枸杞脱落酸生物合成关键酶基因NCED的克隆及表达分析
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国家高技术研究发展计划(863)项目(2007AA10Z189)


Cloning and characterization of 9-cis-epoxycarotenoid dioxygenase gene(NCED)encoding a key enzyme during abscisic acid biosynthesis in Lycium barbarum L
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the National High Technology Research and Development Program of China (863 -2007AA10Z189).

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    摘要:

    脱落酸(abscisicacid, ABA)对植物的生长发育具有独特的调控功能,并在植物适应逆境环境中发挥重要作用。9-顺式环氧类胡萝卜素双加氧酶(NCED)是高等植物中ABA生物合成途径的一个关键酶。根据GenBank中的植物NCED基因的同源序列设计简并引物,通过RT-PCR及RACE技术从枸杞叶片中克隆到一个编码NCED的基因,命名为LbNCED。其cDNA全长为2 316 bp,含有一个1 824 bp的开放阅读框,编码一个含607 氨基酸残基,分子量为67.38 kDa、等电点(pI)为6.43的假定蛋白。其氨基酸序列与番茄(Lycopersicon esculentum)和马铃薯(Solanum tuberosum)的同源性达90%,在N-末端具有一个含15个氨基酸的叶绿体转运肽。Southern 杂交结果表明,该基因在枸杞基因组中以低拷贝形式存在。盐处理和脱水处理的枸杞叶片中LbNCED基因的表达与内源ABA的积累同步变化。

    Abstract:

    Abscisic acid (ABA) regulates the essential physiological and developmental processes of plants and plays imporant roles in plant responses to various environmental stresses. 9-cis-epoxycarotenoid dioxygenase (NCED) is the key regulatory enzyme in the biosynthesis pathway of ABA in higher plants. In the study, a full-lengh cDNA of NCED gene (LbNCED) was fristly isolated and characterized from the leaves of L.barbarum. LbNCED has 2,316 bp, containing a 1,824-bp ORF and encoding 607 amino acids. Comparative and bioinformatics analyses revealed that the homologyamino acida sequence of LbNCED to Lycopersicon esculentum and Solanum tuberosum was 90%. At the N-terminus of the LbNCED located a 15-aa putative chloroplast transit peptide. Southern blot analysis revealed that it was a low-copy gene in the genome of L.barbarum. Real-time Quantitative PCR (RT-QPCR) analysis showed that LbNCED mRNA accumulated most abundantly in leaves. The RT-QPCR analysis revealed that dehydration, salt stress signficantly enhanced LbNCED transcript expression and ABA content accumulation.

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陆平,田跃胜,王名雪,等.枸杞脱落酸生物合成关键酶基因NCED的克隆及表达分析[J].植物遗传资源学报,2013,14(2):303-310.

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  • 收稿日期:2012-02-29
  • 最后修改日期:2012-10-20
  • 录用日期:2013-01-14
  • 在线发布日期: 2013-01-29
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