棉花色素腺体发育相关基因<i>GhNAC201</i>的克隆与表达分析

doi:10.7668/hbnxb.20190587

华北农学报 ›› 2020, Vol. 35 ›› Issue (1): 29-35. doi: 10.7668/hbnxb.20190587

所属专题： 棉花；生物技术；热点文章；

• 作物遗传育种·种质资源·生物技术 • 上一篇下一篇

棉花色素腺体发育相关基因GhNAC201的克隆与表达分析

张曦¹, 钱玉源¹, 刘祎¹, 王广恩¹, 宋世佳², 李晓飞³, 米换房⁴, 崔淑芳¹, 李俊兰¹

1. 河北省农林科学院棉花研究所, 农业部黄淮海半干旱区棉花生物学与遗传育种重点实验室, 国家棉花改良中心河北分中心, 河北石家庄 050051;
2. 河北省农林科学院, 河北石家庄 050051;
3. 河北工程大学, 河北邯郸 056038;
4. 邯郸市农业科学院, 河北邯郸 056001

收稿日期:2019-09-15 出版日期:2020-02-28
通讯作者: 李俊兰(1963-),女,河北清河人,研究员,硕士,主要从事棉花遗传育种与种质资源创新研究。
作者简介:张曦(1980-),女,辽宁辽阳人,助理研究员,在站博士后,主要从事棉花功能基因组研究。
基金资助:
河北省引智及人才培养专项资金（B2018003018）；河北省农林科学院农业科技创新人才队伍建设项目；河北省农林科学院创新工程项目（3-03-07）

Cloning and Expression Analysis of a Cotton Pigment Gland Formation Related Gene GhNAC201

ZHANG Xi¹, QIAN Yuyuan¹, LIU Yi¹, WANG Guang¹, SONG Shijia², LI Xiaofei³, MI Huanfang⁴, CUI Shufang¹, LI Junlan¹

1. Institute of Cotton, Hebei Academy of Agriculture and Forestry Sciences, Key Laboratory of Biology and Genetic Improvement of Cotton in Huanghuaihai Semiarid Area, Ministry of Agriculture, National Cotton Improvement Center Hebei Branch, Shijiazhuang 050051, China;
2. Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051, China;
3. Hebei University of Engineering, Handan 056038, China;
4. Handan Academy of Agricultural Sciences, Handan 056001, China

Received:2019-09-15 Published:2020-02-28

摘要/Abstract

摘要： 为了探究GhNAC201在棉花色素腺体发育中的作用，应用RT-PCR的方法克隆了有腺体棉花中棉所12中GhNAC201基因的编码区序列，并进行生物信息学分析。利用荧光定量PCR对有腺体和无腺体棉花材料不同生育时期、不同组织部位和不同激素处理下GhNAC201基因的表达情况进行分析。结果显示， GhNAC201编码区为948 bp，编码315个氨基酸。GhNAC201蛋白分子量为36.8 ku，理论pI 5.97，二级结构预测存在27.94%的α-螺旋、5.40%的β-转角、18.10%的延伸链和48.57%的无规则卷曲。三维结构预测表明，GhNAC201的26-190氨基酸与水稻胁迫应答NAC1蛋白序列高度匹配，模型维度为X：55.229Å，Y：36.150Å，Z：46.112Å。GhNAC201在30-163氨基酸处存在1个NAM结构域，预测有5个苏氨酸、6个丝氨酸和4个酪氨酸磷酸化位点位于NAM结构域内。亚细胞定位预测GhNAC201可能位于分泌通路或除线粒体、叶绿体之外的其他亚细胞结构。不同物种中GhNAC201同源蛋白序列的相似性较高，存在5个高度保守的基序。GhNAC201与亚洲棉、雷蒙德氏棉中的同源蛋白的基序组成模式完全一致。GhNAC201基因在中棉所12各组织部位的表达量均极显著高于显性和隐性无腺体棉，且受ABA、BR、GA和MeJA诱导极显著上调表达，基因的时空表达模式与腺体密度的时空分布规律也趋于一致。综上所述， GhNAC201与棉花色素腺体发育密切相关，可能通过ABA、BR、GA和JA通路调控腺体发育或棉酚代谢。

关键词: 棉花, NAC转录因子, GhNAC201, 序列分析, 表达分析

Abstract: To investigate the function of GhNAC201 in cotton pigment gland development,RT-PCR was performed to isolate the coding sequence(CDS)of GhNAC201 from glanded cotton Zhongmiansuo 12. Gene expressions of glanded and glandless cotton at different developmental stages,in different tissues and under various hormones treatment were analyzed by qRT-PCR. CDS of GhNAC201 was 948 bp in length and encoded 315 aa,with a theoretical pI 5.97. 27.94% alpha helix,5.40% beta turn,18.10% extended strand and 48.57% random coil were predicted in protein secondary structure. 3D structure of GhNAC201 had been modelled by rice stress-responsive NAC1 with model dimensions of X:55.229 Å,Y:36.150 Å,Z:46.112 Å. GhNAC201 contained a NAM domain at 30-163 aa,and 5 threonine,6 serine and 4 tyrosine phosphorylation sites were predicted within the NAM domain. Subcellular localization prediction showed that GhNAC201 had a certain probability to locate in secretory pathway or other subcellular structures other than mitochondrion and chloroplast. The homology of GhNAC201 homologous proteins in different species was relatively high,and 5 motifs were highly conserved.The motif pattern of GhNAC201 was consistent with its homologous proteins in Gossypium arboretum and G. raimondii. Expression levels of GhNAC201 in the glanded cotton Zhongmiansuo 12 were extremely significantly higher than those in dominant and recessive glandless cotton and could be extremely significantly up-regulated by ABA,BR,GA and MeJA. The spatial-temporal expression pattern of GhNAC201 in Zhongmiansuo 12 tended to be consistent with the spatial-temporal distribution of gland density. In conclusion, GhNAC201 was closely related to the development of pigment glands in cotton,and might regulate gland development or gossypol metabolism through ABA,BR,GA and JA pathways.

Key words: Cotton, NAC transcriptional factor, GhNAC201, Sequence analysis, Expression analysis

中图分类号:

S562
Q78

张曦, 钱玉源, 刘祎, 王广恩, 宋世佳, 李晓飞, 米换房, 崔淑芳, 李俊兰. 棉花色素腺体发育相关基因GhNAC201的克隆与表达分析[J]. 华北农学报, 2020, 35(1): 29-35. doi: 10.7668/hbnxb.20190587.

ZHANG Xi, QIAN Yuyuan, LIU Yi, WANG Guang, SONG Shijia, LI Xiaofei, MI Huanfang, CUI Shufang, LI Junlan. Cloning and Expression Analysis of a Cotton Pigment Gland Formation Related Gene GhNAC201[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(1): 29-35. doi: 10.7668/hbnxb.20190587.

http://www.hbnxb.net/CN/Y2020/V35/I1/29

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棉花色素腺体发育相关基因GhNAC201的克隆与表达分析

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棉花色素腺体发育相关基因GhNAC201的克隆与表达分析

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