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作物学报 ›› 2018, Vol. 44 ›› Issue (02): 236-244.doi: 10.3724/SP.J.1006.2018.00236

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陆地棉钾转运体基因GhHAK5的序列特征及表达分析

晁毛妮1, 温青玉2, 张志勇1, 胡根海1, 张金宝1, 王果1, 王清连1,*()   

  1. 1河南科技学院 / 现代生物育种河南省协同创新中心 / 河南省棉麦分子生态与种质创新重点实验室, 河南新乡 453003
    2河南省农业科学院, 河南郑州 450002
  • 收稿日期:2017-04-25 接受日期:2017-09-10 出版日期:2018-02-12 网络出版日期:2017-10-27
  • 通讯作者: 王清连
  • 基金资助:
    本研究由国家自然科学基金项目(31601347, 31571600), 河南省科技攻关计划项目(172102110022), 河南省高等学校重点科研计划项目(15A210001)和河南科技学院高层次人才计划项目(2014004)资助

Sequence Characteristics and Expression Analysis of Potassium Transporter Gene GhHAK5 in Upland Cotton (Gossypium hirsutum L.)

Mao-Ni CHAO1, Qing-Yu WEN2, Zhi-Yong ZHANG1, Gen-Hai HU1, Jin-Bao ZHANG1, Guo WANG1, Qing-Lian WANG1,*()   

  1. 1 Henan Institute of Science and Technology / Henan Collaborative Innovation Center of Modern Biological Breeding / Henan Key Laboratory for Molecular Ecology and Germplasm Innovation of Cotton and Wheat, Xinxiang 453003, Henan, China
    2 Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2017-04-25 Accepted:2017-09-10 Published:2018-02-12 Published online:2017-10-27
  • Contact: Qing-Lian WANG
  • Supported by:
    This study was supported by the National Natural Science Foundation of China (31601347, 31571600), the Henan Scientific and Technological Research Program (172102110022), Key Research Projects of Henan Higher Education Institutions (15A210001), and High Level Talent Program of Henan Institute of Science and Technology (2014004).

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摘要：

KUP/HAK/KT钾转运体基因家族对植物吸收钾离子发挥重要作用, 鉴定和克隆棉花的钾转运体基因, 对于改良棉花的钾吸收特性, 提高棉花的产量和品质具有重要意义。基于已测序的陆地棉基因组序列, 本研究通过同源克隆的方法鉴定到陆地棉钾转运体基因GhHAK5, 并以陆地棉品种百棉1号为材料对其CDS序列进行扩增。结果表明, GhHAK5基因的CDS全长为2451 bp, 编码816个氨基酸, 分子量和等电点分别为91.23 kD和8.15。GhHAK5蛋白具有KUP/HAK/KT家族基因的保守结构域“K-trans”(Pfam02705)和标志性序列GXXXGDXXXSPLY, 并具有11个跨膜区。在进化上, GhHAK5蛋白与拟南芥AtHAK5亲缘关系最近, 其次是与水稻的OsHAK5, 它们同属Cluster I进化簇。亚细胞定位结果显示, GhHAK5是一个定位于质膜的蛋白, 这与其主要作为钾转运子参与K+吸收的功能是一致的。GhHAK5基因在根中表达量最高, 在茎、叶、花瓣、纤维和花萼中表达量很低, 且其表达受外界低钾环境诱导。本研究结果为进一步了解GhHAK5基因的功能及培育钾高效棉花品种奠定了基础。

关键词: 陆地棉, 钾转运体基因, 亚细胞定位, 低钾, 基因表达

Abstract:

KUP/HAK/KT potassium transporter gene family plays an important role in the absorption of K+ in plants. Identification and cloning of potassium transporter genes in cotton are helpful to improve potassium absorption, and facilitate the increase of yield and quality in cotton. Based on the genome sequence of upland cotton, we identified the potassium transporter gene GhHAK5 by homologous cloning, and amplified its CDS sequence from upland cotton variety Baimian 1. The CDS of GhHAK5 was 2451 bp, encoding a protein with 816 amino acids. The molecular weight and isoelectric point of this protein were 91.23 kD and 8.15, respectively. The GhHAK5 protein contained a conserved domain “K-trans” (Pfam02705) and a symbolic amino acid sequence GXXXGDXXXSPLY of KUP/HAK/KT family gene, and had 11 transmembrane domains. Phylogenetic tree analysis demonstrated that GhHAK5 had a close relationship with AtHAK5, followed by OsHAK5, and they all belong to Cluster I evolutionary cluster. The subcellular localization experiment indicated that GhHAK5 was located in plasma membrane, which is consistent with its function as a potassium transporter involved in K+ uptake. The expression of GhHAK5 was high in roots, but low in leaf, stem, petal, fiber and sepal, and induced by low potassium environment. These results provide a foundation for understanding the function of GhHAK5 and breeding varieties with high potassium efficiency in cotton.

Key words: upland cotton, potassium transporter gene, subcellular localization, low potassium, gene expression

图1

陆地棉GhHAK5蛋白的序列分析 A: GhHAK5的保守域分析; B: GhHAK5中含有的钾转运体标志性序列(用横线表出); C: GhHAK5的跨膜区预测。"

图2

陆地棉GhHAK5蛋白与拟南芥和水稻KUP/HAK/KT家族成员的进化关系分析空心和实心分别代表水稻和拟南芥KUP/HAK/KT家族成员。"

图3

陆地棉GhHAK5蛋白的亚细胞定位 a和d: 荧光下照片; b和e: 明场下照片; c和f: 荧光和明场叠加图。"

图4

陆地棉GhHAK5基因表达水平 A: GhHAK5在陆地棉不同组织中的表达分析(误差线代表3个独立重复的标准差); B: 低钾处理下GhHAK5在根中的表达分析(CK: normal potassium; LK: low potassium; **: P < 0.01; 误差线代表3个独立重复的标准差)。"

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