烟草Myb转录因子<i>NtBL1</i>基因的克隆及其表达分析

doi:10.7668/hbnxb.20190004

摘要/Abstract

摘要： 为揭示Blind基因在普通烟草中的功能，通过同源克隆的方法从普通烟草K326的基因组中克隆得到1个Blind-like基因，即NtBL1，用PROSITE （ExPASy）、Sopma和Swissmodel等软件对NtBL1进行蛋白质结构域分析和高级结构预测，用Mega 7.0和MEME软件对NtBL1及其同源蛋白进行了进化树构建和结构分析，用半定量PCR分析了NtBL1基因在花、叶、根、茎、苗中的表达模式，用qRT-PCR技术分析了ABA、NaCl和PEG处理下NtBL1基因的表达模式。结果表明，NtBL1的开放阅读框长度为1 014 bp，编码337个氨基酸。半定量PCR结果表明，NtBL1在烟草的大部分组织和各时期普遍表达，但是根中的表达量最高。蛋白质结构域分析显示NtBL1是典型的R2R3 Myb家族蛋白，含有2个Myb-type HTH类型的DNA结构域。蛋白质高级结构预测表明，NtBL1蛋白高级结构主要由无规则卷曲和α-螺旋组成。进化和结构分析结果表明，NtBL1基因与番茄Blind等基因的进化距离较近，且具有相似的模体结构。qRT-PCR的结果显示NtBL1基因的表达受ABA的诱导，在处理24 h达到最高表达量；NtBL1基因的表达受NaCl处理的抑制，在24 h出现最低表达量。因此，NtBL1基因可能参与激素和盐胁迫调控下的侧生分生组织的发育。

关键词: 烟草, NtBL1, Myb基因家族, 基因表达

Abstract: In order to discover the function of Blind gene in Nicotiana tabacum, we cloned one Blind-like gene, NtBL1, from K326 by homologue cloning method. The protein domain and structure was predicted by PROSITE(ExPASy), Sopma and Swissmodel, the Phylogeny tree construction and Motif prediction of NtBL1 and its homologues were analyzed separately by Mega 7.0 and MEME, the expression pattern of NtBL1 in different tissues was analyzed by semi-quantitative PCR, and the expression changes of NtBL1 under abiotic stress treatment was detected by qRT-PCR. The results showed that the coding sequence of NtBL1 was 1 014 bp, encoding 337 aa. The expression pattern of NtBL1 in transcription level was generally detected in several organs and growing stages, while it had the highest expression level in root. Bioinformation analysis showed that NtBL1 was a typical R2R3 Myb protein, which contained 2 Myb-type HTH domains. The phylogenetic tree analysis showed that NtBL1 gene was shared lots of motifs with tomato Blind. The results of qRT-PCR showed that the expression level of NtBL1 gene was up-regulated by ABA treatment and the highest expression level appeared in 24 h treatment; The expression level of NtBL1 gene was down-regulated by NaCl treatment and the lowest expression level presented in 24 h treatment. In conclusion, NtBL1 gene might participate in regulation of axillary mersitem development by ABA and NaCl.

Key words: Tobacco, NtBL1, Myb gene family, Gene expression

中图分类号:

Q78
S572.03

刘雅洁, 薛翀, 邱诗蕊, 李想, 唐丽雯, 曾淑华. 烟草Myb转录因子NtBL1基因的克隆及其表达分析[J]. 华北农学报, 2019, 34(6): 26-32. doi: 10.7668/hbnxb.20190004.

LIU Yajie, XUE Chong, QIU Shirui, LI Xiang, TANG Liwen, ZENG Shuhua. Cloning and Expression Analysis of Myb Transcription Factor NtBL1 Gene in Nicotiana tabacum[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(6): 26-32. doi: 10.7668/hbnxb.20190004.

http://www.hbnxb.net/CN/Y2019/V34/I6/26

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烟草Myb转录因子NtBL1基因的克隆及其表达分析

Cloning and Expression Analysis of Myb Transcription Factor NtBL1 Gene in Nicotiana tabacum

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烟草Myb转录因子NtBL1基因的克隆及其表达分析

Cloning and Expression Analysis of Myb Transcription Factor NtBL1 Gene in Nicotiana tabacum

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