CN114868773A - 一种防治果树炭疽病的复合微生物菌剂及其制备方法和应用 - Google Patents
一种防治果树炭疽病的复合微生物菌剂及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种防治果树炭疽病的复合微生物菌剂及其制备方法和应用，将贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液作为有效成分与十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、聚乙二醇、高岭土混合制备成复合微生物菌剂；该复合微生物菌剂活菌存活量高，贮存方便，崩解速度快，对于草莓连作土壤中的对香豆酸和阿魏酸有明显的抑制作用，显著提高草莓植株的株高、根系鲜重，果实增产，增加草莓甜度，并且能够有效的防治草莓炭疽病，能够产生较大的应用价值、经济效益及社会效益，同时充分实现了植物的再次利用，提高了植物的使用价值，绿色环保经济。
Description
技术领域
本发明涉及生防领域，特别是涉及一种防治果树炭疽病的复合微生物菌剂及其制备方法和应用。
背景技术
草莓营养价值丰富，被誉为是“水果皇后”，含有丰富的维生素C、维生素A、维生素E、维生素PP、维生素B1、维生素B2、胡萝卜素、鞣酸、天冬氨酸、铜、草莓胺、果胶、纤维素、叶酸、铁、钙、鞣花酸与花青素等营养物质，深受大家喜爱。随着市场需求量的增加，对于草莓的需求也相应增加，因而草莓种植面积也越来越大。但是草莓长期种植带来的连作障碍是需要克服的一大困难，草莓连作主要表现为生长发育不良、生育期延后、土传病害加重、产量明显下降，而草莓炭疽病是草莓苗期主要的土传病害，给草莓生产带来了严重障碍。草莓的连作障碍是由于其植株根际土壤中含有丰富的酚酸，部分酚酸在低浓度下促进草莓根茎腐炭疽病的发生，而在高浓度下又抑制根系生长。
生物防治方法由于菌株的单一性、病原菌的多样性和大田环境的复杂性，在生产中的防治效果往往不够理想也不够稳定，无法满足实际需求，因此需要复合菌剂这些缺陷。
发明内容
针对上述情况，本发明之目的在于提供一种防治果树炭疽病的复合微生物菌剂及其制备方法，对草莓炭疽病具有较好的防治效果，同时能够克服草莓连作障碍。
其解决的技术方案是：
一种防治果树炭疽病的复合微生物菌剂，包括有效成分：贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液，还包括以下辅助成分：十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、聚乙二醇、高岭土；所述植物提取液来自大蒜、玄参、博落回。
进一步的，所述有效成分中贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液的体积比为：2-15:4-8:5-12:3-5；各发酵液中有效活菌数≥20亿/克。
进一步的，所述复合微生物菌剂中各组分的质量百分比为：10％-40％有效成分、5-10％十二烷基苯磺酸钠、6-13％六聚磷酸钠、5-8％海藻酸钠、2-3％聚乙二醇、高岭土补足至100％。
优选的，所述复合微生物菌剂中各组分的质量百分比为：20％有效成分、8％十二烷基苯磺酸钠、8％六聚磷酸钠、6％海藻酸钠、3％聚乙二醇、高岭土补足至100％。
进一步的，所述植物提取液由以下方法获得：在避光处，将玄参茎叶、大蒜茎叶、博落回果等比例干燥，粉碎过200目筛，加20倍水搅拌浸没，40℃下超声提取1h，然后用去离子水过滤洗涤，40℃下浓缩，最后加入3倍质量的水，得植物提取液。
进一步的，所述复合微生物菌剂由以下方法制备：将有效成分、十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、高岭土按照一定的比例混合均匀，研磨粉碎得到可湿性粉剂，然后加入水和聚乙二醇进行捏合造粒，制成水分散粒剂的初级产物，最后再对初级产物进行挤压造粒和干燥，筛分后得到复合微生物菌剂。
进一步的，所述复合微生物菌剂用于防治草莓炭疽病。
进一步的，用于防治草莓炭疽病的使用方法为：将复合微生物菌剂加水稀释100倍后灌根处理。
本发明的有益技术效果：
(1)本发明使用源自于玄参茎叶、大蒜茎叶、博落回果的植物提取液与贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液复合制备成复合微生物菌剂，充分实现了植物的再次利用，提高了植物的使用价值，绿色环保经济。
(2)本发明的复合微生物菌剂对于草莓连作土壤中的对香豆酸和阿魏酸有明显的抑制作用，同时能显著提高了的草莓植株的株高、根系鲜重，实现果实增产，且果实甜度增加，并且能够有效的防治草莓炭疽病，能够产生较大的应用价值、经济效益及社会效益。
(3)本发明将有效成分与十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、聚乙二醇、高岭土混合，通过合适的配比制备成的复合微生物菌剂活菌存活量高，贮存方便，崩解速度快，因而复合微生物菌剂的使用效果好。
具体实施方式
下面结合具体实施方式对本发明的技术方案作详细描述，但需要强调的是，本发明并不受限于所述的具体实施方式；下述实施例所用微生物菌种可自行分离或通过购买得到；下述实施例所述原料无特殊说明均可通过常规方法制备或购买获得。
实施例1
有效成分的获取：
(1)贝莱斯芽孢杆菌发酵液的制备：挑取贝莱斯芽孢杆菌的单菌落于LB液体培养基，30℃下，180r/min振荡培养18h，得种子液；然后将种子液以4％的接种量接种于LB液体培养基中，在温度为32℃下，200r/min培养48h，得贝莱斯芽孢杆菌发酵液，发酵液中有效活菌数≥20亿/克；其中，所述LB液体培养基含有酵母提取物4g/L、胰蛋白胨12g/L、氯化钠1g/L。
(2)多粘类芽孢杆菌发酵液的制备：挑取多粘类芽孢杆菌的单菌落于液体培养基，28℃下，180r/min振荡培养20h，得种子液；然后将种子液以3％的接种量接种于液体培养基中，在温度为30℃下，210r/min培养54h，得多粘类芽孢杆菌发酵液，发酵液中有效活菌数≥20亿/克；其中，所述液体培养基含有葡萄糖15g/L、酵母提取物8g/L、稻米粉12g/L、小麦粉11g/L、麸皮18g/L、豆粕8g/L、氯化钙0.5g/L、氯化锰0.1g/L、硫酸镁3.8g/L。
(3)短小芽孢杆菌发酵液的制备：挑取短小芽孢杆菌的单菌落于NA液体培养基，30℃下，180r/min振荡培养24h，得种子液；然后将种子液以3％的接种量接种于NA液体培养基中，在温度为30℃下，200r/min培养48h，得短小芽孢杆菌发酵液，发酵液中有效活菌数≥20亿/克；其中，所述NA液体培养基含有3g/L牛肉膏、5g/L蛋白胨、3g/L葡萄糖。
(4)植物提取液的制备：在避光处，将玄参茎叶、大蒜茎叶、博落回果等比例干燥，粉碎过200目筛，加20倍水搅拌浸没，40℃下超声提取1h，然后用去离子水过滤洗涤，40℃下浓缩，最后加入3倍质量的水，得植物提取液。
实施例2
一种防治果树炭疽病的复合微生物菌剂按以下质量百分比包括：15％有效成分、5％十二烷基苯磺酸钠、7％六聚磷酸钠、5％海藻酸钠、2％聚乙二醇、高岭土补足至100％；所述有效成分中贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液的体积比为：2:4:5:3。
实施例3
一种防治果树炭疽病的复合微生物菌剂按以下质量百分比包括：20％有效成分、8％十二烷基苯磺酸钠、8％六聚磷酸钠、6％海藻酸钠、3％聚乙二醇、高岭土补足至100％；所述有效成分中贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液的体积比为：8:6:9:4。
实施例4
一种防治果树炭疽病的复合微生物菌剂按以下质量百分比包括：30％有效成分、10％十二烷基苯磺酸钠、11％六聚磷酸钠、8％海藻酸钠、3％聚乙二醇、高岭土补足至100％；所述有效成分中贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液的体积比为：15:8:12:5。
实施例5
一种防治果树炭疽病的复合微生物菌剂的制备方法：按照实施例2-4中所述比例将有效成分、十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、高岭土混合均匀，研磨粉碎得到可湿性粉剂，然后加入水和聚乙二醇捏合、造粒，制备成水分散粒剂，再对水分散粒剂挤压造粒，最后干燥，筛分后得到复合微生物菌剂。
对比例1
一种防治果树炭疽病的复合微生物菌剂：各组分含量与实施例3相同，不同的是将短小芽孢杆菌发酵液替换成多粘类芽孢杆菌发酵液发酵液，制备方法同实施例5。
对比例2
一种防治果树炭疽病的复合微生物菌剂：各组分含量与实施例3相同，不同的是将有效成分中植物提取液替换成多粘类芽孢杆菌发酵液，制备方法同实施例5。
对比例3
一种防治果树炭疽病的复合微生物菌剂：各组分含量与实施例3相同，不同的是所述植物提取液来自大蒜，植物提取液的制备方法同实施例1，复合微生物菌剂的制备方法同实施例5。
对比例4
一种防治果树炭疽病的复合微生物菌剂：各组分含量与实施例3相同，不同的是将高岭土替换成硅藻土，制备方法同实施例5。
实施例6
复合微生物菌剂对草莓炭疽病的防治实验
试验于河南省商丘市匡城乡的草莓种植大棚中，草莓种植品种：女峰；连作时间：4年；处理：实施例2-4以及对比例1-4所制备的复合微生物菌剂加水稀释100倍后对草莓植株分区等量灌根，另设一个对照处理，施加等量的清水；每个区处理面积为20m2；处理时间：苗期，10d为一周期，施用3次；正常施肥浇水管理。
于草莓开花期在每个区分别随机选取5个50×50cm的取样区域，然后在每个50*50cm的取样区域随机挖取10株草莓植株，通过统计每块地块中该取样草莓植株上的炭疽病斑总数、株高、根鲜重，并检测10株草莓植株的土壤中的酚酸(对香豆酸、阿魏酸)含量；另外，在结果期进行果实产量统计，测试果实的甜度，与空白组对比，获得草莓炭疽病防治率、株高提高率、根鲜重提高率、果实增产率、甜度提升率、酚酸的增加率。根的鲜重，取草莓的地下部分，用水冲洗干净，用吸水纸擦干表面水份后，称重；最终得表1和2：
| 表1 | 对香豆酸增加率/％ | 阿魏酸增加率/％ |
| 实施例2 | -15.3 | -11.6 |
| 实施例3 | -14.6 | -10.2 |
| 实施例4 | -14.7 | -11.5 |
| 对比例1 | -8.5 | -4.8 |
| 对比例2 | 3.7 | 5.1 |
| 对比例3 | -5.3 | -2.7 |
| 对比例4 | -11.4 | -8.6 |
从表1可以看出，实施例2-4制备的复合微生物菌剂对于草莓连作土壤中的对香豆酸和阿魏酸有明显的抑制作用，贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、来自大蒜、玄参、博落回的植物提取液联合作用，抑制效果明显，高岭土的抑制效果稍弱。
从表2可以看出，实施例2-4制备的复合微生物菌剂能有效的防治草莓炭疽病，用于草莓种植连作，显著提高了的草莓植株的株高、根系鲜重，实现果实增产，且果实甜度增加，因此，复合微生物菌剂还能有效的提高草莓的产量和品质。
以上所述是结合具体实施方式对本发明所作的进一步详细说明，不能认定本发明具体实施仅局限于此；对于本发明所属及相关技术领域的技术人员来说，在基于本发明技术方案思路前提下，所作的拓展以及操作方法、数据的替换，都应当落在本发明保护范围之内。
Claims (8)
1.一种防治果树炭疽病的复合微生物菌剂，其特征在于，包括有效成分：贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液，还包括以下辅助成分：十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、聚乙二醇、高岭土；所述植物提取液来自大蒜、玄参、博落回。
2.根据权利要求1所述的复合微生物菌剂，其特征在于，所述有效成分中贝莱斯芽孢杆菌发酵液、多粘类芽孢杆菌发酵液、短小芽孢杆菌发酵液、植物提取液的体积比为：2-15:4-8:5-12:3-5；各发酵液中有效活菌数≥20亿/克。
3.根据权利要求1所述的复合微生物菌剂，其特征在于，所述复合微生物菌剂中各组分的质量百分比为：10％-40％有效成分、5-10％十二烷基苯磺酸钠、6-13％六聚磷酸钠、5-8％海藻酸钠、2-3％聚乙二醇、高岭土补足至100％。
4.根据权利要求1所述的复合微生物菌剂，其特征在于，所述复合微生物菌剂中各组分的质量百分比为：20％有效成分、8％十二烷基苯磺酸钠、8％六聚磷酸钠、6％海藻酸钠、3％聚乙二醇、高岭土补足至100％。
5.根据权利要求1所述的复合微生物菌剂，其特征在于，所述植物提取液由以下方法获得：在避光处，将玄参茎叶、大蒜茎叶、博落回果等比例干燥，粉碎过200目筛，加20倍水搅拌浸没，40℃下超声提取1h，然后用去离子水过滤洗涤，40℃下浓缩，最后加入3倍质量的水，得植物提取液。
6.根据权利要求1-5任一所述的复合微生物菌剂，其特征在于，所述复合微生物菌剂由以下方法制备：将有效成分、十二烷基苯磺酸钠、六聚磷酸钠、海藻酸钠、高岭土按照一定的比例混合均匀，研磨粉碎得到可湿性粉剂，然后加入水和聚乙二醇进行捏合造粒，制成水分散粒剂的初级产物，最后再对初级产物进行挤压造粒和干燥，筛分后得到复合微生物菌剂。
7.根据权利要求1-6任一所述的复合微生物菌剂，其特征在于，所述复合微生物菌剂用于防治草莓炭疽病。
8.根据权利要求7所述的复合微生物菌剂，其特征在于，用于防治草莓炭疽病的使用方法为：将复合微生物菌剂加水稀释100倍后灌根处理。
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