Th2CysPrx基因提高酿酒酵母多种胁迫耐受性研究

doi:10.12302/j.issn.1000-2006.202105023

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南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4): 87-94.doi: 10.12302/j.issn.1000-2006.202105023

Th2CysPrx基因提高酿酒酵母多种胁迫耐受性研究

王媛媛(), 刘百超, 姜波, 王丹妮, 高彩球()

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:2021-05-14 修回日期:2021-09-22 出版日期:2022-07-30 发布日期:2022-08-01
通讯作者: 高彩球
基金资助:
中央高校基本科研业务费专项资金项目(2572020AW09);黑龙江省头雁创新团队计划(林业遗传育种创新研究团队)

Th2CysPrx gene enhanced abiotic stress tolerances of Saccharomyces cerevisiae

WANG Yuanyuan(), LIU Baichao, JIANG Bo, WANG Danni, GAO Caiqiu()

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

Received:2021-05-14 Revised:2021-09-22 Online:2022-07-30 Published:2022-08-01
Contact: GAO Caiqiu

摘要/Abstract

摘要：

【目的】过氧化还原蛋白是生物体调节体内氧化还原系统的一种重要蛋白质,在植物生长代谢中起重要作用。前期研究发现过表达刚毛柽柳Th2CysPrx基因的烟草和柽柳中4种抗氧化酶基因(ThGSTZ1、ThGPX、ThSOD和ThPOD)的表达水平上调, 转基因植株的NaCl胁迫耐受性提高。本实验探究Th2CysPrx基因除了响应NaCl胁迫应答,是否还参与其他盐胁迫和其他胁迫的应答。 【方法】将Th2CysPrx构建到pYES2酵母表达载体上,转化到酿酒酵母INVSC1细胞中,分析比较重组酵母(pYES2-Th2CysPrx)和对照酵母(pYES2)细胞在各种非生物胁迫后的生长情况。【结果】过表达Th2CysPrx基因的重组酵母细胞在低温、KCl和LiCl胁迫后存活率与对照相比差异不显著;但H₂O₂、NaCl或NaHCO₃胁迫后酵母细胞的存活率显著提高。此外,高于0.5 mol/L山梨醇或高温(达到44 ℃),或低于质量分数0.007% CdCl₂胁迫后,过表达Th2CysPrx基因能显著提高重组酵母的细胞存活率。【结论】Th2CysPrx基因可以显著增强酵母细胞对NaCl和NaHCO₃胁迫的耐受能力,以及特定的渗透胁迫、高温胁迫、氧化胁迫和重金属胁迫的耐受能力,但对低温、KCl和LiCl胁迫耐受能力无明显影响。

关键词: 刚毛柽柳, 酵母, 过氧化物还原酶(Prxs), 非生物胁迫

Abstract:

【Objective】 Peroxidoxin is an important protein in organisms for regulation of the redox system in plants, and plays an important role in plant growth and metabolism. Previous research has found that overexpression of the Th2CysPrx gene increased the expression levels of four antioxidant genes (ThGSTZ1, ThGPX, ThSOD and ThPOD), thereby increasing the NaCl stress tolerance of transgenic Arabidopsis and Tamarix hispida. This study explored whether the Th2CysPrx gene is involved in the response to other salts or stresses in addition to NaCl stress. 【Method】 Th2CysPrx was constructed into the pYES2 yeast expression vector and transformed into the INVSC1 cells of Saccharomyces cerevisiae. The cell growth between the recombinant vector and empty vector was compared after applying various abiotic stresses. 【Result】 The survival rate of cells with the overexpressed Th2CysPrx gene was not significantly different from that of the control under cold temperature, KCl or LiCl stress. However, the overexpression of Th2CysPrx gene significantly increased the cell survival rate of yeast cells under oxidative, NaCl and NaHCO₃ stress. In addition, the overexpression of Th2CysPrx gene also significantly improved the cell survival rate in recombinant yeast under stress caused by higher than 0.5 mol/L sorbitol, temperature above 44 ℃, or less than 0.007%(mass fraction) CdCl₂. 【Conclusion】 Th2CysPrx gene can significantly enhance the tolerance of yeast cells not only to NaCl or NaHCO₃ stress, but also to specific osmotic, high-temperature, oxidative, or heavy metal stresses, but exhibits no obvious tolerance to cold temperature, KCl or LiCl stress.

Key words: Tamarix hispida, yeast, peroxidoxin(Prxs), abiotic stress

中图分类号:

Q943

王媛媛,刘百超,姜波,等. Th2CysPrx基因提高酿酒酵母多种胁迫耐受性研究[J]. 南京林业大学学报（自然科学版）, 2022, 46(4): 87-94.

WANG Yuanyuan, LIU Baichao, JIANG Bo, WANG Danni, GAO Caiqiu. Th2CysPrx gene enhanced abiotic stress tolerances of Saccharomyces cerevisiae[J].Journal of Nanjing Forestry University (Natural Science Edition), 2022, 46(4): 87-94.DOI: 10.12302/j.issn.1000-2006.202105023.

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Th2CysPrx基因提高酿酒酵母多种胁迫耐受性研究

Th2CysPrx gene enhanced abiotic stress tolerances of Saccharomyces cerevisiae

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