转BpmiR156基因白桦株系的耐盐性分析

doi:10.3969/j.issn.1000-2006.201908006

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南京林业大学学报（自然科学版） ›› 2020, Vol. 44 ›› Issue (6): 147-151.doi: 10.3969/j.issn.1000-2006.201908006

转BpmiR156基因白桦株系的耐盐性分析

叶查龙(), 颜斌, 申婷婷, 宁坤, 李慧玉^*()

东北林业大学林木遗传育种国家重点实验室,黑龙江哈尔滨 150040

收稿日期:2019-08-04 修回日期:2020-04-07 出版日期:2020-11-30 发布日期:2020-12-07
通讯作者: 李慧玉
基金资助:
东北林业大学大学生创新实验项目(201810225477)

Analysis of salt tolerance in BpmiR156 overexpression Betula platyphylla

YE Zhalong(), YAN Bin, SHEN Tingting, NING Kun, LI Huiyu^*()

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

Received:2019-08-04 Revised:2020-04-07 Online:2020-11-30 Published:2020-12-07
Contact: LI Huiyu

摘要/Abstract

摘要：

【目的】探究植物年龄响应因子miR156的耐盐功能。【方法】以BpmiR156过表达白桦株系和非转基因对照株系为材料,通过PCR及qPCR分析各转基因株系中外源BpmiR156的稳定性及表达情况;同时对转基因及对照株系进行NaCl胁迫处理,调查胁迫后盐害指数、生理指标及生长速度。【结果】各转基因株系中外源BpmiR156已整合到白桦基因组中,且表达量显著高于非转基因对照株系;NaCl胁迫后,转基因株系的H₂O₂、丙二醛(MDA)含量及盐害指数高于对照株系,生长速度变慢。【结论】BpmiR156基因在白桦中稳定表达,BpmiR156基因过表达白桦株系与对照株系相比,在盐胁迫后盐害指数增加,生长速度变慢,膜系统损伤更严重,说明转入miR156基因在一定程度上降低了白桦的耐盐性。

关键词: 白桦, miR156, 转基因, 盐胁迫, 丙二醛, H₂O₂

Abstract:

【Objective】This study aimed to explore the role of miR156, an age-response factor in plants, in Betula platy-phylla (birch) salt tolerance.【Method】Transgenic BpmiR156 overexpression lines and non-transgenic line were generated. The polymerase chain reaction (PCR) and quantitative PCR were used to analyze the stability and expression of the exogenous BpmiR156 in each transgenic line. Under NaCl stress, the salt injury index, physiological index and growth rate of transgenic and non-transgenic lines were analyzed. 【Result】Results indicated that the BpmiR156 was integrated into the genome of the transgenic lines and more highly expressed than those in the non-transgenic line. Following NaCl treatment, the contents of H₂O₂, MDA and the salt injury index were higher in the transgenic lines than those in the non-transgenic line. Moreover, the transgenic lines exhibited a slower growth rate than the non-transgenic line. 【Conclusion】BpmiR156 gene was stably expressed in transgenic birch. Compared with the non-transgenic line, the index of salt injury increased, the growth rate slowed down and the membrane system damage was more serious. This indicates that over-expression of miR156 reduces the salt tolerance of birch.

Key words: Betula platyphylla (birch), miR156, transgenic lines, salt stress, malondialdehyde (MDA), H₂O₂

中图分类号:

S762.3 ⁺6

叶查龙,颜斌,申婷婷,等. 转BpmiR156基因白桦株系的耐盐性分析[J]. 南京林业大学学报（自然科学版）, 2020, 44(6): 147-151.

YE Zhalong, YAN Bin, SHEN Tingting, NING Kun, LI Huiyu. Analysis of salt tolerance in BpmiR156 overexpression Betula platyphylla[J].Journal of Nanjing Forestry University (Natural Science Edition), 2020, 44(6): 147-151.DOI: 10.3969/j.issn.1000-2006.201908006.

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引物名称 primer name	引物序列 primer sequence	用途 using
miR156-F	5'-CACC GTTTCCACATCTGGGAATAAGC3'	PCR 检测 PCR detection
miR156-R	5'-TCAGCAGCTGTTGAAGTTGAAGG3'
pGWB2-F	5'-ACCACGTCTTCAAAGCAAGTGG 3'
pGWB2-R	5'-GGAAATTCCTCTCCTGTCAGACG 3'
qmiR156-F	5'-TTGACAGAAGAGAGTGAGCACACA-3'	qPCR 检测 qPCR detection
qmiR156-R	5'-GCTTGACAGAAGAtAGaGAGCACAA-3'
U6-F	5'-GGTCCTTCGGGACATCCGAT-3'
U6-R	5'-GGGACCATTTCTCGATTTATGCGTG-3'

转BpmiR156基因白桦株系的耐盐性分析

Analysis of salt tolerance in BpmiR156 overexpression Betula platyphylla

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