CN105475360B - 含枯草芽孢杆菌和菌核净的组合物及应用 - Google Patents
含枯草芽孢杆菌和菌核净的组合物及应用 Download PDFInfo
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Abstract
含枯草芽孢杆菌和菌核净的组合物及应用，属于农药领域。所述含枯草芽孢杆菌Pc3和菌核净的组合物，以枯草芽孢杆菌Pc3和菌核净为活性成分，枯草芽孢杆菌Pc3与菌核净的质量比为1000∶1～1∶1000。所述枯草芽孢杆菌Pc3中活孢子数可为0.1～1000亿/g。在枯草芽孢杆菌Pc3和菌核净的组合物中加入农药助剂后即可配得可湿性粉剂、悬乳剂或水乳剂。如向活性成分枯草芽孢杆菌Pc3和菌核净中加入农药助剂并调节pH值为5～7后，配成可湿性粉剂。菌核净可采用原药，枯草芽孢杆菌采用原粉。所述含枯草芽孢杆菌和菌核净的组合物可在制备防治油菜菌核病的农药中应用。
Description
技术领域
本发明属于农药领域，具体是涉及一种含从南极分离的枯草芽孢杆菌(Bacillussp.)Pc3和菌核净的组合物及其在防治油菜菌核病上的应用。
背景技术
由核盘菌(Sclerotinia sclerotiorum)引起的油菜菌核病是油菜生产中的重要病害之一，常年病株率为10％～30％，严重的可达80％以上,病株一般减产10％～70％。油菜是产油效率最高的油料作物之一，目前油菜菌核病的主要防治措施是化学防治、农业防治和生物防治。但至今为止，未发现高效的抗菌核病品种，因此目前对该病害防治主要还是采用化学防治方法，其中使用最多的是苯并咪唑类杀菌剂。随着化学药剂的常年使用，油菜菌核病菌对杀菌剂产生了明显的抗药性，从而使化学药剂的防治效果下降，甚至防治失败。并且随着人们对环境保护、生态平衡、食品安全的重视，传统的化学防治方法已无法满足人们的需求，因此开拓以高效、无公害微生物复配农药为主的综合防治油菜菌核病的技术体系迫在眉睫。
枯草芽孢杆菌属细菌，属生物杀菌剂，是内生芽孢的革兰氏阳性细菌，通过成功定殖至植物根际、体表或体内，同病原菌竞争植物周围的营养，分泌抗菌物质抑制病原菌生长，同时诱导植物防御系统抵御病原菌入侵，从而达到生防的目的。枯草芽孢杆菌杀菌剂克服了传统化学农药污染环境、危害人畜、易产生抗性等缺点，具有选择性强、安全、原料简单等优点,在微生物杀菌剂市场中崭露头角。
本申请人在中国专利ZL201310637219.8公开一种枯草芽孢杆菌Pc3及其在制备防治植物致病真菌发酵上清液中的应用，所述枯草芽孢杆菌(Bacillus subtilis)Pc3的保藏编号为CCTCC NO：M2013470。枯草芽孢杆菌(Bacillus subtilis)Pc3在制备防治植物致病真菌发酵上清液中的应用，防治的植物致病真菌包括：香蕉炭疽菌、宛氏拟青霉、核盘菌、长柄链格孢、绿色木霉、立枯丝核菌和镰刀菌等。在油菜菌核病菌防治的盆栽试验中有较好的生物防治效果，防效可以达到95.6％。枯草芽孢杆菌(Bacillus subtilis)Pc3发酵上清液的制备工艺简单，抑真菌谱广，生防效果好。
枯草芽孢杆菌(Bacillus subtilis)Pc3，从南极分离得到，抑菌谱较广，对多种病原真菌尤其是油菜菌核病菌有很强的抑制作用；生长速度快，适应环境能力强，易于在植物表面定殖，并形成优势种群；能诱导寄主产生抗病性；可分泌多种抗菌物质；对油菜菌核的防治效果较好。该菌株于2013年10月14日保藏于中国典型培养物保藏中心，保藏编号为CCTCC NO：M2013470，中国典型培养物保藏中心的地址为中国，武汉，武汉大学。
菌核净(Dirnethachlon)是一种二甲酞亚胺类杀菌剂，它的有效成分是(N-3,5-二氯苯基)丁二酸亚胺(NDPS)。具有直接杀菌、内渗治疗、抗雄激素作用、残效期长、杀菌谱广、防治效果显著等特点，已被广泛应用于多个类群植物病原真菌，如葡萄孢属、青霉属、核盘菌属、链格孢属、长蠕孢属、丝核菌属、茎点霉属等引起的作物、果树病害防治。对于油菜菌核病、烟草赤腥病、水稻纹枯病、麦类赤霉病、白粉病以及工业防腐具有良好的防效。
发明内容
本发明的第一目的在于针对目前生产上大量使用化学农药防治油菜菌核病、环境污染严重、农药残留超标、油菜品质下降、病原菌抗药性增强，导致防效下降，甚至防治失败的问题，提供一种对油菜菌核病害有较好防治作用和增效作用，不仅能发挥枯草芽孢杆菌Pc3有效控制油菜菌核病的作用，还能降低现有病原菌对菌核净潜在抗药性风险的含枯草芽孢杆菌和菌核净的组合物及应用。
本发明的第二目的在于提供所述含枯草芽孢杆菌和菌核净的组合物在制备防治油菜菌核病的农药中应用。
所述含枯草芽孢杆菌Pc3和菌核净的组合物，以枯草芽孢杆菌Pc3和菌核净为活性成分，枯草芽孢杆菌Pc3与菌核净的质量比为1000∶1～1∶1000；枯草芽孢杆菌Pc3与菌核净的质量比可为500∶1～1∶500，优选为100∶1～1∶100，可进一步优选为50∶1～1∶50，最好为10∶1～1∶10。
所述枯草芽孢杆菌Pc3中活孢子数可为0.1～1000亿/g，优选为0.5～500亿/g，进一步优选为1～250亿/g，最好为1～100亿/g。
在枯草芽孢杆菌Pc3和菌核净的组合物中加入农药助剂后即可配得可湿性粉剂、悬乳剂或水乳剂。如向活性成分枯草芽孢杆菌Pc3和菌核净中加入农药助剂并调节pH值为5～7后，配成可湿性粉剂。菌核净可采用原药，枯草芽孢杆菌采用原粉。农药助剂可选自湿润剂、分散剂、填料、粘结剂、崩解剂等中的至少一种。
所述含枯草芽孢杆菌和菌核净的组合物可在制备防治油菜菌核病的农药中应用。
本发明的组合物降低了油菜菌核病病原菌对菌核净的潜在抗药性风险，将枯草芽孢杆菌Pc3与菌核净组合后，二者之间具有明显的增效作用，同时大大减少了化学农药使用量，从而使该组合物可以有效应用于油菜菌核病的防治，尤其在制备防治油菜菌核病的农药中应用。
具体实施方式
实施例1：枯草芽孢杆菌Pc3菌株芽孢制剂的制备
制备过程：首先将Pc3接入营养琼脂培养基中振荡培养(200r/min,28℃)48h，80℃水浴处理15min，8000r/min离心15min，去上清液获得细菌沉淀，加入10mL过滤除菌的MgSO4溶液(0.lmol/L)和1mL无菌甘油，振荡混匀后再加入lmL浓度为1.5％(w/v)海藻酸钠溶液，然后将上述混合液与50g滑石粉及4g Ca-lignosulphate充分混合，无菌条件下室温保存。经稀释涂板法测定统计，枯草芽孢杆菌Pc3芽孢制剂活芽孢数为1010cfu/g。
实施例2： 20％可湿性粉剂的配制(各组分为质量份)
实施例3：20％可湿性粉剂对盆栽油菜菌核病的防效(％)
将油菜菌核病菌接种到PDA平板的中央，置于25℃培养箱中培养3d，备用。
将枯草芽孢杆菌Pc3(100亿活孢子/克)与菌核净1∶1(质量比)的比例配成20％可湿性粉剂，单独将枯草芽孢杆菌Pc3和菌核净配成上述可湿性粉剂作对比，喷雾，于24h后将准备好的油菜菌核病菌打成直径为0.5cm的菌碟接种油菜叶片的中部，每株接4张叶片，接10株油菜，菌丝块上加脱脂棉保湿，接种5d后调查发病指数，统计防治效果，20％枯草芽孢杆菌Pc3·菌核净可湿性粉剂对盆栽油菜菌核病的防效结果见表1。
表1
盆栽试验结果(表1)表明：20％枯草芽孢杆菌Pc3·菌核净可湿性粉剂对盆栽油菜菌核病的增效比率为28.84％，该复配组合表现出对抑制油菜菌核病菌的生长有增效作用。
实施例4： 20％可湿性粉剂对油菜菌核病田间小区实验的防效(％)
实验油菜品种：浙油18
将枯草芽孢杆菌Pc3(100亿活孢子/克)与菌核净按1∶1(质量比)的比例配成20％可湿性粉剂，单独将枯草芽孢杆菌Pc3和菌核净配成上述可湿性粉剂作对比，于油菜盛花期喷雾，7d后进行第二次防治，用于防治油菜菌核病菌，单独将枯草芽孢杆菌Pc3和菌核净配成上述可湿性粉剂作对比，二次施药后15d调查各个小区发病指数，计算防治效果。20％可湿性粉剂对油菜菌核病田间小区实验的防效结果见表2。
表2
田间小区试验结果(表2)表明：20％枯草芽孢杆菌Pc3·菌核净可湿性粉剂对油菜菌核病的防治增效比率为23.57％，该复配组合表现出对抑制油菜菌核病病原菌的生长有明显的增效作用，与盆栽实验结果一致。
Claims (9)
1.含枯草芽孢杆菌和菌核净的组合物，其特征在于以枯草芽孢杆菌Pc3和菌核净为活性成分，枯草芽孢杆菌Pc3与菌核净的质量比为500∶1～1∶500，所述枯草芽孢杆菌Pc3中活孢子数为0.1～1000亿/g。
2.如权利要求1所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述枯草芽孢杆菌Pc3与菌核净的质量比为100∶1～1∶100，所述枯草芽孢杆菌Pc3中活孢子数为0.5～500亿/g。
3.如权利要求2所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述枯草芽孢杆菌Pc3与菌核净的质量比为50∶1～1∶50，所述枯草芽孢杆菌Pc3中活孢子数为1～250亿/g。
4.如权利要求3所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述枯草芽孢杆菌Pc3与菌核净的质量比为10∶1～1∶10，所述枯草芽孢杆菌Pc3中活孢子数为1～100亿/g。
5.如权利要求1所述含枯草芽孢杆菌和菌核净的组合物，其特征在于在枯草芽孢杆菌Pc3和菌核净的组合物中加入农药助剂，配得可湿性粉剂、悬乳剂或水乳剂。
6.如权利要求5所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述在枯草芽孢杆菌Pc3和菌核净的组合物中加入农药助剂并调节pH值为5～7。
7.如权利要求5所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述菌核净采用原药，枯草芽孢杆菌采用原粉。
8.如权利要求5所述含枯草芽孢杆菌和菌核净的组合物，其特征在于所述农药助剂选自湿润剂、分散剂、填料、粘结剂、崩解剂中的至少一种。
9.如权利要求1所述含枯草芽孢杆菌和菌核净的组合物在制备防治油菜菌核病的农药中应用。
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