CN104694395A - 一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 - Google Patents
一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 Download PDFInfo
- Publication number
- CN104694395A CN104694395A CN201310657681.4A CN201310657681A CN104694395A CN 104694395 A CN104694395 A CN 104694395A CN 201310657681 A CN201310657681 A CN 201310657681A CN 104694395 A CN104694395 A CN 104694395A
- Authority
- CN
- China
- Prior art keywords
- metarhizium anisopliae
- microbial inoculum
- peanut
- solid
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 235000020232 peanut Nutrition 0.000 title claims abstract description 28
- 235000017060 Arachis glabrata Nutrition 0.000 title claims abstract description 27
- 235000010777 Arachis hypogaea Nutrition 0.000 title claims abstract description 27
- 235000018262 Arachis monticola Nutrition 0.000 title claims abstract description 27
- 241000223250 Metarhizium anisopliae Species 0.000 title claims abstract description 26
- 235000013399 edible fruits Nutrition 0.000 title abstract description 10
- 230000000694 effects Effects 0.000 title abstract description 7
- 241001553178 Arachis glabrata Species 0.000 title abstract 5
- 238000000855 fermentation Methods 0.000 title abstract 3
- 230000004151 fermentation Effects 0.000 title abstract 3
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 13
- 239000007787 solid Substances 0.000 claims abstract description 6
- 244000105624 Arachis hypogaea Species 0.000 claims description 23
- 230000002554 disease preventive effect Effects 0.000 claims description 14
- 241000894006 Bacteria Species 0.000 claims description 6
- 238000010563 solid-state fermentation Methods 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
- CBCKQZAAMUWICA-UHFFFAOYSA-N 1,4-phenylenediamine Chemical compound NC1=CC=C(N)C=C1 CBCKQZAAMUWICA-UHFFFAOYSA-N 0.000 claims description 2
- 241000209140 Triticum Species 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 239000002671 adjuvant Substances 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 239000010903 husk Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 238000007790 scraping Methods 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 235000015099 wheat brans Nutrition 0.000 claims description 2
- 201000010099 disease Diseases 0.000 abstract description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 11
- 230000002265 prevention Effects 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 230000007918 pathogenicity Effects 0.000 abstract description 2
- 238000004321 preservation Methods 0.000 abstract 2
- 241000254043 Melolonthinae Species 0.000 abstract 1
- 230000006806 disease prevention Effects 0.000 abstract 1
- 230000000855 fungicidal effect Effects 0.000 abstract 1
- 239000000417 fungicide Substances 0.000 abstract 1
- 244000144972 livestock Species 0.000 abstract 1
- 231100000956 nontoxicity Toxicity 0.000 abstract 1
- 230000001737 promoting effect Effects 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 6
- 241000233866 Fungi Species 0.000 description 5
- 241000238631 Hexapoda Species 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 239000000447 pesticide residue Substances 0.000 description 2
- 230000003449 preventive effect Effects 0.000 description 2
- 230000000452 restraining effect Effects 0.000 description 2
- 241001167556 Catena Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
Classifications
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- 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
- C12R2001/645—Fungi ; Processes using fungi
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及微生物防治领域,特别涉及金龟子绿僵菌菌株、液固双相发酵工艺及发酵菌剂在防治花生果腐病中的应用。包含保藏编号为CGMCCNo.8385的金龟子绿僵菌菌剂1~10万亿孢子/亩穴施于花生,田间防病试验表明对花生果腐病的防治效果为60%以上,增产5~12%,实现了在控制病害的同时,达到保果增产的目的。本发明涉及的金龟子绿僵菌对花生果腐病有良好的防治效果;其次,该菌株还具有促生增产作用;同时,该菌剂的孢子含量高、用量少,且无毒无致病性,对人畜安全,不污染环境,生产成本低。
Description
技术领域
本发明涉及微生物防治领域,特别涉及金龟子绿僵菌菌株、液固双向发酵工艺及发酵菌剂在防治花生果腐病中的应用。
背景技术
花生果腐病为一种花生生长中后期发生的土传病害,是由侵脉新赤壳菌(Neocosmosporasp.)侵染花生果而引起花生果腐烂的一种病害,该病害发病特征如下:(1)发生病变的荚果主要是中后期的秕果和饱果;(2)初步发生时,花生荚果的中果皮和外果皮变为黑褐色斑点,内果皮及种子正常;(3)发病严重时,荚果的整个果皮和种子变黑、腐烂;(4)根部外表皮发黑,而地上部分不变或者更绿。2006~2011年,该病害在我国河北、山东、吉林、河南等花生种植区先后被发现,烂果率在30~100%,这严重损害了当地花生种植户的利益。目前市场上还没有有效的防治该病害药物。因此,提供一种能够有效防治花生果腐病的药物具有重要现实意义。
绿僵菌作为当今世界应用最为广泛的一种昆虫病原真菌,因对昆虫的致病力强、寄主范围广,对人、畜、作物无毒害而受到人们的重视。目前,已经发现金龟子绿僵菌能够寄生的昆虫约有200余种,许多国家对它们在生产、应用和安全性等方面都做了大量细致的研究,对有些害虫已开发出绿僵菌杀虫剂,并收到良好防治效果。近年来,科研人员发现绿僵菌对植物病原菌也有一定的抑制作用,田间试验表明部分绿僵菌制剂对植物病害具有良好的防治效果。因此,急需一种能够实现持续控制花生果腐病和地下害虫,降低花生果农药残留,实现防病虫、保果、增产作用的绿僵菌生物制剂。目前还未见绿僵菌对花生果腐病有防治作用。
发明内容
有鉴于此,本发明提供一株金龟子绿僵菌菌株及其在防治花生果腐病中的应用。该金龟子绿僵菌的保藏编号为CGMCC No.8385。该金龟子绿僵菌菌株能够持续控制花生果腐病,降低花生果农药残留,实现防病、保果、增产。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一株金龟子绿僵菌,其保藏编号为CGMCC No.8385。
该菌株ITS 在GenBank 序列登记号为KF913494。
本发明的金龟子绿僵菌,菌落绒毛状或棉絮状,最初白色,产生孢子时呈绿色;菌丝具分隔和分枝,透明,直径1.5~2.0μm;分生孢子梗很难与菌丝区别,直径约2.0μm,其末端产生瓶形小梗,(8.0~16.0μm)*(1.5~2.2)μm,从瓶梗的末端以向基式连续形成长串链的分生孢子;在同一小梗上,第一层分生孢子后面形成第二层、第三层分生孢子,呈现一系列念珠状排列的孢子;分生孢子为单细胞,长椭圆形,两端钝截形或钝圆,大小变化很大,(5.0~7.0)μm*(2.0~3.0)μm,单个存在时色亮,成堆时呈橄榄绿色,脱落的孢子常聚成菱形孢子团块或壳状结构;平皿对峙实验表明该菌对花生果腐病菌具有明显的抑制作用。
本发明还提供了保藏编号为CGMCC No.8385的金龟子绿僵菌在制备抑制花生果腐病菌药物中的应用。
本发明还提供了一种菌剂,其包括保藏编号为CGMCC No.8385的金龟子绿僵菌及菌剂中可接受的佐剂。
作为优选,所述金龟子绿僵菌的含量≥108 孢子/克。
本发明还提供了上述菌剂的液固双向发酵工艺,包括如下步骤:
步骤1:挑取保藏编号为CGMCC No.8385的金龟子绿僵菌接种于PPDA固体培养基上,于24~30℃培养9~12d,刮取孢子置于研磨器中,用0.05%吐温-80无菌水配制成浓度为1~5×108个/mL的孢子悬液,得第一培养物;
步骤2:吸取2~4mL第一培养物,置于SDAY改良液体培养基中,于25~30℃、160~200 rpm培养70~75h,得第二培养物;
步骤3:将14~16mL第二培养物做为固态发酵的种子液转接至80~100克/袋的固态发酵培养基中,混匀后于25~30℃培养6~9 d后即得。所述固态发酵培养基包括:麦壳1~3重量份,麦麸6~10 重量份,水87~93 重量份。
作为优选,上述菌剂的施用量为1~10万亿孢子/亩。
本发明提供一株金龟子绿僵菌菌株及其在防治花生果腐病中的应用。该金龟子绿僵菌的保藏编号为CGMCC No.8385。包含保藏编号为CGMCC No.8385的金龟子绿僵菌的菌剂按1~10万亿孢子/亩穴施于花生,田间防病试验表明对花生果腐病的防治效果60%以上,增产5~12%,实现了在控制病害的同时,达到保果,增产的目的。本发明涉及的金龟子绿僵菌能对花生果腐病有良好的防治效果;其次,该菌株还具有促生增产作用;同时,该菌剂无毒无致病性,对人畜安全,不污染环境,生产成本低。
Claims (7)
1.一株金龟子绿僵菌,其特征在于,其保藏编号为CGMCC No.8385。
2.根据权利要求1所述的金龟子绿僵菌在制备抑制花生果腐病菌中的应用。
3.一种菌剂,其特征在于,其包括保藏编号为CGMCC No.8385的金龟子绿僵菌及菌剂中可接受的佐剂。
4.根据权利要求3所述菌剂,其特征在于,所述金龟子绿僵菌的含量≥108 孢子/克。
5.根据权利要求3或4所述菌剂的液固双向发酵工艺,其特征在于:包括如下步骤:
步骤1:挑取保藏编号为CGMCC No.8385的金龟子绿僵菌接种于PPDA固体培养基上,于24~30℃培养9~12d,刮取孢子置于研磨器中,用0.05%吐温-80无菌水配制成浓度为1~5×108个/mL的孢子悬液,得第一培养物;
步骤2:吸取2~4mL第一培养物,置于SDAY改良液体培养基中,于25~30℃、160~200 rpm培养70~75h,得第二培养物;
步骤3:将14~16mL第二培养物做为固态发酵的种子液转接至80~100克/袋的固态发酵培养基中,混匀后于25~30℃培养6~9 d后即得。
6.所述固态发酵培养基包括:麦壳1~3重量份,麦麸6~10 重量份,水87~93 重量份。
7.根据权利要求3、4或5任一项所述菌剂,其特征在于,其施用量为1~10万亿孢子/亩。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310657681.4A CN104694395B (zh) | 2013-12-09 | 2013-12-09 | 一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310657681.4A CN104694395B (zh) | 2013-12-09 | 2013-12-09 | 一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104694395A true CN104694395A (zh) | 2015-06-10 |
CN104694395B CN104694395B (zh) | 2017-11-10 |
Family
ID=53341955
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310657681.4A Active CN104694395B (zh) | 2013-12-09 | 2013-12-09 | 一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104694395B (zh) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105255860A (zh) * | 2015-11-23 | 2016-01-20 | 浙江大学 | 一种能使ctab法提取到真菌高质量基因组dna的方法 |
CN106929426A (zh) * | 2016-12-05 | 2017-07-07 | 中国科学院兰州化学物理研究所 | 一种马铃薯专用复合微生物菌剂及其制备方法 |
CN107058190A (zh) * | 2017-06-12 | 2017-08-18 | 北京中农富源生物工程技术有限公司 | 一种高效杀线虫的组合菌剂的制备方法 |
CN110432288A (zh) * | 2019-08-19 | 2019-11-12 | 重庆谷百奥生物研究院有限公司 | 金龟子绿僵菌CQMa421在制备防治芽菜腐烂病的药剂中的应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103131648A (zh) * | 2011-11-30 | 2013-06-05 | 河北农业大学 | 解淀粉芽孢杆菌及其在防治花生果腐病中的应用 |
-
2013
- 2013-12-09 CN CN201310657681.4A patent/CN104694395B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103131648A (zh) * | 2011-11-30 | 2013-06-05 | 河北农业大学 | 解淀粉芽孢杆菌及其在防治花生果腐病中的应用 |
Non-Patent Citations (2)
Title |
---|
RJ MILNER ET AL.: "Preliminary evalution of the use of Metarhizium anisopliae as a mycoinsecticide for control of peanut scarabs", 《CAB DIRECT》 * |
潘汝谦等: "一种值得关注的花生新病害:花生新赤壳菌基腐病", 《植物保护》 * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105255860A (zh) * | 2015-11-23 | 2016-01-20 | 浙江大学 | 一种能使ctab法提取到真菌高质量基因组dna的方法 |
CN105255860B (zh) * | 2015-11-23 | 2018-08-10 | 浙江大学 | 一种能使ctab法提取到真菌高质量基因组dna的方法 |
CN106929426A (zh) * | 2016-12-05 | 2017-07-07 | 中国科学院兰州化学物理研究所 | 一种马铃薯专用复合微生物菌剂及其制备方法 |
CN106929426B (zh) * | 2016-12-05 | 2020-06-09 | 中国科学院兰州化学物理研究所 | 一种用于马铃薯复合微生物菌剂及其制备方法 |
CN107058190A (zh) * | 2017-06-12 | 2017-08-18 | 北京中农富源生物工程技术有限公司 | 一种高效杀线虫的组合菌剂的制备方法 |
CN110432288A (zh) * | 2019-08-19 | 2019-11-12 | 重庆谷百奥生物研究院有限公司 | 金龟子绿僵菌CQMa421在制备防治芽菜腐烂病的药剂中的应用 |
CN110432288B (zh) * | 2019-08-19 | 2021-06-29 | 重庆谷百奥生物研究院有限公司 | 金龟子绿僵菌小孢变种CQMa421在制备防治芽菜腐烂病的药剂中的应用 |
Also Published As
Publication number | Publication date |
---|---|
CN104694395B (zh) | 2017-11-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101638629B (zh) | 一种防治水稻细菌性病害的蛭弧菌及其应用 | |
KR20130038057A (ko) | 복합 미생물에 의한 생약재 발효액을 이용한 생리활성 물질 함유 작물 재배방법 및 그 방법으로 재배된 생리활성 물질 함유 작물 | |
CN102803501B (zh) | 用于制备香豆雌酚的方法 | |
KR20120125042A (ko) | 복합 미생물에 의한 생약재 발효액 제조방법, 그 방법으로 제조된 발효액, 및 그 발효액에 의해 제조된 발효식품 | |
CN105087423A (zh) | 新型芽孢杆菌cjbv及含有其的抗菌组合物 | |
CN106282067B (zh) | 多功能农用复合微生物菌剂及微生态制剂与应用 | |
Luangharn et al. | Antibacterial activity, optimal culture conditions and cultivation of the medicinal Ganoderma australe, new to Thailand | |
CN100406552C (zh) | 一株成团泛菌及其发酵培养方法与应用 | |
KR101138684B1 (ko) | 유용미생물 배양방법 및 이를 이용한 식품 | |
KR101311192B1 (ko) | 꿀벌의 전염병 예방 및 치료 조성물의 제조방법 및 이 방법에 의한 꿀벌용 영양조성물 | |
CN104694395A (zh) | 一株对花生果腐病具有防效的金龟子绿僵菌及其发酵工艺 | |
CN108283310A (zh) | 一种萱草花酵素的制备方法以及酵素原液 | |
CN102021122A (zh) | 一种高效杀虫真菌及其用途 | |
CN100361593C (zh) | 猴头菌发酵茶的生产工艺 | |
CN105420132A (zh) | 一种酿酒酵母及其在制备皮肤外用剂中的应用 | |
KR101276789B1 (ko) | 발효 오일 및 그 발효오일을 포함하는 건강기능식품 | |
CN112889843B (zh) | 一种防治柑橘红蜘蛛的死亡谷芽胞杆菌可湿性粉剂及应用 | |
CN103749542B (zh) | 解淀粉芽孢杆菌井冈霉素复配生物杀菌剂及其应用 | |
KR101892615B1 (ko) | 락토바실러스 사케이 2-6-4 균주 및 이의 용도 | |
CN108690818A (zh) | 一种微生物菌剂及其制备方法 | |
CN108968040A (zh) | 一种番茄酵素饮料及其制备方法 | |
CN103642741A (zh) | 一种复合光合菌剂及其制备方法 | |
CN112931044A (zh) | 白囊耙齿菌的培养基及培养方法 | |
CN103734469A (zh) | 一种饲料添加剂及其制备 | |
CN109526974A (zh) | 一种高效防治水稻稻瘟病的组合物及应用方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |