白桦BpBEE2基因的遗传转化及抗逆性分析

doi:10.7525/j.issn.1673-5102.2019.02.015

植物研究 ›› 2019, Vol. 39 ›› Issue (2): 287-293.doi: 10.7525/j.issn.1673-5102.2019.02.015

白桦BpBEE2基因的遗传转化及抗逆性分析

颜斌, 武丹阳, 李慧玉

东北林业大学林木遗传育种国家重点实验室, 哈尔滨 150040

收稿日期:2018-11-05 出版日期:2019-03-05 发布日期:2019-03-20
通讯作者: 李慧玉 E-mail:lihuiyu2017@126.com
作者简介:颜斌(1992-),男,硕士研究生,主要从事林木遗传育种研究。
基金资助:
国家重点研发计划（2017YFD0600603）

Genetic Transformation and Resistance Analysis of BpBEE2 Gene from Betula platyphylla

YAN Bin, WU Dan-Yang, LI Hui-Yu

State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040

Received:2018-11-05 Online:2019-03-05 Published:2019-03-20
Supported by:
National Key R&D Plan(2017YFD0600603)

摘要/Abstract

摘要： Brassinolide Enhanced Expression2（BEE2）基因属于bHLH转录因子家族，是调控油菜素内酯信号转导的上游调控因子。本研究通过RT-PCR技术克隆BpBEE2基因的全长cDNA序列，构建植物过表达及抑制表达载体，并通过农杆菌介导法进行白桦的遗传转化，对获得的转基因株系进行生长量及盐、旱胁迫分析，结果表明：获得了长度为1 080 bp的全长cDNA序列，成功构建了该基因的过表达及抑制表达载体，并获得了过表达和抑制表达的白桦株系。BpBEE2基因过表达白桦株系的苗高高于对照株系，而抑制表达株系的苗高低于对照株系。同时发现BEE2基因对盐、旱胁迫后对植株的鲜重也产生了影响。说明BpBEE2可能参与了植物的生长发育过程，并且改善了植物的抗旱、耐盐性。

关键词: 白桦, BpBEE2基因, 载体构建, 遗传转化, 抗旱耐盐

Abstract: Brassinolide Enhanced Expression2(BEE2) genes belong to bHLH transcription factors family,and were involved in Brassinolide signal pathway.In this study, BpBEE2 gene was cloned using RT-PCR, and plant over-expression and inhibiting expression vector were constructed. Genetic transformation was operated through agrobacterium mediated, then mass growths of transgenic lines and non-transgenic line were analyzed under natural conditions and salt, drought stress. The length of 1 080 bp cDNA sequence was obtained, over-expression and inhibiting expression vectors were constructed successfully, and transgenic lines were obtained. Seedlings height of over-expression transgenetic line was higher than that of the control line, however, that of inhibiting expression lines was lower than that of the control line. BEE2 gene can ffect fresh weight of plants after salt and drought stress, and BpBEE2 is involved in the process of plant growth and development and plays a key role in drought and salt resistance response.

Key words: Betula platyphylla, BpBEE2 gene, vector construction, genetic transformation, drought and salt resistance

中图分类号:

S792

颜斌, 武丹阳, 李慧玉. 白桦BpBEE2基因的遗传转化及抗逆性分析[J]. 植物研究, 2019, 39(2): 287-293.

YAN Bin, WU Dan-Yang, LI Hui-Yu. Genetic Transformation and Resistance Analysis of BpBEE2 Gene from Betula platyphylla[J]. Bulletin of Botanical Research, 2019, 39(2): 287-293.

参考文献

1. Friedrichsen D M,Nemhauser J,Muramitsu T,et al. Three redundant brassinosteroid early response genes encode putative bHLH transcription factors required for normal growth[J]. Genetics,2002,162(3):1445-1456.
2. Noh S A,Choi Y I,Cho J S,et al. The poplar basic helix-loop-helix transcription factor BEE3-Like gene affects biomass production by enhancing proliferation of xylem cells in poplar[J]. Biochemical & Biophysical Research Communications,2015,462(1):64-70.
3. Cifuentes-Esquivel N,Bou-Torrent J,Galstyan A,et al. The bHLH proteins BEE and BIM positively modulate the shade avoidance syndrome in Arabidopsis seedlings[J]. The Plant Journal,2013,75(6):989-1002.
4. Heisler M G,Atkinson A,Bylstra Y H,et al. SPATULA,a gene that controls development of carpel margin tissues in Arabidopsis,encodes a bHLH protein[J]. Development,2001,128(7):1089-1098.
5. Ling H Q,Bauer P,Bereczky Z,et al. The tomato fer gene encoding a bHLH protein controls iron-uptake responses in roots[J]. Proceedings of the National Academy of Sciences of the United States of America,2002,99(21):13938-13943.
6. Rajani S,Sundaresan V. The Arabidopsis myc/bHLH gene ALCATRAZ enables cell separation in fruit dehiscence[J]. Current Biology,2001,11(24):1914-1922.
7. Seo J S,Joo J,Kim M J,et al. OsbHLH148,a basic helix-loop-helix protein,interacts with OsJAZ proteins in a jasmonate signaling pathway leading to drought tolerance in rice[J]. The Plant Journal,2011,65(6):907-921.
8. Li H M,Sun J Q,Xu Y X,et al. The bHLH-type transcription factor AtAIB positively regulates ABA response in Arabidopsis[J]. Plant Molecular Biology,2007,65(5):655-665.
9. Jiang Y Q,Yang B,Deyholos M K. Functional characterization of the Arabidopsis bHLH92 transcription factor in abiotic stress[J]. Molecular Genetics & Genomics,2009,282(5):503-516.
10. 刘晓月,王文生,傅彬英. 植物bHLH转录因子家族的功能研究进展[J]. 生物技术进展,2011,1(6):391-397. Liu X Y,Wang W S,Fu B Y. Research progress of plant bHLH transcription factor family[J]. Current Biotechnology,2011,1(6):391-397.
11. Wang Y J,Zhang Z G,He X J,et al. A rice transcription factor OsbHLH1 is involved in cold stress response[J]. Theoretical & Applied Genetics,2003,107(8):1402-1409.
12. Yuan X Y,Mao Z J,Gu H Y,et al. Reproductive strategies and seeds behavior of Betula platyphylla Suk. population[J]. Journal of Forestry Research,2000,11(4):241-244.
13. B hlenius H,Huang T,Charbonnel-Campaa L,et al. CO/FT regulatory module controls timing of flowering and seasonal growth cessation in trees[J]. Science,2006,312(5776):1040-1043.
14. 肖素勤,孙振,轩秀霞,等. 用于通路(Gateway)克隆技术的植物表达载体研究进展[J]. 植物科学学报,2012,30(5):528-544. Xiao S Q,Sun Z,Xuan X X,et al. Research progress on plant expression vectors for Gateway cloning technology[J]. Plant Science Journal,2012,30(5):528-544.
15. Ossowski S,Schwab R,Weigel D. Gene silencing in plants using artificial micro RNAs and other small RNAs[J]. The Plant Journal,2008,53(4):674-690.
16. 张瑞萍. 脱水素基因逆境表达模式与白桦遗传转化研究[D]. 哈尔滨:东北林业大学,2009. Zhang R P. Study on expression pattern of ThDHN responding to different abiotic stress and establishment of genetic transformation of Betula platyphylla[D]. Harbin:Northeast Forestry University,2009.
17. 武丹阳,杨洋,李慧玉. 3个白桦BpBEE基因的克隆与表达分析[J]. 浙江农林大学学报,2017,34(1):137-144. Wu D Y,Yang Y,Li H Y. Cloning and expression of three BpBEE genes in Betula platyphylla[J]. Journal of Zhejiang A & F University,2017,34(1):137-144.
18. Moreno J E,Moreno-Piovano G,Chan R L. The antagonistic basic helix-loop-helix partners BEE and IBH1 contribute to control plant tolerance to abiotic stress[J]. Plant Science,2018,271:143-150.
19. Toledo-Ortiz G,Huq E,Quail P H. The Arabidopsis Basic/Helix-Loop-Helix transcription factor family[J]. The Plant Cell,2003,15(8):1749-1770.
20. 许思佳,颜斌,董京祥,等. 白桦BpBEE1基因启动子的表达特性分析[J]. 植物研究,2018,38(2):247-253. Xu S J,Yan B,Dong J X,et al. Analysis of BpBEE1 gene promoter in Betula platyphylla[J]. Bulletin of Botanical Research,2018,38(2):247-253.
21. Li X X,Duan X P,Jiang H X,et al. Genome-wide analysis of basic/helix-loop-helix transcription factor family in rice and Arabidopsis[J]. Plant Physiology,2006,141(4):1167-1184.
22. Zhou J,Li F,Wang J L,et al. Basic helix-loop-helix transcription factor from wild rice(OrbHLH2) improves tolerance to salt-and osmotic stress in Arabidopsis[J]. Journal of Plant Physiology,2009,166(12):1296-1306.

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白桦BpBEE2基因的遗传转化及抗逆性分析

Genetic Transformation and Resistance Analysis of BpBEE2 Gene from Betula platyphylla

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