CN106978359A - One plant of Fusarium solani and its application in the anti-root rot of dendrobium candidum - Google Patents
One plant of Fusarium solani and its application in the anti-root rot of dendrobium candidum Download PDFInfo
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- CN106978359A CN106978359A CN201710358832.4A CN201710358832A CN106978359A CN 106978359 A CN106978359 A CN 106978359A CN 201710358832 A CN201710358832 A CN 201710358832A CN 106978359 A CN106978359 A CN 106978359A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- 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
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Abstract
The invention discloses one plant of Fusarium solani and its application in the anti-root rot of dendrobium candidum.Bacterial strain JSNL007 1, it is identified as Fusarium solani (Fusarium solani), China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, deposit number is CGMCC NO.13686, and preservation date is on March 10th, 2017.Fusarium solani (Fusarium solani) JSNL007 1 of the present invention is notable to dendrobium candidum resistant effect effect, in particular for improving the ability of the anti-root rot of dendrobium candidum, there is certain effect to the use for reducing chemical pesticide, be the important means for ensureing edible safety.The microbial bacterial agent that bacterial strain JSNL007 1 is produced can enable the anti-root rot of dendrobium candidum significantly improve, and condition of culture is simple, easily preserve, it is easy to industrialized production, with good development prospect.
Description
Technical field
The invention belongs to microbial technology field, and in particular to one plant of Fusarium solani and its in the anti-root rot of dendrobium candidum
In application.
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 ".
Due to excessive excavation and environmental destruction, the stem of noble dendrobium is classified as the shielded medicinal material product for being on the brink of extinction by China
One of kind.At present, the market demand of dendrobium candidum is mainly by artificial cultivation, but artificial growth Growth of Dendrobium candidum speed is slow, puts down
Yield poorly, disease occurs more serious.At present, the preventing and treating of disease still relies primarily on agricultural chemicals.From preserving the ecological environment and eat
The angle of product safety is set out, and dendrobium candidum should reduce the usage amount of agricultural chemicals as far as possible as rare Chinese medicine.Orchid is to mycorhiza
Dependence it is stronger, mycorrhizal fungi and the conditional formation homobium of root system of plant are being physiologically one mutually coordinated whole
Body, and during common evolutionary, form specific morphological feature.The most feelings of the correlation of mycorrhizal fungi and host plant
Mutually beneficial under condition, mycorrhizal fungi obtains the required nutriment of growth out of plant, and plant by with symbiotic effects knot
Close, nutrient and moisture needed for preferably also being obtained from soil etc. improve the growth potential of plant, improve the disease-resistant energy of plant
Power.Mycorhiza resource is rationally utilized, the resistance against diseases of dendrobium candidum is improved, the amount of application of chemical pesticide is reduced to a certain extent,
This also complies with the strategy of sustainable development of agricultural.Therefore, mycorhiza correlative study turns into an important side of dendrobium candidum industry
To.
The content of the invention
Goal of the invention：The problem of existing for prior art, the present invention provides the anti-root rot bacterial strain of one plant of dendrobium candidum
JSNL007-1, the resistance against diseases for improving the anti-root rot of dendrobium candidum.It is a further object of the present invention to provide bacterial strain production
Microbial inoculum and its application, the bacterial strain prepare solid microbial the anti-root rot of dendrobium candidum can be enable to significantly improve.
Technical scheme：To achieve these goals, one plant of Fusarium solani (Fusarium as described in the present invention
Solani), the entitled JSNL007-1 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, deposit number
For CGMCC NO.13686, preservation date is on March 10th, 2017.The bacterial strain is in June, 2015 in Jiangxi Province's Yingtan iron sheet
Separation acquisition on the preferable plant of upgrowth situation in stem of noble dendrobium plant area.Bacterial strain JSNL007-1 Main Biologicals are：
Cultivated on PDA plate, mycelia is loose, villiform；Front is canescence, and the back side is milk yellow.
Applications of the bacterial strain JSNL007-1 of the present invention in the anti-root rot of dendrobium candidum.
The microbial bacterial agent of bacterial strain JSNL007-1 productions of the present invention.
Bacterial strain JSNL007-1 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 solani (Fusarium solani) JSNL007-1；
(2) at 27-30 DEG C, cultivated 20~30 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 JSNL007-1 productions of the present invention in the anti-root rot of dendrobium candidum.
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 solani (Fusarium solani) JSNL007-1 of the invention is to dendrobium candidum resistant effect effect
Significantly, in particular for improving the resistance against diseases of the anti-root rot of dendrobium candidum, there is certain effect to the use for reducing chemical pesticide,
Ensure the important means of edible safety.
(2) microbial bacterial agent of Fusarium solani (Fusarium solani) JSNL007-1 productions can resist dendrobium candidum
Root rot can be significantly improved, and condition of culture is simple, easily preserve, it is easy to industrialized production, with good development and application
Prospect.
Brief description of the drawings
Fig. 1 is bacterial strain JSNL007-1 bacterium colony photos of the present invention；
Fig. 2 is the anti-root rot capability analysis figure of bacterial strain JSNL007-1 Mycorrhizal dendrobium candidums of the present invention.
Embodiment
Below in conjunction with drawings and examples, the invention will be further described.
Embodiment 1
In Jiangxi Province's Yingtan dendrobium candidum plant area on the preferable plant of upgrowth situation separation screening to one plant of iron sheet
The anti-root rot bacterial strain JSNL007-1 of the stem of noble dendrobium, the bacterial strain is cultivated on PDA plate, and mycelia is loose, villiform；Front is canescence,
The back side is milk yellow, as shown in Figure 1.
Expand bacterial strain JSNL007-1 ITS areas complete sequence and be sequenced, the rDNA ITS sequences that PCR amplifications are obtained
Complete sequence, as shown in SEQ ID No 1, by the way that discovery is compared on genebank, as a result show bacterial strain JSNL007-1 with
The homology of Fusarium solani (Fusarium solani) bacterial strain is nearest, with bacterial strain Fusarium solani
QK1409120101 and Fusarium solani CDR3P2F2 homologys all reach 99%, and combining form and Physiology and biochemistry are special
Levy, bacterial strain JSNL007-1 is initially identified as Fusarium solani (Fusarium solani), Fusarium solani is named as
JSNL007-1 (Fusarium solani JSNL007-1), belongs to Deuteromycotina knurl seat spore mesh Fusarium.By the bacterial strain
JSNL007-1 is delivered positioned at BeiJing, China, the Chinese microorganism strain preservation conservator of Institute of Microorganism, Academia Sinica
Meeting common micro-organisms center (abbreviation CGMCC) preservation, deposit number is CGMCC NO.13686, and preservation date is in March, 2017
10 days.
Embodiment 2
The microbial bacterial agent of Fusarium solani (Fusarium solani) JSNL007-1 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 JSNL007-1 after cooling, is then manually cultivated；
(2) the specific condition of culture manually cultivated is 27-30 DEG C, daily 8h illumination 16h is dark, cultivated under the conditions of pH6.0
20~30 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 solani (Fusarium solani) JSNL007-1 productions is to dendrobium candidum root rot
Resistance against diseases action effect analysis.
(1) by the cultured solid microbial of embodiment 2, dendrobium candidum seedling cultivation matrix surface, every clump are placed in
The solid fungicide inoculum concentration of dendrobium candidum seedling is 5-6g, and after microbial inoculum, Relative water content maintains 60% with up to
It is few 10 days, to ensure that mycelia survives in the substrate.
(2) Pathogens Causing Root Rot Disease by ultraviolet light and dark processing culture in PDA culture medium, is made germ spore and hangs
Supernatant liquid, is diluted to 2 × 105Individual/mL；
(3) step (1) is under equal conditions managed for examination dendrobium candidum plant, chosen for examination plant top the
Five leaves, wound is manually manufactured with needle point method, in every plant of the blade surface sprinkling pathogen spore suspension 2ml/ of manufacture wound,
Warm indoor temperature is 30-35 DEG C, humidity 95%, and every kind of processing is 100 plants, and sets 3 groups of repetitions；Choose simultaneously rotten without inoculation
Confession examination plant the 5th, the top leaf conduct of the microbial bacterial agent of skin sickle-like bacteria (Fusarium solani) JSNL007-1 productions
Compare (CK), every plant of pathogen spore suspension 2ml/ is sprayed using identical method, and use identical cultural method.
(4) occurring degree of root rot, the statistics incidence of disease, disease index are monitored after being inoculated with daily.
Diseased plant grade scale：Severity Scaling is main to there is Lesion number and diameter according to inoculation blade tow sides, just fixed
Grade scale is：
0：Normally；
Ⅰ：Tow sides have a small amount of scab；
Ⅱ：Tow sides have scab, and onset area accounts for the blade gross area below 1/4；
Ⅲ：Tow sides have scab, and onset area accounts for the 1/4-1/2 of the blade gross area；
Ⅳ：Tow sides have scab, and onset area accounts for blade gross area 1/2-3/4；
Ⅳ：Tow sides have scab, and onset area accounts for the blade gross area more than 3/4.
(total strain number × highest level represents number to disease-sensitive index=∑ (the diseased plant numbers at different levels × level represents numerical value) × 100/
Value)；
Susceptible gene=(susceptible strain tree/inoculation strain tree) × 100%.
It is not inoculated with being planted for examination for the microbial bacterial agent that Fusarium solani (Fusarium solani) JSNL007-1 is produced
Strain and the susceptible gene and the result of disease-sensitive index of microbe inoculation microbial inoculum, as shown in Figure 2.
Shown by Fig. 2, in 30 days after artificial infection pathogen spore, be inoculated with Fusarium solani (Fusarium
Solani) the dendrobium candidum plant susceptible gene and disease-sensitive index of the solid fungicide of JSNL007-1 productions, which are significantly lower than, is not inoculated with
The dendrobium candidum plant of Fusarium solani (Fusarium solani) JSNL007-1 microbial inoculums.
SEQUENCE LISTING
<110>Jiangsu Polytechnic College of Agriculture and Forestry
<120>One plant of Fusarium solani and its application in the anti-root rot of dendrobium candidum
<130> 2017
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 502
<212> DNA
<213>Fusarium solani（Fusarium solani）
<400> 1
aagttgggtg ttttacggca tggccgcgcc gctctccagt tgcgaggtgt tagctactac 60
gcaatggaag ctgcggcggg accgccactg tatttgaggg acggcgtgtg cccacagggg 120
gcttccgccg atccccaacg ccaggcccgg gggcctgagg gttgtaatga cgctcgaaca 180
ggcatgcccg ccagaatact ggcgggcgca atgtgcgttc aaagattcga tgattcactg 240
aattctgcaa ttcacattac ttatcgcatt tcgctgcgtt cttcatcgat gccagagcca 300
agagatccgt tgttgaaagt tttaatttat ttgcttgttt actcagaaaa acattataaa 360
aacagagtta ggggtcctct ggcgggggcg gcccgttgtt acagggccgt ctgttcccgc 420
cgaagcaacg ttttaggtat gttcacaggg ttgatgagtt gtataactcg gtaatgatcc 480
ctccgcaggt caccctacgg aa 502
Claims (8)
1. one plant of Fusarium solani (Fusarium solani), the entitled JSNL007-1 of bacterial strain has been preserved in China Microbiological bacterium
Preservation administration committee common micro-organisms center is planted, deposit number is CGMCC NO.13686, and preservation date is March 10 in 2017
Day.
2. applications of the bacterial strain JSNL007-1 in the anti-root rot of dendrobium candidum described in claim 1.
3. the microbial bacterial agent that bacterial strain JSNL007-1 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 solani (Fusarium solani) JSNL007-1；
(2) at 27-30 DEG C, cultivated 20~30 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
101.3 kPa condition of high voltage under sterilize 20 minutes.
6. application of the microbial bacterial agent in the preventing and treating anti-root rot of dendrobium candidum described in a kind of utilization claim 3.
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 (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108485986A (en) * | 2018-03-13 | 2018-09-04 | 上海市园林科学规划研究院 | A kind of cinnamomum camphora microorganism mix bacterium agent and its application |
| CN108517305A (en) * | 2018-04-18 | 2018-09-11 | 山东省果树研究所 | One kind having the active perfectly round streptomycete LZH-48 of antagonism pears root rot and its application |
| CN109136327A (en) * | 2018-10-08 | 2019-01-04 | 华中农业大学 | A method of identification citrus plant Resistance To Root Rot Disease |
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| 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 |
| CN107418906A (en) * | 2017-03-13 | 2017-12-01 | 青岛九和宜生生物科技有限公司 | A kind of composite bacteria agent and its application for improving color green pepper yield and anti-root rot |
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2017
- 2017-05-19 CN CN201710358832.4A patent/CN106978359B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN107418906A (en) * | 2017-03-13 | 2017-12-01 | 青岛九和宜生生物科技有限公司 | A kind of composite bacteria agent and its application for improving color green pepper yield and anti-root rot |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108485986A (en) * | 2018-03-13 | 2018-09-04 | 上海市园林科学规划研究院 | A kind of cinnamomum camphora microorganism mix bacterium agent and its application |
| CN108517305A (en) * | 2018-04-18 | 2018-09-11 | 山东省果树研究所 | One kind having the active perfectly round streptomycete LZH-48 of antagonism pears root rot and its application |
| CN109136327A (en) * | 2018-10-08 | 2019-01-04 | 华中农业大学 | A method of identification citrus plant Resistance To Root Rot Disease |
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