白桦BpMYB2基因及其启动子克隆、表达分析

doi:10.3969/j.issn.1000-2006.2014.03.005

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南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (03): 24-28.doi: 10.3969/j.issn.1000-2006.2014.03.005

白桦BpMYB2基因及其启动子克隆、表达分析

张楠,崔志远,孙丹,王超^*

东北林业大学,林木遗传育种与生物技术国家重点实验室,黑龙江哈尔滨 150040

出版日期:2014-05-15 发布日期:2014-05-15
基金资助:
收稿日期:2013-07-01 修回日期:2013-10-09
基金项目:国家高技术研究发展计划(2011AA100202); 中央高校基本科研业务费专项资金项目(DL11EA02)
第一作者:张楠,硕士生。*通信作者:王超,副教授。E-mail:clock7993@126.com。
引文格式:张楠,崔志远,孙丹,等. 白桦BpMYB2基因及其启动子克隆、表达分析[J]. 南京林业大学学报:自然科学版,2014,38(3):24-28.

Cloning and expression analysis of the BpMYB2 gene and its promoter in Betula platyphylla

ZHANG Nan,CUI Zhiyuan,SUN Dan,WANG Chao^*

State Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Northeast Forestry University, Harbin 150040,China

Online:2014-05-15 Published:2014-05-15

摘要/Abstract

摘要： 克隆了1条白桦MYB转录因子基因,命名为BpMYB2。实时定量PCR分析表明BpMYB2基因在白桦不同器官和组织内的表达量不同,在直立木木质部中表达量最高,其次是人工弯曲处理2周的对应木、应拉木、花序,在叶和芽中表达量极低; 利用染色体步移技术,获得长度为1 284 bp的启动子序列。启动子进行PLACE分析发现,序列中含有TATA box、ARR1AT、WRKY71OS等元件。构建了由BpMYB2启动子驱动GUS报告基因在植物中的表达载体,命名为BpMYB2-1301,利用农杆菌介导法瞬时转化白桦幼苗,并进行组织化学染色。结果表明:BpMYB2的启动子具有启动子活性,能够驱动GUS基因在白桦中表达; GUS瞬时表达信号在茎、叶柄中较强,在叶片中较弱,说明BpMYB2基因与白桦木质部发育相关。

Abstract: In this study, a MYB transcription factor named BpMYB2 was cloned from Betula platyphylla. The realtime PCR analysis showed that the expression of BpMYB2 was various in different tissues, and it expressed in the highest level in normal wood, then the second level in opposite wood, and then in tension wood and flower, while the least expression were in leaves and buds. A 1 284 bp BpMYB2 promoter was cloned by genome walking method, and the promoter included some cis-elements such as TATA box, ARR1AT, WRKY71OS by PLACE analysis. Further, the BpMYB2 promoter was inserted to pCAMBIA1301 vector under control of the 35S promoter to generate the BpMYB2-1301 recombinant construct, which was transient expressed in B. platyphylla seedlings by Agrobacterium tumefaciens mediated method. The histochemical GUS staining showed that the BpMYB2 promoter could drive the expression of GUS gene in B. platyphylla, and expressed in high level in stems and petioles while in low level in leaves. In conclusion, BpMYB2 is important forxylem development and lay basis for utilizing the transcription factor to regulate the xylem development in B. platyphylla.

中图分类号:

Q781
S722

张楠,崔志远,孙丹,等. 白桦BpMYB2基因及其启动子克隆、表达分析[J]. 南京林业大学学报（自然科学版）, 2014, 38(03): 24-28.

ZHANG Nan,CUI Zhiyuan,SUN Dan,WANG Chao. Cloning and expression analysis of the BpMYB2 gene and its promoter in Betula platyphylla[J].Journal of Nanjing Forestry University (Natural Science Edition), 2014, 38(03): 24-28.DOI: 10.3969/j.issn.1000-2006.2014.03.005.

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白桦BpMYB2基因及其启动子克隆、表达分析

Cloning and expression analysis of the BpMYB2 gene and its promoter in Betula platyphylla

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