抗旱耐盐菌剂的制备及其对甘草种子萌发的影响

doi:10.13560/j.cnki.biotech.bull.1985.2020-0120

摘要/Abstract

摘要： 制备高效抗逆微生物菌剂并研究其对旱盐胁迫下甘草生长及生理生化特性的影响。采用单因素和正交试验优化短小芽孢杆菌发酵培养基;单因素试验结合Box-Behnken试验优化发酵工艺并获得高效抗逆微生物菌剂;通过水培发芽试验验证其对旱盐胁迫下甘草生长的影响及其机制。结果显示,优化后的发酵培养基为：麦芽糖4.0%,大豆蛋白胨3.0%,KH₂PO₄0.01%,其余为水;优化后的发酵工艺为：发酵时间42 h、发酵温度34.0℃、初始pH 7.00、摇床转速230 r/min接种量2.0%、装液量100 mL/250 mL;接种菌剂后,干旱、盐胁迫下甘草幼苗发芽及生长指标均有不同程度的提高;盐胁迫下接种菌剂显著提高了抗氧化物质谷胱甘肽（Glutathione,GSH）、抗坏血酸（Ascorbate,AsA）及渗透调节物质（可溶性糖、可溶性蛋白）含量;干旱、盐胁迫下接种菌剂显著提高碳氮代谢酶活性,干旱胁迫下硝酸还原酶（Nitrate reductase,NR）活性提高77%,盐胁迫下蔗糖转化酶（Invertase,INV）活性提高445.5%。所得短小芽孢杆菌菌剂通过提高抗氧化物质、渗透调节物质含量及调节碳氮代谢酶活性,提高甘草的抗旱耐盐性,进而促进其生长。

关键词: 微生物菌剂, 抗旱耐盐性, 抗氧化, 渗透调节, 碳氮代谢

Abstract: The objective of this work is to prepare the highly effective stress-resistant microbial agent and to study their effects on the growth,development and physiological and biochemical characteristics of licorice under drought and salt stress. Single factor and orthogonal experiments were used to optimize the fermentation medium of Bacillus pumilus. Single factor experiments were combined with the Box-Behnken test to optimize the fermentation process,and a highly effective stress-resistant microbial agent was obtained. The hydroponic germination test was used to verify its effects on licorice growth under drought and salt stress and its mechanism. Results showed that the optimized fermentation medium was as：maltose 4.0%,soy peptone 3.0%,KH₂PO₄ 0.01%,and the rest was water. The optimized fermentation process was as：fermentation time 42 h,fermentation temperature 34℃,initial pH 7.00,shaking speed 230 r/min,the inoculation amount 2.0%,and the liquid content 100 mL/250 mL. The germination and growth indexes of licorice seedlings under drought and salt stress were improved to varying degrees. The inoculated agent significantly increased the contents of antioxidants（glutathione and ascorbate）and osmotic regulators（soluble sugars and soluble proteins）. Inoculation with bacterial agents under drought and salt stress significantly increased the activities of carbon and nitrogen metabolism enzymes. The activity of nitrate reductase increased by 77% under drought stress,and the activity of sucrose invertase increased by 445.5% under salt stress. In conclusion,the B. pumilus obtained in this study may improve the drought resistance and salt tolerance of licorice by increasing the content of antioxidants,osmoregulatory substances and regulating the activities of carbon and nitrogen metabolism enzymes,thereby promote its growth.

Key words: microbial agents, drought and salt tolerance, antioxidant, osmotic adjustment, carbon and nitrogen metabolism

张晓佳, 卢亚军, 张文晋, 张瑜, 崔高畅, 郎多勇, 张新慧. 抗旱耐盐菌剂的制备及其对甘草种子萌发的影响[J]. 生物技术通报, 2020, 36(9): 180-193.

ZHANG Xiao-jia, LU Ya-jun, ZHANG Wen-jin, ZHANG Yu, CUI Gao-chang, LANG Duo-yong, ZHANG Xin-hui. Preparation of Drought-resistant and Salt-tolerant Bacteria and Its Effect on Germination of Licorice Seeds[J]. Biotechnology Bulletin, 2020, 36(9): 180-193.

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