CN105861376A - 一种粘质沙雷氏菌及其分离方法和应用 - Google Patents
一种粘质沙雷氏菌及其分离方法和应用 Download PDFInfo
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- CN105861376A CN105861376A CN201610288569.1A CN201610288569A CN105861376A CN 105861376 A CN105861376 A CN 105861376A CN 201610288569 A CN201610288569 A CN 201610288569A CN 105861376 A CN105861376 A CN 105861376A
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- serratia marcescens
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- serratia
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/425—Serratia
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/06—Coating or dressing seed
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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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Genetics & Genomics (AREA)
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- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Environmental Sciences (AREA)
- Biomedical Technology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Soil Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明属于微生物技术领域,具体涉及一种粘质沙雷氏菌Serratia marcescens及其分离方法和应用,所述粘质沙雷氏菌保藏于中国典型培养物保藏中心,保藏编号CCTCC M 2015634,保藏日期2015年10月22日。本发明通过将田间自然死亡的蛴螬进行虫体表面消毒,在无菌条件下将其匀浆,加无菌水涂布于NA培养基平板上培养,挑取单菌落纯化获得粘质沙雷氏菌菌株TC‑1;本发明粘质沙雷氏菌作为生物杀虫剂、种子包衣剂和肥料添加剂使用,减少农业生产中农药残留、靶生物抗药性及面源污染等问题。
Description
技术领域
本发明属于微生物技术领域,具体涉及一种粘质沙雷氏菌及其分离方法和应用。
背景技术
微生物源农药(microbial insecticide)是指利用微生物活体及其代谢产物,用来杀虫、灭鼠、灭菌、除草等的制剂,主要包括真菌源、细菌源、病毒源、线虫源及抗生素类农药。微生物源杀虫剂具有低毒、污染轻、部分药物选择性高等优点,符合农药从传统的有机化学物质向“环境和谐农药”或“生物合理性农药”转化的发展趋势,受到国内外研究者及使用者的青睐。
粘质沙雷氏菌(Serratia marcescens)是一种重要的昆虫致病菌,并作为害虫的病原微生物受到生物防治工作者的高度关注。人们已从为害植物地上器官的多种有害昆虫体内分离获得了具有杀虫活性的粘质沙雷氏菌
中国专利CN104109649A公开了一种粘质沙雷氏菌NlM280及作为杀虫剂的应用,粘质沙雷氏菌NlM280发酵培养获得的菌悬液或菌悬液离心后的上清液可应用于褐飞虱或二化螟的防治;对褐飞虱采用注射和饲喂该菌菌悬液或发酵上清液的结果表明,其菌悬液注射对褐飞虱的致死率2天可达100%,饲喂菌悬液3天致死率为79.8%;该菌株发酵液的上清液在55℃以下处理均不影响其致病效果,注射发酵上清液致死率2天时可达97.2%,饲喂发酵上清液效果滞后,6天时致死率达84.2%。菌悬液和发酵上清液喷施水稻后饲喂二化螟,对二化螟的致死率4天时可达80.0%以上。中国专利105331566A公开了一种本发明涉及一种粘质沙雷氏菌S-JS1及其在害虫防治中的应用,粘质沙雷氏菌S-JS1固体培养菌体或发酵培养获得的菌悬液均可应用于褐飞虱、甜菜夜蛾和斜 纹夜蛾的防治;对褐飞虱采用喷雾该菌菌悬液的结果表明,其菌悬液8×109cfu/ml对褐飞虱3龄若虫、长翅成虫和短翅成虫的校正死亡率5天可达67.47%、63.58%和78.28%;添加100μl浓度为108cfu/ml的该菌株发酵液到饲料块(1×1×1cm)中,甜菜夜蛾和斜纹夜蛾3龄幼虫取食5天后,校正死亡率分别为45.64%和32.20%;9天后校正死亡率分别为76.36%和62.06%。但是,上述专利粘质沙雷氏菌对地下害虫的防治作用不明显。
发明内容
为克服上述缺陷,本发明的第一目的在于提供一种粘质沙雷氏菌(Serratia marcescens)。
本发明的第二目的还在于提供一种粘质沙雷氏菌的分离方法。
本发明的第三目的还在于提供一种粘质沙雷氏菌的应用。
为实现上述目的,本发明采用如下技术方案:
一种粘质沙雷氏菌,保藏于中国典型培养物保藏中心(CCTCC),地址:湖北省武汉市武昌区八一路珞珈山武汉大学,保藏编号CCTCC
NO:M2015634,保藏日期2015年10月22日。
一种上述的粘质沙雷氏菌的分离方法,包括以下步骤:
采集自田间自然死亡的蛴螬浸入70-75%乙醇10-15s,然后浸入0.08-0.12%升汞3-5min,进行虫体表面消毒,在无菌条件下将其匀浆,加无菌水3-6ml,取200μL,涂布于NA培养基平板上,培养24-48h,挑取单菌落纯化获得粘质沙雷氏菌菌株TC-1;
所述的NA培养基组成为:牛肉膏3.0g,蛋白胨5.0g,酵母粉3.0g,蔗糖10.0g,琼脂粉15g,蒸馏水1000mL。
一种粘质沙雷氏菌作为生物杀虫剂的应用。
优选地,所述的粘质沙雷氏菌作为蛴螬杀虫剂的应用。
一种粘质沙雷氏菌作为种子包衣剂的应用。
本发明的积极有益效果:
1.本发明粘质沙雷氏菌对植物地下害虫有致病性,作为蛴螬杀虫剂杀虫效果好;菌株TC-1对蛴螬有明显的毒性,当浓度达1×109cfu/ml时,校正死亡率达97.14%,半致死浓度为1.62×108cfu/ml;菌株TC-1具有杀线活性,72h秀丽线虫的致死率达93.3%;同时对小菜蛾,甜菜夜蛾、棉铃虫致死作用明显,处理96h矫正死亡率分别是72.2%、94.6%、83.8%;而且家蚕对本发明菌株TC-1有拒食反应,说明菌株TC-1可以防止害虫对蔬菜的损害。
2.本发明粘质沙雷氏菌作为生物杀虫剂、种子包衣剂和肥料添加剂使用,减少农业生产中农药残留、靶生物抗药性及面源污染等问题。
附图说明
图1为本发明菌株TC-1的扫描电镜图;
图2为本发明菌株TC-1在NA培养基上48h的菌落图;
图3为本发明菌株TC-1的16S rDNA序列PCR扩增电泳图;
图4为本发明基于16S rDNA序列构建的菌株TC-1的系统发育树。
具体实施方式
下面结合一些具体实施方式,对本发明进一步说明。
一种粘质沙雷氏菌,保藏于中国典型培养物保藏中心(CCTCC),保藏编号CCTCC M 2015634,保藏日期2015年10月22日。
实施例1粘质沙雷氏菌的分离及回接感染实验
(1)采集自田间自然死亡的蛴螬浸入70%乙醇10s,然后浸入0.1%升汞4min,进行虫体表面消毒,在无菌条件下将其匀浆,加无菌水5ml,取200μL,涂布于NA培养基平板上,培养24h,挑取单菌落纯化获得细菌菌株TC-1;
所述的NA培养基组成为:牛肉膏3.0g,蛋白胨5.0g,酵母粉3.0g,蔗糖10.0g,琼脂粉15g,蒸馏水1000mL,pH值7.0,121℃高压灭菌25min;
(2)将菌株TC-1接种到NB培养基中,在28℃180rpm下摇床培养2d,获得菌株TC-1细菌悬浮液,将细菌悬浮液浓度调至109cfu/ml,加入到土壤中,混拌均匀,使土壤含菌量达到1×109cfu/g,以土豆块为饲料喂养蛴螬,观察并收集死虫,对死虫进行病原菌的再分离,方法同步骤(1)所述;
所述的NB培养基的组成:牛肉膏3.0g,蛋白胨5.0g,酵母粉3.0g,蔗糖10.0g,蒸馏水1000mL,pH值7.0,121℃高压灭菌25min。
结果分析:根据科赫氏法则,用分离的病原菌株TC-1纯培养物回接感染植食性蛴螬,蛴螬与菌体接触48h后开始死亡,84h达到死亡高峰,幼虫感病后行动迟缓不活跃,基本停食,蛴螬死亡后体表变红,身体逐渐软化。从感病而死的蛴螬体中重新分离培养,所得的再分离细菌在培养性状、生理生化等方面都与原接种的菌株TC-1一致,据此确认蛴螬为菌株TC-1感染致死。
实施例2菌株的鉴定
1.菌株的形态分析:菌株TC-1接种于NA培养基上于28℃恒温培养48h,观察单菌落形态、大小、色泽:菌株TC-1呈短杆状(参见图1),无芽孢,有荧光,革兰氏染色阴性,周生鞭毛,具有运动性。在NA培养基上48h后菌落呈圆形,光滑湿润,边沿整齐,表面隆起(参见图2),玫红色;
2.菌株的生理生化测定:对菌株TC-1进行乳糖、木胶糖、阿拉伯糖、棉籽糖、蜜二糖、山梨醇、西蒙氏枸橼酸盐、胆汁七叶苷、丙二酸盐、苯丙氨酸脱氨酸、侧金盏花醇、鸟氨酸、赖氨酸脱羧酶、赖氨酸、精氨酸脱羧酶、甲基红、硫化氢、尿素等28种生理生化指标测定。
测定方法:按杭州天和微生物试剂有限公司生产的肠杆菌科细菌生化试剂盒说明书进行测定及结果判定。
生理生化鉴定结果表明(见下表1):菌株TC-1兼性好氧;甲基红试验、氧化酶、尿素及硫化氢反应阴性;苯丙氨酸脱羧酶、赖氨酸脱酸酶、鸟氨酸脱酸酶及精氨酸脱羧酶反应呈阳性;该菌能利用蔗糖、葡萄糖、木糖、甘露醇、山梨醇产酸,不能利用阿拉伯糖、乳糖、蜜二糖作为唯一碳源;能利用侧金盏花醇、西蒙氏枸橼酸盐、胆汁七叶苷,可使明胶水解。
表1菌株TC-1的生理生化特性
检测项目 | 反应 | 检测项目 | 反应 |
氧化酶 | - | 苯丙氨酸脱羧酶 | + |
革兰氏染色 | - | 赖氨酸脱酸酶 | + |
运动性 | + | 鸟氨酸脱酸酶 | + |
硫化氢 | - | 精氨酸脱羧酶 | + |
甘露醇 | + | 胆汁七叶苷 | - |
尿素 | - | 甲基红试验 | - |
吲哚试剂 | + | 蔗糖 | + |
蜜二糖 | - | 葡萄糖产酸 | + |
山梨醇 | + | 葡萄糖磷酸盐胨 | - |
DNase | + | 棉籽糖 | - |
乳糖 | - | 阿拉伯糖 | - |
侧金盏花醇 | + | 明胶液化 | + |
木糖 | + | 西蒙氏枸橼酸盐 | + |
注:+阳性;-阴性。
3.16S rDNA序列测定:将菌株TC-1接种到NB培养基中,在28℃180rpm下摇床培养2d,获得菌株TC-1细菌悬浮液,1000rpm离心10min,取上清,进行DNA提取,采用引物primer F27和primer R1492对菌株TC-1的基因组DNA进行16S rDNA的PCR扩增。
50μL扩增反应体系所加试剂:引物2μL,模板1μL,dNTP(含Mg2+)4μL,ddH2O 37.5μL,Taq聚合酶0.5μL,10×Buffer 5μL。PCR反应条件:94℃预变性5min;94℃变性60s,53℃退火30s,72℃延伸60s,共30个循环,最后72℃延伸10min。扩增产物经1.0%琼脂糖凝胶电泳检测,送至上海英骏生物技术有限公司测序。
结果表明:测得菌株TC-1的16S rDNA为1500bp(参见图3),以16S rDNA同源性为基础构建了相关种属的细菌系统发育树(参见图4),菌株TC-1(GenBank登录号KF700093)和粘质沙雷氏菌Serratia marcescens NR041980聚在同一分支,两者的最大相似性达100%。根据16S rDNA同源性比较解,结合菌株的形态学观察和生理学特征,将菌株TC-1鉴定为粘质沙雷氏菌(Serratia marcescens)。
实施例3对蛴螬毒力测定
从NA斜面上挑取一环菌株TC-1,接种到50mL NB液体培养基上,在28℃下摇床(180rpm)培养12h进行菌种活化,从活化菌体悬浮液中取1ml接种到250ml NB液体培养基中,28℃摇床(180rpm)培养48h后,稀释至1×109cfu/ml浓度后备用。
采用拌土接种法:根据试虫数量和养虫和容量预计所需土壤量称取土壤,配制菌株TC-1菌体悬浮液(1×109cfu/ml),稀释一定浓度加入到土壤中,混拌均匀,使土壤含菌量达到1×109cfu/g、1/2×109cfu/g、1/4×109cfu/g、1/8×109cfu/g、和1/16×109cfu/g,土壤含水量约17%。饲养盒中放入1头蛴螬,加土豆块为饲料,填装菌土,每处理试虫数60头,3次重复;以清水处理土壤为对照。处理后置于室温下饲养,每3d检查1次,根据不同批次试虫存活情况共检查7~10次,记录死亡虫数,结果见表2。
死亡率(%)=死亡虫数/供试虫数×100;
矫正死亡率(%)=(处理死亡率-对照死亡率)/(1-对照死亡率)×100。
表2菌株TC-1对蛴螬的毒力
毒力方程:Y=-9.8855+2.067X r=0.9747。
由表2可知:菌株TC-1对蛴螬有明显的毒性。当浓度达1×109cfu/ml时,校正死亡率达97.14%,半致死浓度为1.62×108cfu/ml。
实施例4对秀丽线虫毒力测定
在低营养无机盐培养基平板上铺一层无菌玻璃纸,将菌株TC-1涂布在玻璃纸上,待细菌长满平板,在玻璃纸中间加入50μl约1000条的秀丽线虫悬浮液,平板被分成20个小区,36h后随机选择5个小块,在解剖镜下观察数秀丽线虫的死亡情况。每一处理3个重复,每次观察不少于30条线虫,记录总虫数及死亡的虫数,并用头发刺激线虫,4h后再观察记录死亡的虫数(秀丽线虫有假死现象),每24h统计一次,结果见表3。
结果分析:菌株TC-1具有杀线活性,72h对秀丽线虫的致死率达93.3%,死亡的线虫虫体为红色,从死亡的线虫体内分离致病菌TC-1。
实施例5对家蚕毒力测定
新鲜的桑叶,清水洗净,晾干。实验组喷洒浓度为1×109cfu/ml的菌株TC-1的菌体悬浮液(叶的正面喷,且确定叶片每一地方都喷到),再晾干,喂食家蚕; 对照组喂食不含菌株TC-1的桑叶。每隔24h观察一次,并记录蚕的死活情况,结果见下表3。
死亡率(%)=死亡虫数/供试虫数×100;
矫正死亡率(%)=(处理死亡率-对照死亡率)/(1-对照死亡率)×100。
结果分析:用喷过菌株TC-1的桑叶喂蚕,48h内陆续有家蚕幼虫死亡,96h后死亡率到达高峰86.6%,144h后,实验组全部死亡。而对照组,在前120h内死亡率为1.6%。对死亡的蚕进行病原菌的分离,未分离到红色的粘质沙雷氏菌,说明家蚕死亡原因是拒食,说明菌株TC-1可以防止害虫对蔬菜的损害。
实施例6对小菜蛾、甜菜夜蛾、棉铃虫的毒力测定
虫饵料切成0.2×0.2×0.2cm3,用浓度为1×109cfu/ml的菌株TC-1的菌体悬浮液浸泡10min,控干水分。实验组供试害虫喂食菌液处理过的饵料,对照组喂食同等大小未用菌液处理的饵料。每一处理设置4个重复,每一重复20条虫子。每隔24h观察一次,并记供试害虫的死活情况。棉铃虫有猎杀同类的现象,故每只单独培养,结果见下表3。死亡率(%)=死亡虫数/供试虫数×100;
矫正死亡率(%)=(处理死亡率-对照死亡率)/(1-对照死亡率)×100。
表3菌株TC-1对部分靶生物的毒力
注:-:未观察;+:化蛹,CK:清水对照。
结果分析:菌株TC-1浸泡后的饵料喂食小菜蛾、甜菜夜蛾、棉铃虫连续观察6天,供试昆虫陆续都有死亡现象,尤其甜菜夜蛾,在48h内死亡率达92.5%,小菜蛾和棉铃虫的死亡高峰在72h-96h。
实验组小菜蛾96h死亡率达75.3%;甜菜夜蛾,实验组进食缓慢,对刺激不敏感,48h死亡率达92.5%,而对照死亡率只有7.5%;棉铃虫实验组与对照组进食正常,48h后,实验组虫子对刺激不敏感,且陆续有虫子死亡,144h后,死亡率达92.5%,且从死亡的棉铃虫尸体内,能分离到相应的红色的TC-1。
实施例7花生专用包衣剂
2015年4月,用粘质沙雷氏菌TC-1包衣花生。处理方法如下:利用NB培养基(牛肉膏3.0g,蛋白胨5.0g,酵母粉3.0g,蔗糖10.0g,蒸馏水1000mL,pH值7.0,121℃高压灭菌25min)培养粘质沙雷氏菌TC-1 36h,离心,弃上清,留菌体,然后菌体稀释至109cfu/mL,与花生种子混匀,一穴播种2粒,宽行30cm,窄行20cm,株距15cm,以清水和农药毒死蜱处理的花生作为对照,结果参见表4。
表4粘质沙雷氏菌包衣花生的效果
出苗率(%) | 产量(Kg/亩) | 增产率(%) | |
菌TC-1 | 88.19 | 456.33 | 62.98 |
农药毒死蜱 | 87.00 | 442 | 57.86 |
菌TC-1+药 | 92.63 | 447 | 59.64 |
清水 | 53.25 | 280 |
注:增产率=(A-B)/B,A为菌株TC-1,农药,菌株TC-1+农药处理的产量;B为清水对照处理的产量。
由表4可知:粘质沙雷氏菌TC-1处理的花生出苗率和产量显著高于清水对照,粘质沙雷氏菌TC-1处理的花生增产率达62.98%,高于农药处理毒死蜱结果,每亩比毒死蜱处理的花生增产14Kg(每亩增收约100元),因此,本发明采用粘滞沙雷氏菌代替农药处理花生种子,既环保有保证农民的收入。
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<110> 南阳师范学院
<120> 一种粘质沙雷氏菌及其分离方法和应用
<130> /
<160> 1
<170> PatentIn version 3.5
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<211> 1448
<212> DNA
<213> 天然
<400> 1
gtggcatacg aagcttacca tgcaagtcga gcggtagcac aggggagctt
gctccctggg 60
tgacgagcgg cggacgggtg agtaatgtct gggaaactgc ctgatggagg
gggataacta 120
ctggaaacgg tagctaatac cgcataacgt cgcaagacca aagaggggga
ccttcgggcc 180
tcttgccatc agatgtgccc agatgggatt agctagtagg tggggtaatg
gctcacctag 240
gcgacgatcc ctagctggtc tgagaggatg accagccaca ctggaactga
gacacggtcc 300
agactcctac gggaggcagc agtggggaat attgcacaat gggcgcaagc
ctgatgcagc 360
catgccgcgt gtgtgaagaa ggccttcggg ttgtaaagca ctttcagcga
ggaggaaggt 420
ggtgagctta atacgctcat caattgacgt tactcgcaga agaagcaccg gctaactccg
480
tgccagcagc cgcggtaata cggagggtgc aagcgttaat cggaattact
gggcgtaaag 540
cgcacgcagg cggtttgtta agtcagatgt gaaatccccg ggctcaacct
gggaactgca 600
tttgaaactg gcaagctaga gtctcgtaga ggggggtaga attccaggtg
tagcggtgaa 660
atgcgtagag atctggagga ataccggtgg cgaaggcggc cccctggacg
aagactgacg 720
ctcaggtgcg aaagcgtggg ggagcaaaca ggattagata ccctggtagt
ccacgctgta 780
aacgatgtcg atttggaggt tgtgcccttg aggcgtggct tccggagcta
acgcgttaaa 840
tcgaccgcct ggggagtacg gccgcaaggt taaaactcaa atgaattgac
gggggcccgc 900
acaagcggtg gagcatgtgg tttaattcga tgcaacgcga agaaccttac
ctactcttga 960
catccagaga acttagcaga gatgctttgg tgccttcggg aactctgaga
caggtgctgc 1020
atggctgtcg tcagctcgtg ttgtgaaatg ttgggttaag tcccgcaacg
agcgcaaccc 1080
ttatcctttg ttgccagcgg ttcggccggg aactcaaagg agactgccag
tgataaactg 1140
gaggaaggtg gggatgacgt caagtcatca tggcccttac gagtagggct
acacacgtgc 1200
tacaatggcg tatacaaaga gaagcgacct cgcgagagca agcggacctc
ataaagtacg 1260
tcgtagtccg gattggagtc tgcaactcga ctccatgaag tcggaatcgc
tagtaatcgt 1320
agatcagaat gctacggtga atacgttccc gggccttgta cacaccgccc
gtcacaccat 1380
gggagtgggt tgcaaaagaa gtaggtagct taaccttcgg gagggcgcta
ccacttggat 1440
cagtcggc
1448
Claims (5)
1.一种粘质沙雷氏菌(Serratia marcescens),其特征在于,所述粘质沙雷氏菌保藏编号为CCTCC M 2015634。
2.一种权利要求1所述粘质沙雷氏菌的分离方法,其特征在于,包括以下步骤:
采集自田间自然死亡的蛴螬浸入70-75%乙醇10-15s,然后浸入0.08-0.12%升汞3-5min,进行虫体表面消毒,在无菌条件下将其匀浆,加无菌水3-6ml,取200μL涂布于NA培养基平板上,培养24-48h,挑取单菌落纯化获得粘质沙雷氏菌菌株TC-1;
所述的NA培养基组成为:所述的NA培养基组成为:牛肉膏3.0g,蛋白胨5.0g,酵母粉3.0g,蔗糖10.0g,琼脂粉15g,蒸馏水1000mL。
3.一种权利要求1所述的粘质沙雷氏菌作为生物杀虫剂的应用。
4.根据权利要求3所述的应用,其特征在于,所述的粘质沙雷氏菌作为蛴螬杀虫剂的应用。
5.一种权利要求1所述的粘质沙雷氏菌作为种子包衣剂的应用。
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CN107736381A (zh) * | 2017-11-24 | 2018-02-27 | 江苏省农业科学院 | 一种用于防治刺吸性害虫的微生物与化学杀虫剂复配组合 |
CN108179123A (zh) * | 2017-12-28 | 2018-06-19 | 山东农业大学 | 一株白眉野草螟罹病虫体来源的粘质沙雷氏菌及其应用 |
CN110241100A (zh) * | 2019-07-03 | 2019-09-17 | 南阳师范学院 | 粘质沙雷氏菌高产几丁质酶的合成培养基及其发酵方法 |
CN111357770A (zh) * | 2020-05-12 | 2020-07-03 | 广东省生物资源应用研究所 | 粘质沙雷氏菌在制备白蚁拒食剂中的应用 |
CN113088467A (zh) * | 2021-04-09 | 2021-07-09 | 北京师范大学 | 一种沙雷氏菌菌株及其应用 |
CN114181870A (zh) * | 2021-12-30 | 2022-03-15 | 华南农业大学 | 一株具有广谱毒性的粘质沙雷氏菌SfSm-1及其应用 |
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CN107736381A (zh) * | 2017-11-24 | 2018-02-27 | 江苏省农业科学院 | 一种用于防治刺吸性害虫的微生物与化学杀虫剂复配组合 |
CN108179123A (zh) * | 2017-12-28 | 2018-06-19 | 山东农业大学 | 一株白眉野草螟罹病虫体来源的粘质沙雷氏菌及其应用 |
CN108179123B (zh) * | 2017-12-28 | 2021-04-06 | 山东农业大学 | 一株白眉野草螟罹病虫体来源的粘质沙雷氏菌及其应用 |
CN110241100A (zh) * | 2019-07-03 | 2019-09-17 | 南阳师范学院 | 粘质沙雷氏菌高产几丁质酶的合成培养基及其发酵方法 |
CN111357770A (zh) * | 2020-05-12 | 2020-07-03 | 广东省生物资源应用研究所 | 粘质沙雷氏菌在制备白蚁拒食剂中的应用 |
CN111357770B (zh) * | 2020-05-12 | 2020-08-28 | 广东省生物资源应用研究所 | 粘质沙雷氏菌在制备白蚁拒食剂中的应用 |
WO2021057308A1 (zh) * | 2020-05-12 | 2021-04-01 | 广东省科学院动物研究所 | 粘质沙雷氏菌在制备白蚁拒食剂中的应用 |
CN113088467A (zh) * | 2021-04-09 | 2021-07-09 | 北京师范大学 | 一种沙雷氏菌菌株及其应用 |
CN114181870A (zh) * | 2021-12-30 | 2022-03-15 | 华南农业大学 | 一株具有广谱毒性的粘质沙雷氏菌SfSm-1及其应用 |
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