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作物学报 ›› 2015, Vol. 41 ›› Issue (09): 1445-1453.doi: 10.3724/SP.J.1006.2015.01445

• 研究简报 • 上一篇    下一篇

谷子转录因子基因SiNAC45在拟南芥中对低钾及ABA的响应

王二辉1, 2, *, 胡利芹2, *, 薛飞洋1, 2, 李微微2, 徐兆师2, 李连城2, 周永斌2, 马有志2, 刁现民2, 贾冠清2, 陈明2, *, 闵东红1, *   

  1. 1 西北农林科技大学农学院 / 旱区作物逆境生物学国家重点实验室, 陕西杨凌712100; 2 中国农业科学院作物科学研究所 / 农作物基因资源与基因改良国家重大科学工程 / 农业部麦类生物学与作物遗传育种重点实验室, 北京100081
  • 收稿日期:2015-03-27 出版日期:2015-09-12 网络出版日期:2015-09-12
  • 通讯作者: 闵东红, E-mail:mdh2493@126.com, Tel: 13609123593; 陈明, E-mail:chenming02@caas.cn, Tel: 13683360891
  • 作者简介:第一作者联系方式: E-mail:wangerhui191132@163.com, Tel: 13366320737 **同等贡献(Contributed equally to this work)
  • 基金资助:
    本研究由国家转基因生物新品种培育重大专项(2014ZX08002-003B), 陕西省农业科技攻关项目(2013K02-01), 陕西省科技统筹创新工程计划项目(2014KTZB02-01-01)和北京市科技计划项目(Z14110000231418)资助

Overexpression of Millet Transcription Factor Gene SiNAC45 to Response of Low Potassium Stress and ABA Treatment in Transgenic Arabidopsis

WANG Er-Hui1, 2, **, HU Li-Qin2, **, XUE Fei-Yang1, 2, LI Wei-Wei2, XU Zhao-Shi2, LI Lian-Cheng2, ZHOU Yong-Bin2, MA You-Zhi2, DIAO Xian-Min2, JIA Guan-Qing2, CHEN Ming2, *, MIN Dong-Hong1, *   

  1. 1 College of Agronomy, Northwest A＆F University / State Key Laboratory of Arid Region Crop Adversity Biology, Yangling 712100, China; 2 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility For Crop Gene Resource and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crop, Ministry of Agriculture, Beijing 100081, China
  • Received:2015-03-27 Published:2015-09-12 Published online:2015-09-12

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摘要： NAC (nascent polypeptide-associated complex)转录因子在植物生长发育和非生物胁迫响应等过程中发挥重要的调控作用, 目前, 关于NAC转录因子参与耐低钾胁迫的研究报道很少。本研究在前期工作对低钾胁迫的转录组测序基础上, 对筛选出的一个低钾胁迫下表达量上调的NAC类转录因子基因SiNAC45进行了深入研究。结果表明, SiNAC45基因全长1383 bp, 编码461个氨基酸, 分子量为50.7 kD, 等电点为6.92。SiNAC45在20~100个氨基酸之间有一段NAM保守结构域。系统进化树结果显示, SiNAC45位于NAC基因家族的第1亚族。基因表达谱分析显示, SiNAC45主要在谷子根部表达, 并且能够被低钾和ABA诱导表达。亚细胞定位结果显示SiNAC45定位于细胞核。基因功能分析结果显示, 在不同浓度低钾处理下, 和野生型拟南芥相比, SiNAC45转基因拟南芥的根长和植株鲜重显著增加, 说明过表达SiNAC45可以提高转基因植物对低钾胁迫的抗性。下游基因表达分析结果显示, 在SiNAC45转基因植物中两种重要的钾离子转运体基因AKT1和HAK1的表达显著提高, 证明SiNAC45通过调控植物钾离子转运体基因的表达影响植物对低钾胁迫的耐性。种子萌发试验结果显示, SiNAC45转基因拟南芥与野生型拟南芥相比对ABA的敏感性降低, 说明SiNAC45可能负调ABA信号。

关键词: 谷子, NAC转录因子, 低钾胁迫, ABA反应, 基因功能分析

Abstract: NAC (nascent polypeptide-associated complex) like transcription factors play important role in plant growth and development, abiotic stress response, and other processes. Currently, few researches reported NAC like transcription factors involving in tolerance to low potassium stress. In this study, we found and researched a NAC like transcription factor gene SiNAC45 on the basis of transcriptome sequence of millet under low potassium stress which had been completed in previous work. The result show that the full-length of SiNAC45 is 1383 bp, encoding 461 amino acids, with molecular weight and isoelectric point of 50.7 kD and 6.92, respectively. There is a conserved NAM domain between 20-100 amino acids of SiNAC45. The phylogenetic tree showed that SiNAC45 belonged to the first subfamily of NAC gene family. The gene expression profile results indicated SiNAC45 mainly expressed in roots and was induced by ABA and low potassium treatment. The protein subcellular localization results of SiNAC45 revealed that it was localized in the nucleus. Gene functional analysis showed that under treatment with different concentrations of potassium, root length and fresh weight of SiNAC45 transgenic Arabidopsis significantly increased compared with those of wild-type Arabidopsis, and there was no significant difference in the number of lateral roots between transgenic and wild-type Arabidopsis, indicating that overexpressing of SiNAC45 in transgenic plants can enhance tolerance to low potassium stress. Expression analysis of downstream gene showed that expression of two important potassium transporter genes AKT1 and HAK1 increased significantly in SiNAC45 transgenic plants, indicating that SiNAC45 affects the tolerance to low potassium stress of plants by regulating the expression of potassium transporter gene. Seed germination test results showed that Arabidopsis carrying SiNAC45 decreased the sensitivity to ABA compared with wild-type Arabidopsis, indicating that SiNAC45 maybe negatively regulate ABA signal pathway.

Key words: Millet, NAC transcription factor, Low potassium stress, ABA response, Gene function analysis

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