小麦β胡萝卜素异构酶基因定位克隆及表达分析
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中央高校基本科研业务费专项(2014YB032)


Cloning, Mapping and Expression Analysis of βcarotene Isomerase Encoding Genes in Wheat (Triticum aestivum L.)
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    摘要:

    β胡萝卜素异构酶(D27)是独脚金内酯(SLs)合成通路的第一个酶。该研究以普通小麦‘中国春’为材料,通过RTPCR克隆了小麦β胡萝卜素异构酶对应cDNA(TaD27),并对其在不同组织和低磷胁迫下的表达进行分析。结果显示:(1)克隆获得2个高度同源cDNA片段,分别定位于7A、7D 染色体的长臂,命名为TaD277ALTaD277DL;序列分析发现,7B染色体长臂上还存在另一个同源基因TaD277BL。(2)3个TaD27基因均包含7个外显子,编码区长度分别为828 bp(7AL)、840 bp(7BL) 和843 bp(7DL) ,编码蛋白的N端均含有叶绿体转运肽;进化分析表明,植物D27蛋白主要聚集在3个进化枝中,小麦、水稻和玉米等单子叶植物的D27高度同源,聚集在同一进化分枝。(3)组织表达分析表明,TaD27基因在叶、叶鞘和茎中的表达量较高,在幼穗中的表达量相对较低,在根中的表达量最低;低磷胁迫下,TaD27在根中的表达量先下降后升高,胁迫至6~12 h时,表达量达到最低,96 h时基本恢复至初始状态,而在叶中表达量则是先升高后下降,6 h达到峰值,12 h基本恢复到初始水平,随后表达量继续下降。

    Abstract:

    βcarotene isomerase (D27) is the first enzyme involved in strigolactones biosynthesis. In this study, the cDNAs encoding βcarotene isomerase (TaD27) were cloned from wheat cultivar Chinese Spring by RTPCR. Further, their expression in different tissues and under low phosphate stress were analysed by Realtime Quantitative PCR. The result showed that: (1) Two TaD27 cDNAs were cloned, which was maped on the long arms of 7A and 7D chromosome, respectively (named TaD277AL and TaD277DL). In addition, we found another locus located on chromosome 7BL encoding the third homolog (TaD277BL) by sequence analysis. (2) Each of the three TaD27s contains 7 exons composing an ORF of 828 bp (7AL), 840 bp(7BL) or 843 bp (7DL), respectively and encodes a protein containing a chloroplast transit peptide at Nterminal. Phylogenetic analyses indicated that D27s in plants clustered into three independent clades. TaD27s share a high level of identity with orthologs in other monocots such as Oryza sativa and Zea mays, as a result, they are clustered in the same clade. (3) The expression level of TaD27 is relatively high in leaves, sheaths and stems, lower in young panicles, and lowest in roots. Under low phosphate stress the expression of TaD27 in roots is down regulated to the bottom in 6-12 h firstly, and then it rises to the normal level after 96 h. On the contrary, the expression of TaD27 in leaves is up regulated in early stage and the peak is at 6h, then it continuously drops to a relatively low point at 96 h.

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张 朋,贾 笑,邓 帅,等.小麦β胡萝卜素异构酶基因定位克隆及表达分析[J].西北植物学报,2016,36(7):1315-1320

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  • 在线发布日期: 2016-08-16
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