CN114946886A - 一种哈茨木霉可分散油悬剂及其应用 - Google Patents
一种哈茨木霉可分散油悬剂及其应用 Download PDFInfo
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- -1 sodium alkyl benzene Chemical class 0.000 claims abstract description 8
- 235000012424 soybean oil Nutrition 0.000 claims abstract description 8
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/38—Trichoderma
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
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Abstract
本发明公开了一种哈茨木霉可分散油悬剂及其应用,涉及生物农药领域,按照重量百分比包含:哈茨木霉孢子粉2%、大豆油20%、凡士林15%、烷基苯磺酸钠20%、有机膨润土8%、木质素磺酸盐35%。本发明将哈茨木霉孢子分散到色拉油中依靠表面活性剂形成的稳定、分散、悬浮液体,有利于增强孢子制剂在田间自然条件下的定殖能力,提高防治效果,延长储存稳定性及货架期。
Description
技术领域
本发明涉及生物农药领域,尤其涉及一一种哈茨木霉可分散油悬剂及其应用。
背景技术
哈茨木霉是木霉菌的一种,属半知菌亚门,丝孢纲,丛梗抱目,丛梗抱科,是一种具有重要的生防价值的真菌,主要通过竞争作用、抗生作用、重寄生作用、诱导抗性等作用机制来预防由腐霉菌、立枯丝核菌、镰刀菌、核盘菌等病原菌引起的多种植物病害,具有保护和治疗双重作用。同时能够有效改善作物根系生长环境,促进弱苗根系发达和老化根系复苏,提高作物光合作用效率,增强作物长势和抗病抗逆能力,对于多种因素导致的作物生长不良,也有良好的预防和改善作用。
哈茨木霉菌剂主要为孢子制剂,制剂的商品化进展十分缓慢,可湿性粉剂、颗粒剂是常用的剂型,目前市面上的产品存在诸多问题。可湿性粉剂、颗粒剂是孢子、载体、润湿剂、分散剂、稳定剂等一系列助剂,按比例经充分混合粉碎后,达到一定粉粒细度的固体剂型。可湿性粉剂悬浮率偏低,容易沉淀成团使得喷洒过程中不均匀而影响药效,加工、应用中也存在严重的卫生和安全问题。颗粒剂生产效率低,载体用量大,有效含量低,药效低,使用时相对不安全。孢子制剂应用时易受温度、湿度等外界因素的影响可湿性粉剂、颗粒剂中添加的助剂与哈茨木霉孢子长时间接触也会影响其活力,使得孢子制剂在田间自然条件下定殖能力差、防治效果不稳定,储存稳定性差、货架期短。
可分散油悬剂是孢子分散在油类中依靠表面活性剂形成高分散稳定的悬浮液体制剂。可分散油悬浮剂粒径小,悬浮率高、分散性好、黏附性好,对靶标具有良好的亲和性,便于机械化施用等优势。在植物表面的铺展和粘着力都比较强,耐雨水冲刷,增效作用明显,持效期比较长,对人畜低毒。孢子表面的油膜起到保护膜的作用,并维持孢子低的含水量,减少真菌孢子与环境中氧气的接触,油膜中含有的抗氧化剂进一步降低孢子与氧的作用,使真菌孢子代谢处于低的水平,有效延长了孢子贮存时间、降低贮存成本。油孢子与稀释液混合加水后,油包裹真菌孢子均匀分散在水中,可直接提供孢子萌发所需要的水分,增加油孢子的抗低湿能力。同时在实际田间应用中混剂中的油还可以破坏寄主表皮脂类结构,增加表皮水分蒸发,提高真菌孢子周围环境湿度,为孢子萌发持续提供水分,从而降低孢子对环境湿度的依赖性。可分散油悬浮剂成功克服了真菌孢子难以在干燥环境中使用、杀虫效果受环境中湿度影响的缺陷。相对湿度很低的环境中利于孢子萌发,高温环境下又能延长孢子的寿命,还有利于孢子对疏水基质的吸附如昆虫体壁或植物表面,具有巨大的应用潜力。
发明内容
本发明的一种哈茨木霉菌菌株,所述的菌株为哈茨木霉菌;保藏名称为哈茨木霉菌Trichoderma harzianum,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址:北京市朝阳区北辰西路1号院3号,保藏编号CGMCC No.21471,保藏时间2021年3月17日
本发明针对现有技术的不足,本发明提供了一种哈茨木霉可分散油悬剂。所述的哈茨木霉可分散油悬剂,将哈茨木霉孢子分散到色拉油中依靠表面活性剂形成的稳定、分散、悬浮液体。
为实现上述目的,本发明提供了一种哈茨木霉可分散油悬剂,按照重量百分比包含:哈茨木霉孢子粉2%、大豆油20%、凡士林15%、烷基苯磺酸钠20%、有机膨润土8%、木质素磺酸盐35%。
进一步地,所述哈茨木霉孢子粉的制备方法,具体过程如下:
步骤一:摇瓶种子制备:将保存于4℃冰箱的斜面菌种,接种于液体摇瓶培养基,30℃、200rpm,恒温震荡培养48h;
摇瓶种子培养基:葡萄糖 2%、酵母粉 0.5%、豆饼粉 1.5%、磷酸二氢钾 0.3%、MgSO4·7H2O 0.2%
步骤二:将培养好的摇瓶种子培养基,接种于一级种子发酵罐,28-30℃、200rpm通风搅拌培养24h,通风比1vvm;
培养基 葡萄糖2%、酵母粉 0.5%、豆饼粉 1.5%、磷酸二氢钾 0.3%、MgSO4·7H2O0.2%
步骤三:固体发酵:将培养好的一级种子罐发酵液,接种于固体发酵培养基,25-30℃,培养5-7天;
固体发酵培养基:麸皮42%、稻壳57%、硫酸铵0.5%、磷酸氢二钾0.3%、磷酸二氢钾0.2%
步骤四:烘干:将发酵好的固体物料,在55-60℃,烘干使物料水分降低的10%以下;
步骤五:筛分:将烘干好的哈茨木霉固体物料,用200目振动筛进行孢子和固体颗粒的分离,得到哈茨木霉孢子粉。
进一步地,所述油悬剂加工过程,在环境温度不高于30℃的条件下,分别将油悬剂的实施例各物质加入搅拌机中,搅拌0.5-1小时,混合均匀即得到哈茨木霉分散油悬剂。
与现有技术相比,本发明的有益效果为:有利于增强孢子制剂在田间自然条件下的定殖能力,提高防治效果,延长储存稳定性及货架期。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明的一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1:
一种哈茨木霉可分散油悬剂,按照重量百分比包含:哈茨木霉孢子粉2%、大豆油20%、凡士林15%、烷基苯磺酸钠20%、有机膨润土8%、木质素磺酸盐35%。
实施例2:
一种哈茨木霉可分散油悬剂,按照重量百分比包含:哈茨木霉孢子粉10%、大豆油76%、烷基苯醚磺酸盐5%、十二烷基苯磺酸钠5%、烷基酚聚醚硫酸盐3%、有机膨润土1%。
实施例3:
一种哈茨木霉可分散油悬剂,按照重量百分比包含:哈茨木霉孢子粉10%、大豆油40%、黄原胶15%、农乳0201B # 25%、MorwetD-425 10%。
一、哈茨木霉孢子粉的制备
哈茨木霉菌依次经过摇瓶菌种、一级种子液、固体发酵、烘干、筛分,得到哈茨木霉菌孢子粉,具体过程如下:
1、摇瓶种子制备:将保存于4℃冰箱的斜面菌种,接种于液体摇瓶培养基,30℃、200rpm,恒温震荡培养48h;
摇瓶种子培养基:葡萄糖:2% 酵母粉:0.5% 豆饼粉:1.5%,磷酸二氢钾:0.3%
MgSO4.7H2O:0.2%
2、将培养好的摇瓶种子培养基,接种于一级种子发酵罐,28-30℃、200rpm通风搅拌培养24h,通风比1vvm;
培养基:葡萄糖:2% 酵母粉:0.5% 豆饼粉:1.5%,磷酸二氢钾:0.3% MgSO4·7H2O:0.2%
3、固体发酵:将培养好的一级种子罐发酵液,接种于固体发酵培养基,25-30℃,培养5-7天;
固体发酵培养基:麸皮42%、稻壳57%、硫酸铵0.5%、磷酸氢二钾0.3%、磷酸二氢钾0.2%
4、烘干:将发酵好的固体物料,在55-60℃,烘干24h,使物料水分降低的10%以下;
5、筛分:将烘干好的哈茨木霉固体物料,用200目振动筛进行孢子和固体颗粒的分离,得到哈茨木霉孢子粉;
二、油悬剂加工过程
在环境温度不高于30℃的条件下,分别将油悬剂三个实施例各物质加入搅拌机中,搅拌0.5-1小时,混合均匀即得到哈茨木霉分散油悬剂。
三、萌发率测定
将哈茨木霉孢子油悬剂制成 1×108个孢子/ml 悬浮液分装于PCR管中,常温存放1个月、3个月、6个月、9个月、12个月后检测哈茨木霉孢子的活性,考察孢子的储存稳定性,采用平板计数法,记录孢子萌发率。每一处理设三个重复。以低温(4℃)密封保存的干孢子萌发率为对照。
26℃保存条件下的油悬剂中孢子萌发情况
同列数据后标小写字母表示其差异达显著水平(P<0.05),字母相同差异不显著,字母不同差异显著。
常温存放 1个月的萌发率都在 90%以上,存放时间越长萌发率越低,存放 12 个月实施例3孢子的萌发率81.65%,高于低温密封保存干孢子的萌发率。
四、分散性测定
将1×1010个孢子/ml的哈茨木霉油悬浮剂分别装入 250ml 的烧杯内,在 26℃条件下,搅拌,制成悬浮液,测量分层高度,通过计算分层的比例来评价分散效果。哈茨木霉实施例3油悬剂悬浮率最高。
五、哈茨木霉油悬剂对黄瓜灰霉病的防治效果盆栽试验
孢子悬浮液的制备:将黄瓜灰霉菌在 PDA 培养基上 28℃ 培养一周,待病原真菌长出大量孢子后,用含有机硅的去离子水从培养基上洗脱、用血球计数板计数将孢子浓度调整为106个/ml备用。
黄瓜灰霉病盆栽试验:将孢子悬浮液喷施于培养好的黄瓜幼苗叶片表面,24h后将三个实施例菌悬液50倍稀释液匀喷施于黄瓜叶面至全部润湿,每个处理 3 盆,4 次重复,用清水处理作对照。喷雾处理后 7 天,分级调查各处理发病情况,每处理至少调查10 片叶,计算各处理的病情指数和防治效果。
哈茨木霉油悬剂对黄瓜灰霉病菌的盆栽药效试验
病情指数=【(各病级病叶数×代表级值)/(总叶数×发病最高级的代表级值)】×100
防治效果=【(对照病情指数-处理病情指数)/对照病情指数】×100
病情指数分级标准为:0 级:无病斑;1 级:病斑占整个叶面积的 5%以下;3 级:病斑占整个叶面积的 5%~15%;5 级:病斑占整个叶面积的 15%~25%;7 级:病斑占整个叶面积的 25%~50%;9 级:病斑占整个叶面积的 50%以上。
油悬剂实施例3对黄瓜灰霉病的防治效果最好。
哈茨木霉油悬剂对辣椒疫霉的防治效果病盆栽试验:
孢子悬浮液的制备:将辣椒疫霉病菌在 PDA 培养基上 28℃ 培养一周,待病原真菌长出大量孢子后,用含有机硅的去离子水从培养基上洗脱、用血球计数板计数将孢子浓度调整为106个/ml备用。
辣椒疫霉病盆栽试验:将培养好的辣椒幼苗先接种10 mL的106个/mL辣椒疫霉孢子悬浮液,1d后将三个实施例菌悬液50倍稀释液匀喷施于辣椒幼苗,每个处理 3 盆,4 次重复,用清水处理作对照,30 d分级调查各处理发病情况,每处理15株幼苗,计算各处理的病情指数和防治效果。
以上仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (6)
1.一种哈茨木霉可分散油悬剂,其特征在于,按照重量百分比包含:哈茨木霉孢子粉2%、大豆油20%、凡士林15%、烷基苯磺酸钠20%、有机膨润土8%、木质素磺酸盐35%。
2.一种哈茨木霉可分散油悬剂,其特征在于,按照重量百分比包含:哈茨木霉孢子粉10%、大豆油76%、烷基苯醚磺酸盐5%、十二烷基苯磺酸钠5%、烷基酚聚醚硫酸盐3%、有机膨润土1%。
3.一种哈茨木霉可分散油悬剂,其特征在于,按照重量百分比包含:哈茨木霉孢子粉10%、大豆油40%、黄原胶15%、农乳0201B # 25%、MorwetD-425 10%。
4.根据权利要求1-3中任意一条所述的一种哈茨木霉可分散油悬剂,其特征在于,所述哈茨木霉孢子粉的制备方法,具体过程如下:
步骤一:摇瓶种子制备:将保存于4℃冰箱的斜面菌种,接种于液体摇瓶培养基,30℃、200rpm,恒温震荡培养48h;
摇瓶种子培养基:葡萄糖 2%、酵母粉 0.5%、豆饼粉 1.5%、磷酸二氢钾 0.3%、 MgSO4·7H2O 0.2%;
步骤二:将培养好的摇瓶种子培养基,接种于一级种子发酵罐,28-30℃、200rpm通风搅拌培养24h,通风比1vvm;
培养基 葡萄糖2%、酵母粉 0.5%、豆饼粉 1.5%、磷酸二氢钾 0.3%、MgSO4·7H2O 0.2%;
步骤三:固体发酵:将培养好的一级种子罐发酵液,接种于固体发酵培养基,25-30℃,培养5-7天;
固体发酵培养基:麸皮42%、稻壳57%、硫酸铵0.5%、磷酸氢二钾0.3%、磷酸二氢钾0.2%;
步骤四:烘干:将发酵好的固体物料,在55-60℃,烘干使物料水分降低的10%以下;
步骤五:筛分:将烘干好的哈茨木霉固体物料,用200目振动筛进行孢子和固体颗粒的分离,得到哈茨木霉孢子粉。
5.根据权利要求1-3中任意一条所述的一种哈茨木霉可分散油悬剂,其特征在于,所述油悬剂加工过程,在环境温度不高于30℃的条件下,分别将油悬剂的配方各物质加入搅拌机中,搅拌0.5-1小时,混合均匀即得到哈茨木霉分散油悬剂。
6.一种如权利要求1-5中任意一条所述的哈茨木霉可分散油悬剂在黄瓜灰霉病或辣椒疫霉病菌中的应用。
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