Cloning and Expression Analyses of NAC Transcription Factor Gene in Bothriochloa ischaemum

doi:10.11733/j.issn.1007-0435.2013.03.027

›› 2013, Vol. 21 ›› Issue (3): 590-597.DOI: 10.11733/j.issn.1007-0435.2013.03.027

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Cloning and Expression Analyses of NAC Transcription Factor Gene in Bothriochloa ischaemum

FANG Zhi-hong¹, WANG Xue-min², LI Jun², DONG Jie², GAO Hong-wen², DONG Kuan-hu¹

1. College of Animal Science and Technology, Shanxi Agricultural University, Taigu, Shanxi Province 030801, China;
2. Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China

Received:2013-03-19 Revised:2013-04-21 Online:2013-06-15 Published:2013-06-05

白羊草NAC转录因子基因的克隆及表达分析

方志红¹, 王学敏², 李俊², 董洁², 高洪文², 董宽虎¹

1. 山西农业大学动物科技学院, 山西太谷 030801;
2. 中国农业科学院北京畜牧兽医研究所, 北京 100193

通讯作者: 董宽虎
作者简介:方志红(1983- ),女,内蒙古赤峰人,博士-硕士研究生,研究方向为饲料作物生产与利用,E-mail:fangzhihong2007@126.com
基金资助:
教育部高等学校博士学科点专项科研基金项目(20101403110002);山西省科技公关项目(20120311011-1);山西省科技基础条件平台建设项目(2012091004-0101)资助

Abstract

Abstract: NAC transcription factors are specific to plants and play an important role in a diverse set of developmental processes. Using RT-PCR and RACE technology with a pair of degenerate primer designed base on the sequence of the homologous gene from other plants, a cDNA of NAC gene was cloned from Bothriochloa ischaemum, named BiNAC. Sequence analysis showed that the full-length of cDNA sequence was 1549 bp including an 1125 bp open reading frame which encoded a 374-amino-acid polypeptide and had the typical characteristics of NAC family. Phylogenetic tree analysis indicated that NAC gene belonged to the ATAF subfamily, and the BiNAC gene of B. ischaemum was closely related to that of Sorghum bicolor with 94% homology. Real-time PCR results revealed that the expression of BiNAC was the most abundant in stem and the least in root. The expression level of BiNAC gene was increased in leaves with NaCl stress. Meanwhile, the plant expression plasmid pBI121-BiNAC-GUS was constructed successfully, which will lay a foundation for the functional analysis of BiNAC gene in the future.

Key words: Bothriochloa ischaemum, NAC transcription factor, Expression analysis, Gene sequence

摘要： NAC转录因子是植物特有的转录因子并在植物的生长发育过程中发挥重要的作用,根据其他植物NAC转录因子基因的氨基酸保守序列设计简并引物,通过RT-PCR和RACE技术从白羊草(Bothriochloa ischaemum)中获得了全长cDNA,命名为BiNAC。序列分析表明,该cDNA片段全长为1549 bp,开放阅读框1125 bp,编码374个氨基酸,具有典型的NAC类蛋白的结构特征。进化树分析表明,该蛋白属于ATAF亚族,与高粱(Sorghum bicolor)亲缘关系最近,同源性达到94%。Real-time PCR 检测结果表明,BiNAC基因在茎中的表达量最高,根中表达量最少,并且受NaCl胁迫表达上调。同时成功构建了植物表达载体pBI121-BiNAC-GUS,为进一步开展NAC基因的功能研究创造了条件。

关键词: 白羊草, NAC转录因子, 表达分析, 基因序列

CLC Number:

Q943.2

FANG Zhi-hong, WANG Xue-min, LI Jun, DONG Jie, GAO Hong-wen, DONG Kuan-hu. Cloning and Expression Analyses of NAC Transcription Factor Gene in Bothriochloa ischaemum[J]. , 2013, 21(3): 590-597.

方志红, 王学敏, 李俊, 董洁, 高洪文, 董宽虎. 白羊草NAC转录因子基因的克隆及表达分析[J]. , 2013, 21(3): 590-597.

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Cloning and Expression Analyses of NAC Transcription Factor Gene in Bothriochloa ischaemum

白羊草NAC转录因子基因的克隆及表达分析

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