CN109294954B - 一种淀粉酶链霉菌及其应用 - Google Patents
一种淀粉酶链霉菌及其应用 Download PDFInfo
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- CN109294954B CN109294954B CN201811242781.XA CN201811242781A CN109294954B CN 109294954 B CN109294954 B CN 109294954B CN 201811242781 A CN201811242781 A CN 201811242781A CN 109294954 B CN109294954 B CN 109294954B
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Abstract
本发明提供一种淀粉酶链霉菌及其应用，所述淀粉酶链霉菌命名为(St reptomyces diastaticus)FJAT‑31547，所述淀粉酶链霉菌(Streptomyces dia staticus)FJAT‑31547，于2018年3月22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，地址为北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.15479，该菌株的16S rRNA序列如SEQ ID NO.1所示，该菌株可用于防治番茄土传真菌和细菌性枯萎病。本发明菌株发酵产物能够抗尖孢镰刀菌引起番茄枯萎病、青枯雷尔氏菌引起番茄青枯病，且防治效果显著、选择性强，对今后从微生物资源中筛选和开发具有广谱抗菌活性物质具有重要指导意义，也为番茄土传真菌和细菌病害的生物防治提供菌株资源。
Description
【技术领域】
本发明涉及微生物技术领域，具体涉及一种淀粉酶链霉菌及其应用。
【背景技术】
随着化学农药大量且长期的应用，有害生物抗药性问题日渐突显，给人类健康及环境安全带来隐患。生物防治因具有高效性、强选择性、不易产生有害生物抗药性，对人畜低毒等特点，已成为研究热点。放线菌是最早发现具有生防效果的微生物，其中应用于生物防治的主要是链霉菌属Streptomyc es、诺卡菌属Nocardia、游动放线菌Actinoplanes、小单孢菌属Micromonos pora等。放线菌生防制剂，具有无毒、无残留、对环境友好、防病持效期长等优点，在生物防治中具有广阔的应用前景。利用放线菌防治植物病害，已有许多成功事例。Kemira(1989)利用灰绿链霉菌S.griseovidis的活细胞制剂防治镰孢菌Fusariumsp.、疫霉菌Phytophthora sp.和丝核菌Rhizoctonia sp.等一些常见的土传病原菌。MarkA.Roberts(2001)分离、筛选出700株具有抑制和杀灭多种植物病原菌能力的放线菌，尤其对真菌引起的根部病害效果显著。任迁琪等(2016)从辣椒根际土中分离筛选出10株对西瓜枯萎病菌具有拮抗作用的放线菌，其中一株链霉菌属效果最佳，盆栽防效66.7％。郭敬华等(2007)通过试验证明玫瑰黄链霉菌对黄瓜白粉病防效达88.26％。
番茄是世界性的经济作物，我国是全球最重要的的番茄生产国和出口国，种植面积已超过2000万亩。随着种植规模化，特别是设施栽培技术的发展，使得番茄连作现象普遍存在。由连作引起的土传病害已严重影响番茄产量和品质，其中尖镰孢菌Fusariumoxysporum引起的枯萎病和青枯雷尔氏菌Ralstonia solanacearum引起的青枯病发病最为严重，破坏性最大，二者均为侵染维管束的病害，其阻断维管束的运输功能，影响水分和营养的传导，造成植株萎蔫和死亡。目前对番茄土传病害主要依赖化学药剂，虽然对番茄的增产起了重要作用，但同时也造成了环境污染。因此生物防治被认为是一条有效的、可持续发展的植物病害防治途径。放线菌所产生抗生素及其次级代谢产物对植物病原菌的防治起到重要的作用。目前，已报道了一些具有自主知识产权的生防放线菌，但针对番茄土传病害的高效生防放线菌资源还十分有限。
【发明内容】
本发明要解决的技术问题，在于提供一种淀粉酶链霉菌及其应用，该菌株发酵产物能够抗尖孢镰刀菌引起番茄枯萎病、青枯雷尔氏菌引起番茄青枯病，且防治效果显著、选择性强。
本发明是这样实现的：
一种淀粉酶链霉菌，所述淀粉酶链霉菌命名为(Streptomyces diastaticus)FJAT-31547，所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547，于2018年3月22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，地址为北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.15479。
进一步地，所述所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547的16S rRNA序列如SEQ ID NO.1所示。
进一步地，所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547用于防治尖孢镰刀菌引起的番茄枯萎病、青枯雷尔氏菌引起的番茄青枯病。
进一步地，所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547用于制备尖孢镰刀菌引起的番茄枯萎病、青枯雷尔氏菌引起的番茄青枯病害生物防治菌剂产品。
本发明具有如下优点：
本发明是来自番茄根系土壤的具有广谱抗菌活性的淀粉酶链霉菌(Streptomycesdiastaticus)FJAT-31547，该菌株的发酵产物能够抗尖孢镰刀菌(Fusarium oxysporum)引起番茄枯萎病、青枯雷尔氏菌(Ralstonia solanacearum)引起番茄青枯病，且防治效果显著。因此本发明对番茄土传真菌和细菌病害的防治具有高效性、强选择性，同时不易产生有害生物抗药性，对人畜低毒，对今后从微生物资源中筛选和开发具有广谱抗菌活性物质具有重要指导意义，也为番茄土传真菌和细菌病害的生物防治提供菌株资源。
【附图说明】
下面参照附图结合实施例对本发明作进一步的说明。
图1为本发明实施例1中菌株FJAT-31547的菌落形态图。
图2为本发明实施例1中菌株FJAT-31547的菌体扫描电镜图。
图3为本发明实施例1中的菌株FJAT-31547基于16S rRNA序列构建系统发育树。
【具体实施方式】
本发明涉及一种淀粉酶链霉菌，所述淀粉酶链霉菌命名为(Streptomyc esdiastaticus)FJAT-31547，所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547，于2018年3月22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，地址为北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.15479。
所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547的16S rRN A序列如SEQ ID NO.1所示，在GenBank上的登入号为MH084838。
所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547用于防治番茄土传真菌和细菌性枯萎病。
所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547用于制备番茄土传真菌和细菌性枯萎病害生物防治菌剂产品。
以下结合具体实施例对本发明作进一步的说明。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施案例1：菌株FJAT-31547的分离和鉴定
(1)菌株的分离：从福建莆田东阳村番茄种植地收集土壤样品，称取土壤样品10g，按10-1、10-2、10-3、10-4、10-5和10-6进行系列梯度稀释，获得稀释液200μL涂布于高氏1号培养基(可溶性淀粉20g，KNO31 g,NaCl 0.5g,K2HPO40.5 g,MgSO4·7H2O 0.5g,FeSO4·7H2O0.01g，蒸馏水定容至1L，pH 7.3，121℃，灭菌30min)，30℃、培养7d；培养获得的菌株重新划线接种于高氏1号培养基培养基上，并置于30℃条件下培养7d，将培养获得的菌株命名为菌株FJAT-31547。
(2)菌株的形态鉴定：将菌株FJAT-31547接种于高氏1号培养基，30℃培养7d，观察菌落的形态特征，挑取单菌落进行扫描电镜样品制备。样品经2.5％戊二醛固定过夜后，经0.1mol/L磷酸缓冲液(pH 7.0)洗涤3次(每次15min)，用1％的锇酸溶液固定样品1-2h，用梯度浓度30％-50％-70％-80％-90％-95％乙醇对样品进行脱水处理(每级15min)再用100％的乙醇处理两次(每次20min)，用环氧丙烷处理两次(每次20min)，用CO2临界点干燥仪，用EIKO离子溅射仪镀膜，处理好的样品在JSM-6380LV扫描电镜中观察菌体形态和照相。
结果如下，菌株FJAT-31547在高氏1号培养基生长7d后，菌落表面绒粉状，中间为粉红色，色素透入培养基，边缘呈放射状，气生菌丝为白色，基内菌丝为粉红色(如图1所示)。在8000倍扫描电镜下，菌株FJAT-31547气生菌丝比较直，无螺旋状结构，有分枝状，菌丝断裂形成孢子，孢子呈成串生长(如图2所示)。这些特征与链霉菌一致。
(3)菌株的生理生化测定：采用API 20E和API 50CH对菌株FJAT-31547进行生理生化测定。将菌株FJAT-31547在高氏1号固体培养基上活化后，挑取单菌落转接于高氏1号液体培养基中，30℃、170rpm振荡培养48h，获得菌悬液。API 20E测定：用量程为1000μL的枪，将菌悬液加入到API 20E试剂条各微量小管内，在试验精氨酸、赖氨酸、鸟氨酸、硫化氢和脲素小杯内加入矿物油以形成厌氧环境。盖上培养盒，于35℃培养24h后，参考说明表判读其结果。API 50CH测定：将菌悬液加到试剂条上的50个微生化管中，最后用无菌液体石蜡封管静置35℃培养，24h后观察试验结果。试剂条变为黄色(对七叶灵检测结果为黑色)为阳性反应，变为绿色(七叶灵检测除外)为弱阳性反应，不变色为阴性反应，以48h观察到的结果作为最终结果。
结果如下：能够发酵乳糖，分解精氨酸，能够利用梓檬酸盐，分解尿酸，液化明胶，还原硝酸盐，其余都为阴性反应(具体结果见下表1)。在API50CH试验中，对甘油、D-阿拉伯糖、L-阿拉伯糖、核糖、D-木糖、半乳糖、葡萄糖、果糖、甘露糖、鼠李糖、肌醇、甘露醇、α-甲基-D-甘露糖甙、N-乙酰-葡糖胺、熊果甙、七叶灵、纤维二糖、乳糖、蔗糖、海藻糖、淀粉、肝糖、龙胆二糖、D-来苏糖、D-阿拉伯糖醇、葡萄糖酸盐为阳性反应，其余为阴性(具体结果见下表2)。
表1菌株FJAT-31547的API 20E试验结果
注：“+”为阳性反应,“-”为阴性反应
表2菌株FJAT-31547的API 50CH试验结果
注：“+”为阳性反应,“-”为阴性反应
(4)菌株的16S rRNA测定：采用细菌通用16S rRNA对菌株FJAT-31547进行分子鉴定。具体步骤如下：
a、菌株活化：用接种环将菌株FJAT-31547划线于高氏1号固体培养基上，置于30℃恒温培养箱中培养7d；
b、制备种子液：将步骤a所获得的菌株FJAT-31547的单菌落接种于20mL的高氏1号液体培养基中，30℃、170r/min振荡培养5d，得种子液；
c、基因组DNA的提取：取种子液1mL于已灭菌的1.5mL离心管中，并按照Promega试剂盒说明书操作以提取菌株FJAT-31547的基因组DNA；
d、PCR鉴定：以提取所获得的菌株FJAT-31547的基因组DNA为模板，利用16S rRNA通用引物27F(5′-GAGTTTGAT CCT GGC TCAG-3′，如SEQ ID NO.2所示)和1492R(5′-ACGGCTACCTTG TTACGACTT-3′，如SEQ ID NO.3所示)进行PCR反应；
PCR反应体系(25μL)：ddH2O 9.5μL，Mix 12μL，上下游引物各1μL，DNA 1.5μL；
PCR反应程序为：94℃预变性5min；接着94℃变性30s，55℃退火60s，72℃复性90s，30个循环；最后72℃延伸10min；
PCR产物的检测：将PCR产物点样于1.5％的琼脂糖凝胶(含染料GV-1)，以100bpDNA Ladder Marker作为标准分子量，80V电压、1倍TA E缓冲液中电泳40min，用凝胶成像系统观察结果。
测序：将PCR产物送至铂尚生物技术有限公司进行测序。
16S rRNA序列(如SEQ ID NO.1所示)分析：将所得序列在细菌序列比对网站EZtaxon-e.ezbiocloud.net上进行序列比对分析后，初步判断得出该菌株的分类地位。选择相关的参考菌株序列，用软件Mega 6.0构建N-J(N eighbor-joining)系统发育树，确定菌株的分类地位。
结果如下：FJAT-31547的16S rRNA基因序列长度为1396bp，与NCBI基因库序列比对结果显示与淀粉酶链霉菌(Streptomyces diastaticus subsp.Ardesiacus)同源性达99.64％。基于16S rRNA基因序列构建了系统发育树(如图3所示)，结果表明，菌株FJAT-31547与淀粉酶链霉菌(Streptomyces diastaticus subsp.Ardesiacus)的模式菌株NRRL-1773(T)的相似性为92％。
综上，分离得到的菌株FJAT-31547为淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547。
实施案例2：菌株FJAT-31547的抑菌谱测定
(1)菌株FJAT-31547对10种病原真菌的抑菌效果
供试病原真菌菌株：10种病原真菌，由福建省农业科学院农业生物资源研究所提供。菌株具体信息见下表3。
培养基配方：马铃薯蔗糖固体培养基：马铃薯200.0g，蔗糖20.0g，琼脂18.0g,蒸馏水定容至1L，pH为7.2；半固体马铃薯蔗糖培养基：马铃薯200.0g，蔗糖20.0g，琼脂9g，蒸馏水定容至1L；马铃薯蔗糖液体培养基：马铃薯200.0g，蔗糖20.0g，蒸馏水定容至1L，pH为7.2；上述培养基用于病原真菌的培养。用于淀粉酶链霉菌FJAT-31547的高氏1号液体培养基：可溶性淀粉20g，氯化钠0.5g，硫酸亚铁0.01g，硝酸钾1g，磷酸氢二钾0.5g，硫酸镁0.5g，蒸馏水定容至1L，pH为7.2；高氏1号固体培养基在液体培养基的基础上加入15g/L琼脂。
菌株的活化和菌悬液制备：供试的病原真菌活化于马铃薯蔗糖固体培养基，FJAT-31547活化于高氏1号固体培养基。挑取病原真菌单菌落于马铃薯蔗糖液体培养基、FJAT-31547于高氏1号液体培养基中，于30℃、170r/min摇床振荡培养，FJAT-31547培养7d，真菌培养5d，将FJAT-31547和各真菌的培养液用无菌水稀释，制成浓度为108CFU/mL的菌悬液，4℃保存备用。
抑菌圈法检测抑菌效果：将1mL病原真菌的菌悬液与9mL 50℃的半固体马铃薯蔗糖培养基混合后，倒入已制备好的马铃薯蔗糖固体培养基中，制成含病原真菌的双层培养基；待培养基凝固后，用1mL枪头在平板上打孔，在孔内加入200μL过0.22μm滤膜的放线菌菌悬液，以水为阴性对照，0.025mg/mL潮霉素为阳性对照。30℃生物培养箱培养培养3-5d。观察抑菌透明圈的产生，拍照并测量透明圈直径。
结果分析：淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547对雪霉叶枯病菌、大豆拟茎点霉菌、青霉菌、尖孢镰刀菌、腐皮镰刀菌、白僵菌、黑曲霉有抑制作用，其中对尖孢镰刀菌抑菌效果最好，抑菌圈直径达21.27mm。FJAT-31547对黄曲霉、烟曲霉、链格孢菌无抑制作用(具体结果见下表3)。
表3菌株FJAT-31547对10种病原真菌的抑菌效果
(2)菌株FJAT-31547对12种病原细菌的抑菌效果
供试病原细菌：12种病原细菌，由福建省农业科学院农业生物资源研究所提供。病原细菌具体信息见下表4。
培养基配方：LB液体培养基：胰蛋白胨10g，氯化钠10g，酵母粉5g,蒸馏水定容至1L，pH为7.2。LB固体培养基是在液体培养基的基础上加入1.5g/L琼脂，LB半固体培养基是在液体培养基的基础上加入0.9g/L琼脂。
菌株活化和菌悬液：供试的病原细菌活化于LB固体培养基，30℃培养2d，菌株FJAT-31547活化于高氏1号固体培养基，30℃培养7d。分别挑取病原细菌单菌落于LB液体培养基、菌株FJAT-31547单菌落于高氏1号液体培养基中，于30℃、170r/min摇床振荡培养，病原细菌培养1d，FJAT-31547培养7d，将FJAT-31547和各细菌的培养液用无菌水稀释，制成浓度为108CFU/mL的菌悬液，4℃保存备用。
抑菌圈法检测抑菌效果：将1mL病原细菌菌悬液与9mL 50℃的LB半固体培养基混合后，倒入已制备好的LB固体培养基中，制成含病原细菌的双层培养基；待培养基凝固后，用1mL枪头在平板上打孔，在孔内加入200μL过0.22μm滤膜的放线菌菌悬液，以水为阴性对照，0.025mg/mL潮霉素为阳性对照，30℃培养1-2d。观察抑菌透明圈的产生，拍照并测量透明圈直径。
结果分析：由表4可知，淀粉酶链霉菌FJAT-31547对大肠杆菌、青枯雷尔氏菌、沙门氏菌、根癌农杆菌、氟氏柠檬酸杆菌、软腐果胶杆菌、辣椒软腐菌和辣椒疮痂菌有抑制作用，对青枯雷尔氏菌抑菌效果最强，抑菌圈达21.57mm。对阴沟肠杆菌、生癌肠杆菌、铜绿假单胞菌、燕麦食酸菌西瓜亚种无抑制作用。
表4菌株FJAT-31547对12种病原真菌的抑菌效果
实施案例3：菌株FJAT-31547对番茄土传病害的防治效果
(1)菌株FJAT-31547对番茄细菌性青枯病的防治效果
供试番茄品种：购自福建农科农业良种开发有限公司的“农科180”番茄种子
菌悬液制备：将菌株FJAT-31547于高氏1号固体培养基上活化后，挑取单菌落于高氏1号液体培养基中，30℃、170r/min摇床振荡培养7d，将培养所得菌液用无菌水稀释，制成浓度为107CFU/mL的菌悬液，待用；将现有公认的青枯雷尔氏菌标准菌株GMI1000经TTC平板(蛋白胨10.0g，水解酪蛋白1.0g，葡萄糖5.0g，2，3，5氯化三苯基氮唑0.05g，蒸馏水定容至1L，pH值7.2)活化后，接种于SPA液体培养基(蔗糖20g，蛋白胨5g，KH2PO40.5 g，MgSO40.025g，蒸馏水定容至1L，pH值7.2)中，于170r/min、30℃下培养48h，将培养所得菌液用无菌水稀释至107CFU/mL，得到菌株GMI1000的菌悬液，待用；
试验处理：设置实验组和阳性对照组，在实验组中，将菌株FJAT-31547的菌悬液伤根接种于5-6叶龄的番茄盆栽苗上，3d后再接种菌株GMI1000的菌悬液，两次接种的接种量均为100mL/株，共接种30盆；阳性对照组以清水代替FJAT-31547的菌悬浮液，即将清水伤根接种于5-6叶龄的番茄盆栽苗上，3d后再接种菌株GMI1000的菌悬液；各处理设3个重复，将接种后的番茄苗放置于光照培养箱(每天光培养12h、暗培养12h，温度30±1℃，相对湿度90％)，每天观察番茄植株的发病情况，统计处理番茄植株的发病率和防效。
发病率＝(发病株数/接种株数)×100％
防效＝[(对照组发病率-实验组发病率)/对照组发病率]×100％
试验结果：阳性对照组在接种菌株GMI1000后第4d植株开始发病，番茄植株出现萎蔫症状，并随时间推移日益严重，至第10d发病率达100％；而实验组在接种菌株GMI1000第10d，植株发病率为23.08％，防效为76.92％(见下表5)。
表5菌株FJAT-31547番茄青枯病的防治效果
(2)菌株FJAT-31547对番茄真菌性枯萎病的防治效果
供试番茄品种：农科180
菌悬液制备：将菌株FJAT-31547于高氏1号固体培养基上活化后，挑取单菌落于高氏1号液体培养基中，30℃、170r/min摇床振荡培养7d，将培养所得菌液用无菌水稀释，制成浓度为107CFU/mL的菌悬液，待用；将番茄枯萎病菌(尖孢镰刀菌，Fusarium oxysporum)经马铃薯蔗糖固体培养基活化后，接种于马铃薯蔗糖液体培养基中，于170r/min、30℃下培养5d，将培养所得菌液用无菌水稀释，制成浓度为107CFU/mL菌悬液，待用；
试验处理：设置实验组和阳性对照组，在实验组中，将菌株FJAT-31547的菌悬液伤根接种于5-6叶龄的番茄盆栽苗上，3d后再接种番茄枯萎病菌菌悬液，两次接种的接种量均为100mL/株，共接种30盆；阳性对照组将清水伤根接种于5-6叶龄的番茄盆栽苗上，3d后再接种番茄枯萎病菌菌悬液；各处理设3个重复，将接种后的番茄苗放置于光照培养箱(每天光培养12h、暗培养12h，温度30±1℃，相对湿度90％)，每天观察番茄植株的发病情况，统计处理番茄植株的发病率和防效。
发病率＝(发病株数/接种株数)×100％
防效＝[(对照组发病率-实验组发病率)/对照组发病率]×100％
试验结果：阳性对照组在接种番茄枯萎病菌后第15d植株开始发病，番茄植株出现萎蔫症状，并随时间推移日益严重，至第28d发病率达100％；而实验组先接种菌株FJAT-31547，3d后再接种番茄枯萎病菌的番茄，在接种番茄枯萎病菌第28d，植株发病率为病19.41％，防效为80.59％(见下表6)。
表6菌株FJAT-31547番茄枯病的防治效果
本发明是来自番茄根系土壤的具有广谱抗菌活性的淀粉酶链霉菌(Streptomycesdiastaticus)FJAT-31547，该菌株的发酵产物能够抗尖孢镰刀菌(Fusarium oxysporum)引起番茄枯萎病、青枯雷尔氏菌(Ralstonia solanacearum)引起番茄青枯病，且防治效果显著、选择性强。对今后从微生物资源中筛选和开发具有广谱抗菌活性物质具有重要指导意义，也为番茄土传真菌和细菌病害的生物防治提供菌株资源。
虽然以上描述了本发明的具体实施方式，但是熟悉本技术领域的技术人员应当理解，我们所描述的具体的实施例只是说明性的，而不是用于对本发明的范围的限定，熟悉本领域的技术人员在依照本发明的精神所作的等效的修饰以及变化，都应当涵盖在本发明的权利要求所保护的范围内。
序列表
<110> 福建省农业科学院农业生物资源研究所
<120> 一种淀粉酶链霉菌及其应用
<130> 100
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1396
<212> DNA
<213> （Streptomyces diastaticus）
<400> 1
tgcagtcgaa cgatgaacca ccttcgggtg gggattagtg gcgaacgggt gagtaacacg 60
tgggcaatct gccctgcact ctgggacaag ccctggaaac ggggtctaat accggatact 120
gacctgccaa ggcatcttgg cgggtcgaaa gctccggcgg tgcaggatga gcccgcggcc 180
tatcagcttg ttggtgaggt aatggctcac caaggcgacg acgggtagcc ggcctgagag 240
ggcgaccggc cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg 300
gaatattgca caatgggcga aagcctgatg cagcgacgcc gcgtgaggga tgacggcctt 360
cgggttgtaa acctctttca gcagggaaga agcgaaagtg acggtacctg cagaagaagc 420
gccggctaac tacgtgccag cagccgcggt aatacgtagg gcgcaagcgt tgtccggaat 480
tattgggcgt aaagagctcg taggcggctt gtcgcgtcgg ttgtgaaagc ccggggctta 540
accccgggtc tgcagtcgat acgggcaggc tagagttcgg taggggagat cggaattcct 600
ggtgtagcgg tgaaatgcgc agatatcagg aggaacaccg gtggcgaagg cggatctctg 660
ggccgatact gacgctgagg agcgaaagcg tggggagcga acaggattag ataccctggt 720
agtccacgcc gtaaacggtg ggcactaggt gtgggcaaca ttccacgttg tccgtgccgc 780
agctaacgca ttaagtgccc cgcctgggga gtacggccgc aaggctaaaa ctcaaaggaa 840
ttgacggggg cccgcacaag cggcggagca tgtggcttaa ttcgacgcaa cgcgaagaac 900
cttaccaagg cttgacatac accggaaagc atcagagatg gtgcccccct tgtggtcggt 960
gtacaggtgg tgcatggctg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca 1020
acgagcgcaa cccttgtccc gtgttgccag caactcttcg gaggttgggg actcacggga 1080
gaccgccggg gtcaactcgg aggaaggtgg ggacgacgtc aagtcatcat gccccttatg 1140
tcttgggctg cacacgtgct acaatggccg gtacaatgag ctgcgatacc gcaaggtgga 1200
gcgaatctca aaaagccggt ctcagttcgg attggggtct gcaactcgac cccatgaagt 1260
cggagtcgct agtaatcgca gatcagcatt gctgcggtga atacgttccc gggccttgta 1320
cacaccgccc gtcacgtcac gaaagtcggt aacacccgaa gccggtggcc caaccccttg 1380
tgggagggag cttcga 1396
<210> 2
<211> 19
<212> DNA
<213> （人工序列）
<400> 2
gagtttgatc ctggctcag 19
<210> 3
<211> 21
<212> DNA
<213> （人工序列）
<400> 3
acggctacct tgttacgact t 21
Claims (3)
1.一种淀粉酶链霉菌，其特征在于：所述淀粉酶链霉菌命名为(Strep tomycesdiastaticus)FJAT-31547，所述淀粉酶链霉菌(Streptomyces diastat icus)FJAT-31547，于2018年3月22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，地址为北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC No.15479；
所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547的16S rRN A序列如SEQID NO.1所示。
2.一种淀粉酶链霉菌的应用，其特征在于：所述淀粉酶链霉菌是权利要求1中所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547，用于防治尖孢镰刀菌引起的番茄枯萎病、青枯雷尔氏菌引起的番茄青枯病。
3.一种淀粉酶链霉菌的应用，其特征在于：所述淀粉酶链霉菌是权利要求1中所述淀粉酶链霉菌(Streptomyces diastaticus)FJAT-31547，用于制备尖孢镰刀菌引起的番茄枯萎病、青枯雷尔氏菌引起的番茄青枯病害生物防治菌剂产品。
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