具有除草活性的生防菌株GD-9发酵条件优化及菌剂制备
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摘要
为了明确具有除草活性的生防菌株GD-9发酵过程中各因子的配比和最优条件,采用单因素试验对菌株最适碳源、氮源、载体、固态发酵基质进行了筛选,应用正交试验设计研究了碳源、氮源、初始含水量、接种量、初始pH、培养时间、培养温度7种因素对菌株活菌数的影响。试验结果表明菌株GD-9最佳固态发酵条件为:氮源NaNO_3 58.4 mg/g,碳源葡萄糖48.4 mg/g,培养基质最适初始含水量为258 mg/g,最适接种量为0.26 mL/g,初始pH 7.6,最佳培养时间147.8 h,最佳培养温度30.7℃,最适宜的载体为黏土,分散剂为聚乙烯醇,稳定剂为膨润土,润湿剂为糊精。通过发酵试验结果制备生防菌株GD-9的固体菌剂,该研究为菌剂的商品化生产奠定基础。
In order to clarify the proportion and optimal conditions of various factors in the fermentation process of biocontrol strain GD-9 with herbicidal activity, the single factor experiment was performed to screen the optimal carbon source, nitrogen source, carrier and solid fermentation substrate of the strain. The orthogonal experiment design was used to study the influences of carbon source, nitrogen source, initial water content, inoculation amount, initial pH value, culture time and culture temperature on the colony number of the strain. The experimental results showed that the optimal solid state fermentation conditions of strain GD-9 were N-source NaNO_3 58.4 mg/g, C-source glucose 48.4 mg/g, 258 mg/g of initial water content of the culture medium, 0.26 mL/g of inoculation amount, the initial pH 7.6, 147.8 h and 30.7℃ of culture time and temperature, respectively. The optimal carrier, dispersant, stabilizer and wetting agent were clay, polyvinyl alcohol, bentonite and dextrin. The solid microbial inoculum of biocontrol strain GD-9 was prepared through fermentation test results. This study laid a foundation for the commercial production of the microbial inoculum.
In order to clarify the proportion and optimal conditions of various factors in the fermentation process of biocontrol strain GD-9 with herbicidal activity, the single factor experiment was performed to screen the optimal carbon source, nitrogen source, carrier and solid fermentation substrate of the strain. The orthogonal experiment design was used to study the influences of carbon source, nitrogen source, initial water content, inoculation amount, initial pH value, culture time and culture temperature on the colony number of the strain. The experimental results showed that the optimal solid state fermentation conditions of strain GD-9 were N-source NaNO_3 58.4 mg/g, C-source glucose 48.4 mg/g, 258 mg/g of initial water content of the culture medium, 0.26 mL/g of inoculation amount, the initial pH 7.6, 147.8 h and 30.7℃ of culture time and temperature, respectively. The optimal carrier, dispersant, stabilizer and wetting agent were clay, polyvinyl alcohol, bentonite and dextrin. The solid microbial inoculum of biocontrol strain GD-9 was prepared through fermentation test results. This study laid a foundation for the commercial production of the microbial inoculum.
引文
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[2] 崔新倩.花生田杂草化学防除现状及趋势[J].农药科学与管理,2011,14(12):156-158.
[3] 黄石旺,刘正日,陈和春,等.现代农业发展中除草剂的危害及对策[J].现代农业科技,2010(14):28-29.
[4] CUTLER H G,Perspectives on discovery of microbial phytotoxins with herbicide activity [J].Weed Technology,1988(2):525-532.
[5] 丁建清.生物防治——杂草综合防治的重要内容[J].杂草学报,1995,9(1):60-64.
[6] 陈燕芳,丁伟,丁吉林,等.天然产物除草剂研究进展[J].杂草科学,2007(2):1-5.
[7] LI Yongquan,SUN Ziling,ZHUANG Xiaofeng,et al.Research progress on microbial herbicides [J].Crop Protection,2003,22(2):247-252.
[8] 仉欢,王开运.微生物除草剂研究进展[J].杂草科学,2010(2):1-8.
[9] 藤森岭.微生物源农药杂草防除[J].农业技术,1993,48(1):18-21.
[10] ZHANG Jinlin,ZHANG Lihui,LIU Yingchao,et al.The herbicidal activity of mutant isolates from Botrytis cinerea[J].Agricultural Sciences in China,2006,5(8):622-628.
[11] 马娟,董金皋.微生物除草剂与生物安全[J].植物保护,2006,32(1):9-12.
[12] 涂璇.辣椒疫霉生防菌筛选及生防菌剂应用研究[D].杨凌:西北农林科技大学,2004.
[13] 郭亮,刑晓旭,郑学明,等.重铬酸钾溶液对细菌活性的影响[J].山东畜牧兽医,2009,30(2):4-8.
[14] 曹红娣.农药可湿性粉剂生产工艺的改进[J].上海农业科技,2005(6):77-79.
[15] 张剑,董晔欣,张金林,等.一株具有高除草活性的真菌菌株[J].菌物学报,2008,27(5):645-651.