CN106135287A - 含二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂和生防菌的菌药组合物、制剂及应用 - Google Patents
含二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂和生防菌的菌药组合物、制剂及应用 Download PDFInfo
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- CN106135287A CN106135287A CN201610493201.9A CN201610493201A CN106135287A CN 106135287 A CN106135287 A CN 106135287A CN 201610493201 A CN201610493201 A CN 201610493201A CN 106135287 A CN106135287 A CN 106135287A
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- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A—HUMAN NECESSITIES
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
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- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/08—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having one or more single bonds to nitrogen atoms
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- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/08—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having one or more single bonds to nitrogen atoms
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Landscapes
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Abstract
本发明公开了一种含二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂和生防菌的菌药组合物、制剂及应用，属于植物保护和农药学领域。该菌药组合物由二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂和生防菌菌粉组成，以质量比计，二硝基苯胺类杀菌剂:二硫代氨基甲酸酯类杀菌剂:生防菌菌粉＝1～10:1～10:1～10，菌粉中活菌数不低于106cfu/g。生防菌优选为解淀粉芽孢杆菌ZY‑9‑13，保藏编号：CGMCC No.6162。二硝基苯胺类的氟啶胺可使病原菌呼吸作用中的氧化磷酸化解偶联，阻碍能量合成，抑制病原菌的菌丝生长、孢子形成与萌发，二硫代氨基甲酸酯类杀菌剂为多作用位点抑制剂，这两类杀菌剂与解淀粉芽孢杆菌配合使用对烟草根结线虫病、烟草黑胫病具有优异的防治效果。
Description
技术领域
本发明涉及一种含二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂和生防菌的菌药组合物，同时还涉及包含该菌药组合物的制剂及应用，属于植物保护和农药学领域。
背景技术
烟草黑胫病(Tobacco black shank)是一种世界范围的毁灭性土传病害，其病原菌为疫霉菌烟草变种[Phytophthora parasitica var nicotianae(Breda de Hann)Tuker]。烟草疫霉在烟株苗期至成株期均可侵染，主要侵染烟草茎秆，也可侵染叶片。烟草黑胫病在茎基形成水渍状黑斑，皮层组织变黑凹陷，并沿茎部上下扩展，髓部黑褐色坏死并干缩呈碟片状(万维肖,何孝兵,曲平治,等.烟草黑胫病早期预防技术[J].植物医生,2014(02):50-50)。叶片染病后初为水浸状暗绿色小斑，后扩大为中央黄褐色坏死，边缘有轮纹的膏药状黑斑，全株萎蔫(孙计平,李雪君,吴照辉,等.烟草黑胫病的研究进展[J].湖北农业科学,2011,50(16):3253-3256)。烟草疫霉以菌丝体与厚壁孢子随病株残体在土壤或堆肥中越冬，成为次年的初侵染源。病株上产生的孢子囊或释放出来的游动孢子通过地面流水或随风雨进行再侵染(王万能,肖崇刚.烟草黑胫病的综合防治及其研究进展[J].南方农业学报,2003(2):42-43)。随着20世纪80年代后期我国烟草种植面积的不断扩大，烟草黑胫病迅速扩展。烟草黑胫病平均发病率为10％～20％，发病率高的田块达75％，严重的甚至造成绝收(陈瑞泰,朱贤朝,王智发,等.全国16个主产烟省(区)烟草侵染性病害调研报告[J].中国烟草科学,1997(04):1-7)。
化学防治是烟草黑胫病防控的重要措施，其中甲霜灵、氟啶胺、代森锰锌等农药是生产中使用较为广泛的药剂。甲霜灵是苯基酰胺类内吸性杀菌剂，80年代中期在我国生产使用，在卵菌病害防治中发挥着重要作用，其作用机制为抑制病原菌的聚合酶活性，干扰RNA合成。氟啶胺(SHIRLAN，IKF-1216)是由日本石原产业公司开发的2,6-二硝基苯胺类广谱杀菌剂，通过抑制孢子形成与萌发、菌丝生长等抵制感染，耐雨水冲刷、持效期长，对灰葡萄孢引起的多种灰霉病有特效，同苯并咪唑类、二羧酰亚胺类及目前市场上已有的杀菌剂无交互抗性，对交链孢属、疫霉属、单轴霉属、核盘菌属和黑星菌属等病菌引起的病害亦有良好的活性。但是随着生产中的普遍应用，化学杀菌剂的抗性问题也随之发生。王万能等研究表明，化学药剂处理后烟草黑胫病的菌丝体能引起交配型的变化，进而产生抗药性突变菌株，甚至某些菌株的抗药性可以稳定遗传，其中以甲霜灵为代表苯基酰胺类杀菌剂最为突出，而且抗性水平很高(王万能,肖崇刚.烟草黑胫病的综合防治及其研究进展[J].广西农业科学,2003,(2):42-44)。而化学药剂复配可以减少一种杀菌剂成分的用量，延缓抗药性的发生，同时降低生产成本，是植物病害防治中的重要措施之一。
烟草黑胫病的化学防治是病害综合防治中的重要措施，目前其复配药剂也有相关报道和专利。如公布号CN105265448A的发明专利公开的一种用于防治烟草根腐、黑胫病的杀菌剂，主要由甲霜灵和恶霉灵组成，辅以甲醇、乳化剂后兑水使用即可；公布号CN104430438A的发明专利公开的一种杀菌组合物，其有效成分为恶唑菌酮和霜脲氰，二者的重量比为80:1～1:80，适用于防治由病原真菌引起的植物病害，尤其是烟草黑胫病及瓜果、蔬菜等作物上的霜霉病、疫病等；公布号CN103478163A的发明专利公开了一种含苯噻菌胺的杀菌农药组合物，除苯噻菌胺外还包括乙磷铝，二者的重量比为1:40～60:1，该组合物可用于防治烟草黑胫病、作物疫病及霜霉病等。
生防菌株是天然存在的生物活体，具有良好的环境相容性。利用生防菌株防治烟草黑胫病具有防效高、安全性好、成本低等优点。公告号CN101220335B的发明专利公开的一种防治烟草黑胫病的菌株，名称为淡紫拟青霉ZY-19-2，保藏编号CCTCC NO:M207196，该菌株对烟草黑胫病菌有较强的抑制作用，可定殖在烟草植株根际抵抗病菌的浸染；公告号CN103614319B的发明专利也公开了一种用于防治烟草黑胫病的阿氏芽孢杆菌J-6，保藏编号CCTCC NO:M2013475，该菌株能诱导烟叶拮抗黑胫病从而发挥疫霉防治效果；公告号CN102925393B的发明专利公开的一种兼抗根结线虫和烟草疫霉的烟草内生菌，名称为解淀粉芽孢杆菌ZY-9-13，保藏编号CGMCC NO:6162，该菌株对烟草根结线虫和黑胫病均具有防治效果，可与烟草育苗基质混合使用或在烟草植株移栽时施入根际周围，生防效果显著。但是，上述专利均未涉及杀菌剂与生防菌剂联用的内容。
可持续发展的农业生产系统要求减少农药的投入，而生物、化学协同防治病害可减轻对环境的压力，延缓对化学药剂产生抗性，使药效更稳定和持久(尹敬芳.生物-化学协同防治辣椒疫病菌药合剂初步研究[D].中国农业大学,2006)。已有研究表明，生防菌剂和化学药剂复配对多种病原菌均有良好的防治效果。如黄蓝状菌与甲霜锰锌复配可防治小麦赤霉病(毛雪琴,彭志荣,邱海萍,等.生防菌株MT-06发酵条件及复配杀菌剂对小麦赤霉病的防效[J].浙江农业科学,2013(07):821-824)，枯草芽胞杆菌与戊唑醇复配可防治蚕豆枯萎病(陈志谊,任海英,刘永锋,等.戊唑醇和枯草芽孢杆菌协同作用防治蚕豆枯萎病及增效机理初探[J].农药学学报,2002(4):40-44)，枯草芽胞杆菌与苯醚甲环唑、嘧菌酯等复配可防治苹果轮纹病(周寒.杀菌剂和生防菌协调控制苹果采后轮纹病及作用机制[D].安徽农业大学,2009)，井冈霉素与枯草芽胞杆菌协同防治水稻纹枯病(陈志谊,刘永锋,陆凡.井冈霉素和生防菌Bs-916协同控病作用及增效机理[J].植物保护学报,2003,30(4):429-434.)等等，但是以上菌药复配的防治靶标均不涉及烟草疫霉。公布号CN104381303A的发明专利公开了一种用于防治真菌病害的菌药组合物，由核黄素与枯草芽孢杆菌Tpb55菌粉按照质量比1.21～2.34:97.96～98.79复配而成，相较单独施用枯草芽孢杆菌Tpb55菌粉，该菌药组合物能显著提高烟草黑胫病、赤星病等病害防治功效，增强植株抗病性，但是也仅达到与化学药剂相当的防治效果。
发明内容
本发明的目的是提供一种含二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂和生防菌的菌药组合物，该组合物对烟草根结线虫病、烟草黑胫病等具有优异的防治效果，防效显著优于使用单一种类的杀菌剂、生防菌菌剂以及上述任一种类杀菌剂与生防菌菌剂的组合，且可延缓化学抗药性的产生。
同时，本发明还提供一种包含上述菌药组合物的菌药制剂。
最后，本发明再提供一种上述菌药组合物、菌药制剂在防治烟草根结线虫病和/或烟草黑胫病中的应用。
为了实现以上目的，本发明所采用的技术方案是：
菌药组合物，由二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂和生防菌菌粉组成，以质量比计，二硝基苯胺类杀菌剂:二硫代氨基甲酸酯类杀菌剂:生防菌菌粉＝1～10:1～10:1～10，菌粉中活菌数不低于106cfu/g。
所述二硝基苯胺类杀菌剂选自氟啶胺等。
所述二硫代氨基甲酸酯类杀菌剂选自代森锌、代森铵、代森锰、代森联、代森锰锌、甲基代森锌、丙森锌、福美双、福美铁、福美锌等中的一种或多种。如单独选用代森锌、代森锰锌、丙森锌或福美双，或者选用代森锰与丙森锌、甲基代森锌与福美双的组合，亦或者选用代森锌、丙森锌与福美锌的组合。相较使用单一一种二硫代氨基甲酸酯类杀菌剂，杀菌剂复配(两种及以上杀菌剂配合使用)具有相加作用或增效作用。并且，复配杀菌剂与生防菌联合使用对烟草根结线虫病、烟草黑胫病等的防治效果优于单独使用化学杀菌剂或生防菌菌剂。同时，复配杀菌剂中各组分配比可以任意选取，如代森锰锌与丙森锌按照质量比1～10:1～10复配，又如代森锌与甲基代森锌、福美锌按照质量比1～10:1～10:1～10复配，等等。由于组合形式多样，此处不一一列举。
优选的，菌药组合物中二硝基苯胺类杀菌剂为氟啶胺，二硫代氨基甲酸酯类杀菌剂选自代森锌、代森铵、代森锰、代森联、代森锰锌、甲基代森锌、丙森锌、福美双、福美铁、福美锌等中的任意一种。二硫代氨基甲酸酯类杀菌剂中代森锌、代森锰锌、福美双等的杀菌机理相同或相似，因此从中选择任意一种与氟啶胺复配，复配杀菌剂再与生防菌联合使用，不仅能简化配方，同时对烟草根结线虫病、烟草黑胫病等也能达到良好的防治效果。其中，氟啶胺可使病原菌呼吸作用中的氧化磷酸化解偶联，阻碍能量合成，抑制病原菌的菌丝生长、孢子形成与萌发。二硫代氨基甲酸酯类杀菌剂为多作用位点抑制剂，不易产生抗性。
所述生防菌菌粉可参照专利(公告号CN102925393B)中方法制备。其中，生防菌优选为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)ZY-9-13，保藏编号：CGMCC No.6162，保藏时间：2012年05月29日，保藏单位：中国微生物菌种保藏管理委员会普通微生物中心，单位地址：北京市朝阳区北辰西路1号院3号。该菌株兼抗烟草根结线虫和烟草疫霉，尤其能抑制烟草黑胫病菌菌丝生长及孢子萌发，在烟草根际较好的定殖。参见专利“一种兼抗根结线虫和烟草疫霉的烟草内生菌及其应用”(公告号：CN102925393B)。经试验验证，二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂对解淀粉芽孢杆菌(CGMCC No.6162)的生长影响微弱，杀菌剂与菌株之间的相容性良好，可以进行菌药组合。
所述菌药组合物可采用常规方法制备成任意一种适于农业使用的剂型，如可湿性粉剂、悬浮剂、水分散颗粒剂、乳油、水乳剂、微乳剂、水剂、微胶囊剂、可溶性粉剂等。制剂中菌药组合物为有效成分，其含量为制剂总质量的0.1～99.9％。
菌药制剂，包含上述菌药组合物，以及一种或多种农药学上可接受的助剂。所述助剂包括润湿分散剂、稳定剂、防冻剂、崩解剂、增稠剂、消泡剂、有机溶剂、载体等，以及其他有益于有效成分在制剂中稳定发挥药效的物质，是农药制剂中常用的或允许使用的各种成分。助剂的具体选择和用量可根据配方要求通过常规试验确定。
所述润湿分散剂可选自十二烷基硫酸钠、十二烷基磺酸钠、十二烷基苯磺酸钠、十二烷基苯磺酸钙、木质素磺酸钙、木质素磺酸钠、月桂酸硫酸钠、烷基聚氧乙烯醚磺酸盐、烷基酚聚氧乙烯醚甲醛缩合物、聚氧乙烯聚氧丙烯嵌段共聚物、月桂醇聚氧乙烯醚硫酸钠、烷基萘甲醛缩合物磺酸盐、拉开粉、十二烷基聚氧乙烯醚磷酸酯、羟甲基纤维素等中的任意一种或多种。
所述稳定剂可选自邻苯三酚、BHT、BHA、柠檬酸、环氧大豆油、亚麻油、丁香酚、蓖麻油、橄榄油、对苯二酚、水杨酸、山梨酸钠、丁基缩水甘油醚、苯基缩水甘油醚、甲苯基缩水甘油醚、聚乙烯基乙二醇二缩水甘油醚、二萘酚、一萘酚中的任意一种或多种。
所述防冻剂可选自甘油、丙二醇、乙二醇、尿素、氯化钠中的任意一种。
所述崩解剂可选自硫酸铵、氯化钙、氯化镁、氯化铝、膨润土、磷酸二氢胺、尿素等中的任意一种或多种。
所述增稠剂可选自黄原胶、阿拉伯胶、聚乙烯醇、羧甲基纤维素、甲基纤维素、杂多糖、可溶性淀粉、蔗糖、葡萄糖、海藻酸钠、丙烯酸系聚合物、聚乙烯吡咯烷酮、松香、石膏、硫酸钠、骨胶、大豆蛋白、糊精、硅酸、硅酸铝镁等中的任意一种或多种。
所述消泡剂可选自硅油类消泡剂、硅酮类消泡剂、C10～20饱和脂肪酸类消泡剂、C8～10脂肪醇类消泡剂中的任意一种。
所述有机溶剂可选自甲苯、二甲苯、烷基苯、异丙醇、丁醇、乙二醇、二甘醇、三甘醇、丙二醇、丙三醇、山梨醇、苯甲醇、环己醇、丙酮、甲基异丁基酮、环己酮、二甲基甲酰胺、吡咯烷酮、二氯甲烷、氯仿、四氯化碳、植物油等中的任意一种或多种。
所述载体可选自石英、粘土、蒙脱土、二氧化硅、硅藻土、浮石、石膏、滑石、膨润土、高岭土、陶土、白炭黑、方解石、大理石、海泡石、白云石、凹凸棒土等中的任意一种或多种。
菌药组合物的应用，具体为菌药组合物在防治烟草根结线虫和/或烟草黑胫病中的应用。
菌药制剂的应用，具体为菌药制剂在防治烟草根结线虫和/或烟草黑胫病中的应用。
本发明的有益效果：
本发明中菌药组合物由二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，其中二硝基苯胺类的氟啶胺可使病原菌呼吸作用中的氧化磷酸化解偶联，阻碍能量合成，抑制病原菌的菌丝生长、孢子形成与萌发，二硫代氨基甲酸酯类杀菌剂为多作用位点抑制剂，且不易产生抗性，这两类杀菌剂与解淀粉芽孢杆菌配合使用对烟草根结线虫病、烟草黑胫病具有优异的防治效果，防效显著优于使用单一种类的杀菌剂、生防菌菌剂以及上述任一种类杀菌剂与生防菌菌剂的组合，且可延缓化学抗药性的产生，特别适用于田间烟草黑胫病的防治。
具体实施方式
下述实施例仅对本发明作进一步详细说明，但不构成对本发明的任何限制。
实施例1
菌药组合物，由氟啶胺、代森锌和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:代森锌:解淀粉芽孢杆菌菌粉＝1:1:1，菌粉中活菌数不低于106cfu/g。
实施例2
菌药组合物，由氟啶胺、代森锰和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:代森锰:解淀粉芽孢杆菌菌粉＝10:1:1，菌粉中活菌数不低于106cfu/g。
实施例3
菌药组合物，由氟啶胺、代森锰锌和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:代森锰锌:解淀粉芽孢杆菌菌粉＝1:6:9，菌粉中活菌数不低于106cfu/g。
实施例4
菌药组合物，由氟啶胺、甲基代森锌和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:甲基代森锌:解淀粉芽孢杆菌菌粉＝5:5:1，菌粉中活菌数不低于106cfu/g。
实施例5
菌药组合物，由氟啶胺、丙森锌和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:丙森锌:解淀粉芽孢杆菌菌粉＝8:3:1，菌粉中活菌数不低于106cfu/g。
实施例6
菌药组合物，由氟啶胺、福美双和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:福美双:解淀粉芽孢杆菌菌粉＝10:10:1，菌粉中活菌数不低于106cfu/g。
实施例7
菌药组合物，由氟啶胺、福美锌和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:福美锌:解淀粉芽孢杆菌菌粉＝1:1:10，菌粉中活菌数不低于106cfu/g。
实施例8
菌药组合物，由氟啶胺、代森锰锌、福美双和解淀粉芽孢杆菌(CGMCC No.6162)菌粉组成，以质量比计，氟啶胺:代森锰锌:福美双:解淀粉芽孢杆菌菌粉＝4:3:7:9，菌粉中活菌数不低于106cfu/g。
实施例9
菌药制剂，由以下质量份数的原料制成：氟啶胺40份，代森锌10份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉5份(菌粉中活菌数不低于106cfu/g)，十二烷基磺酸钠8份，月桂醇基硫酸钠3份，十二烷基硫酸钠5份，乙基纤维素5份，蔗糖5份，硅藻土12份，凹凸棒土12份；制备方法为：按照质量份数准确取各原料，粉碎后过筛，混合均匀，得到可湿性粉剂。
实施例10
菌药制剂，由以下质量份数的原料制成：氟啶胺11份，代森锰14份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉24份(菌粉中活菌数不低于106cfu/g)，木质素磺酸钙8份，十二烷基硫酸钠3份，蔗糖4份，白炭黑15份，硅藻土18份；制备方法同实施例9。
实施例11
菌药制剂，由以下质量份数的原料制成：氟啶胺30份，甲基代森锌24份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉6份(菌粉中活菌数不低于106cfu/g)，十二烷基磺酸钠10份，月桂醇基硫酸钠8份，木质素磺酸钙5份，乙基纤维素5份，蔗糖5份，高岭土20份；制备方法同实施例9。
实施例12
菌药制剂，由以下质量份数的原料制成：氟啶胺5份，代森锰锌6份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉44份(菌粉中活菌数不低于106cfu/g)，十二烷基萘磺酸钠6份，拉开粉8份，硫酸钠3份，黄原胶5份，蔗糖5份，高岭土22份；制备方法为：按照质量份数准确取各原料，混合均匀后造粒，干燥、筛分，得到悬浮剂。
实施例13
菌药制剂，由以下质量份数的原料制成：氟啶胺8份，丙森锌48份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉8份(菌粉中活菌数不低于106cfu/g)，十二烷基萘磺酸钠12份，拉开粉3份，淀粉3份，硫酸钠2份，黄原胶5份，蔗糖5份，膨润土12份，硅藻土12份；制备方法同实施例12。
实施例14
菌药制剂，由以下质量份数的原料制成：氟啶胺25份，福美锌13份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉27份(菌粉中活菌数不低于106cfu/g)，十二烷基萘磺酸钠12份，拉开粉5份，硫酸钠3份，蔗糖8份，膨润土18份，硅藻土8份；制备方法同实施例12。
实施例15
菌药制剂，由以下质量份数的原料制成：氟啶胺20份，福美双20份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉20份(菌粉中活菌数不低于106cfu/g)，聚乙烯吡咯烷酮5份，十二烷基酚聚氧乙烯醚3份，十二烷基苯磺酸钠3份，环氧乙烷5份，乙基纤维素8份，凹凸棒土22份；制备方法为：按照质量份数准确取各原料，混合均匀后超微气流粉碎，造粒，得到水分散颗粒剂。
实施例16
菌药制剂，由以下质量份数的原料制成：氟啶胺18份，代森锰锌18份，福美双12份，解淀粉芽孢杆菌(CGMCC No.6162)菌粉6份(菌粉中活菌数不低于106cfu/g)，木质素磺酸钠8份，二丁基萘磺酸钠3份，十二烷基苯磺酸钠4份，聚乙烯吡咯烷酮5份，淀粉3份，蔗糖3份，硅藻土18份；制备方法同实施例15。
在本发明的其他实施例中，还可将菌药制剂制成水乳剂、水剂、微胶囊剂等剂型，助剂采用本领域常规助剂即可。
在本发明的其他实施例中，还可从二硫代氨基甲酸酯类杀菌剂中任选多种与氟啶胺复配，复配杀菌剂再与生防菌组合或制成菌药制剂。
对比例1
菌药制剂，由以下质量份数的原料制成：氟啶胺50份，解淀粉芽孢杆菌(CGMCCNo.6162)菌粉5份(菌粉中活菌数不低于106cfu/g)，其他助剂及制备方法同实施例9。
对比例2
菌药制剂，由以下质量份数的原料制成：代森锌50份，解淀粉芽孢杆菌(CGMCCNo.6162)菌粉5份(菌粉中活菌数不低于106cfu/g)，其他助剂及制备方法同实施例9。
上述各实施例及对比例中的解淀粉芽孢杆菌(CGMCC No.6162)菌粉参照专利(公告号CN102925393B)具体实施方式中方法制备成可湿性粉剂。
1、菌药组合物的协同增效作用测定
采用菌丝生长速率法测定烟草黑胫病菌对杀菌剂、解淀粉芽孢杆菌和菌药组合物的药剂敏感性。表1中杀菌剂原药(未加解淀粉芽孢杆菌)的供试浓度为3.50μg/mL，表2中二硝基苯胺类杀菌剂原药(未加解淀粉芽孢杆菌)的供试浓度为1.00μg/mL，表3中二硫代氨基甲酸酯类杀菌剂原药(未加解淀粉芽孢杆菌)的供试浓度为10.00μg/mL；解淀粉芽孢杆菌(CGMCC No.6162)菌悬液的浓度为1010cfu/mL，菌药组合物(各类杀菌剂原药对应浓度+解淀粉芽孢杆菌(CGMCC No.6162)1010cfu/mL)，复配形式具体见下表1～3。将直径5mm菌碟置于含药的燕麦培养基中央，每药剂处理重复3次，以无药处理为对照，采用十字交叉法测量菌落直径，计算不同处理对烟草黑胫病菌的抑制率，评价菌药组合物对黑胫病菌的抑制作用，结果见下表1～3。
采用Abbott数学模型计算菌药组合物对病原菌的抑制效果(Colby S,Calculating synergistic and antagonistic responses of herbicide combinations[J].Weeds,1967)。
药剂对烟草黑胫病菌的理论抑制率计算公式如下：
E＝(X1+X2+…)-X1·X2·…/100；
式中：E代表药剂对烟草黑胫病菌的预期(理论)抑制率，X1、X2…分别代表药剂A、药剂B…对烟草黑胫病菌的抑制率。
评价方法为：期望(理论)抑制率与实际测定抑制率相比较，当理论值＜实际值时，表现为增效作用；当理论值＝实际值时，表现为相加作用；当理论值＞实际值时，表现为拮抗作用。
表1二硝基苯胺类、二硫代氨基甲酸酯类杀菌剂与解淀粉芽孢杆菌对菌丝生长的抑制效果
表2二硝基苯胺类杀菌剂与解淀粉芽孢杆菌对烟草黑胫病菌菌丝生长的抑制效果
表3二硫代氨基甲酸酯类杀菌剂与解淀粉芽孢杆菌对烟草黑胫病菌菌丝生长的抑制效果
从表1～3可以看出，含二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂和解淀粉芽孢杆菌(CGMCC No.6162)的菌药组合物对烟草黑胫病菌的抑制效果显著优于使用单一种类的杀菌剂、解淀粉芽孢杆菌以及上述任一种类杀菌剂与解淀粉芽孢杆菌的组合。表明本发明中菌药组合物各组分之间具有协同增效作用。
2、菌药制剂对烟草黑胫病菌的温室药效试验
通过温室接种烟草黑胫病菌，并喷施实施例9～16及对比例1～2制备的菌药制剂，具体方法如下：参照董瑜的方法制备菌谷接种物，将20个直径5mm的菌饼加入500g灭菌的谷子中，28℃培养14d，混合均匀后备用(董瑜.防治烟草黑胫病缓释型杀菌剂研究[D].中国农业科学院,2015)；将中烟100烟苗种植在10×10cm的营养钵中，营养土与蛭石的比例为1:1，待烟苗长至6～7片真叶时，进行药剂灌根处理，每个营养钵加药液50mL，每个处理5株烟苗，以不施加药剂、加50mL清水的处理作为对照。试验设三次重复，施药3d后于烟株根部接种菌谷10g/株，施药7d后进行病害调查，按照以下公式计算防治效果：
防治效果(％)＝(对照病情指数-处理病情指数)/对照病情指数×100％。
烟草黑胫病分级标准参照中华人民共和国烟草行业标准(YC/T39-1996)，试验结果见下表4。表4中，72％甲霜·锰锌可湿性粉剂购自浙江禾本科技有限公司，500g/L氟啶胺悬浮剂购自日本石原产业株式会社，50％福美双可湿性粉剂购自山西科力科技有限公司，解淀粉芽孢杆菌(CGMCC No.6162)菌剂参照专利(公告号CN102925393B)具体实施方式中方法制备，以上均为对照组。
表4菌药制剂对烟草黑胫病的防治效果
从表4可以看出，实施例9～16中菌药制剂对烟草黑胫病均具有优异的防治效果，防效达81％以上，优于市售的化学药剂、解淀粉芽孢杆菌菌剂及对比例1～2中菌药制剂，为黑胫病的防治提供了新型备选药剂，通过与市售药剂轮换使用，可降低抗药性的产生。
Claims (8)
1.菌药组合物，其特征在于：由二硝基苯胺类杀菌剂、二硫代氨基甲酸酯类杀菌剂和生防菌菌粉组成，以质量比计，二硝基苯胺类杀菌剂:二硫代氨基甲酸酯类杀菌剂:生防菌菌粉＝1～10:1～10:1～10，菌粉中活菌数不低于106cfu/g。
2.根据权利要求1所述的菌药组合物，其特征在于：所述二硝基苯胺类杀菌剂为氟啶胺。
3.根据权利要求1所述的菌药组合物，其特征在于：所述二硫代氨基甲酸酯类杀菌剂选自代森锌、代森铵、代森锰、代森联、代森锰锌、甲基代森锌、丙森锌、福美双、福美铁、福美锌中的一种或多种。
4.根据权利要求1～3中任一项所述的菌药组合物，其特征在于：所述生防菌为解淀粉芽孢杆菌ZY-9-13，保藏编号：CGMCC No.6162。
5.包含如权利要求1～4中任一项所述菌药组合物的菌药制剂。
6.根据权利要求5所述的菌药制剂，其特征在于：还包含一种或多种农药学上可接受的助剂。
7.如权利要求1～4中任一项所述菌药组合物在防治烟草根结线虫和/或烟草黑胫病中的应用。
8.如权利要求5或6所述的菌药制剂在防治烟草根结线虫和/或烟草黑胫病中的应用。
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