杀线虫解淀粉芽胞杆菌Sneb709与费氏中华根瘤菌Sneb183共培养发酵条件优化  

Optimization of Fermentation Conditions for Co-culture of Nematocidal Bacillus amyloliquefaciens Sneb709 and Sinorhizobium fredii Sneb183

作　　者：李佳娣 赵劲捷 范海燕 朱晓峰[1] 王媛媛[2] 刘晓宇[3] 段玉玺[1] 陈立杰[1] LI Jiadi;ZHAO Jinjie;FAN Haiyan;ZHU Xiaofeng;WANG Yuanyuan;LIU Xiaoyu;DUAN Yuxi;CHEN Lijie(College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,China;College of Biological Science and Technology,Shenyang Agricultural University,Shenyang 110866,China;College of Science,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[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

出　　处：《中国生物防治学报》2021年第4期772-785,共14页Chinese Journal of Biological Control

基　　金：国家重点研发计划(2017YFD0201104);辽宁省教育厅科学研究经费项目(LSNQN201902);辽宁省重点研发项目(2019JH2/10200012)。

摘　　要：根结线虫可引起作物发生严重的土传病害,生物防治是持续控制此类病害的有效措施。本实验室前期筛选获得多株杀线虫活性的芽胞杆菌和根瘤菌,本研究将解淀粉芽胞杆菌Sneb709与费氏中华根瘤菌Sneb183进行共培养后发现,共培养发酵液对南方根结线虫2龄幼虫J2活性显著提高,因此本文利用响应面法对其发酵条件进行优化,获得更好的生防效果。以J2的校正死亡率为指标,采用单因素法、最陡爬坡法和响应面法设计并优化共培养菌株的发酵液培养基成分以及培养条件,确定共培养发酵液的最适培养基成分为:山梨糖1.98 g/L、黄豆粉1.06 g/L、花生饼粉2.08 g/L;最佳发酵条件为:温度30.4°C、初始pH 6.49、装液量103.15 mL。使用优化条件对两个菌株进行共培养后,其发酵液对J2的毒杀活性显著提高,提升至84.17%,使用发酵罐验证,共培养发酵液的毒杀活性提升了24.72%。该结果为共培养菌株有效的活性产物分离提取等后续研究及杀线虫生防菌的产业化提供了理论依据。Root-knot nematodes can cause serious soil-borne diseases of crops,and biological control is an effective measure to sustainability control such diseases.Several strains of Bacillus and Rhizobium with nematicidal activity were obtained in previous work of our laboratory.In this study,Bacillus amyloliquefaciens Sneb709 and Sinorhizobium fredii Sneb183 were co-cultured,and the co-culture fermentation broth had significantly improved activity against Meloidogyne incognita second-stage juveniles(J2).To obtain better biocontrol effects,the fermentation conditions were optimized by response surface methodology.Taking the corrected mortality rate of J2 as an indicator,the single factor method,steepest climbing method and response surface method are used to design and optimize the fermentation broth composition and culture conditions of the co-cultured strains.The most suitable medium composition of the co-culture fermentation broth was determined as follows:sorbose 1.98 g/L,soy flour 1.06 g/L,peanut meal 2.08 g/L.The optimal fermentation conditions were as follows:temperature 30.4℃,initial pH 6.49 and liquid volume 103.15 mL.After the two strains were co-cultured with optimized conditions,the toxic activity of the fermentation broth on J2 was significantly improved,with the mortality of 84.17%.After verification in the fermenter,the toxic activity of the co-culture fermentation broth increased by 24.72%.This result provides a theoretical basis for subsequent studies on separation and extraction of active products of co-cultured strains and the industrialization of nematocidal biocontrol bacteria.

关 键 词：根结线虫 芽胞杆菌 根瘤菌 共培养 发酵条件 

分 类 号：S476[农业科学—农业昆虫与害虫防治]