CN106399131A - Penicillium rubens strain and application thereof - Google Patents
Penicillium rubens strain and application thereof Download PDFInfo
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- 241001280501 Penicillium rubens Species 0.000 title claims abstract description 16
- 241000196324 Embryophyta Species 0.000 claims abstract description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 241000894006 Bacteria Species 0.000 claims abstract description 13
- 235000020232 peanut Nutrition 0.000 claims abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- 238000002360 preparation method Methods 0.000 claims description 25
- 239000000047 product Substances 0.000 claims description 22
- 239000000284 extract Substances 0.000 claims description 14
- 244000061456 Solanum tuberosum Species 0.000 claims description 13
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 13
- 238000000855 fermentation Methods 0.000 claims description 13
- 230000004151 fermentation Effects 0.000 claims description 13
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 9
- 244000105624 Arachis hypogaea Species 0.000 claims description 9
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 9
- 235000018262 Arachis monticola Nutrition 0.000 claims description 9
- 239000000706 filtrate Substances 0.000 claims description 8
- 230000012010 growth Effects 0.000 claims description 7
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- 238000002791 soaking Methods 0.000 claims description 6
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- 239000008272 agar Substances 0.000 claims description 5
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- 244000005700 microbiome Species 0.000 claims description 5
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- 108010080698 Peptones Proteins 0.000 claims description 4
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- 230000029553 photosynthesis Effects 0.000 abstract 1
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- 238000000034 method Methods 0.000 description 22
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- 241000228143 Penicillium Species 0.000 description 6
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- IICCLYANAQEHCI-UHFFFAOYSA-N 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',4',5',7'-tetraiodospiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound O1C(=O)C(C(=C(Cl)C(Cl)=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 IICCLYANAQEHCI-UHFFFAOYSA-N 0.000 description 5
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- 229930187593 rose bengal Natural products 0.000 description 5
- 229940081623 rose bengal Drugs 0.000 description 5
- STRXNPAVPKGJQR-UHFFFAOYSA-N rose bengal A Natural products O1C(=O)C(C(=CC=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 STRXNPAVPKGJQR-UHFFFAOYSA-N 0.000 description 5
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- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000371966 Penicillus <bivalve> Species 0.000 description 2
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
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- 239000000463 material Substances 0.000 description 2
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- 238000005406 washing Methods 0.000 description 2
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- 244000241257 Cucumis melo Species 0.000 description 1
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- 241000209510 Liliopsida Species 0.000 description 1
- 244000038157 Palaquium xanthochymum Species 0.000 description 1
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- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
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- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- ZGBSOTLWHZQNLH-UHFFFAOYSA-N [Mg].S(O)(O)(=O)=O Chemical compound [Mg].S(O)(O)(=O)=O ZGBSOTLWHZQNLH-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
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- 235000019674 grape juice Nutrition 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
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- 230000002262 irrigation Effects 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 230000002073 mitogenetic effect Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000000618 nitrogen fertilizer Substances 0.000 description 1
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
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- 238000009928 pasteurization Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
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- 238000003672 processing method Methods 0.000 description 1
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- 235000013311 vegetables Nutrition 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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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
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- 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
- C12R2001/80—Penicillium
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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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- Chemical Kinetics & Catalysis (AREA)
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- Agronomy & Crop Science (AREA)
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Abstract
The invention discloses a penicillium rubens strain which is named as penicillium rubens (PR1), collected in China General Microbiological Culture Collection Center (CGMCC for short). The address is Institute of Microbiology, Chinese Academy of Sciences, #3, Yard 1, Beichen West Road, Chaoyang District of Beijing; the strain collection number is CGMCC NO. 13189. According to the invention, the penicillium rubens strain is separated and purified from wild species of peanuts for the first time. The endophyte can have a mutualism relation with plants, and the plants provide products of photosynthesis, water and mineral substances for the endophyte; meanwhile, the endophyte produces secondary metabolites to stimulate plant growth and development, and the insect pest injury resistance, bacteria resistance, stress resistance, resistance of hosts on environmental stress and the like can be improved. A novel biological function fertilizer developed by combining the secondary metabolites and novel fertilizers has the function advantages, cost advantages, fertilizer reduction and effect increase advantages, and energy conservation and environment friendliness advantages, the current industry development requirements are met, and the novel fertilizer level in China is greatly improved.
Description
Technical field
The present invention relates to one plant is produced dark purple mould and its application and in particular to one plant has growth promotion and makees to multiple kinds of crops
Product dark purple mould, belongs to biological technical field.
Background technology
Endophyte of plant (Endophyte) is certain phase or whole stage moves in the tissue and organ of health plant
The fungi in portion or bacterium.But unlike pathogen has obvious illness, endophyte of plant does not show any obvious symptom, and general
Store-through is in higher plant, woody, herbaceous plant, all has endophyte in monocotyledon and dicotyledon.Recent decades
Research show, the alcohol extracting thing of some endophyte of plant has and promotes plant growth, improve drought resisting, increase absorption of fertilizer etc.
Effect, causes the concern of botanists.Research shows, at least tens kinds endophytes of every kind of wild plant exist.Estimate roughly
Meter, about as many as 400,000 kinds of the species of endophyte of plant.From plant evolution angle analysis, the diversity of plant also necessarily leads to
The diversity of its endophyte.So, these endophytes are particularly likely that the treasure-house of valuable compounds therefrom.Become biological at present
Potential microbial pesticide, yield increasing fungus or be used as potential biological and ecological methods to prevent plant disease, pests, and erosion carrier bacterium in preventing and treating.
In recent years, China's fertilizer production and consumption are increased with speed faster always, but soil degradation, utilization rate of fertilizer are low
And resource, the energy and the manpower thereby resulting in greatly wastes and is always the huge problem that China's agricultural production faces.
Penicillium (Penicillium) bacterial classification is the important microbial resources of a class, is the group of microbial diversity
Become part, its distribution extensively, is often distributed in soil, air, food, saprophytic timber etc., with human lives, research and production
It is closely related, and provides microorganism resource for mycology research, agricultural and biotechnology industry sustainable development, there is important economy
Meaning.
Produce a kind of mould fungi belonging to Penicillium of dark purple, at present it is studied less, reported the country of this kind have Britain,
The U.S., Belgium, China, substratess include mouldy "Hami" melon, unfermentable grape juice, beer bottle cap etc..For producing, dark purple is mould
The research of bacterium application is then more rare, and it is a kind of Bacteria Culture pollution that the product penicillin that Fleming finds produces the mould P.rubens of dark purple
Thing (Houbraken et al.2011).And have and promote mould to have not yet to see report as the product dark purple of growth function.
Content of the invention
For above-mentioned prior art, it is an object of the invention to provide one plant is produced dark purple mould and its application.
For achieving the above object, the present invention adopts following technical proposals：
According to the first aspect of the invention, provide one plant of product dark purple mould, be named as the product mould (Penicillium of dark purple
Rubens) PR1, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address
For：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica), culture presevation number is CGMCC
NO.13189.
This bacterial strain is to separate to obtain from Brazilian wild-type peanut sample, and bacterial strain cultivates 7d, diameter 27-29mm, bacterium on PDA
The face that falls is flat；Quality is velvet-like or and slightly cotton-shaped；Conidium structure produces in a large number, conidium face green or celadon；Bacterium
Filament white；No diffusate；Reverse side yellow green or faint yellow；Sclerotium is no.Falx stem 36-120 × 3.0-4.0 μm, colourless, wall light
Sliding；Penicillus three is verticillate, occasionally has two-wheel raw verticillate with four；Bottle stalk ampoule shape, 6.9-9.1 × 2.7-3.7 μm, stalk neck is short；Mitogenetic
Spore is spherical, subsphaeroidal or oval, 3.2-4.2 × 2.7-3.5 μm, and wall is smooth；Conidia chain assumes loose cylinder.
According to the second aspect of the invention, provide a kind of plant induced resistance agent, its active component is that above-mentioned product dark purple is mould
Mycelial ethanol extract after the fermentation of (Penicillium rubens) PR1.
The present invention also provides the preparation method of above-mentioned plant induced resistance agent, including：Above-mentioned product dark purple of fermenting is mould
(Penicillium rubens) PR1, zymotic fluid centrifugal filtration obtains mycelial step；And mycelium is adopted second
The step that alcohol is extracted.
In above-mentioned preparation method, the raw material of the fermentation medium that fermentation adopts consists of：Potato extract 1000ml, ferment
Female cream 1.0g, peptone 3.0g, glucose 15.0g, agar 17.0g.
The preparation method of described potato extract is：Take peeled potatoes 200g, be cut into small pieces, plus distilled water 1000 milli
Rise and boil 30 minutes, filter off potato ball, filtrate is complemented to 1000 milliliters.
In above-mentioned preparation method, the condition of fermentation is：Inoculum concentration is 5-15% (percentage by volume), fermented and cultured temperature is
20-30 DEG C, cultivate 4-6 days；Preferably, the condition of fermentation is：Inoculum concentration is that 10%, fermented and cultured temperature is 25 DEG C, cultivates 5
My god.
In above-mentioned preparation method, the operation of alcohol extract is：Mycelium is dried, pulverizes, add ethanol cold soaking 20-30h,
Then ultrasonic extraction 0.5-1.5h；Repeat to extract 2-4 time, merging filtrate, obtain final product.
The operation of described alcohol extract is：Mycelium is dried at 50 DEG C, pulverizes, add ethanol cold soaking 12 hours, so
Ultrasonic extraction 2h afterwards；Repeat to extract 2 times, merging filtrate.
The addition of described ethanol is the 15-30% of mycelium (after being dried) weight；Extract the amount phase adding ethanol every time
With.
According to the third aspect of the invention we, mould (Penicillium rubens) PR1 of above-mentioned product dark purple and/or plant are provided
Inducer is promoting crop growth or is promoting the application in increasing crop yield.
In above-mentioned application, described crops are peanut, cowpea and radish.
Beneficial effects of the present invention：
(1) system of the present invention separates from Brazilian wild-type peanut first, is purified into a kind of product dark purple trichoderma strain.This endophyte can
It is in mutualism relation with plant, plant is supplied to endophyte photosynthate, water and mineral matter；Meanwhile, endophyte produces
Raw secondary metabolite stimulates growth and development of plants, improves insect pest, Anti-bacterium, the opposing to environment-stress of resistance and host
Power etc..Its secondary metabolite is combined exploitation new bio functional form fertilizer (slow-release or control-release fertilizer, fertilizer, water with new-type fertilizer
Conventional fertilizers such as molten fertilizer, nitrogenous fertilizer etc.) there is functionality advantage, cost advantage, fat-reducing synergy advantage, energy-conserving and environment-protective advantage, meet and work as
Front industry development requirement, will increase substantially China's new-type fertilizer level.
(2) present invention, can be effectively to the product mould generation of dark purple by carrying out alcohol extract to the mycelium after fermentation
Secondary metabolite is enriched with, and improves it to the growth promotion of crops and production-increasing function.
Brief description
Fig. 1：The colonial morphology of PR1 bacterial strain.
Fig. 2：The hypha form of PR1 bacterial strain and spore shape (40 × 0.65 and 100 × 1.4).
Fig. 3：The phylogenetic tree that bacterial strain PR1 is set up by ITS sequence analysis.
Specific embodiment
Involved instrument, reagent, material etc. in following embodiments, unless otherwise noted, are in prior art existing
Conventional instrument, reagent, material etc., can be either commercially available by regular.Involved experimental technique in following embodiments, inspection
Survey method etc., unless otherwise noted, is existing normal experiment method, detection method etc. in prior art.
Embodiment 1：The separation of bacterial strain and identification
1. the separation of bacterial strain and screening：
(1) separation of bacterial strain
Use aseptic water washing again after being rinsed well with running water after Brazilian wild-type peanut sample collection, be cut into after drying
0.5cm segment, is placed in 70-75% ethanol disinfection 3-5min, washes 18 minutes with 10% pasteurization liquid again, finally after rinsing well
With aseptic water washing 4 times, dry standby after surface moisture.
The above-mentioned root sterilized aseptically is inoculated on rose bengal medium flat board, is placed in 25 DEG C of degree cultures
After case is cultivated 5 days, you can see that the edge that sample cuts through grows mycelia, repeatedly separate through flat board (rose bengal medium), pure
Change, finally obtain one plant of bacterium, numbering is PR1；
Rose bengal medium formula and preparation method are as follows：Peptone 5g, glucose 10g, potassium dihydrogen phosphate 1g, sulfuric acid
Magnesium (MgSO4 7H2O) 0.5g, agar 20g, 1/3000 rose-bengal solution 100mL, distilled water 1000mL, chloramphenicol 0.1g.
After above-mentioned each composition adds dissolving in distilled water, then plus rose-bengal solution.Another a small amount of ethanol dissolves chloramphenicol, adds culture
In base, after packing, 121 DEG C of sterilizing 20min.
2. the form of bacterial strain PR1 and Biological characteristics
Bacterial strain PR1 is cultivated 7d, diameter 27-29mm in room temperature, PDA, bacterium colony face is flat；Quality is velvet-like or and slightly wads a quilt with cotton
Shape；Conidium structure produces in a large number, conidium face green or celadon；White mycelium；No diffusate；Reverse side yellow green
Or it is faint yellow；Sclerotium is no.Falx stem 36-120 × 3.0-4.0 μm, colourless, wall is smooth；Penicillus three is verticillate, occasionally have two-wheel raw and
Four is verticillate；Bottle stalk ampoule shape, 6.9-9.1 × 2.7-3.7 μm, stalk neck is short；Conidium is spherical, subsphaeroidal or oval, 3.2-
4.2 × 2.7-3.5 μm, wall is smooth；Conidia chain assumes loose cylinder.This bacterium has the feature of Penicillium, tentatively sentences
Break as producing dark purple mould (Penicillium rubens).The colonial morphology of this bacterium is as shown in figure 1, bacterium under different enlargement ratio
The hypha form of strain and spore shape as shown in Figure 2 (left figure enlargement ratio is 40 × 0.65, right figure enlargement ratio is 100 ×
1.4).
3. the molecular biology identification of bacterial strain PR1
With ITS1 and ITS4 as primer, fungi PR1 bacterial strain detached from wild-type peanut root is expanded, and right
Amplified production is sequenced；
ITS1：TCCGTAGGTGAACCTGCGG；
ITS4：TCCTCCGCTTATTGATATGC.
The sequence of amplified production is as shown in SEQ ID NO.1, specific as follows：
Based on blast search, choose and the research immediate kind of strain sequence or hithermost evolutionary branching representative strain
Sequence compare, pass through ML algorithm using Mega6.0 software, set up using 1000 achievement results of bootstrap and evolve
Tree (as shown in Figure 3).
Result shows PR1 fungi and reaches 99% with the homology similitude producing dark purple mould (Penicillium rubens), ammonia
The similitude of base acid sequence has also reached 99%.
By Morphological Identification and molecular biology identification, final qualification result bacterial strain PR1 is that product dark purple is mould
(Penicillium rubens).
Screening being separated the Strain Designation obtaining is to produce dark purple mould (Penicillium rubens) PR1, in being preserved in
(abbreviation CGMCC, address is state's Microbiological Culture Collection administration committee common micro-organisms center：Chaoyang District, Beijing City North Star west
Road 1 institute 3, Institute of Microorganism, Academia Sinica), culture presevation number is CGMCC NO.13189.
Embodiment 2：The preparation of plant induced resistance agent
Concrete preparation method is as follows：
1. fermented and cultured：
Mould (the Penicillium of product dark purple by embodiment 1 separating and preserving being saved at 4 DEG C on test tube slant
Rubens) the bacterial classification of PR1, is connected in flat board PDA culture medium, 25 DEG C cultivate 6 days, with card punch agar dig block be inoculated in equipped with
The 250mL triangular flask of 50mL seed culture fluid (PDA liquid medium), in 25 DEG C, rotary shaker under 180r/min is cultivated 3 days
As seed, inoculated equipped with the 500mL triangular flask of 150mL fermentation medium with 10% amount, cultivate 5 days under similarity condition,
Terminate fermentation, obtain seed liquor.
PDA culture medium is filled a prescription：Potato 200g, glucose 20g, agar 20g, distilled water 1000ml.
Fermentative medium formula：Potato extract 1.0L, yeast extract 1.0g, peptone 3.0g, glucose 15.0g, fine jade
Fat 17.0g.
The preparation of potato extract：Take 200 grams of peeled potatoes, be cut into small pieces, plus 1000 milliliters of distilled water boils 30
Minute, filter off potato ball, filtrate is complemented to 1000 milliliters.
2. alcohol extract：
By zymotic fluid centrifugal filtration, obtain mycelium, mycelium is dried at 60 DEG C, pulverizes, add ethanol cold soaking 24 little
When, then ultrasonic extraction 1h；Repeat to extract 3 times, merging filtrate, obtain final product plant induced resistance agent.The addition of described ethanol is mycelia
The 20% of body weight；The amount extracting addition ethanol every time is identical.
Embodiment 3：Application in peanut growth-promoting and volume increase for the plant induced resistance agent
1. test method：
Process 1：Seed dressing (plant induced resistance agent of 0.1 gram of embodiment 2 preparation mixes 30 kilograms of seeds).
Process 2：(plant induced resistance agent of 0.01 gram of embodiment 2 preparation mixes 30 kilograms of seeds, in blooming for seed dressing plus once spraying
The plant induced resistance agent of 0.02 gram of embodiment 2 preparation of later stage converts 15 kg of water dilution sprayings).
Process 3：(plant induced resistance agent of 0.01 gram of embodiment 2 preparation mixes 30 kilograms of seeds for seed dressing plus secondary spraying.In blooming
The plant induced resistance agent of 0.02 gram of embodiment 2 preparation of later stage converts 15 kg of water dilution sprayings；Harvest front 1 month second spraying).
Each process is repeated 3 times.
Outdoor cropping, 20 centimetres of spacing in the rows, line-spacing is 50 centimetres.50 kilograms of certain brand composite fertilizer special applied by every mu of base.
Survey product method：Often processing growth selection, to harvest 2 meters of ridges than more uniform ridge long, takes peanut and dries belt leather and weighs
Weigh with shelling.
2. result of the test：
Table 1：Volume increase result (the unit to blackcurrant pigment for the plant induced resistance agent：Kilogram)
| Process | Gross weight | Volume increase (mean value) | Grain weight | Volume increase (mean value) |
| CK | 0.79 | / | 0.57 | / |
| Process 1 | 0.875 | 10.8% | 0.645 | 13.1% |
| Process 2 | 1.065 | 34.8% | 0.795 | 39.5% |
| Process 3 | 0.885 | 12.0% | 0.65 | 14.0% |
Test result indicate that, process 2 amount of increase in production maximum, gross weight and net weight respectively reach 34.8% and 39.5%, place
The amount of increase in production of reason 1 and process 2 is close.Survey product process can be seen that the peanut processing 2 and grain is all significantly greater than at other
Reason, crude fruit is less, and comparison is ripe later, and immature water particle is relatively more.
Embodiment 4：Application in cowpea growth-promoting and volume increase for the plant induced resistance agent
1. test method：
Using method：After emerging latter 10 days, the plant induced resistance agent of 0.02 gram of embodiment 2 preparation converts 30 kg of water pouring roots, every plant
1000 milliliters, the plant induced resistance agent of 0.02 gram of embodiment 2 preparation of Post flowering converts 15 kg of water dilution sprayings.Spray after harvesting every time
Once.Cultivation mid-term is converted 30 kg of water with plant induced resistance agent prepared by 0.02 gram of embodiment 2 and is topdressed once with reference to drip irrigation.
Planting type：Spring greenhouse production.Seeding row spacing is 20 × 60 centimetres, 2 plants of every cave.It is special multiple that certain brand applied by every mu of base
50 kilograms of Hefei.
Survey product method：Harvest every time and weigh, the yield adding up initial 5 results adds up as output standard.
2. result of the test：
First five time harvest than comparison amount of increase in production 45%, process in addition to yield index, respectively process the quantity podding big
The index such as little is limited apparently higher than comparison, later stage by frame height, and yield composition is affected, and later stage yield does not count.?
The harvest yield of cowpea early stage can be significantly improved in production process, be conducive to the early yield of agricultural product to be formed, due to early stage vegetable
The price of dish is higher, so having dual the effect of increasing income.
Embodiment 5：Application in radish growth-promoting and volume increase for the plant induced resistance agent
1. test method：
Processing method：Divide 2 concentration for the treatment of, the plant induced resistance agent of respectively 0.02 gram embodiment 2 preparation and 0.01 gram of reality
The plant induced resistance agent applying example 2 preparation is watered 15 kilograms of sprayings.With spray after releasing true leaf of emerging, it is defined by soaking root, 20
Spray after it once, envelope is sprayed once behind ridge again.
Cultural method：In outdoor cropping after entering the hottest period of the summer.Seeding row spacing is 20 × 30 centimetres, and individual plant is cultivated.Three repetitions.Every mu of base
Apply rich 50 kilograms of board composite fertilizer special of friend.
Survey product method：Results are to extract radish, remove blade, radish of weighing.
2. result of the test：
Each yield that processes is respectively 10.3 kilograms of 0.02 gram of every kettle water process, 11.7 kilograms of 0.01 gram of every kettle water process, right
It is 7.5 kilograms according to yield.Increase production 37.3% and 56% respectively, the radish quality simultaneously processing substantially is improved, and has fruit
Sweet crisp mouthfeel, pungent very weak, can directly eat raw, and compare very pungent, be difficult to eat raw.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any be familiar with those skilled in the art in the technical scope of present disclosure, technology according to the present invention scheme and its invention
Design in addition equivalent or change, all should be included within the scope of the present invention.
SEQUENCE LISTING
<110>Tai'an Yu Da bio tech ltd
<120>One plant is produced dark purple mould and its application
<130> 2016
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 677
<212> DNA
<213>Bacterial strain PR1
<400> 1
ttgatatgct taagttcagc gggtatccct acctgatccg aggtcaacct ggataaaaat 60
ttgggttgat cggcaagcgc cggccgggcc tacagagcgg gtgacaaagc cccatacgct 120
cgaggaccgg acgcggtgcc gccgctgcct ttcgggcccg tcccccggga tcggaggacg 180
gggcccaaca cacaagccgt gcttgagggc agaaatgacg ctcggacagg catgcccccc 240
ggaataccag ggggcgcaat gtgcgttcaa agactcgatg attcactgaa tttgcaattc 300
acattacgta tcgcatttcg ctgcgttctt catcgatgcc ggaaccaaga gatccgttgt 360
tgaaagtttt aaataattta tattttcact cagactacaa tcttcagaca gagttcgagg 420
gtgtcttcgg cgggcgcggg cccgggggcg taagcccccc ggcggccagt taaggcgggc 480
ccgccgaagc acaaggtaaa ataaacacgg gtgggaggtt ggacccagag ggccctcact 540
cggtaatgat ccttccgcag gttcacctac ggaagccagg ggggctccaa aaagcgctgg 600
actacctgat ccgaggtcac ctggataaaa atttgggttg atcggcaagc gccggccggg 660
cctacagagc gggtgac 677
Claims (10)
1. one plant of product dark purple is mould, is named as product dark purple mould (Penicillium rubens) PR1, has been preserved in China Microbiological bacterium
Plant preservation administration committee common micro-organisms center, abbreviation CGMCC, address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, culture presevation number is CGMCC NO.13189.
2. a kind of microbial inoculum, its active component is sending out of product dark purple mould (Penicillium rubens) PR1 described in claim 1
Mycelial ethanol extract after ferment.
3. the preparation method of the microbial inoculum described in claim 2 is it is characterised in that include：Above-mentioned product dark purple of fermenting is mould
(Penicilliumrubens) PR1, zymotic fluid centrifugal filtration obtains mycelial step；And mycelium is adopted ethanol
The step extracted.
4. preparation method as claimed in claim 3 is it is characterised in that the raw material of the fermentation medium of fermentation employing consists of：
Potato extract 1000ml, yeast extract 1.0g, peptone 3.0g, glucose 15.0g, agar 17.0g.
5. preparation method as claimed in claim 4 is it is characterised in that the preparation method of described potato extract is：Remove
Skin potato 200g, is cut into small pieces, plus 1000 milliliters of distilled water boils 30 minutes, filters off potato ball, filtrate is complemented to
1000 milliliters.
6. preparation method as claimed in claim 3 is it is characterised in that the condition of fermentation is：Inoculum concentration is 5-15%, fermentation training
Foster temperature is 20-30 DEG C, cultivates 4-6 days；Preferably, the condition of fermentation is：Inoculum concentration is 10%, fermented and cultured temperature is 25
DEG C, cultivate 5 days.
7. preparation method as claimed in claim 3 is it is characterised in that the operation of alcohol extract is：By dry for mycelium, powder
Broken, add ethanol cold soaking 20-30h, then ultrasonic extraction 0.5-1.5h；Repeat to extract 2-4 time, merging filtrate, obtain final product；Preferably
, the operation of described alcohol extract is：Mycelium is dried at 60 DEG C, pulverizes, add ethanol cold soaking 24 hours, then ultrasonic
Extract 1h；Repeat to extract 3 times, merging filtrate.
8. preparation method as claimed in claim 7 is it is characterised in that the addition of described ethanol is the 15- of mycelium weight
30%；The amount extracting addition ethanol every time is identical.
9. mould (Penicillium rubens) PR1 of the product dark purple described in claim 1 and/or microbial inoculum are promoting crop growth
Or the application in promotion increasing crop yield.
10. application as claimed in claim 9 is it is characterised in that described crops are peanut, cowpea and radish.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109957517A (en) * | 2019-04-22 | 2019-07-02 | 南京工业大学 | A kind of A Si Penicillium notatum and its microbial inoculum and application |
| CN115067363A (en) * | 2022-08-22 | 2022-09-20 | 山东蓬勃生物科技有限公司 | Application of extract of mycelium of penicillium erythraeum PR1 |
| CN117617269A (en) * | 2023-10-18 | 2024-03-01 | 山东蓬勃生物科技有限公司 | Application of Penicillium erythrorhizon PR1 mycelium extract as plant immunity inducer |
-
2016
- 2016-12-16 CN CN201611169743.7A patent/CN106399131B/en active Active
Non-Patent Citations (2)
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| WIRAT PHUWIWAT等: "The effect of Penicillium notatum on plant growth.", 《FUNGAL DIVERSITY》 * |
| 李长林等: "分离自海藻的青霉属三个中国新记录种", 《菌物学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109957517A (en) * | 2019-04-22 | 2019-07-02 | 南京工业大学 | A kind of A Si Penicillium notatum and its microbial inoculum and application |
| CN109957517B (en) * | 2019-04-22 | 2019-12-03 | 南京工业大学 | Aspecium, microbial inoculum and application thereof |
| CN115067363A (en) * | 2022-08-22 | 2022-09-20 | 山东蓬勃生物科技有限公司 | Application of extract of mycelium of penicillium erythraeum PR1 |
| CN115067363B (en) * | 2022-08-22 | 2023-01-10 | 山东蓬勃生物科技有限公司 | Application of extract of mycelium of penicillium erythraeum PR1 |
| CN117617269A (en) * | 2023-10-18 | 2024-03-01 | 山东蓬勃生物科技有限公司 | Application of Penicillium erythrorhizon PR1 mycelium extract as plant immunity inducer |
| CN117617269B (en) * | 2023-10-18 | 2024-04-12 | 山东蓬勃生物科技有限公司 | Application of Penicillium erythrorhizon PR1 mycelium extract as plant immunity inducer |
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