CN114736828A - 一种生防菌株及其在植物病害防治中应用 - Google Patents
一种生防菌株及其在植物病害防治中应用 Download PDFInfo
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- CN114736828A CN114736828A CN202210447456.7A CN202210447456A CN114736828A CN 114736828 A CN114736828 A CN 114736828A CN 202210447456 A CN202210447456 A CN 202210447456A CN 114736828 A CN114736828 A CN 114736828A
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Abstract
本发明提供了一种生防菌ZWP15菌株,该菌株于2021年9月1日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.23331,对丁香假单胞杆菌猕猴桃致病变种(Pseudomonas syringae pv.actinidiae)具有拮抗作用。ZWP15菌株为猕猴桃细菌性溃疡病生物防治提供重要的材料积累,可开发成生物菌剂以代替常规铜制剂,用于防治猕猴桃细菌性溃疡病。
Description
技术领域
本发明属于生物资源利用技术领域,涉及一种微生物菌株及其生物活性,具体涉及一种对猕猴桃溃疡病具有拮抗作用的生防菌株。
背景技术
丁香假单胞猕猴桃致病变种(Pseudomonas syringaepv. actinidiae,Psa)引起的猕猴桃细菌性溃疡病(Kiwifruit bacterial canker)是一种严重威胁猕猴桃健康生长和其产业发展的毁灭性病害。由于该病可在短期内爆发造成大面积树体死亡,且具有适生范围广、致病性强、流行规律复杂、危害部位多等特点。多年来,我国对于该病害的防治主要以化学防治为主,但是长期盲目过量施用化学农药已经造成耐药菌株的出现,致使土壤农药残留过高,生态环境破坏、人类健康隐患等问题日益严重。随着社会的发展和物质水平的提高,人们对温饱的需求转变成了对生活品质的追求,大众更青睐于健康绿色的果品,因此生物防治在病害防治中占据越来越重要的地位。其中微生物杀菌剂因其对人畜无毒、对环境无污染、抑菌范围广、稳定、安全高效等特性,满足了人们的需求,所以将具有拮抗作用的有益微生物开发成微生物菌剂,已成为一种重要的生防策略和产业界的迫切需要。
发明内容
本发明要解决的技术问题:猕猴桃细菌性溃疡病(又称为猕猴桃溃疡病)是猕猴桃最具毁灭性的病害之一,目前防治高度依赖铜制剂类化学药剂,但其长期大量使用的健康和环境风险日益受到重视;再者,市场推广使用的微生物菌剂品种有限,且对猕猴桃细菌性溃疡病的防治效果难以满足预期。
土壤是微生物的大本营,很多性能优良的生防菌株都是从土壤中分离获得的。戴尔福特菌(Delftia)是土壤中一类重要的资源性微生物种类,也是与人类有着密切关系且应用广泛的一种微生物。目前该菌主要应用在制备重金属吸附剂、回收水体或吸附土壤中的重金属、修复重金属污染水体或重金属污染土壤等方面。本发明旨在探寻生物农药的新来源,从土壤中分离筛选出一种戴尔福特菌,首次报道其对猕猴桃细菌性溃疡病菌具有显著的拮抗作用,并预判其在农业生产中将具有重要的潜在应用价值。
为解决现有技术存在的问题,本发明分离筛选、鉴定戴尔福特菌(Delftia),并测定该菌对猕猴桃细菌性溃疡病防效的技术思路如下。
(1)从发生猕猴桃细菌性溃疡病的猕猴桃果园中采集健康植株根际土土样,分离寻找生防菌。
(2)生防菌株的拮抗效果测定:通过平板对峙试验,筛选出对Psa具有抑菌效果的菌株。
(3)生防菌株的防效测定:利用离体枝条有伤接种、叶盘真空渗透接种等测定方法,探究生防菌株的室内防效。
(4)生防菌株的鉴定:提取菌株DNA进行PCR扩增,产物经测序后将测序结果与NCBI数据库中相关菌株的16S rDNA序列进行BLAST比对分析,利用MEGA6软件以邻接法构建系统发育树,初步确定拮抗菌株的分类地位;然后通过扫描电子显微镜观察菌株形态。
基于上述技术思路,本发明获得一种戴尔福特菌(Delftia)ZWP15菌株(本发明简称为为ZWP15菌株),采集于陕西省周至县中旺村猕猴桃园健康“秦美”猕猴桃植株的根际土,保藏信息如下。
分类命名:Delftia lacustris ZWP15
保藏时间:2021年9月1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
保藏单位地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所
保藏编号:CGMCC No.23331
通过平板对峙试验获知,所述ZWP15菌株对丁香假单胞杆菌猕猴桃致病变种(Pseudomonas syringae pv. actinidiae)具有拮抗作用。ZWP15菌株及发酵液均可对猕猴桃细菌性溃疡病菌Pseudomonas syringae pv. Actinidiae产生有效的抑制作用。由此,基于本发明的技术效果,本领域普通技术人员,结合现有技术,可以开发出一种丁香假单胞杆菌猕猴桃致病变种(Pseudomonas syringae pv. actinidiae)抑制剂,该抑制剂的活性成分为ZWP15菌株的菌体,或ZWP15菌株的发酵液,或ZWP15菌株的菌体和发酵液的混合物。
本发明首次揭示ZWP15菌株对猕猴桃细菌性溃疡病菌的拮抗活性,并通过室内抑制试验和田间防效试验得以证实。本发明所述ZWP15菌株生长繁殖快,适应能力强,对环境友好,对人畜无毒无害,可作为一种较好的生防资源进行挖掘。因此,本发明进一步提出ZWP15菌株在猕猴桃溃疡病防治中的应用,弥补ZWP15菌株用于猕猴桃病害防治的技术空白。本发明所述ZWP15菌株的菌体及发酵液均可用于猕猴桃溃疡病的防治。
鉴于所述ZWP15菌株对丁香假单胞杆菌猕猴桃致病变种显著的抑菌特性,以及在农业生产应用的安全性,本发明还提供一种生防菌剂,所述生防菌剂的活性成分为ZWP15菌株的菌体,或ZWP15菌株的发酵液,或ZWP15菌株的菌体和发酵液的混合物。该生防菌剂用于猕猴桃溃疡病的防治。
作为生防菌剂的优选实施方式,所述生防菌剂加工成农业上可接受的制剂后施用,用于防治猕猴桃溃疡病。本领域技术人员应当知晓,本发明所提供的生防菌剂通常以制剂(比如可湿性粉剂、悬浮剂、膏状涂抹剂等)的形式在农业上使用,除含有ZWP15菌株的菌体和/或发酵液外,还包括农业上可接受的载体或助剂。
示例性地,ZWP15菌株的菌体,或其发酵液干燥物(冻干粉),可以制备成常规的可湿性粉剂或粉剂,本领域技术人员很熟悉使用相应的载体或助剂完成本发明。可使用的分散剂如聚羧酸盐类(TERSPERSE 2700、T36、GY-D06等)、木质素磺酸盐类(Ufoxane 3A、Borresperse NA、Borresperse CA-SA等)、萘和烷基萘甲醛缩合物磺酸盐类(NNO、MF、Morwet D-425、Tamol NN、TERSPERSE2020等)、拉开粉BX(二丁基萘磺酸钠)、EO-PO嵌段聚醚类、烷基酚聚氧乙烯基醚磷酸酯、烷基酚聚氧乙烯基醚甲醛缩合物硫酸盐(SOPA),中一种或多种;润湿剂如硫酸盐类(K-12)、磺酸盐类(ABS-Na、BX、Terwet 1004等)、复合润湿剂(Morwet EFW)中一种或多种;填料如硅藻土、高岭土、轻钙、滑石粉、白炭黑、凹凸棒土、陶土、硫酸铵、尿素、蔗糖、葡萄糖、玉米淀粉、硫酸钠、多聚磷酸钠等一种或多种。
示例性地,ZWP15菌株的发酵液或发酵液干燥物(冻干粉)可以制备成常规的悬浮剂。制备悬浮剂可选用的分散剂如聚羧酸盐、木质素磺酸盐、烷基萘磺酸盐(扩散剂NNO)、TERSPERSE 2020(美国亨斯迈公司HUNTSMAN出品,烷基萘磺酸盐类)中一种或多种;乳化剂如BY(蓖麻油聚氧乙烯醚)系列乳化剂(BY-110、BY-125、BY-140)、农乳700#(通用名:烷基酚甲醛树脂聚氧乙烯醚)、农乳2201、斯盘-60#(通用名:山梨醇酐单硬脂酸酯)、吐温-60#(通用名:失水山梨醇单硬脂酸酯聚氧乙烯醚)、农乳1601#(通用名:苯乙基苯酚聚氧乙烯聚氧丙烯醚)、TERSPERSE 4894(美国亨斯迈公司出品)中的一种或多种;润湿剂如烷基酚聚氧乙烯基醚甲醛缩合物硫酸盐、烷基酚聚氧乙烯醚磷酸酯、苯乙基酚聚氧乙烯基醚磷酸酯、烷基硫酸盐、烷基磺酸盐、萘磺酸盐、TERSPERSE 2500(美国亨斯迈公司出品)中一种或多种;增稠剂如白炭黑、聚乙烯醇、膨润土、硅酸镁铝中一种或多种;安全剂MON13900;防冻剂如乙二醇、丙二醇、甘油、尿素、无机盐类如氯化钠中一种或多种;分散介质如大豆油、菜籽油、棉籽油、玉米油、蓖麻油、棕榈油、环氧大豆油、油酸甲酯及其甲酯化油类、柴油、机油、矿物油,及酯类溶剂如邻苯二甲酸二甲酯、邻苯二甲酸二丁酯、乙酸乙酯、苯甲酸甲酯中一种或多种。
示例性地,ZWP15菌株的菌体,或其发酵液或发酵液干燥物(冻干粉)可以制备成常规的颗粒剂。本领域技术人员可以选用以硅藻土、高岭土、轻钙、滑石粉、白炭黑、凹凸棒土、陶土等为材料制成的颗粒母核,采用圆盘造粒技术在颗粒母核表面粘附(吸附)ZWP15菌株的菌体,或其发酵液或发酵液干燥物(冻干粉)。在另一优选的实施例中,本领域技术人员可以将所述颗粒剂制备成具有多层结构的核壳颗粒,每一层所含成分可以相同或不同。
需要说明的是,上述制剂及其所选用载体或助剂的举例,仅是本发明技术方案具体实施方式的阐述或示例性指引。本领域技术人员基于现有技术,可以对制剂剂型和载体或助剂类别进行适应性拓展和有选择性的使用。
与现有技术相比,本发明“生防菌株及其在植物病害防治中应用”具有下述的有益效果或优点。
(1)本发明从土壤微生物资源中筛选出一种戴尔福特(Delftia)ZWP15菌株,首次揭示其对猕猴桃细菌性溃疡病菌具有显著的拮抗作用,为用于猕猴桃细菌性溃疡病防治的新型生物菌剂的开发,为猕猴桃细菌性溃疡病生物防治提供重要的材料积累和理论借鉴。
(2)本发明所述ZWP15菌株具有稳定的天然来源,发酵过程简便易控,发酵液制备成本较低,对人畜无毒无害,对农作物无药害,绿色环保,为新型生物菌剂的开发提供了有利条件,在农业生产中的植物病害防治领域具有重要的潜在应用价值。
(3)通过抑菌试验和田间药效获知,本发明所述ZWP15菌株防治猕猴桃细菌性溃疡病效果显著,其对猕猴桃细菌性溃疡病的防效与铜制剂相当,是一种优异的猕猴桃细菌性溃疡病生物防治微生物。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将本发明实施例中所涉及附图做简单的说明,显而易见地,所列出的附图仅仅是本发明实施例的一部分内容的呈现。
图1是本发明所述ZWP15菌株的皿内形态图。
图2是本发明所述ZWP15菌株对猕猴桃细菌性溃疡病菌的皿内抑菌活性图。
图3是本发明所述ZWP15菌株的显微结构图。
图4是基于邻接法构建的本发明所述ZWP15菌株的系统发育树。
图5是本发明所述ZWP15菌株对猕猴桃细菌性溃疡病菌的离体枝条防效试验结果示意图,a:阳性对照,只滴加猕猴桃细菌性溃疡病菌液;b:先滴加猕猴桃细菌性溃疡病菌液,后滴加ZWP15菌液;c:先滴加ZWP15菌液,后滴加猕猴桃细菌性溃疡病菌液;d:阴性对照,滴加无菌水。
图6是本发明所述ZWP15菌株对猕猴桃细菌性溃疡病菌的离体叶片防效试验结果示意图,a:阳性对照,只滴加猕猴桃细菌性溃疡病菌液;b:先滴加猕猴桃细菌性溃疡病菌液,后滴加ZWP15菌液;c:先滴加ZWP15菌液,后滴加猕猴桃细菌性溃疡病菌液。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合实施例对本发明做进一步详细说明。应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明的保护范围。
实施例1
本实施例描述ZWP15菌株的分离、纯化过程及鉴定结果。
1、菌株的分离、纯化
土样采集于陕西省周至县中旺村猕猴桃园。在猕猴桃细菌性溃疡病发病“秦美”猕猴桃园中,采集其中未发病且生长健壮植株的根际土壤(距地表15-20cm)。取土时,抖落根部的大块土壤,之后将黏附于根系表面的土壤再进行抖落并用塑料自封袋收集保存,作为待用样品。
采用稀释涂布法分离土壤样品中的目标生防菌。取不同稀释浓度的土壤悬液适量,分别在固体LB、PDA以及高氏一号培养基上涂布均匀,每种培养基、每个浓度设置2-3个重复。将平板置于28℃恒温培养箱中,连续培养48h。
挑选形态各异的菌落分别进行纯化,即当平板上长出单菌落时,将形态不同的单菌落用接种环挑出至新的固体培养基上,再培养。对纯化后的菌株进行平板对峙,3次重复,筛选对猕猴桃细菌性溃疡病病原菌具有明显拮抗作用的菌株。
在PDA培养基上分离筛选得到ZWP15菌株,ZWP15菌株放置于装有50%甘油的冻存管中(1:1)在-80℃条件下保存。
2、菌株16S rDNA序列鉴定
按照细菌基因组总DNA提取试剂盒(江苏康为世纪生物科技股份有限公司,CW0552)操作提取菌株DNA;选择通用引物对菌株的16S rDNA基因序列进行PCR扩增,扩增产物通过1.2%琼脂糖凝胶电泳检测,确定目标条带;按照回收试剂盒(Thermo Scientific,GeneJET胶回收试剂盒)操作进行胶回收;回收产物与载体相连接后克隆DNA片段;按照试剂盒(南京擎科生物科技有限公司,质粒小提试剂盒T-PM0201)操作提取质粒,送至生工生物工程股份有限公司测序。
将测序结果复制到NCBI数据库,BLAST比对得到多株相关菌株的16S rDNA序列,筛选出之中同源性较高的16S rDNA序列,下载为.fasta格式文件,通过MEGA6软件对选中序列进行调整校正,保存为.mass文件,返回MEGA6软件生成系统发育树,从而确定拮抗菌株的分类地位。
构建的系统发育树如图4所示。通过在NCBI中进行最大同源性比较,发现16S rDNA测序结果与Delftia lacustris 16S rDNA同源性达到99.93%,该菌株鉴定为戴尔福特菌(Delftia)ZWP15菌株。
3、菌体形态鉴定
利用扫描电子显微镜观察菌体形态。将-80℃甘油冻存管中保存的ZWP15菌株活化于PDA平板培养基,2d后挑选平板上合适部位菌落,用手术刀片切成约4mm×6mm的小块制备样品;将样品4℃浸渍于4%戊二醛过夜固定;用pH6.8,浓度0.1M的PBS缓冲液浸洗10分钟,30%-90%乙醇各15-20分钟重复漂洗4次;100%乙醇30分钟重复3次脱水;放入二氧化碳临界点干燥仪(Emitech,英国)中干燥;置于离子溅射仪(HITACHI,日本)喷金;制好的样品在S-4800场发射扫描电子显微镜10.0kV电压下进行观察、拍照,得到ZWP15菌株形态。如图3所示,ZWP15菌体呈杆状,单生。
实施例2
本实施例描述ZWP15菌株的皿内抑菌活性试验。
在PDA平板上划线活化ZWP15菌株,如图1所示,在PDA平板上ZWP15菌株的菌落较小,菌落为圆形,白色,周围有较浅色晕圈。用打孔器制成菌饼,放置于混好猕猴桃细菌性溃疡病菌菌液的LB培养基平板中央,28°C培养24小时后用十字交叉法测量抑菌圈直径(图2),试验设2个重复,取其平均值,抑菌圈直径达31mm。由此可以得出,ZWP15菌株对对猕猴桃细菌性溃疡病菌具有拮抗作用。
实施例3
本实施例描述ZWP15菌株的室内抑菌活性试验。
1、离体枝条有伤接种
采集当年生猕猴桃健康枝条,用0.6% NaClO溶液表面消毒20min,无菌水冲洗3次,晾干;截成三段适当长度的短枝条,每段制造1个伤口,伤口处滴加10μL菌液。每个处理5-10根枝条,重复3次。接种处理后的材料置于人工气候培养箱,20d记录结果。
处理a:阳性对照,在伤口处只滴加猕猴桃细菌性溃疡病病原菌液。
处理b:在伤口处先滴加猕猴桃细菌性溃疡病病原菌液,24h后再滴加ZWP15菌液。
处理c:在伤口处先滴加ZWP15菌液,24h后再滴加猕猴桃细菌性溃疡病病原菌液。
处理d:阴性对照,在伤口处只滴加无菌水。
猕猴桃细菌性溃疡病病原菌菌悬液的配制:病原菌在LB培养基平板上进行划线活化,24h后挑取单菌落于装有5mL LB液体培养基的试管中,28℃恒温振荡培养箱摇培24h,取2mL于离心管中,8000rpm离心3 min,倒掉上清液,加入1mL灭菌蒸馏水,涡旋振荡制成菌悬液。使用紫外分光光度计测定浓度,波长设置600nm,最终试验所用菌液浓度为OD=0.1(108 cfu/mL)。
ZWP15菌悬液的配制:菌株ZWP15在PDA培养基平板上进行划线活化,24h后挑取单菌落于装有5mL LB液体培养基的试管中,28℃恒温振荡培养箱摇培24h,取2mL于离心管中,8000rpm离心3min,倒掉上清液,加入1mL灭菌蒸馏水,涡旋振荡制成菌悬液。浓度同猕猴桃细菌性溃疡病病原菌菌悬液。
试验结果如图5所示,在离体条件下,阳性对照(处理a)枝条发病较重,处理b枝条病斑长度小于阳性对照,处理c枝条几乎未发病,经统计学分析,其“预防”效果显著(P<0.001)。本试验说明,ZWP15菌株对猕猴桃细菌性溃疡病菌具有拮抗作用,ZWP15菌株可以用于防治猕猴桃细菌性溃疡病。
2、叶盘真空渗透接种
采集长势一致的猕猴桃叶片,用0.6% NaClO溶液表面消毒10min,无菌水冲洗3次,放置晾干;使用打孔器避开叶脉制叶盘,叶盘置于菌液中真空渗透(0.1MPa,30s,反复3次)。每个处理5-10个叶盘,试验重复3次;无菌水清洗3次,置于水琼脂平板。接种处理后的材料置于人工气候培养箱,3d记录结果。本试验所用猕猴桃细菌性溃疡病病原菌菌悬液、ZWP15菌悬液同上。
处理a:阳性对照,只渗透猕猴桃细菌性溃疡病病原菌液。
处理b:先渗透猕猴桃细菌性溃疡病病原菌液,12h后再渗透ZWP15菌液。
处理c:先渗透ZWP15菌液,12h后再渗透猕猴桃细菌性溃疡病病原菌液。
处理d:阴性对照,只渗透无菌水。
试验结果如图6所示,在离体条件下,阳性对照(处理a)叶片发病较重,处理b叶片病斑面积小于阳性对照,处理c叶片几乎未发病,经统计学分析,其“预防”效果显著(P<0.001)。本试验说明,ZWP15菌株对猕猴桃细菌性溃疡病菌具有拮抗作用,ZWP15菌株可以用于防治猕猴桃细菌性溃疡病。
实施例4
本实施例描述ZWP15菌株发酵液的田间防效试验。
ZWP15菌株发酵液的制备方法:将在-80°C冰箱保存的ZWP15菌株接种在PDA培养基,于28°C培养2d后,用打孔器取3个单菌落的菌饼,将菌饼接入种子培养基(LB培养基:胰蛋白胨10g/L,酵母粉5g/L,NaCl 10g/L)中,于28°C,180rpm/min培养24h后,按5%的接种量将种子液接入发酵培养基(胰蛋白胨10g/L,葡萄糖8g/L,NaCl 8g/L)中,于28°C,180 rpm/min发酵96h收集发酵液。
在陕西省眉县营头镇,选取土壤肥力、栽培管理水平中等的猕猴桃果园,4-5年生徐香品种为试验对象;分别于秋季采果后(10月中下旬)、冬季落叶前(11月上旬)、春季开花前(翌年3月上旬)采用喷淋枝干的方式施用ZWP15菌株发酵液5倍稀释液、46%氢氧化铜稀释1000倍、枯草芽孢杆菌可湿性粉剂(德强生物股份有限公司)100倍稀释液,共计3个处理,以常规管理为对照。于翌年春季病害发生高峰3月中下旬,调查记载各处理发病情况,计算药剂防效。田间防效试验结果,见表1。
病株率%=发病株数/调查总株数×100。
防效%=(对照病株率-处理病株率)/对照病株率×100。
表1,不同药剂田间防效试验结果
药剂与处理 | ZWP15发酵液稀释5倍 | 46%氢氧化铜稀释1000倍 | 枯草芽孢杆菌可湿性粉剂稀释100倍液 |
防效% | 74.23 | 70.21 | 48.06 |
由表1呈现的统计结果获知,ZWP15菌株发酵液稀释5倍对猕猴桃细菌性溃疡病表现出较好的防治效果,防效达74.23%,与46%氢氧化铜稀释1000倍的防效相当,明显高于枯草芽孢杆菌可湿性粉剂稀释100倍液的防治效果。
以上所述,仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,都应涵盖在本发明的保护范围之内。
序列表
<110> 西北农林科技大学深圳研究院
<120> 一种生防菌株及其在植物病害防治中应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1407
<212> DNA
<213> ZWP15菌株(2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 1
cgagttccat gcagtcgaac ggtaacaggt cttcggacgc tgacgagtgg cgaacgggtg 60
agtaatacat cggaacgtgc ccagtcgtgg gggataacta ctcgaaagag tagctaatac 120
cgcatacgat ctgaggatga aagcggggga ccttcgggcc tcgcgcgatt ggagcggccg 180
atggcagatt aggtagttgg tgggataaaa gcttaccaag ccgacgatct gtagctggtc 240
tgagaggacg accagccaca ctgggactga gacacggccc agactcctac gggaggcagc 300
agtggggaat tttggacaat gggcgaaagc ctgatccagc aatgccgcgt gcaggatgaa 360
ggccttcggg ttgtaaactg cttttgtacg gaacgaaaaa gctccttcta atacaggggg 420
cccatgacgg taccgtaaga ataagcaccg gctaactacg tgccagcagc cgcggtaata 480
cgtagggtgc gagcgttaat cggaattact gggcgtaaag cgtgcgcagg cggttatgta 540
agacagatgt gaaatccccg ggctcaacct gggaactgca tttgtgactg catggctaga 600
gtacggtaga gggggatgga attccgcgtg tagcagtgaa atgcgtagat atgcggagga 660
acaccgatgg cgaaggcaat cccctggacc tgtactgacg ctcatgcacg aaagcgtggg 720
gagcaaacag gattagatac cctggtagtc cacgccctaa acgatgtcaa ctggttgttg 780
ggaattagtt ttctcagtaa cgaagctaac gcgtgaagtt gaccgcctgg ggagtacggc 840
cgcaaggttg aaactcaaag gaattgacgg ggacccgcac aagcggtgga tgatgtggtt 900
taattcgatg caacgcgaaa aaccttaccc acctttgaca tggcaggaag tttccagaga 960
tggattcgtg ctcgaaagag aacctgcaca caggtgctgc atggctgtcg tcagctcgtg 1020
tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc ttgtcattag ttgctacatt 1080
cagttgagca ctctaatgag actgccggtg acaaaccgga ggaaggtggg gatgacgtca 1140
agtcctcatg gcccttatag gtggggctac acacgtcata caatggctgg tacagagggt 1200
tgccaacccg cgagggggag ctaatcccat aaaaccagtc gtagtccgga tcgcagtctg 1260
caactcgact gcgtgaagtc ggaatcgcta gtaatcgcgg atcagcatgc cgcggtgaat 1320
acgttcccgg gtcttgtaca caccgcccgt cacaccatgg gagcgggtct cgccagaagt 1380
aggtagccta accgcaagga gggcact 1407
Claims (8)
1.一种生防菌ZWP15菌株,其特征在于,所述ZWP15菌株于2021年9月1日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.23331。
2.根据权利要求1所述的生防菌ZWP15菌株,其特征在于,所述ZWP15菌株对丁香假单胞杆菌猕猴桃致病变种(Pseudomonas syringae pv. actinidiae)具有拮抗作用。
3.生防菌ZWP15菌株在猕猴桃溃疡病防治中的应用,其特征在于,所述ZWP15菌株于2021年9月1日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCCNo.23331。
4.根据权利要求3所述的应用,其特征在于,所述ZWP15菌株用于防治猕猴桃细菌性溃疡病。
5.根据权利要求3所述的应用,其特征在于,所述ZWP15菌株的发酵液用于防治猕猴桃细菌性溃疡病。
6.一种生防菌剂,其特征在于,所述生防菌剂的组分包括ZWP15菌株,或包括ZWP15菌株的发酵液,或包括ZWP15菌株和发酵液的混合物。
7.根据权利要求6所述的生防菌剂,其特征在于,所述生防菌剂加工成农业上可接受的制剂后施用。
8.根据权利要求6所述的生防菌剂,其特征在于,所述生防菌剂用于防治猕猴桃细菌性溃疡病。
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