CN106967624A - One plant of Fusarium oxysporum and its application in officinal dendrobium stem plantation - Google Patents

One plant of Fusarium oxysporum and its application in officinal dendrobium stem plantation Download PDF

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Publication number
CN106967624A
CN106967624A CN201710358315.7A CN201710358315A CN106967624A CN 106967624 A CN106967624 A CN 106967624A CN 201710358315 A CN201710358315 A CN 201710358315A CN 106967624 A CN106967624 A CN 106967624A
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Prior art keywords
fusarium oxysporum
jsnl008
dendrobium candidum
bacterial strain
plant
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CN201710358315.7A
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CN106967624B (en
Inventor
徐超
张红岩
杨鹤同
席刚俊
汪善锋
郑凯
史俊
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Jiangsu Polytechnic College of Agriculture and Forestry
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Jiangsu Polytechnic College of Agriculture and Forestry
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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
    • C12N1/14—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
    • C12R2001/77—Fusarium
    • 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
    • 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

Abstract

The invention discloses one plant of Fusarium oxysporum and its application in officinal dendrobium stem plantation.Bacterial strain JSNL008 2, it is identified as Fusarium oxysporum (Fusarium oxysporum), China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, deposit number is CGMCC NO.13689, and preservation date is on March 10th, 2017.2 couples of Fusarium oxysporum (Fusarium oxysporum) JSNL008 of the present invention promotes Growth of Dendrobium candidum effect significantly, has certain effect to the use for reducing chemical fertilizer, it is ensured that the important means of edible safety.The microbial bacterial agent that bacterial strain JSNL008 2 is produced simultaneously can remarkably promote Growth of Dendrobium candidum, and condition of culture is simple, easily preserve, it is easy to industrialized production, with good development prospect.

Description

One plant of Fusarium oxysporum and its application in officinal dendrobium stem plantation
Technical field
The invention belongs to microbial technology field, and in particular to one plant of Fusarium oxysporum and its in officinal dendrobium stem plantation Using.
Background technology
Dendrobium (Dendrobium) is the second largest category of orchid family, and herbaceos perennial, the whole world amounts to more than 1500 kinds. 《Chinese Pharmacopoeia》The stem of noble dendrobium recorded is divided into two classes, the i.e. stem of noble dendrobium and dendrobium candidum, the latter's dendrobium candidum by version in 2010 (D.officinale Kimuraet Migo) is also extraordinary to nourish and the effect that takes care of health in addition to having good medicinal efficacy, It is extremely precious Chinese medicine, have the laudatory title of " gold in medicine ".
Dendrobium candidum leaf area is small, and rate of photosynthisis is low, requires harsh to growing environment, grows nonparasitically upon another plant on big trunk and stone more Between wall, crack of stone, therefore grow very slow.Further, since excessive excavation and environmental destruction, the stem of noble dendrobium are classified as by China It is on the brink of one of shielded medicinal material kind of extinction.The in imminent danger and demand status of the stem of noble dendrobium causes numerous domestic R＆D institution to the stem of noble dendrobium Resource conducts a research, and these researchs have taken in terms of Dendrobium Plants, biological characteristics, tissue cultures and cultivation domestication is found out Obtained many achievements.But the cultivation of the current stem of noble dendrobium is produced still based on field planting pattern, in order to meet the market demand, stone is improved Dry measure used in former times yield, blindly using chemical fertilizer and agricultural chemicals, has a strong impact on its product quality.How effective development organic farming pattern, carry High product quality turns into the bottleneck of the industry benign development.
Research shows that Seeds of Dendrobium Candidum lacks endosperm tissue, and the development of embryo is no more than globular stage, and its is sexual numerous Difficulty is grown, need to could be sprouted with mycosymbiosis.Adult orchid forms one kind that endo-mycorrhiza is nature with mycosymbiosis Universal phenomenon, nutriment necessary to mycorrhizal fungi provides growth, breeding for orchid, has obvious growth-promoting to plant Effect.Mycorhiza resource is rationally utilized, the speed of growth of dendrobium candidum is improved, the administration of chemical fertilizer is reduced to a certain extent Amount, this also complies with the strategy of sustainable development of agricultural.Therefore, mycorhiza correlative study is important as one of dendrobium candidum industry Direction.
The content of the invention
Goal of the invention：The problem of existing for prior art, the present invention provides one plant of Fusarium oxysporum JSNL008-2, right Dendrobium candidum growth-promoting functions effect is notable.It is a further object of the present invention to provide the microbial inoculum of bacterial strain production, prepared by the bacterial strain Solid microbial can remarkably promote Growth of Dendrobium candidum.
Technical scheme：To achieve these goals, one plant of Fusarium oxysporum (Fusarium as described in the present invention Oxysporum), the entitled JSNL008-2 of bacterial strain, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms Center, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode：100101, protect It is CGMCC NO.13689 to hide numbering, and preservation date is on March 10th, 2017.The bacterial strain is in March, 2016 in Jiangxi Province's Yingtan Separation acquisition on the preferable plant of upgrowth situation in dendrobium candidum plant area of city.Bacterial strain JSNL008-2 principal biologicals are special Property is：Cultivated on PDA plate, mycelia is milk yellow, growth is radial, invests substratess surface；Reverse side milk yellow, non-pigment Deposition.
Applications of the bacterial strain JSNL008-2 of the present invention in officinal dendrobium stem plantation.
The microbial bacterial agent of bacterial strain JSNL008-2 productions of the present invention.
Bacterial strain JSNL008-2 of the present invention produces the preparation method of microbial bacterial agent, by made by following steps：
(1) by culture matrix of cotton seed hulls bacteria residue, ammonium nitrate be that external source nitrogen source, sucrose are external source carbon source, be prepared into solid Culture medium, sterilizing is inoculated with Fusarium oxysporum (Fusarium oxysporum) JSNL008-2；
(2) at 28-30 DEG C, cultivated 20~25 days under the conditions of daily 8h illumination 16h dark, pH6.0；
(3) step (2) cultured solid microbial is cut into 1cm3The block of size, it is standby.
Wherein, step (1) described sterilising conditions are sterilizing under 121 DEG C and 101.3 kPas of condition of high voltage 20 minutes.
Application of the microbial bacterial agent of bacterial strain JSNL008-2 productions of the present invention in iron in officinal dendrobium stem plantation.
Wherein, the application is concretely comprised the following steps：Solid microbial inoculation is placed in dendrobium candidum seedling cultivation base Matter surface, the solid fungicide inoculum concentration of every clump of dendrobium candidum seedling is 5-6g.
Further, the dendrobium candidum seedling cultivation matrix relative water content maintains more than 60% at least 10 days.With Ensure that mycelia survives in the substrate.
Beneficial effect：Compared with prior art, it is an advantage of the invention that：
(1) Fusarium oxysporum (Fusarium oxysporum) JSNL008-2 of the invention is to promoting Growth of Dendrobium candidum Effect significantly, has certain effect to the use for reducing chemical fertilizer, is the important means for ensureing edible safety.
(2) microbial bacterial agent of Fusarium oxysporum (Fusarium oxysporum) JSNL008-2 productions can be remarkably promoted Growth of Dendrobium candidum, and condition of culture is simple, easily preserve, it is easy to industrialized production, with good development prospect.
Brief description of the drawings
Fig. 1 is bacterial strain JSNL008-2 bacterium colony photos of the present invention；
Fig. 2 is the growth-promoting capability analysis figure of bacterial strain JSNL008-2 Mycorrhizal dendrobium candidums of the present invention.
Embodiment
Below in conjunction with drawings and examples, the invention will be further described.
Embodiment 1
Separation screening promotes to one plant on the preferable plant of upgrowth situation in Jiangxi Province's Yingtan dendrobium candidum plant area Growth of Dendrobium candidum bacterial strain JSNL008-2, the bacterial strain is cultivated on PDA plate, and mycelia is milk yellow, and growth is radial, attached In substratess surface；Reverse side milk yellow, non-pigment deposition, as shown in Figure 1.Expand bacterial strain JSNL008-2 ITS areas complete sequence simultaneously It is sequenced, the rDNA ITS sequences complete sequence that PCR amplifications are obtained is as shown in SEQ ID No 1, by enterprising in genebank Row, which is compared, to be found, as a result shows bacterial strain JSNL008-2 and Fusarium oxysporum (Fusarium oxysporum) bacterial strain homology Recently, with bacterial strain Fusarium oxysporum ELRF 7 and Fusarium oxysporum FJAT-31103 homologys most Height, combining form and physiological and biochemical property, Fusarium oxysporum (Fusarium is initially identified as by bacterial strain JSNL008-2 Oxysporum), Fusarium oxysporum JSNL008-2 (Fusarium oxysporum JSNL008-2) is named as, belongs to Fungi Imperfecti Subphylum knurl seat spore mesh Fusarium.Bacterial strain JSNL008-2 is delivered positioned at BeiJing, China, Institute of Microorganism, Academia Sinica China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC) preservation, deposit number is CGMCC NO.13689, preservation date is on March 10th, 2017.
Embodiment 2
The microbial bacterial agent of Fusarium oxysporum (Fusarium oxysporum) JSNL008-2 productions：
(1) by culture matrix of cotton seed hulls bacteria residue, ammonium nitrate be that external source nitrogen source, sucrose are external source carbon source, be prepared into solid Culture matrix loads in autoclave bags, in being sterilized 20 minutes under 121 DEG C and 101.3 kPas of condition of high voltage, treats culture matrix Aseptically Arbuscular Mycorrhizal Fungi fungi JSNL008-2 after cooling, is then manually cultivated；
(2) the specific condition of culture manually cultivated is 28-30 DEG C, daily 8h illumination 16h is dark, cultivated under the conditions of pH6.0 20~25 days,
(3) step (2) cultured solid microbial is cut into 1cm3The block of size, it is standby.
Embodiment 3
The microbial bacterial agent of Fusarium oxysporum (Fusarium oxysporum) JSNL008-2 productions is to promoting dendrobium candidum Growth effect analysis.
(1) by the cultured solid microbial of embodiment 2, dendrobium candidum seedling cultivation matrix surface is placed in, is cultivated Dendrobium candidum selects bore 15cm nutritive cube, and each nutritive cube cultivates 1 clump of (5 plants) dendrobium candidum.Growing way base is selected during inoculation This consistent dendrobium candidum plant, and carry out the measure of height of seedling and ground diameter.The solid fungicide inoculum concentration of every clump of dendrobium candidum seedling For 5-6g, after microbial inoculum, Relative water content maintains more than 60% at least 10 days, with ensure mycelia in the substrate into It is living.
(2) 6 months after being inoculated with, the Mycorrhizal degree detecting of dendrobium candidum root system is carried out, it is ensured that the Fusarium oxysporum of inoculation (Fusarium oxysporum) JSNL008-2 is combined with dendrobium candidum root, forms mycorhiza.12 months after inoculation, enter The growth-promoting effect analysis of row dendrobium candidum Mycorrhizal plant is as shown in table 1 and Fig. 2.Choose without inoculation Fusarium oxysporum The confession examination plant of the microbial bacterial agent of (Fusarium oxysporum) JSNL008-2 productions, is only inoculated with equivalent cotton seed hulls bacteria residue Dendrobium candidum plant be adjoining tree (CK), and use identical cultural method.
Growth-promoting effects of the JSNL008-2 of table 1 to dendrobium candidum plant
In artificial infection Fusarium oxysporum (Fusarium oxysporum) JSNL008-2 it can be seen from table 1 and Fig. 2 The solid fungicide of production is after 12 months, is inoculated with the dendrobium candidum plant heights of seedling of JSNL008-2 microbial inoculums, fresh weight apparently higher than not being inoculated with The dendrobium candidum plant of JSNL008-2 microbial inoculums.
SEQUENCE LISTING
<110>Jiangsu Polytechnic College of Agriculture and Forestry
<120>One plant of Fusarium oxysporum and its application in officinal dendrobium stem plantation
<130> 2017
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 517
<212> DNA
<213>Fusarium oxysporum（Fusarium oxysporum）
<400> 1
gatatgctta agttcagcgg gtattcctac ctgatccgag gtcaacattc agaagttggg 60
gtttaacggc gtggccgcga cgattaccag taacgagggt tttactacta cgctatggaa 120
gctcgacgtg accgccaatc aatttgagga acgcgaatta acgcgagtcc caacaccaag 180
ctgtgcttga gggttgaaat gacgctcgaa caggcatgcc cgccagaata ctggcgggcg 240
caatgtgcgt tcaaagattc gatgattcac tgaattctgc aattcacatt acttatcgca 300
ttttgctgcg ttcttcatcg atgccagaac caagagatcc gttgttgaaa gttttgattt 360
atttatggtt ttactcagaa gttacatata gaaacagagt tttaggggtc ctctggcggg 420
ccgtcccgtt ttaccgggag cgggctgatc cgccgaggca acaagtggta tgttcacagg 480
ggtttgggag ttgtaaactc ggtaatgatc cctccgc 517

Claims (8)

1. one plant of Fusarium oxysporum (Fusarium oxysporum), the entitled JSNL008-2 of bacterial strain has been preserved in Chinese micro- life Thing culture presevation administration committee common micro-organisms center, the preservation time is on March 10th, 2017, and deposit number is CGMCC NO.13689。
2. applications of the bacterial strain JSNL008-2 in officinal dendrobium stem plantation described in claim 1.
3. the microbial bacterial agent that bacterial strain JSNL008-2 described in a kind of utilization claim 1 is produced.
4. the preparation method of the microbial bacterial agent described in a kind of claim 3, it is characterised in that by made by following steps：
(1) by culture matrix of cotton seed hulls bacteria residue, ammonium nitrate be that external source nitrogen source, sucrose are external source carbon source, be prepared into solid culture Base, sterilizing is inoculated with Fusarium oxysporum (Fusarium oxysporum) JSNL008-2；
(2) at 28-30 DEG C, cultivated 20~25 days under the conditions of daily 8h illumination 16h dark, pH6.0；
(3) step (2) cultured solid microbial is cut into 1cm3The block of size, it is standby.
5. preparation method according to claim 3, it is characterised in that step (1) described sterilising conditions be 121 DEG C and Sterilized 20 minutes under 101.3 kPas of condition of high voltage.
6. a kind of application of the microbial bacterial agent described in utilization claim 3 in officinal dendrobium stem plantation.
7. application according to claim 6, it is characterised in that the application is concretely comprised the following steps：By solid microbe bacterium Agent inoculation is placed in dendrobium candidum seedling cultivation matrix surface, and the solid fungicide inoculum concentration of every clump of dendrobium candidum seedling is 5-6g.
8. application according to claim 6, it is characterised in that the dendrobium candidum seedling cultivation matrix relative water content dimension Hold more than 60% at least 10 days.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109182144A (en) * 2018-09-29 2019-01-11 江苏农林职业技术学院 The fungal elicitor and the preparation method and application thereof for promoting purple dendrobium seed to sprout
CN114806887A (en) * 2022-03-28 2022-07-29 云南大学 Fungus strain for promoting germination of dendrobium chrysosma and application

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103503924A (en) * 2013-08-29 2014-01-15 中国农业科学院蔬菜花卉研究所 Preparation method and applications of fusarium oxysporum inoculant
CN104195055A (en) * 2014-09-10 2014-12-10 江苏农林职业技术学院 Fusarium oxysporum and application thereof in resisting leaf spot disease of dendrobium officinale
CN105219656A (en) * 2015-09-25 2016-01-06 江苏农林职业技术学院 A kind of Fusarium oxysporum and the application in promotion Growth of Dendrobium candidum thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103503924A (en) * 2013-08-29 2014-01-15 中国农业科学院蔬菜花卉研究所 Preparation method and applications of fusarium oxysporum inoculant
CN104195055A (en) * 2014-09-10 2014-12-10 江苏农林职业技术学院 Fusarium oxysporum and application thereof in resisting leaf spot disease of dendrobium officinale
CN105219656A (en) * 2015-09-25 2016-01-06 江苏农林职业技术学院 A kind of Fusarium oxysporum and the application in promotion Growth of Dendrobium candidum thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109182144A (en) * 2018-09-29 2019-01-11 江苏农林职业技术学院 The fungal elicitor and the preparation method and application thereof for promoting purple dendrobium seed to sprout
CN114806887A (en) * 2022-03-28 2022-07-29 云南大学 Fungus strain for promoting germination of dendrobium chrysosma and application
CN114806887B (en) * 2022-03-28 2023-09-05 云南大学 Fungus strain for promoting germination of Jinsha Jiang Danhu and application

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