过量表达拟南芥<em>CAT</em>提高烟草对气体甲醛的吸收和抗性

doi:10.13523/j.cb.20150506

中国生物工程杂志

2015, Vol. 35

Issue (5): 41-48 DOI: 10.13523/j.cb.20150506

研究报告

过量表达拟南芥CAT提高烟草对气体甲醛的吸收和抗性

韩双, 杨志丽, 陈丽梅

昆明理工大学生命科学与技术学院生物工程技术研究中心昆明 650500

Over Expression of Arabidopsis CAT Improved the Absorption and Tolerance of Gas HCHO in Tobacco

HAN Shuang, YANG Zhi-li, CHEN Li-mei

Biotechnology Research Center, College of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China

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摘要：

为提高植物对甲醛的吸收和耐受,利用拟南芥CAT的编码区构建植物过表达载体pk₂-PrbcS-^*T-CAT,在野生型(WT)烟草叶绿体中过量表达CAT产生2个表达量不同的转基因株系(C1、C3)。用10ppm、20ppm和40ppm气体甲醛处理WT和转基因烟草,分析过量表达CAT的转基因烟草对甲醛的吸收效率和耐受性。结果表明,转基因烟草对气体甲醛的吸收量明显高于WT,且随着处理浓度的升高,转基因株系叶片的可溶性总糖、总蛋白质和总叶绿素含量都显著高于WT;转基因株系叶片氧化损伤指标H₂0₂、丙二醛(MDA)和羰基化蛋白(PC)的含量都显著低于野生型。转基因烟草(C1)的PM H⁺-ATPase和氢泵活性在40ppm处理后为WT的3倍和2.5倍,C1的气孔导度为WT的2.67倍。这些结果表明,在烟草中过量表达拟南芥CAT能降低甲醛进入烟草细胞引起的氧化损伤,提高烟草对甲醛的脱毒能力,增强烟草对甲醛的吸收和抗性。揭示气体甲醛胁迫时质膜H⁺-ATPase和氢泵活性影响叶片气孔的导度,这可能是转基因烟草甲醛吸收效率提高的原因之一。

关键词： 过氧化氢酶; 过量表达; 转基因烟草; 气体甲醛胁迫; 甲醛吸收

Abstract:

To improve the absorption and tolerance of HCHO in plants, the plant expression vector pk₂-PrbcS-^*T-CAT was constructed using Arabidopsis thaliana CAT (catalase) coding region. Overexpression of CAT in wild-type (WT) tobacco chloroplasts produced two transgenic lines (C1, C3) which had different expression quantity. Using 10ppm, 20ppm and 40ppm gas HCHO treated WT and transgenic tobacco, and then analyzed the HCHO absorption efficiency and tolerance of CAT transgenic tobacco. The results showed that the absorption quantity of gas HCHO by transgenic tobacco was higher than wild type (WT), and with the increasing concentration, total soluble glucide, total protein and total chlorophyll content in leaves of the transgenic lines were significantly higher than WT. The oxidative damage index of H₂O₂, malondialdehyde (MDA) and protein carbonyl (PC) which content in transgenic lines leaves were significantly lower than the WT. The PM H⁺-ATPase and hydrogen pump activity of transgenic tobacco (C1) was 3 times and 2.5 times than WT after 40 ppm HCHO treatment. Stomatal conductance of C1 is 2.67 times than WT. These results indicated that overexpression of Arabidopsis CAT could reduce oxidative damage in tobacco cells which induced by HCHO, and improved the ability of tobacco to HCHO detoxification and enhanced the absorption and resistance of tobacco to HCHO. It was revealed that the stomatal conductance affected by PM H⁺-ATPase and hydrogen pump activity when gas HCHO stressed tobacco, which might be one of the causes of higher HCHO absorption efficiency of transgenic tobacco.

Key words: Catalase Overexpression Transgenic tobacco Gas formaldehyde stress Formaldehyde-uptake

收稿日期: 2015-01-13 出版日期: 2015-05-25

ZTFLH:

Q-33

基金资助:

国家自然科学基金资助项目(30970263)

通讯作者: 陈丽梅 E-mail: chenlimeikm@126.com

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引用本文:

韩双, 杨志丽, 陈丽梅. 过量表达拟南芥CAT提高烟草对气体甲醛的吸收和抗性[J]. 中国生物工程杂志, 2015, 35(5): 41-48.

HAN Shuang, YANG Zhi-li, CHEN Li-mei. Over Expression of Arabidopsis CAT Improved the Absorption and Tolerance of Gas HCHO in Tobacco. China Biotechnology, 2015, 35(5): 41-48.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150506 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I5/41

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