岩土高效溶蚀菌株Bt NL-11发酵条件优化及应用效果分析

王凌剑; 贾赵辉; 张金池; 唐兴港; 孙昕; 孟苗婧; 刘鑫

doi:10.12302/j.issn.1000-2006.202206022

王凌剑, 贾赵辉, 张金池, 唐兴港, 孙昕, 孟苗婧, 刘鑫

南京林业大学学报（自然科学版） ›› 2024, Vol. 48 ›› Issue (3) : 71-80.

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南京林业大学学报（自然科学版） ›› 2024, Vol. 48 ›› Issue (3) : 71-80. DOI: 10.12302/j.issn.1000-2006.202206022

专题报道Ⅱ:土壤生态修复理论与技术研究(执行主编张金池)

岩土高效溶蚀菌株Bt NL-11发酵条件优化及应用效果分析

王凌剑 ¹^,² ,
贾赵辉 ¹ ,
张金池 ¹^,^* ,
唐兴港 ¹ ,
孙昕 ¹ ,
孟苗婧 ¹ ,
刘鑫 ¹^,²

作者信息 +

Optimization for fermentation conditions and analysis of application effect for high efficiency dissolution strain Bt NL-11 from Bacillus thuringiensis

WANG Lingjian ¹^,² ,
JIA Zhaohui ¹ ,
ZHANG Jinchi ¹^,^* ,
TANG Xinggang ¹ ,
SUN Xin ¹ ,
MENG Miaojing ¹ ,
LIU Xin ¹^,²

Author information +

文章历史 +

摘要

【目的】为科学治理废弃矿山边坡,探究土壤细菌永久绿化法在生态修复中的应用与推广,对分离筛选出的高效溶蚀菌株进行发酵条件优化及应用效果分析。【方法】从南京幕府山风化岩壁土壤中分离出多种溶蚀微生物,并从中挑选出1种表现突出的溶蚀细菌菌株NL-11[经16S rRNA鉴定为苏云金芽孢杆菌(Bacillus thuringiensis,Bt)]进行发酵条件优化,利用盆栽试验观测其应用效果。首先利用单因素和Plackett Burman(PB)试验筛选出影响菌株生长的3个主要因素,即装液量、培养温度和时间;在此基础上使用最陡爬坡路径逼近最大响应区域;再利用Box-Behnken试验设计及响应面分析法进行回归分析;最后通过比较预测值与实测值验证模型可靠性。利用优化结果制备菌液,将制备好的菌液调节为低(10 cfu/mL)、中(1×10⁵ cfu/mL)和高(1×10⁹ cfu/mL)3种菌液浓度拌入基质(分别为T1、T2、T3处理)进行盆栽试验,设置不加菌液的处理为空白对照(CK),研究不同浓度菌液对矿物风化、植物和根系生长的促进作用。【结果】模型准确可靠,菌株NL-11的最佳发酵培养条件为:装液量19.51 mL,接种量2%(体积分数),初始pH 7.0,培养温度30.30 ℃,培养时间22.07 h,在此优化条件下发酵液中活菌数达到1.47×10¹⁰ cfu/mL,是未优化前的2.03倍。盆栽试验结果表明,菌株NL-11能够促进矿物风化,以高浓度菌液效果最显著;NL-11能够促进矿质养分溶解,以高浓度菌液效果最显著;NL-11能够促进植物及根系生长,以中浓度效果最显著。【结论】通过优化试验显著提高了菌株NL-11的发酵活菌产量,为菌株在边坡治理中的应用提供技术支持,综合评价菌株的应用效果并考虑生产成本等因素,喷播实践中的合适菌液浓度为1×10⁵ cfu/mL。

Abstract

【Objective】 This study aimed to scientifically manage abandoned mine slopes, explore the application and promotion of the soil bacteria permanent greening method in restoration, optimize the fermentation conditions, and analyze the application effect of the isolated and screened high-efficiency solubilizing bacteria.【Method】 A variety of solubilizing microorganisms were isolated from the weathered rock wall soil in Nanjing Mufu Mountain, and a prominent solubilizing strain, NL-11, identified as Bacillus thuringiensis by 16S rRNA, was selected to optimize fermentation conditions, and its application effect was observed with the potting test. The three main factors affecting the growth of the strain (liquid volume, temperature, and time), were screened using the univariate and Plackett Burman tests; on this basis, the steepest climbing path was used to approximate the maximum response area; then, the Box-Behnken experimental design was used and the response surface analysis method was used for regression analysis. Finally, model reliability was verified by comparing the predicted values with the measured values. The optimized results were used to prepare the bacterial solution, and then adjusted to low (10 cfu/mL), medium (1 × 10⁵ cfu/mL), and high (1 × 10⁹ cfu/mL) concentrations and mixed into the substrate (T1, T2, and T3 treatments, respectively) for the pot experiments, and the treatment without the bacterial solution was set as a blank control (CK) to study the effects of the different bacterial solution concentrations on mineral weathering and plant and root growth.【Result】 The model was accurate and reliable, and the optimal fermentation culture conditions for NL-11 were as follows: a liquid volume of 19.51 mL, an inoculum level of 2%, an initial pH of 7.0, a temperature of 30.30 ℃, and a time of 22.07 h. The number of viable bacteria in the fermentation broth under these optimized conditions reached 1.47 × 10¹⁰ cfu/mL, which was 2.03 times higher than that before optimization. The results of the pot tests showed that strain NL-11 could promote mineral weathering, and the effect of the high concentration of the bacterial solution was the most significant. Furthermore, strain NL-11 could promote the dissolution of mineral nutrients, and the effect of the high concentration of the bacterial solution was the most significant. Strain NL-11 could also promote plant and root growth, and the effect of the medium concentration was the most significant. 【Conclusion】 The optimization test significantly improved the production of fermentation of the live bacteria of strain NL-11 and provided technical support for the application of the strain in the management of slopes. The suitable concentration of the bacterial solution in spraying practice is 1 × 10⁵ cfu/mL by the comprehensive evaluation of the application effect and consideration of the production cost and other factors.

导出引用

王凌剑, 贾赵辉, 张金池, 等. 岩土高效溶蚀菌株Bt NL-11发酵条件优化及应用效果分析[J]. 南京林业大学学报（自然科学版）. 2024, 48(3): 71-80 https://doi.org/10.12302/j.issn.1000-2006.202206022

WANG Lingjian, JIA Zhaohui, ZHANG Jinchi, et al. Optimization for fermentation conditions and analysis of application effect for high efficiency dissolution strain Bt NL-11 from Bacillus thuringiensis[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2024, 48(3): 71-80 https://doi.org/10.12302/j.issn.1000-2006.202206022

中图分类号： X751;S732;Q89

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