn.1000-7091.2013.02.009.

DENG Xiao-yun, DAI Hong-yi, LIANG Mei-xia. Cloning and Expression Analysis of Ent-Kaurene Synthasegene MdKS in Apple(Malus domestica Borkh)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(2): 46-51. doi: 10.3969/j.issn.1000-7091.2013.02.009.

http://www.hbnxb.net/CN/Y2013/V28/I2/46

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苹果内根-贝壳杉烯合成酶基因的克隆及表达分析

Cloning and Expression Analysis of Ent-Kaurene Synthasegene MdKS in Apple(Malus domestica Borkh)

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