响应面法优化根结线虫生防真菌Snef5的发酵工艺

doi:10.16409/j.cnki.2095-039x.2016.04.012

中国生物防治学报 ›› 2016, Vol. 32 ›› Issue (4): 503-510.DOI: 10.16409/j.cnki.2095-039x.2016.04.012

响应面法优化根结线虫生防真菌Snef5的发酵工艺

杨传旭¹, 赵迪⁴, 谭卓¹, 朱晓峰¹, 刘晓宇³, 段玉玺¹, 王媛媛², 陈立杰¹

1. 沈阳农业大学植物保护学院, 沈阳 110866;
2. 沈阳农业大学生物科学技术学院, 沈阳 110866;
3. 沈阳农业大学理学院, 沈阳 110866;
4. 沈阳农业大学分析测试中心, 沈阳 110866

收稿日期:2016-01-21 出版日期:2016-08-08 发布日期:2016-07-29
通讯作者: 陈立杰, 王媛媛
作者简介:杨传旭,女,硕士研究生,E-mail:1206034989@qq.com
基金资助:
国家自然科学基金（31471748，31171823）；沈阳农业大学天柱山学者资助

Ferment Optimization of Biocontrol Fungus Snef5 against Meloidogyne incognita by Response Surface Methodology

YANG Chuanxu¹, ZHAO Di⁴, TAN Zhuo¹, ZHU Xiaofeng¹, LIU Xiaoyu³, DUAN Yuxi¹, WANG Yuanyuan², CHEN Lijie¹

1. College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China;
2. College of Biological Science and Technology, Shenyang Agricultural University, Shenyang 110866, China;
3. College of Science, Shenyang Agricultural University, Shenyang 110866, China;
4. Analytical Instrumentation Center, Shenyang Agricultural University, Shenyang 110866, China

Received:2016-01-21 Online:2016-08-08 Published:2016-07-29

摘要/Abstract

摘要： 为提高生防真菌Simplicillium chinense Snef5的杀线虫活性，本研究采用响应面法对其发酵条件进行优化。首先，采用Plackett?Burman法筛选出有显著效应的3个因素：初始pH值、蔗糖和MgSO₄·7H₂O用量，而后通过Box?Benhnken设计和响应面分析法对其进行分析，获得了优化的最佳初始pH值为6.74，蔗糖含量4.19%，MgSO₄·7H₂O含量0.04%。经摇瓶和发酵罐发酵验证，该理论预测值与实测值无显著差异；优化后的发酵液杀线虫活性与原始发酵液相比，摇瓶发酵与30 L发酵罐产出的发酵液对南方根结线虫2龄幼虫的致死率分别提高了20.6%和21.3%，显示出较好的优化效果。试验结果对生防真菌的发酵及实际应用具有重要指导作用。

关键词: 生防真菌Snef5, 响应面法, 南方根结线虫, 发酵条件优化

Abstract: Simplicillium chinense strain Snef5 was screened as unique biological control fungi to plant nematodes in our previous research. In order to enhance its nematicidal activity, optimization of fermentation conditions were investigated for this fungus using response surface methodology. First, through Plackett-Burman screening experiments, three factors including the initial pH, the concentration of sucrose and MgSO₄·7H₂O in the broth appeared to have significant effects on the fermentation. According to the results of Box-Benhnken design and response surface analysis, the optimal conditions for strain Snef5 fermentation were obtained with the initial pH 6.74, 4.19% sucrose and 0.04% MgSO₄·7H₂O in the medium. Fermentation experiments with shake flasks and 30 L fermentor verified that there was no statistical difference between the nematicidal activity forecasted by the model and that obtained in the tests. After optimizing the fermentation broth, Snef5 enhanced its nematicidal activity by exhibition of increasing the second juvenile (J2) mortality rate of Meloidogyne incognita. Compared to the original fermentation broth, it showed 20.6% enhancement in the J2 mortality rate when using optimized broth in the flasks and 21.3% when using in the 30 L cylinders, respectively. Our research should have important implications for biocontrol fungi fermentation and application.

Key words: biocontrol fungus Simplicillium chinense Snef5, response surface methodology, Meloidogyne incognita, fermentation optimization

中图分类号:

S476

杨传旭, 赵迪, 谭卓, 朱晓峰, 刘晓宇, 段玉玺, 王媛媛, 陈立杰. 响应面法优化根结线虫生防真菌Snef5的发酵工艺[J]. 中国生物防治学报, 2016, 32(4): 503-510.

YANG Chuanxu, ZHAO Di, TAN Zhuo, ZHU Xiaofeng, LIU Xiaoyu, DUAN Yuxi, WANG Yuanyuan, CHEN Lijie. Ferment Optimization of Biocontrol Fungus Snef5 against Meloidogyne incognita by Response Surface Methodology[J]. journal1, 2016, 32(4): 503-510.

参考文献

[1] 段玉玺. 植物线虫学[M]. 北京: 科学出版社, 2011, 1-2.
[2] Box G E P, Wilson K B. On the experimental attainment of optimum conditions[J]. Journal of The Royal Statistical Socienty Serries B, 1951, 13(1): 1-20.
[3] Miller A, Sitter R R. Using the folded-over 12-run placket-burman design to consider interactions[J]. Technometrics, 2001, 43(1): 44-54.
[4] Ambat P, Ayyanna C. Optimizing medium constituents and fermentation conditions for citric acid production from palmyra jaggery using response surface methods[J]. Journal of Microbiology and Biotechnology, 2001, 17: 331-335.
[5] Trupkin S, Levin S, Forchiassin F. Optimization of a culture medium for ligninolytic enzyme production and synthetic dye decolorization using response surface methodology[J]. Journal of Industrial Microbiology Biotechnology, 2003, 30: 682-690.
[6] 花玉鹏, 文才艺, 刘伟成, 等. 响应曲面法优化利迪链霉菌A02发酵培养基[J]. 中国生物防治学报, 2011, 27(4): 520-527.
[7] 陆继臣, 迟乃玉, 窦少华, 等. 响应面法优化灰霉病生防菌CNY-04培养条件[J]. 微生物学通报, 2003, 40(8): 1414-1422.
[8] 赵迪, 刘彬, 李玲玉, 等. 白僵菌及其伴生菌发酵液对线虫的毒力研究[J]. 农药学学报, 2013, 15(2): 178-182.
[9] Zhao D, Liu B, Li L, et al. Simplicillium chinense, as a biological control agent against plant parasitic nematodes[J]. Biocontrol Science and Techology, 2013, 23(8): 980-986.
[10] 陈立杰, 刘彬, 段玉玺, 等. 白僵菌发酵液对不同种类线虫生物活性的影响[J]. 沈阳农业大学学报, 2008, 39(3): 305-308.
[11] Chen S Y, Dickson D W. A technique for determining live second-stage juveniles of Heterodera glycines[J]. Journal of Nematology, 2000, 32(1): 117-121.
[12] 常亚飞, 陈守文, 喻子牛. 苏云金素发酵培养基的优化设计[J]. 中国生物防治, 2006, 22(3): 190-194.
[13] Cho J H, Kim Y B, Kim E K. Optimization of culture media for Bacillus species by statistical experimental design methods[J]. Korean Journal of Chemical Engineering, 2009, 26(3): 754-759.
[14] 张建华, 梁昌聪, 刘磊. 淡紫拟青霉E16液体发酵工艺研究[J]. 河南农业科学, 2014, 43(11): 87-92.
[15] 曾庆飞, 李传浩, 黄惠琴, 等. 根结线虫拮抗放线菌菌株DA07118的筛选与鉴定及其发酵条件的优化[J]. 中国生物防治, 2009, 25(3): 255-259.
[16] 刘亮山, 林茂松, 鄢小宁, 等. 生防链霉菌JH108-2的发酵培养基优化[J]. 植物病理学报, 2006, 36(3): 259-266.
[17] 王永娟, 张杰, 孔繁芳, 等. 嗜线虫致病杆菌HB310培养基筛选和培养条件优化[J]. 环境科学学报, 2014, 36(6): 997-1003.

响应面法优化根结线虫生防真菌Snef5的发酵工艺

Ferment Optimization of Biocontrol Fungus Snef5 against Meloidogyne incognita by Response Surface Methodology

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