慈竹BeSWEET42和BeSWEET1a2基因的克隆和表达分析
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四川省应用基础研究项目(18YYJC0920);


Cloning and Expression Analysis of BeSWEET42 and BeSWEET1a2 from Bambusa emeiensis
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    摘要:

    糖外排转运蛋白(sugar will eventually be exported transporters, SWEET)在植物光合同化产物的运输中主要参与韧皮部糖的装载过程。该研究基于慈竹转录组库筛选出9条完整的BeSWEET序列,对其进行生物信息学分析,以慈竹茎和嫩叶的cDNA为模板对9条BeSWEET序列中的BeSWEET42和BeSWEET1a2进行基因克隆,采用实时荧光定量(qRTPCR)分析其在叶肉、叶脉、根、茎中的表达水平以及外源糖诱导下的表达变化。结果表明:(1)系统进化分析显示,9条BeSWEET序列被分为4大类群,其中BeSWEET42与水稻SWEET4近缘,聚类到Clade Ⅱ,具有2个MtN3保守结构域;BeSWEET1a2与水稻SWEET1a和SWEET1b近缘,聚类到Clade Ⅰ。(2)序列分析显示,慈竹BeSWEET42和BeSWEET1a2分别编码256个和222个氨基酸,分别具有7个和5个跨膜结构域。(3)亚细胞定位预测显示:慈竹BeSWEET42和BeSWEET1a2均定位于质膜。(4)qRTPCR结果显示,慈竹BeSWEET42和BeSWEET1a2在叶肉、叶脉、根和茎中均有表达,分别在根和叶脉中最为显著,推测其可能协作参与了糖类从源到库的转运。(5)在外源己糖诱导下慈竹BeSWEET42和BeSWEET1a2均显著上调表达,显示出对己糖的偏好性。该研究结果为进一步探讨慈竹BeSWEET蛋白调控糖转运的生物学功能奠定了基础。

    Abstract:

    SWEET (sugar will eventually be exported transporters) protein participates in the loading of phloem sugars. In this study, 9 BeSWEET sequences were screened based on the transcriptome library of Bambusa emeiensis and analyzed by bioinformatics; Genes of BeSWEET42 and BeSWEET1a2 among the 9 BeSWEET sequences were cloned with the cDNA of the stem and young leaf of B. emeiensis. Real time quantitative expression analysis (qRTPCR) was used to analyze its expression levels in mesophyll, veins, roots, stems and the expression changes of BeSWEET gene induced by exogenous sugar. The results showed that: (1) phylogenetic tree analysis showed that 9 BeSWEET sequences were divided into four clades. BeSWEET42 which has two MtN3 conserved domains is homologous to rice SWEET4, clustered into Clade Ⅱ. BeSWEET1a2 is homologous to rice SWEET1a and SWEET1b, and clustered into Clade Ⅰ. (2) Sequence analysis showed that BeSWEET42 and BeSWEET1a2 encode 256 and 222 amino acids, respectively, and have 7 and 5 transmembrane domains, respectively. (3) The prediction of subcellular localization indicates that both genes are located in the plasma membrane. (4) qRTPCR analysis shows that BeSWEET42 and BeSWEET1a2 are mainly expressed in roots and veins, respectively. It is speculated that they may cooperate in the transport of carbohydrates from source to sink. (5) Both BeSWEET42 and BeSWEET1a2 were significantly upregulated under the induction of hexose, showing a preference for hexose. Our study provides the insight for understanding of biological functions of B. emeiensis BeSWEET proteins in regulating sugar transport.

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欧 倩,王博雅,杜恬恬,等.慈竹BeSWEET42和BeSWEET1a2基因的克隆和表达分析[J].西北植物学报,2020,40(11):1833-1839

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  • 在线发布日期: 2020-12-11
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