CN103436452B - Acremonium persicinum JX524288 and application thereof - Google Patents
Acremonium persicinum JX524288 and application thereof Download PDFInfo
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Abstract
The invention relates to acremonium persicinum JX524288 with a collection number of CCTCC NO (China Center For Type Culture Collection Number): M2013200. The invention further provides an application of fermentation liquid of the strain in antagonizing pathogenic fungi and promoting soybean germinating.The acremonium persicinum JX524288 has a broad-spectrum plant pathogenic fungus anagonism function, can be used for preventing and controlling diseases in crop, can avoid harms caused by application of chemical reagents as a metabolite of the strain is relatively safe to plant seeds and has a function of promoting the soybean seed germinating, and has wide application prospects in agricultural production.
Description
Technical field
The invention belongs to microorganism field, relate to the mould JX524288 of strain peachiness top spore and an application thereof, particularly a strain can wide spectrum antagonizing pathogenic fungi and the mould JX524288 of peachiness top spore and the application thereof that promote soybean germination.
Background technology
Plant diseases is one of natural disaster of serious harm agriculture production, according to Food and Argriculture OrganizationFAO, estimates, the grain drop in production causing because of disease every year is in the world more than 10%.Use chemical bactericide can effectively control causing harm of Plant diseases, but there is contaminate environment, destroy the problems such as the eubiosis, drug residue in chemical bactericide, especially since the 1950's, global environment suffers unprecedented damage and pollution, < < Agenda 21 > > has been passed through in United Nations Conference on Environment and Development in 1992, established the strategic position of Sustainable development in global economy and society, therefore, the biological control of Plant diseases more and more comes into one's own.
It is that some antagonistic microbe starts for the inhibition of soil-borne pathogen from sanfprd (1926) report soil that the science of biological control is recorded.The research work of China's plant biological and ecological methods to prevent plant disease, pests, and erosion starts from the initial stage fifties, and since the seventies, the research and practice of biological control of diseases is very active, and development rapidly.In recent years, the documents and materials of relevant biological control both at home and abroad sharply increased, and the production practice of biological control are also expanding.
Biological control microorganism from saprophytic soil microorganisms to plant materials microecosystem the utilization of microorganism; The gas of the kind of preventing and treating disease from kind of biography and soil-borne disease to overground part passes and worm biography disease.Due to biological control have advantages of environment, ecology and human health safer, therefore, exploitation and the research of countries in the world attention Biocontrol microorganism very.
Current research has found that many microorganisms have biological and ecological methods to prevent plant disease, pests, and erosion function, on producing the fungi of widespread use have that wood is mould, chaetomium, yeast, Paecilomyces lilacinus, Verticillium chlamydosporiun and mycorrhizal fungi etc.Li Li has reported the mould growth-inhibiting effect to cabbage caterpillar of top spore, and Song Liwen has reported the mould restraining effect to aphid of top spore, but top spore is mould, in suppressing plant pathogenic fungi, have not been reported.
Summary of the invention
The first object of the present invention is to provide the mould JX524288 of a strain peachiness top spore, and when having the antagonizing pathogenic fungi effect of wide spectrum, its metabolite has certain promoter action to Germination of Soybean Seed, for biocontrol of plant disease provides new Microbial resources.
The second object of the present invention is to provide the purposes of the mould JX524288 of above-mentioned peachiness top spore.
The present invention is achieved through the following technical solutions:
One, strain peachiness top spore mould (Acremonium persicinum) JX524288, its deposit number is CCTCC NO:M2013200.
The imperfect stage of this microorganism belongs to imperfect fungi door Deuteromycota, hyphomycetes Hyphomycetes, Moniliales Moniliales, light color Cordycepps Moliliaceae.
In campus is selected in grand celebration, on hillside, with spade, take the soil sample that depth of soil is about 4-10cm, pack in sampling paper bag, paper bag mark is got well and take back laboratory and carry out the separation of fungi in soil and obtain.Spore mould bacterium colony mycelia in solid Cha Shi culture medium flat plate in peachiness of the present invention top is carpet shape, and bacterium colony surface is pale pink, microscopic examination demonstration, and the conidial fructification of this bacterial strain is bottle stalk, bottom obstructing, expands by bottle, alternate, spore raw on bottle stalk top.
The phylogenetic systematics of the mould fungal bacterial strain of peachiness of the present invention top spore is identified.By fungi universal primer ITS4 and ITS5, utilize pcr amplification to go out the rRNA gene of 546bp, this sequence is contrasted by the nucleotide sequence of collecting in blast program and GenBank, result shows, similarity is at more than 99% 30 the bacterial strain nucleotide sequences that have, mainly top spore mould (Acremonium), ascomycetes (Ascomycota), Cordyceps (Cordyceps), the sequence of strains A cremonium persicinum FN706545 and Antagonistic Fungus is closely similar, and length differs 1 base.The result of identify shows, this bacterial strain is peachiness top spore mould (Acremonium persicinum).
The sequence of bacterial strain of the present invention compares with clustalx software to the sequence of 20 bacterial strains similar to it in GenBank, by Neighbor Joining method in MEGA4 program, set up phylogenetic tree again, find, the sibship of Antagonistic Fungus and Acremonium persicinum FN706545 is nearest, reach 99%, verified that this bacterial strain is peachiness top spore mould (Acremonium persicinum).
The culture condition of this fungal bacterial strain is: on PDA substratum, the growth conditions of its mycelia and spore is best, and aerial hyphae growth is flourishing, and sporulation quantity is more.Take Cha Shi substratum as basic medium, and the suitableeest carbon source of mycelial growth is that sucrose, nitrogenous source are Tryptones; When pH8.5, temperature are 30 ℃, can improve mycelial growth rate, add mineral ion Zn
2+favourable to mycelial growth.
Two, the application of above-mentioned peachiness top spore mould (Acremonium persicinum) JX524288 on antagonizing pathogenic fungi and promotion soybean germination.
Adopt the positively effect of technique scheme: peachiness of the present invention top spore mould (Acremonium persicinum) JX524288 fungal bacterial strain has the effect of the antagonistic plant pathogenic fungi of wide spectrum, can be used for farm crop and prevents and treats disease; Simultaneously because the metabolite of this bacterial strain is comparatively safe to plant seed, and there is the effect that promotes Germination of Soybean Seed, therefore, can overcome and use the harm that chemical reagent brings, in agriculture production, have broad application prospects.
Accompanying drawing explanation
Fig. 1 is the colonial morphology figure of bacterial strain on solid medium;
Fig. 2 is conidiophore and conidial microscopic morphology figure of peachiness of the present invention top spore mould (Acremonium persicinum) JX524288 bacterial strain;
Fig. 3 is the aspect graph that mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and the face-off of rice sheath blight disease silk core germ are cultivated;
Fig. 4 is the Phylogenetic dendrogram of peachiness of the present invention top spore mould (Acremonium persicinum) JX524288 bacterial strain 18S rRNA sequence;
Fig. 5 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and sickle-like bacteria is cultivated;
Fig. 6 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and Exserohilum turcicum is cultivated;
Fig. 7 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and sclerotium blight of sunflower bacterium is cultivated;
Fig. 8 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and julienne potatoes pyrenomycetes is cultivated;
Fig. 9 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and soybean sclerotinia crown rot bacterium is cultivated;
Figure 10 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and fusarium graminearum is cultivated;
Figure 11 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and fusarium moniliforme is cultivated;
Figure 12 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and soybean root rot sickle-like bacteria is cultivated;
Figure 13 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and fusarium avenaceum is cultivated;
Figure 14 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and corn circle pinta bacterium is cultivated;
Figure 15 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and cucumber fusarium axysporum is cultivated;
Figure 16 is the aspect graph that the face-off of mould (Acremonium persicinum) the JX524288 bacterial strain of peachiness of the present invention top spore and tomato early blight bacterium is cultivated;
Figure 17 is the impact of spore mould (Acremonium persicinum) JX524288 fermented liquid in different concns peachiness top on soybean seeds percentage of germination;
Figure 18 is the impact of spore mould (Acremonium persicinum) JX524288 fermented liquid in different concns peachiness top on soybean seeds germinating energy;
Figure 19 is the impact of spore mould (Acremonium persicinum) JX524288 fermented liquid in different concns peachiness top on soybean seeds germination index;
Peachiness top spore involved in the present invention mould (Acremonium persicinum) JX524288, in in May, 2013 preservation center that 14 Patent Office of the People's Republic of China or international monopoly tissue are admitted carried out patented procedure preservation, depositary institution's full name is Chinese Typical Representative culture collection center, referred to as CCTCC, depositary institution address: China. Wuhan. Wuhan University, deposit number: CCTCC NO:M2013200.
Embodiment
Below in conjunction with embodiment and comparative example, technical scheme of the present invention is described further, but should not be construed as limitation of the present invention:
Embodiment 1
The separation of the present embodiment explanation bacterial strain.
Choose in campus place representative on hillside, with spade, take the soil sample that depth of soil is about 4-10cm, pack in sampling paper bag, the soil in paper bag should not have larger particles, paper bag mark is got well and take back the separation that fungi in soil is carried out in laboratory.Take the soil sample that 5g dries in the shade, put in sterilized mortar and grind, till soil sample is ground to form to uniform particle.With sterilized water, soil sample is diluted to 10
-1, 10
-2, 10
-3, 10
-4, 10
-5soil supension doubly, draws the about 0.1mL of different dilution soil supensions and is coated on PDA substratum, establishes three repetitions for every group, and 28 ℃ of thermostat containers are secretly cultivated.After bacterium colony grows, the different single bacterium colony of picking size, form, color and luster culture purified again on PDA flat board.
Wherein, the prescription of PDA flat board is: potato 200g, glucose 20g, agar 20g, water 1000ml.
The phytopathogen rhizoctonia of preserving in the refrigerator of laboratory (Rhizoctonia solani) is transferred to PDA flat board to be activated from inclined-plane, 28 ℃ of cultivations, after rhizoctonia covers with flat board, the punch tool that is 5mm with diameter breaks into some bacterium an official document or notes by rhizoctonia flat board, with in inoculating needle picking bacterium an official document or note access PDA flat board, and apart from rhizoctonia bacterium an official document or note 3cm place access from the separated fungi bacterium an official document or note obtaining in the middle of soil, distance after 28 ℃ of cultivation 2-4d between the interior fungi of measured soil and pathogenic fungi, screens the Antagonistic Fungus in soil.
Restraining effect is strong and weak to be represented with "+"." +++ " represents that restraining effect is very strong, and antibacterial radius length is greater than 6.0mm; " ++ " represents restraining effect medium tenacity, and antibacterial radius length is between 2.0mm and 6.0mm; "+" represents that restraining effect is weak or do not have activity, antibacterial radius to be less than 2.0mm or without antibacterial radius, pathogenic bacteria grows normally and covered Antagonistic Fungus.The soil sample gathering is carried out to separation, according to differences such as the color of the bacterium colony growing on flat board and forms, carry out separation and Culture, for colony colour and form, estimate similar bacterial strain, a picking one strain is preserved.By the method, it is stronger that separation has obtained a strain restraining effect, pathogenic bacteria mycelial growth inhibition rate reached to 75% bacterial strain.Morphological observation and microscopic examination by Antagonistic Fungus bacterium colony show, as depicted in figs. 1 and 2, the bacterium colony mycelia of this bacterial strain is carpet shape, and bacterium colony surface is pale pink, and conidial fructification is bottle stalk, and expand bottle stalk bottom, alternate, and spore raw on bottle stalk top.In the rhizoctonia thalline edge nearest with it, the mycelia of rhizoctonia is sparse and thin, and color is thin out, as shown in Figure 3, proves effectively antagonism rhizoctonia of this bacterial strain.By fungi universal primer ITS4 and ITS5, utilize pcr amplification to go out the rRNA gene of 546bp, this sequence is contrasted by the nucleotide sequence of collecting in blast program and GenBank, result shows, similarity is at more than 99% 30 the bacterial strain nucleotide sequences that have, mainly top spore mould (Acremonium), ascomycetes (Ascomycota), Cordyceps (Cordyceps), the sequence of strains A cremonium persicinum FN706545 and Antagonistic Fungus is closely similar, and length differs 1 base.The result of identify shows, this bacterial strain is peachiness top spore mould (Acremonium persicinum).The sequence of bacterial strain compares with clustalx software to the sequence of 20 bacterial strains similar to it in GenBank, by Neighbor Joining method in MEGA4 program, set up phylogenetic tree again, find, the sibship of Antagonistic Fungus and Acremonium persicinum FN706545 is nearest, reach 99%, verified that this bacterial strain is peachiness top spore mould (Acremonium persicinum).The qualification result of phylogenetic systematics is identical with morphologic qualification result, and Phylogenetic dendrogram as shown in Figure 4.
Embodiment 2
The mensuration of the present embodiment explanation bacterial strain antimicrobial spectrum.
What according to embodiment 1, obtain has the effective strain of stronger antagonistic ability to rhizoctonia, adopts mycelial growth to suppress method it is carried out to antimicrobial spectrum mensuration.Mycelial growth suppresses method: the various phytopathogens of preserving in the refrigerator of laboratory are transferred on PDA flat board and are activated, in 28 ℃ of thermostat containers, cultivate 4-6d, the aseptic punch tool that is 5mm with diameter breaks into bacterium an official document or note of the same size at pathogenic bacteria thalline marginal position respectively, with inoculating needle picking bacterium an official document or note, access respectively in PDA flat board, at the bacterium an official document or note apart from bacterium an official document or note 3mm place access Antagonistic Fungus, every group is repeated for 3 times, is placed in after 28 ℃ of thermostat containers are cultivated 3-5d and measures respectively the antibacterial radius between Antagonistic Fungus and various pathogenic bacteria.
Restraining effect is strong and weak to be represented with "+"." +++ " represents that restraining effect is very strong, and antibacterial radius length is greater than 6.0mm; " ++ " represents restraining effect medium tenacity, and antibacterial radius length is between 2.0mm and 6.0mm; "+" represents that restraining effect is weak or do not have activity, antibacterial radius to be less than 2.0mm or without antibacterial radius, pathogenic bacteria grows normally and covered Antagonistic Fungus.Fungal bacterial strain of the present invention, all has inhibition ability to the growth of a large amount of plant pathogenic fungi mycelia, and bacteriostasis is stronger, as shown in table 1, and this bacterial strain resists the face-off figure of multiple pathogenic fungi as shown in Fig. 5 to Figure 16.
Spore mould antimicrobial spectrum in table 1 peachiness top is measured
Note: " +++ " represents that restraining effect is very strong, and antibacterial radius length is greater than 6.0mm; " ++ " represents restraining effect medium tenacity, and antibacterial radius length is between 2.0mm and 6.0mm; "+" represents that restraining effect is weak or do not have activity, antibacterial radius to be less than 2.0mm or without antibacterial radius, pathogenic bacteria grows normally and covered Antagonistic Fungus.
Embodiment 3
The present embodiment illustrates the cultivation of this bacterial strain.
With Cha Shi substratum (prescription: SODIUMNITRATE 3g, dipotassium hydrogen phosphate 1g, magnesium sulfate (MgSO
47H
2o) 0.5g, Repone K 0.5g, ferrous sulfate 0.01g, agar 15g, distilled water 1000mL) be minimum medium, take respectively 20g glucose, sucrose, lactose, maltose, Zulkovsky starch, seminose and Mierocrystalline cellulose and replace the sucrose in Cha Shi substratum, make the substratum containing different carbon sources.Equally, take respectively 3g urea, SODIUMNITRATE, ammonium sulfate, peptone, soy peptone, Tryptones, yeast extract replace the NaNO in Cha Shi substratum
3, make the substratum containing different nitrogen sources.
With hydrochloric acid and the sodium hydroxide solution of 1mol/L, the pH value of nutrient solution is adjusted to respectively to 4.0,5.0,6.0,7.0,8.0,8.5,9.0, then adds agar powder, (121 ℃, 20min), make the substratum of different pH values in packing sterilizing.The thermostat container that is placed in respectively culture temperature and is 25 ℃, 28 ℃, 30 ℃, 32 ℃ and 35 ℃ is cultivated.Take Cha Shi substratum as minimum medium, add respectively the MnSO of 0.01g
4, CuSO
4and ZnSO
4make the substratum containing different mineral ions, cultivate 6d and start to measure every group of mycelial growth diameter, every 24h, measure once.Calculate the mycelial growth rate of bacterial strain of the present invention under different culture condition.
Found that, bacterial strain of the present invention be take Cha Shi substratum as basic medium, and the suitableeest carbon source of mycelial growth is that sucrose, nitrogenous source are Tryptones; When pH8.5, temperature are 30 ℃, can improve mycelial growth rate, add mineral ion Zn
2+favourable to mycelial growth.
Embodiment 4
The present embodiment illustrates the stability test of the metabolite of this bacterial strain.
First prepare Cha Shi substratum, with the mould spore inoculating of inoculating needle scraping peachiness top spore in substratum, be placed on shaking table 28 ℃, 160rpm shaking culture, spore mould fermented liquid in cultured peachiness top is processed to 30min under 121 ℃ of conditions, then use nuclepore membrane filter filtration sterilization, with undressed bacteria-free filtrate and sterilized water, compare, adopt pipe an official document or note method to measure its bacteriostasis.Fermented liquid is cultivated after 15d, standing depositing, be 3 months and 6 months storage time, with sterilized water, compares, and adopts pipe an official document or note method, draws fermented supernatant fluid cause of disease indicator is carried out to bacteriostasis detection.Fermented liquid is cultivated after 15d, with liquid-transfering gun, draw fermented supernatant fluid 30mL, be divided into 3 parts, pour in watch-glass, be placed on respectively under the fluorescent lamp of 40w and natural visible ray and irradiate, irradiation duration is 4h, 8h and 12h, after handling, utilizes millipore filtration to carry out filtration sterilization, adopts pipe an official document or note method to detect its bacteriostatic activity.Experimental result discovery, the metabolite of bacterial strain of the present invention is almost constant at bacteriostasis after high temperature, illumination and long-time preservation, and all, more than 90%, stability is stronger.
Embodiment 5
The present embodiment illustrates that this bacterial strain has the effect that promotes Germination of Soybean Seed.
Select fullly, plant skin complete, 50 of soybean seeds of uniform size (agriculture 29 of pacifying), use distilled water flushing seed-coat, then, with 0.1% mercuric chloride sterilization 8min, aseptic water washing for several times, to being covered with in the culture dish of three metafiltration paper, add equivalent sterilized water, filter paper is soaked.Seed after sterilization is evenly placed on the filter paper in culture dish, and evenly sprays the treatment solution 10mL of different concns in each ware, with substratum and fermenation raw liquid, compare, each processes three repetitions, the constant temperature culture of 28 ℃.From the 2d cultivating, set time every day is recorded the seed number sprouted in each culture dish, and (germination standard is that radicle stretches out hilum partial-length and surpasses the vertical footpath of seed half is standard, requirement radicle growth is normal, radicle bending in the shape of a spiral coiling person will not add up), and supplementary 10mL treatment solution, the seed 4g that gets cultivation 3,4,5d, puts into the freezing preservation of centrifuge tube, for enzyme assay experiment (label concentration and date).After 7d, from each culture dish, get at random 20 grain germination seeds.Statistics percentage of germination, germinating energy, germination index, as shown in Figure 17-19.Method of calculation are as follows:
Germination rate (Gr)=(subnumber is planted experimentally in germinating seed number/confession) * 100%;
Subnumber * 100% is planted experimentally in germinating seed number/confession when germinating energy (Gp)=day, chitting piece number reached the highest;
Germination index (Gi)=Σ Gt/Dt;
In formula, Gt is the sprouting number of t days; Dt is germination number of days.
Find, the metabolite of bacterial strain of the present invention is to soybean seeds safety, and has the effect that promotes seed germination.When fermented liquid concentration is 60%, seed germination rate is the highest, can reach 85%; And fermented liquid is when 60% this concentration, and the germinating energy of seed and germination index also reach maximum, and germinating energy and germination index are respectively 43% and 18%.
Comparative example 1
This comparative example is for illustrating the effect of this bacterial classification.
Quarter Design: test adopts plot experiment, repeats for 3 times, 6 ridges, every processing community, and every ridge 10m is capable long.Establish altogether 3 processing, contrast (water logging kind), the mould fermented liquid seed soaking of 60% top spore and the seed soaking of 3 ‰ derosal, seed soaking time is 6h.
Seed treatment: choose soybean seeds, with 75% alcohol surface sterilization 2min, sterile water wash 5 times, airing, continues to employ seed soaking.Seed after seed soaking is by the distance sowing of 20/m.
Soil treatment: to soybean community ridging, (spore suspension concentration is 10 by the soybean Fusarium oxysporum An Mei community 200mL spore suspension after numerous with the expansion of wheat substratum in ridge with hoe
7/ mL) inoculation is mixed with soil after inoculation at once.Be seeded in mixed soybean ridge, Routine Management after soybean emerges, does not apply fertilizer and agricultural chemicals.
Survey item and method: the soybean seedling 30d that grows after emerging, investigation disease index, soybean root disease incidence adopts the disease scale standard of (1987) such as Xin Huipu to carry out classification, disease index and preventive effect; After results, measure the indexs such as plant height and output.
Disease index=[∑ (diseased plant number * onset grade)/(summation * the highest onset grade is counted in strain)] * 100
Root rot grade scale is as follows:
0 grade: main root, fibrous root is sound, and without scab, root nodule is many; 1 grade: on main root, have fragmentary scab, but not in flakes, on fibrous root without scab; 2 grades: on main root, scab in flakes, but is less than 1/4 of root girth, and fibrous root is fallen ill slightly; 3 grades: on main root, scab is greater than 1/4 of girth, but be less than 1/2, fibrous root scab is more, but not in flakes; 4 grades: on main root, scab is greater than 1/2 of girth, but be less than 3/4, fibrous root is fallen ill slightly; 5 grades: whole root all has scab to surround, butt rot, the nearly nothing of fibrous root.
Experimental result is found, through the community of bacterial strain seed soaking of the present invention root rot in seedling stage incidence, with derosal seed soaking district, is similar to, and preventive effect can reach 70% left and right, and after results, plant height and cell production are improved.As shown in table 2 and table 3.
The prevention effect of the different seed soakings of table 2 to root rot
Claims (2)
1. strain peachiness top spore mould (Acremonium persicinum) JX524288, its deposit number is CCTCC NO:M2013200.
2. the application of the fermented liquid of peachiness claimed in claim 1 top spore mould (Acremonium persicinum) JX524288 on antagonizing pathogenic fungi and promotion soybean germination.
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| CN103897992B (en) * | 2014-04-30 | 2017-08-04 | 中国农业科学院棉花研究所 | The endogenetic fungus CEF 193 of cotton and its application |
| CN104449742A (en) * | 2014-11-27 | 2015-03-25 | 黑龙江八一农垦大学 | Greenhouse tomato soil biological modifier |
| MX2019010349A (en) * | 2017-03-01 | 2019-11-21 | Indigo Ag Inc | Endophyte compositions and methods for improvement of plant traits. |
| CN114540204B (en) * | 2022-02-22 | 2023-06-23 | 南宁师范大学 | Acremonium persicum H1-1 and screening method and application thereof |
| CN114574369B (en) * | 2022-03-08 | 2023-04-07 | 吉林农业大学 | Acremonium persicinum MR-47 and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5462386A (en) * | 1977-10-26 | 1979-05-19 | Kirin Brewery | Production of alkali protease |
| CN101970639A (en) * | 2008-03-14 | 2011-02-09 | 安斯泰来制药株式会社 | Microorganism producing cyclic compound |
| CN103205367A (en) * | 2013-05-10 | 2013-07-17 | 国家海洋局第三海洋研究所 | Acremonium persicinum hansfordii S2915 and preparation method of antifungal active protein of acremonium persicinum hansfordii S2915 |
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2013
- 2013-08-05 CN CN201310335286.4A patent/CN103436452B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5462386A (en) * | 1977-10-26 | 1979-05-19 | Kirin Brewery | Production of alkali protease |
| CN101970639A (en) * | 2008-03-14 | 2011-02-09 | 安斯泰来制药株式会社 | Microorganism producing cyclic compound |
| CN103205367A (en) * | 2013-05-10 | 2013-07-17 | 国家海洋局第三海洋研究所 | Acremonium persicinum hansfordii S2915 and preparation method of antifungal active protein of acremonium persicinum hansfordii S2915 |
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