CN114009451B - 深海放线菌11791在制备防治农牧业鳞翅目害虫药物中的应用 - Google Patents
深海放线菌11791在制备防治农牧业鳞翅目害虫药物中的应用 Download PDFInfo
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Abstract
本发明公开了深海放线菌11791在制备防治农牧业鳞翅目害虫药物中的应用。本发明通过实验发现，深海放线菌Streptomyces niveus SCSIO 11791的发酵产物对鳞翅目害虫小菜蛾杀灭效果，因此可以用于该类害虫引起的农作物的病虫害防治。本发明产品与部分商品化学农药的效果相当或者略好，如环氧虫啶。本发明生产成本低，绿色环保，是开发新型天然药物的理想资源。本发明生产周期短，发酵制备较容易，易于扩大生产。
Description
技术领域：
本发明属于生物技术领域，具体涉及一种深海放线菌Streptomyces sp.SCSIO11791及其发酵制剂在制备防治农牧业鳞翅目害虫药物中的应用。
背景技术：
有鳞翅目(Lepidoptera)昆虫，属有翅亚纲下的一目。全世界已知约20万种，中国已知约8000余种。该目为昆虫纲中仅次于鞘翅目的第2个大目，广泛分布于各地农牧生产区域，尤其以热带地区种类最为丰富。绝大多数鳞翅目种类的幼虫为各类栽培作物的害虫，体形稍大的常食尽叶片或钻蛀枝干，为农业生产的一大危害，如常见的小菜蛾、玉米螟、草地夜贪蛾等。高效、低毒的选择性防治鳞翅目害虫，对提高农业产量、提高农产品的质量都大有帮助。
海洋微生物、尤其是深海微生物，因起生长环境特殊，具有更多结构新颖活性物质的生产潜力，将其所产活性物质或其发酵制剂应用于农牧业的生产，不但有利于发展绿色、高效的农牧业，也是未来安全、绿色生物农药的一个重要发展方向。
发明内容：
本发明的目的是提供深海放线菌Streptomyces sp.SCSIO 11791在制备防治农业鳞翅目害虫药物中的应用。
本发明通过实验发现，深海放线菌Streptomyces sp.SCSIO 11791的发酵制剂对农业鳞翅目害虫有较强的毒力，因此可以用于鳞翅目害虫的防治。
因此，本发明的第一个目的是提供深海放线菌Streptomyces sp.SCSIO 11791在制备防治农牧业鳞翅目害虫药物中的应用。
优选，是深海放线菌Streptomyces sp.SCSIO 11791的发酵制剂在制备防治农牧业鳞翅目害虫药物中的应用。
进一步优选，所述的深海放线菌Streptomyces sp.SCSIO 11791的发酵制剂是其发酵产物的液体和(或)固形物部分。
优选，所述的深海放线菌Streptomyces sp.SCSIO 11791的发酵产物是将深海放线菌Streptomyces sp.SCSIO 11791接种到发酵培养基进行发酵培养获得的发酵产物，所述的发酵培养基按质量百分数计，包括可溶性淀粉0.5-1％，黄豆粉0.2-1％，葡萄糖2.0％，酵母提取粉0.2％，细菌学蛋白胨0.2％，KH2PO4 0.05％，MgSO4·7H2O 0.05％，粗海盐0.3-3.0％，CaCO3 0.2％，NaCl 0.4％，余量为水，pH 7.2-7.4；固体培养基发酵还要在液体培养基中添加1.5-2.0％的琼脂粉。
优选，所述的接种到发酵培养基进行发酵培养，是28℃下、以200rpm的转速培养或固体平板静置培养6-8天。
优选，所述的鳞翅目害虫是小菜蛾。
本发明的第二个目的是提供一种防治农牧业鳞翅目害虫药物，其含有深海放线菌Streptomyces sp.SCSIO 11791或其发酵制剂作为活性成分。
与现有技术相比，本发明具有以下有益效果：
(1)本发明产品与部分商品化学农药的效果相当或者略好，如环氧虫啶。
(2)本发明生产成本低，节能环保，是开发新型天然药物的理想资源
(3)本发明生产周期短，生产制备较容易，易于扩大生产。
本发明的深海放线菌Streptomyces sp.SCSIO 11791保藏于中国科学院南海海洋研究所海洋微生物中心，地址：广东省广州市海珠区新港西路196号，中科院南海海洋研究所。本发明的深海放线菌Streptomyces sp.SCSIO 11791公开于文献Chlorinated bis-indole alkaloids from deep-sea derived Streptomyces sp.SCSIO 11791withantibacterial and cytotoxic activities，Song,Yongxiang、Yang,Jiafan、Yu,Jianchen、Li,Jie、Yuan,Jie、Wong,Nai-Kei Ju,Jianhua，2020,73(8)，542-547，本申请人也持有，并保证自申请日起20年内向公众提供。
具体实施方式：
以下实施例是对本发明的进一步说明，而不是对本发明的限制。
实施例1：
一、材料与方法
1、材料
1.1种子培养基：
发酵培养基的配制方法如下：所述的发酵培养基按质量百分数计，葡萄糖0.4％，酵母提取粉0.4％，麦芽提取粉1％，海盐3％，pH 7.2，琼脂粉1.8％，溶剂为水。按照配方将称量好的上述物质溶解于水中、混匀，以每瓶200mL分装于500mL的锥形瓶中，于115℃灭菌30分钟，作为平板活化菌种的种子培养基。9cm直径的平板，每块约20mL培养基，划线接种深海放线菌Streptomyces sp.SCSIO 11791，待5-7天菌种长出后做为菌种(种子培养液)转接。
2、方法
2.1发酵制剂的制备
(1)放大发酵培养基配方：发酵培养基按质量百分数计，液体培养基包括可溶性淀粉0.5％，黄豆粉0.5％，葡萄糖2.0％，酵母提取粉0.2％，细菌学蛋白胨0.2％，KH2PO40.05％，MgSO4·7H2O 0.05％，粗海盐3.0％，CaCO3 0.2％，NaCl 0.4％，余量为水，pH 7.2-7.4；具体配制方法是将各成分溶于水中混用，固体培养基发酵还要在液体培养基中添加质量分数1.8％的琼脂粉，于115℃灭菌20分钟，作为放大发酵培养基。
(2)发酵培养：
在无菌条件下，将培养好的种子培养液分别接种于放大发酵培养基中，每1000ml锥形瓶(含约200mL放大发酵培养基)接种一瓶种子培养液10％(体积分数)，以200rpm的转速、在28℃下、摇床培养7-9天得海洋放线菌Streptomyces sp.SCSIO 11791发酵产物。将上述发酵产物冷冻干燥，得发酵粉剂，即为11791发酵制剂。
实施例2：发酵制剂对农业鳞翅目害虫的毒力测定
1供试药剂
实验药剂编号
2实验方法
(1)将环氧虫啶配制成终浓度为180ppm的水溶液，11791发酵制剂配制成浓度为质量分数5％的水溶液。
(2)将甘蓝叶片用清水洗净晾干，选取大小适中的叶片浸于不同浓度的药液中，10s后取出，于室温晾干后，将叶片置于铺有滤纸的培养皿中。选取小菜蛾3龄幼虫作为试虫，用毛笔将其轻轻挑到叶片上，每皿10头，每个处理重复3次。以水作为空白对照。用保鲜膜封口、加盖于(25±1)℃光照培养箱中保持120小时后检查死亡率。
(3)死亡判断标准：用拨针轻触虫体无反应或反应不协调者为死亡。
(4)对试验结果进行统计分析。
3结果与分析
表1样品在120h对小菜蛾胃毒测定结果
结果分析：由表1对小菜蛾胃毒测定结果可知，阳性对照药物在180ppm的环氧虫啶和稀释20倍的11791发酵制剂杀虫率分别为79.31％，85.19％。而空白对照虫子死亡率仅为0％。由此可见，11791发酵制剂在稀释20倍的情况下，具有良好的杀虫活性，其效果甚至好于180ppm的阳性对照药物环氧虫啶。
4结论
深海放线菌Streptomyces sp.SCSIO 11791发酵所产发酵制剂在直接稀释20倍时对农业鳞翅目害虫小菜蛾有较好防治效果，具有进一步深入试验研究的价值和应用的市场潜力。
Claims (4)
1.深海放线菌Streptomyces sp. SCSIO 11791的发酵制剂在制备防治农牧业鳞翅目害虫药物中的应用，所述的鳞翅目害虫是小菜蛾。
2.根据权利要求1所述的应用，其特征在于，所述的深海放线菌Streptomyces sp.SCSIO 11791的发酵制剂是其发酵产物的液体和/或固形物部分。
3. 根据权利要求2所述的应用，其特征在于，所述的深海放线菌Streptomyces sp.SCSIO 11791的发酵产物是将深海放线菌Streptomyces sp. SCSIO 11791接种到发酵培养基进行发酵培养获得的发酵产物，所述的发酵培养基按质量百分数计，包括可溶性淀粉0.5-1%，黄豆粉0.2-1%，葡萄糖2.0%，酵母提取粉0.2%，细菌学蛋白胨0.2%，KH2PO4 0.05%，MgSO4·7H2O 0.05%，粗海盐 0.3-3.0%，CaCO3 0.2%，NaCl 0.4%，余量为水，pH 7.2-7.4；固体培养基发酵还要在液体培养基中添加1.5-2.0%的琼脂粉。
4. 根据权利要求3所述的应用，其特征在于，所述的接种到发酵培养基进行发酵培养，是28℃下、以200 rpm的转速培养或固体平板静置培养6-8天。
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