CN108823107A - A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding - Google Patents
A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding Download PDFInfo
- Publication number
- CN108823107A CN108823107A CN201810768050.2A CN201810768050A CN108823107A CN 108823107 A CN108823107 A CN 108823107A CN 201810768050 A CN201810768050 A CN 201810768050A CN 108823107 A CN108823107 A CN 108823107A
- Authority
- CN
- China
- Prior art keywords
- white
- mushroom
- chestnut mushroom
- chestnut
- artp
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- 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
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
Abstract
The present invention relates to a kind of white chestnut mushroom kinds and selection using ARTP induced-mutation technique breeding, belong to edible mushroom breeding of new variety field.Technical solution is:It is named as and moves white M-520, breeding forms on the basis of No. 1 bacterial strain in chestnut mushroom Qianxi, and the bacterial strain of the kind is registered on the books number in the preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center:CGMCC No.15876.10 μ L chestnut mushroom protoplast suspension slide glasses are put into ARTP mutagenesis instrument fixed bit and postponed by white chestnut mushroom textual study ARTP induced-mutation technique breeding, are 8 SLM in throughput, and the mutagenic treatment time is set as under the conditions of 30-40s, can get mutant.Beneficial effects of the present invention:Cap is white, and chestnut mushroom aromatic flavour;It is still white after processing is cooked, provides a kind of new food materials for the various delicious dish of production color.
Description
Technical field
The present invention relates to a kind of white chestnut mushroom kinds and selection using ARTP induced-mutation technique breeding, belong to edible mushroom
Breeding of new variety field.
Background technique
Chestnut mushroom, scientific name grifola frondosus, also known as:Polyporus frondosus, grifola frondosa, thousand Buddhist bacterium, chestnut mushroom, lotus flower mushroom etc.;It is a kind of treasure
Dilute food, the dual-purpose fungi of medicine belong to Basidiomycotina, Hymenomycetes, Aphyllophorales, Polyporaceae, set flower Pseudomonas.Chestnut mushroom cultivation in China's is ground
Studying carefully starts from last century the eighties, and emphasis key scientific and technological projects during being Hebei Department of Science and Technology of Shanxi Province " eight or five " -- chestnut mushroom is imitative
Wildness cultural technique research.Chestnut mushroom cultivation Remarkable Progress On Electric Artificial progress in 1992, imitates wild artificial cultivation chestnut mushroom and just succeeds.
Currently, the artificial cultivation kind of chestnut mushroom mainly has a Qianxi 1, the kinds such as Qianxi 3, cap color is Dark grey to greyish black
Color, kind is more single, without white(Light color)Kind.
Summary of the invention
The object of the present invention is to provide a kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding, bacterium
Lid is white, and chestnut mushroom aromatic flavour;It is still white after processing is cooked, is provided for the various delicious dish of production color
A kind of new food materials solve the above problem existing for prior art.
Object of the present invention is to what is realized by following technical solution:
White chestnut mushroom kind provided by the invention, is on the basis of No. 1 bacterial strain in chestnut mushroom Qianxi, using ARTP induced-mutation technique breeding
It forms, which is named as " moving white M-520 ", and the bacterial strain of the kind is preserved in China Committee for Culture Collection of Microorganisms
Common micro-organisms center(Abbreviation CGMCC, address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), collection registers on the books
Number:CGMCC No.15876.
The white chestnut mushroom kind, soil covering culture 16~50cm of whole strain diameter weigh 1~10kg;2~5cm of bacteria cover diameter, bacterium
Leaf 2~6mm of thickness, for mushroom type shaped like chrysanthemum, mushroom color is white;The kind belongs to middle warm type kind, and misshapen mushroom is few, and change of tide is fast, fiber
It is few.
The selection of the white chestnut mushroom kind is hanged 10 μ L chestnut mushroom protoplasts using ARTP induced-mutation technique breeding
Liquid slide glass is put into ARTP mutagenesis instrument fixed bit and postpones, and is 8 SLM in throughput, the mutagenic treatment time is set as 30-40 s condition
Under, it can get mutant.
The white chestnut mushroom textual study ARTP induced-mutation technique breeding, comprises the following steps:
1. preparing, following media is spare, preparing experiment drug:
(1)PDA culture medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 20 g of agar, water 1000 mL, pH are natural;
(2)Improve PDA culture medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, 3 g of peptone, phosphorus
3 g of acid dihydride potassium, vitamin B10.05 g, 20 g of agar, water 1000 mL, pH are natural;
(3)Regeneration culture medium:Potato(Remove the peel liquor)200 g, 109.306 g of mannitol, 4 g of glucose, 4 g of maltose, ferment
4 g of female powder, 4 g of peptone, 15 g of agar, water 1000 mL, pH are natural;
(4)Sawdust liquor culture medium:100 g of sawdust, potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, egg
White 3 g of peptone, 3 g of potassium dihydrogen phosphate, vitamin B10.05 g, 20 g of agar, water 1000 mL, pH are natural;
(5)Fluid nutrient medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, 3 g of peptone, di(2-ethylhexyl)phosphate
3 g of hydrogen potassium, vitamin B10.05 g, water 1000 mL, pH are natural;
Preparing experiment drug:
Glycerol, methanol, glucose, potassium dihydrogen phosphate, magnesium sulfate, vitamin B1, it is agar, peptone, maltose, yeast powder, sweet
Reveal alcohol, gel reservoir, TEMED, ammonium persulfate, sucrose, bromophenol blue, acetic acid -1- naphthalene ester, acetic acid -2- naphthalene ester, Fast Blue RR salt,
Tris(Trishydroxymethylaminomethane), glycine, acetone, ultrapure water, distilled water, alcohol, lywallzyme, lywallzyme is by the micro- life in Guangdong
Object is produced;Other reagents are the analytical reagents of market purchase;
2. test apparatus and agent prescription:
ARTP mutagenesis instrument is mutation breeding system, buys in the market, thinks Biotechnology Co., Ltd's production of radically reforming.
Agent prescription
(1)2% enzyme solution is prepared:100 mg lywallzymes are accurately weighed, are dissolved in the mannitol of 5 mL, 0.6 M, 0.22 μm of micropore filter
Film filtration sterilization(Matching while using);
(2)Homeo-osmosis agent(0.6 M mannitol):109.306 g mannitol are accurately weighed, are dissolved in 800 mL ultrapure waters, most
Constant volume is to 1000 mL afterwards;
(3)Gel reservoir:The gel reservoir that the raw work in Shanghai is sold(Acr/Bis=29:1);
(4)Glue buffer is concentrated(0.5 mol/L, pH 6.8):6.06 g Tris are accurately weighed, 80 mL ultrapure waters, pH are dissolved in
6.8 are adjusted to hydrochloric acid, last constant volume to 100 mL;
(5)Separation gel buffer(1.5 mol/L, pH 8.9):18.16 g Tris are accurately weighed, 80 mL ultrapure waters, pH are dissolved in
8.9 are adjusted to hydrochloric acid, last constant volume to 100 mL;
(6)Electrode buffer(0.025 mol/L Tris, 0.2 mol/L glycine):Accurately weigh 3.028 g Tris,
14.414 g glycine are dissolved in 800 mL ultrapure waters, last constant volume to 1000 mL;
(7)10% AP solution:0.1 g ammonium persulfate is accurately weighed, 1.0 mL ultrapure waters are dissolved in;
(8)TEMED:The raw work in Shanghai is sold;
(9)6.5 phosphate buffer of pH:10.69 g sodium dihydrogen phosphates, 11.3 g disodium hydrogen phosphates are accurately weighed, 800 mL are dissolved in
Ultrapure water, last constant volume to 1000 mL;
(10)Esterase isozyme dyeing liquor:133.4 mg Fast Blue RR salt are accurately weighed, 200 mL, 0.1 mol/L, pH 6.5 are dissolved in
Phosphate buffer in, filtering is stand-by, and 66.7 mg acetic acid -1- naphthalene esters, 66.7 mg acetic acid -2- naphthalene esters are weighed before use, are dissolved in 4
In mL acetone, then poured into above-mentioned filtrate dropwise, it is stirring while adding to mix it uniformly to dye.
3. the mycelial culture of chestnut mushroom
The bacterial strain of chestnut mushroom Qianxi 1, buys in the market, such as:The chestnut mushroom of Qianxi County Lin Zhongbao Biotechnology Co., Ltd production is moved
Western No. 1 bacterial strain, Qianxi 1 is chestnut mushroom bacterial strain commodity common name;
The strain inoculated of chestnut mushroom Qianxi 1 activate 7 days in improvement PDA culture medium, with the punch of 5 mm of diameter from activating bacterium
It is punched on the edge of kind, on 5 pieces of inoculation liquid medium withins of picking, 25 DEG C of constant temperature stationary culture 10 days, shake manually sooner or later daily
It shakes once, it is spare;
4. the preparation of No. 1 protoplast in chestnut mushroom Qianxi
(1)The fresh mycelium for collecting above-mentioned culture is washed 2 times with four layers of filtered through gauze with the mannitol of 0.6 M, then is used
Filter paper sops up liquid extra in mycelium, and mycelium is collected into the EP pipe of 10 mL;
(2)The metering of the lywallzyme of 1 mL is added by every 100 mg fresh mycelia, lywallzyme, 34 DEG C of water are added in mycelia
Bath, 2 h of time, every 30 min rock once;
(3)After digesting 1 h, the preparation of protoplast is observed;
(4)After 2 h, the mannitol that isometric 0.6 M is added terminates enzymatic hydrolysis, is filtered with 400 mesh cell sieves, filtrate is collected into
In EP pipe, centrifugation(Room temperature, 3000 rpm, 10 min), obtain protoplast;
(5)Obtained protoplast is washed 2 times with the mannitol of 0.6 M, is centrifuged(Room temperature, 3000 rpm, 10 min), discard
Supernatant, then suspended with the mannitol of 0.6 M, obtain protoplast suspension;
(6)Protoplast is counted using blood counting chamber, by protoplast suspension concentration dilution to 1 × 107A/mL;
5. protoplast ARTP mutagenesis and regeneration
(1)Open ARTP mutagenesis instrument(Helium is opened after first switch power supply)Pre-sterilization sterilizes before being tested;
(2)Seven metal sample slide glasses are placed in each 30 s of calcination of alcolhol burner flame envelope respectively in super-clean bench, are put into sterilized
It is cooling in glass plate, take 10 μ L protoplast suspensions in slide glass;
(3)The plate equipped with processing specimen slides will be placed and move to ARTP mutagenesis system operation storehouse, put slide glass with aseptic nipper
To corresponding hole location, adjusting knob rises microscope carrier, until slide glass is at 2 mm of flow ports, and closes door;
(4)Throughput and processing are set, clicks start button, starts to process sample;Throughput is 8 SLM, mutagenic treatment time
It is set as 40 s;
(5)Sample treatment finishes, and is put slide glass into the EP pipe equipped with 1 mL, 0.6 M mannitol with aseptic nipper;EP pipe is existed
1 min is shaken on oscillator, the protoplast suspension being attached on slide glass is eluted in liquid, obtains the primary of mutagenic treatment
Plastid suspension;
(6)Protoplast suspension after above-mentioned mutagenesis is diluted with 0.6 M mannitol, 100 μ L dilutions is taken to be coated on again
On raw culture medium, mark, 25 DEG C of constant temperature incubations cultivate the bacterial strain regenerated;
6. the screening of double-core mutagenesis
The bacterial strain regenerated is chosen one by one, is placed in improvement PDA culture medium and cultivates, sediments microscope inspection is used in picking mycelia film-making
Observation, using clamp connection as standard, determines single double-core of bacterial strain, and the mycelium for having clamp connection is double-core mutagenesis mycelia;
7. the antagonistic experiment of mutagenesis
The double-core mutagenic strain screened is activated on improvement PDA culture medium plate with the bacterial strain of parent's chestnut mushroom Qianxi 1;
Use punch(5 mm of diameter)It punches, is inoculated on PDA culture medium plate on the edge of strain, face-off growth;Each processing
It repeats three times;After 25 DEG C are cultivated 15 days, result is observed;
8. esterase isozyme verifies the sub- affiliation of mutagenesis
(1)The culture of mycelia
By the double-core mutagenic strain of screening acquisition, the strain inoculated of chestnut mushroom Qianxi 1 in improvement PDA culture medium, 25 DEG C of constant temperature incubations
15 days;
(2)The preparation of esterase isozyme
The mycelia of culture is scraped with blade, is collected into 2 mL EP pipes, the concentration that 800 μ L dilute 4 times is added in every 1 g tissue
Glue buffer is put into one evening of frost in -80 DEG C of ultra low temperature freezers;It then takes out and is placed in mortar, quartz sand is added and is fully ground
(Whole process carries out on ice);Lapping liquid is centrifuged(4 DEG C, 10000 rpm, 10 min), take same volume supernatant and
Sucrose solution(40%)Mixing, adds a small amount of 0.1% bromophenol blue solution, is mixed with oscillator, be put into 4 DEG C of refrigerators and save;
(3)Electrophoresis and enzyme spectrum analysis
Using discontinuous Horizontal starch gel electrophoresis(PAGE), electrode buffer is the Tris-Gly that pH is 8.3;
Resolving gel concentration is 7.5%, pH 8.9;Concentration gum concentration is 4%, pH 6.8;Electrophoresis and dyeing are carried out according to a conventional method;
According to different strains enzyme band mobility after electrophoresis(Rf)Difference, number, the depth of the width of enzyme band, color of enzyme band can
To analyze the difference between different strains;
Enzyme band mobility(Rf)=enzyme is with migration distance/indicator migration distance;
Will dyeing enzyme band be divided into 0=do not have;1=shallower;2=medium;3=relatively deep, the esterase isozyme information of each bacterial strain is counted, is used
2.1 Software of Fuzzy Clustering Analysis of NTSYSpc Version is calculated the genetic similarity between bacterial strain, and is lost using the building of UPGMA method
Pass Cluster tree;
9. the fruiting of mutagenesis screens
Screening obtains Protoplast Mutation and the bacterial strain of starting strain chestnut mushroom Qianxi 1 carries out cultivation fruiting;
Medium for original variety:Chestnut bits 80%, wheat bran skin 8%, gypsum and sugar is each 1%, sandy loam or loam 10%, mass ratio;
Culture material formula:Chestnut bits 70%, wheat bran 20%, the raw-soil 8%, gypsum 1%, sugar 1%, mass ratio;
Planting type:Imitate wild fruiting --- after cultivating bacteria bag mycelia purseful, polybag is sloughed, bacteria stick is fitly arranged in thing
In the furrow first dug, appropriate gap is stayed between bacteria stick, bacteria stick gap and around banket, surface is covered with 1~2 centimetre of soil layer.
ARTP is atmospheric pressure at room plasma(Atmospheric and Room Temperature Plasma)Letter
Claim, can generate under atmospheric pressure temperature between 25-40 DEG C, have high activity particle(It is former including the helium in excitation state
Son, oxygen atom, nitrogen-atoms, OH free radical etc.)The plasma jet of concentration.
Preservation explanation
It is recommended that systematic name:Grifola frondosus;
Latin name:
Grifola frondosa
Join the biomaterial of evidence:Move white M-520
Preservation mechanism:China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is referred to as:CGMCC
Address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date:On June 25th, 2018
Collection is registered on the books number:CGMCC No.15876
Beneficial effects of the present invention:Cap is white, and chestnut mushroom aromatic flavour;Be still white after processing is cooked, for production color,
Delicious dish fragrant, that taste is various provides a kind of new food materials.
Figure of description
Fig. 1 is the protoplast of No. 1 strain in chestnut mushroom of embodiment of the present invention Qianxi;
Fig. 2 is the strain protoplast ARTP mutagenesis lethality curve of chestnut mushroom of embodiment of the present invention Qianxi 1;
Fig. 3 is that induction mutation of bacterium of chestnut mushroom of embodiment of the present invention Qianxi 1 regenerates bacterium colony;
Fig. 4 is that the antagonism of mutagenesis of embodiment of the present invention screens;
Fig. 5 is 16 chestnut mushroom mutagenic strain Isoenzyme Patterns of Esterase figures of the embodiment of the present invention;
Fig. 6 is 16 chestnut mushroom mutagenic strain Isoenzyme Patterns of Esterase ideographs of the embodiment of the present invention;
Fig. 7 is 16 chestnut mushroom mutagenic strain esterase isozyme dendrograms of the embodiment of the present invention;
Fig. 8 is the chestnut mushroom fruiting figure of prior art Qianxi 1;
Fig. 9 is that the embodiment of the present invention moves white M-520 and the chestnut mushroom fruiting figure of prior art Qianxi 1 compares, and bottom right white is this
Inventive embodiments move white M-520;
Figure 10 is that the embodiment of the present invention moves white M-520 chestnut mushroom fruiting figure.
Specific embodiment
The present invention will be further described by the following examples.
White chestnut mushroom kind provided by the invention, is that breeding forms on the basis of No. 1 bacterial strain in chestnut mushroom Qianxi, the kind quilt
It is named as " moving white M-520 ", the bacterial strain of the kind is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart(Abbreviation CGMCC, address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), collection registers on the books number:CGMCC
No.15876。
The white chestnut mushroom kind, soil covering culture 16~50cm of whole strain diameter weigh 1~10kg;2~5cm of bacteria cover diameter, bacterium
Leaf 2~6mm of thickness, for mushroom type shaped like chrysanthemum, mushroom color is white;The kind belongs to middle warm type kind, and misshapen mushroom is few, and change of tide is fast, fiber
It is few.
The selection of the white chestnut mushroom kind is hanged 10 μ L chestnut mushroom protoplasts using ARTP induced-mutation technique breeding
Liquid slide glass is put into ARTP mutagenesis instrument fixed bit and postpones, and is 8 SLM in throughput, the mutagenic treatment time is set as 30-40 s condition
Under, it can get mutant.
The white chestnut mushroom textual study ARTP induced-mutation technique breeding, comprises the following steps:
1, it is spare to prepare following media:
(1)PDA culture medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 20 g of agar, water 1000 mL, pH are natural;
(2)Improve PDA culture medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, 3 g of peptone, phosphorus
3 g of acid dihydride potassium, vitamin B10.05 g, 20 g of agar, water 1000 mL, pH are natural;
(3)Regeneration culture medium:Potato(Remove the peel liquor)200 g, 109.306 g of mannitol, 4 g of glucose, 4 g of maltose, ferment
4 g of female powder, 4 g of peptone, 15 g of agar, water 1000 mL, pH are natural;
(4)Sawdust liquor culture medium:100 g of sawdust, potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, egg
White 3 g of peptone, 3 g of potassium dihydrogen phosphate, vitamin B10.05 g, 20 g of agar, water 1000 mL, pH are natural;
(5)Fluid nutrient medium:Potato(Remove the peel liquor)200 g, 20 g of glucose, 1.5 g of magnesium sulfate, 3 g of peptone, di(2-ethylhexyl)phosphate
3 g of hydrogen potassium, vitamin B10.05 g, water 1000 mL, pH are natural;
Preparing experiment drug:
Glycerol, methanol, glucose, potassium dihydrogen phosphate, magnesium sulfate, vitamin B1, it is agar, peptone, maltose, yeast powder, sweet
Reveal alcohol, gel reservoir, TEMED, ammonium persulfate, sucrose, bromophenol blue, acetic acid -1- naphthalene ester, acetic acid -2- naphthalene ester, Fast Blue RR salt,
Tris(Trishydroxymethylaminomethane), glycine, acetone, ultrapure water, distilled water, alcohol, lywallzyme, lywallzyme is by the micro- life in Guangdong
Object is produced;Other reagents are the analytical reagents of market purchase;
2, test apparatus and agent prescription:
ARTP mutagenesis instrument is mutation breeding system, buys in the market, thinks Biotechnology Co., Ltd's production of radically reforming;
Agent prescription
(1)2% enzyme solution is prepared:100 mg lywallzymes are accurately weighed, are dissolved in the mannitol of 5 mL, 0.6 M, the filter of 0.22 um micropore
Film filtration sterilization(Matching while using);
(2)Homeo-osmosis agent(0.6 M mannitol):109.306 g mannitol are accurately weighed, are dissolved in 800 mL ultrapure waters, most
Constant volume is to 1000 mL afterwards;
(3)Gel reservoir:The gel reservoir that the raw work in Shanghai is sold(Acr/Bis=29:1);
(4)Glue buffer is concentrated(0.5 mol/L, pH 6.8):6.06 g Tris are accurately weighed, 80 mL ultrapure waters, pH are dissolved in
6.8 are adjusted to hydrochloric acid, last constant volume to 100 mL;
(5)Separation gel buffer(1.5 mol/L, pH 8.9):18.16 g Tris are accurately weighed, 80 mL ultrapure waters, pH are dissolved in
8.9 are adjusted to hydrochloric acid, last constant volume to 100 mL;
(6)Electrode buffer(0.025 mol/L Tris, 0.2 mol/L glycine):Accurately weigh 3.028 g Tris,
14.414 g glycine are dissolved in 800 mL ultrapure waters, last constant volume to 1000 mL;
(7)10% AP solution:0.1 g ammonium persulfate is accurately weighed, 1.0 mL ultrapure waters are dissolved in;
(8)TEMED:The raw work in Shanghai is sold;
(9)6.5 phosphate buffer of pH:10.69 g sodium dihydrogen phosphates, 11.3 g disodium hydrogen phosphates are accurately weighed, 800 mL are dissolved in
Ultrapure water, last constant volume to 1000 mL;
(10)Esterase isozyme dyeing liquor:133.4 mg Fast Blue RR salt are accurately weighed, 200 mL, 0.1 mol/L, pH 6.5 are dissolved in
Phosphate buffer in, filtering is stand-by, and 66.7 mg acetic acid -1- naphthalene esters, 66.7 mg acetic acid -2- naphthalene esters are weighed before use, are dissolved in 4
In mL acetone, then poured into above-mentioned filtrate dropwise, it is stirring while adding to mix it uniformly to dye;
3, the mycelial culture of chestnut mushroom
The bacterial strain of chestnut mushroom Qianxi 1, buys in the market, such as:The chestnut mushroom of Qianxi County Lin Zhongbao Biotechnology Co., Ltd production is moved
Western No. 1 bacterial strain, Qianxi 1 is chestnut mushroom bacterial strain commodity common name;
The strain inoculated of chestnut mushroom Qianxi 1 activate 7 days in improvement PDA culture medium, with the punch of 5 mm of diameter from activating bacterium
It is punched on the edge of kind, on 5 pieces of inoculation liquid medium withins of picking, 25 DEG C of constant temperature stationary culture 10 days, shake manually sooner or later daily
It shakes once, it is spare;
4, the preparation of No. 1 protoplast in chestnut mushroom Qianxi
Mycelial culture is carried out first;The strain inoculated of chestnut mushroom Qianxi 1 is activated 7 days in improvement PDA culture medium, uses diameter
The punch of 5 mm is punched from the edge of activated spawn, on 5 pieces of inoculation liquid medium withins of picking, 25 DEG C, and stationary culture 10
It, rocks primary manually sooner or later daily;
The preparation of protoplast
(1)The fresh mycelium for collecting above-mentioned culture is washed 2 times with four layers of filtered through gauze with the mannitol of 0.6 M, then is used
Filter paper sops up liquid extra in mycelium, and mycelium is collected into the EP pipe of 10 mL;
(2)The metering of the lywallzyme of 1 mL is added by every 100 mg fresh mycelia, lywallzyme, 34 DEG C of water are added in mycelia
Bath, 2 h of time, every 30 min rock once;
(3)After digesting 1 h, the preparation of protoplast is observed;
(4)After 2 h, the mannitol that isometric 0.6 M is added terminates enzymatic hydrolysis, is filtered with 400 mesh cell sieves, filtrate is collected into
In EP pipe, centrifugation(Room temperature, 3000 rpm, 10 min), obtain protoplast;
(5)Obtained protoplast is washed 2 times with the mannitol of 0.6 M, is centrifuged(Room temperature, 3000 rpm, 10 min), discard
Supernatant, then suspended with the mannitol of 0.6 M, obtain protoplast suspension;
(6)Protoplast is counted using blood counting chamber, by protoplast suspension concentration dilution to 1 × 107A/mL;
Protoplast ARTP mutagenesis and regeneration
(1)Open ARTP mutagenesis instrument(Helium is opened after first switch power supply)Pre-sterilization sterilizes before being tested;
(2)Seven metal sample slide glasses are placed in each 30 s of calcination of alcolhol burner flame envelope respectively in super-clean bench, are put into sterilized
It is cooling in glass plate, take 10 μ L protoplast suspensions in slide glass;
(3)The plate equipped with processing specimen slides will be placed and move to ARTP mutagenesis system operation storehouse, put slide glass with aseptic nipper
To corresponding hole location, adjusting knob rises microscope carrier, until slide glass is at 2 mm of flow ports, and closes door;
(4)Throughput and processing are set, clicks start button, starts to process sample;Throughput is 8 SLM, processing time difference
It is set as:0 s,10 s,20 s,30 s,40 s,50 s,60 s;
(5)Sample treatment finishes, and is put slide glass into the EP pipe equipped with 1 mL, 0.6 M mannitol with aseptic nipper;EP pipe is existed
1 min is shaken on oscillator, the protoplast suspension being attached on slide glass is eluted in liquid, forms the protoplast of mutagenesis
Suspension;
(6)The protoplast suspension of above-mentioned mutagenesis is diluted with 0.6 M mannitol, 100 μ L dilutions is taken to be coated on regeneration
It on culture medium, marks, 25 DEG C of constant temperature incubations cultivate the bacterial strain regenerated;
5, the screening of double-core mutagenesis
The bacterial strain regenerated is chosen one by one, is placed in improvement PDA culture medium and cultivates, sediments microscope inspection is used in picking mycelia film-making
Observation, using clamp connection as standard, determines single double-core of bacterial strain, and the mycelium for having clamp connection is double-core mutagenesis mycelia;
6, the antagonistic experiment of mutagenesis
The double-core mutagenic strain screened is activated on improvement PDA culture medium plate with the bacterial strain of parent's chestnut mushroom Qianxi 1;
Use punch(5 mm of diameter)It punches, is inoculated on PDA culture medium plate on the edge of strain, face-off growth;Each processing
It repeats three times;After 25 DEG C are cultivated 15 days, result is observed;
7, the sub- esterase isozyme experiment of mutagenesis
The culture of mycelia
By the double-core mutagenic strain of screening acquisition, the strain inoculated of chestnut mushroom Qianxi 1 in improvement PDA culture medium, 25 DEG C of constant temperature incubations
15 days;
The extraction of esterase isozyme
The mycelia of culture is scraped with blade, is collected into 2 mL EP pipes, the concentration that 800 μ L dilute 4 times is added in every 1 g tissue
Glue buffer is put into one evening of frost in -80 DEG C of ultra low temperature freezers;It then takes out and is placed in mortar, quartz sand is added and is fully ground
(Whole process carries out on ice);Lapping liquid is centrifuged(4 DEG C, 10000 rpm, 10 min), take same volume supernatant and
Sucrose solution(40%)Mixing, adds a small amount of 0.1% bromophenol blue solution, is mixed with oscillator, be put into 4 DEG C of refrigerators and save;
Electrophoresis and enzyme spectrum analysis
Using discontinuous Horizontal starch gel electrophoresis(PAGE), electrode buffer is the Tris-Gly that pH is 8.3;
Resolving gel concentration is 7.5%, pH 8.9;Concentration gum concentration is 4%, pH 6.8;Electrophoresis and dyeing are carried out according to a conventional method;
According to different strains enzyme band mobility after electrophoresis(Rf)Difference, number, the depth of the width of enzyme band, color of enzyme band can
To analyze the difference between different strains;
Enzyme band mobility(Rf)=enzyme is with migration distance/indicator migration distance;
Will dyeing enzyme band be divided into 0=do not have;1=shallower;2=medium;3=relatively deep, the esterase isozyme information of each bacterial strain is counted, is used
2.1 Software of Fuzzy Clustering Analysis of NTSYSpc Version is calculated the genetic similarity between bacterial strain, and is lost using the building of UPGMA method
Pass Cluster tree;
8, the fruiting of mutagenesis
Screening obtains Protoplast Mutation and the bacterial strain of starting strain chestnut mushroom Qianxi 1 carries out cultivation fruiting;
Medium for original variety:Chestnut bits 80%, wheat bran skin 8%, gypsum and sugar is each 1%, sandy loam or loam 10%, mass ratio;
Culture material formula:Chestnut bits 70%, wheat bran 20%, the raw-soil 8%, gypsum 1%, sugar 1%, mass ratio;
Planting type:Imitate wild fruiting --- after cultivating bacteria bag mycelia purseful, polybag is sloughed, bacteria stick is fitly arranged in thing
In the furrow first dug, appropriate gap is stayed between bacteria stick, bacteria stick gap and around banket, surface is covered with 1~2 centimetre of soil layer.
A kind of white chestnut mushroom kind using ARTP induced-mutation technique breeding, is cultivated using the above method;;Soil covering culture
16~50cm of whole strain diameter, weighs 1~10kg;2~5cm of bacteria cover diameter, bacterium leaf 2~6mm of thickness, for mushroom type shaped like chrysanthemum, mushroom color is white
Color;The kind belongs to middle warm type kind, and misshapen mushroom is few, and change of tide is fast, and fiber is few.
Experimental result of the embodiment of the present invention and analysis
1, the preparation of 1 strain protoplast of chestnut mushroom Qianxi
After digesting 2 h, the protoplast liberation situation of starting strain chestnut mushroom Qianxi 1 is observed, referring to Fig.1.The transparent nothing of protoplast
Color is the spherical shape of sharp outline, not of uniform size.
2, the strain protoplast ARTP mutagenesis of chestnut mushroom Qianxi 1 and regeneration
The protoplast ARTP mutagenesis lethality of chestnut mushroom Qianxi 1
Mutagenesis lethality curve is referring to Fig. 2.When handling time 0 s to 30 s, the lethality of protoplast is ramped, to processing
The lethality of lethality up to 75.7%, 40 s are 80.0% when 30 s of time, and 50 s lethalities are 84.4%, and 60 s lethalities are close
100%。
According to lethality curve, influence variation of the plasma to protoplast is greatly, unstable when 0 s to 30 s, to 40
A possibility that tend towards stability stabilization when s, and lethality is larger, and the protoplast survived is more, screens mutant strain is big, experiment
Choosing 40 s is the mutagenesis optimization process time.
The Protoplast mutagenesis and regeneration result of chestnut mushroom Qianxi 1
According to lethality curve, 40 s is selected to carry out repeating experiment, the protoplast suspension dilution 3 that 40 s of mutagenic treatment is obtained
Times, it draws 100 μ L and is spread evenly across in regeneration culture, 25 DEG C of cultures.After cultivating 7 day time, protoplast starts to regenerate
Bacterium colony, regenerated plate is referring to Fig. 3.The single colonie come will be regenerated to choose, improvement PDA culture medium plate is transferred to, expands culture.
Picking colony 100 altogether.
3, the screening of single double-core of No. 1 induction mutation of bacterium in chestnut mushroom Qianxi
The nucleated mycelium observed under the microscope has clamp connection, and mycelia is sturdy, glossy.It is screened, is selected by single double-core
Double-core bacterial strain 51 out, number M401-M536.By antagonistic experiment, make further screening.
4, the antagonistic experiment of No. 1 induction mutation of bacterium in chestnut mushroom Qianxi
By 51 double-core bacterial strains and the progress antagonistic experiment of starting strain chestnut mushroom Qianxi 1, by between observation mutagenesis and parent
Antagonism filters out the bacterial strain that antagonism line can be generated with parents, referring to Fig. 4.16 mutagenic strains are obtained altogether.
5, the sub- esterase isozyme experiment of chestnut mushroom mutagenesis
The different enzyme band of 13 mobilities is detected altogether for 16 chestnut mushroom mutagenic strains of examination and the bacterial strain of starting strain Qianxi 1.From
Clustering figure can be seen that the affiliation of mutagenic progeny Yu starting strain Qianxi 1.Wherein M410, M412, M419,
M513, M517, M518 difference are smaller, similarity factor 1.00, can tentatively regard as same strain.M415 and the bacterial strain of Qianxi 1
Affiliation is closer, should be same bacterial strain.
It is final to obtain 11 kinds of mutagenic strain by esterase isozyme cluster analysis result.Respectively:M407,M410,
M413、M417、M418、M423、M425、M512、M520、M529、M533。
Referring to attached drawing 5,1-17 swimming lane respectively represents Qianxi 1, M407, M408, M410, M411, M412, M415,
M419、M423、M425、M512、M513、M517、M518、M520、M529、M533
Referring to attached drawing 6,1-17 swimming lane respectively represents Qianxi 1, M407, M408, M410, M411, M412, M415, M419,
M423、M425、M512、M513、M517、M518、M520、M529、M533
Referring to attached drawing 7,1-17 swimming lane respectively represents Qianxi 1, M407, M408, M410, M411, M412, M415, M419,
M423、M425、M512、M513、M517、M518、M520、M529、M533
Fruiting experiment main result
The chestnut mushroom yield and morphological feature of bacterium bag during healthy bacterium bag quantity and fruiting after strains tested bacterium germination are recorded, in detail
See that chestnut mushroom fruiting experiment record data are shown in Table 1.
Wherein, M520 biological transformation ratio is 103.6%, and fructification sports white, other mutagenesis are still grey, dark brown
Color., M520 is the present invention, is known as moving white M-520.
The chestnut mushroom Qianxi of prior art 1, fresh goods 15~60cm of diameter, weighs 1~10kg.Stem branch is more, bacteria cover diameter 2
~7cm, bacterial context 2~7mm of thickness, tube length≤1.5mm;For cap Dark grey to grey black, bacterial context, tube are white.Good toughness
It is non-breakable, crisp in taste, aromatic flavour.Referring to attached drawing 8.
Chestnut mushroom of the present invention(Move white M-520)Piece type is medium-and-large-sized, and soil covering culture 16~50cm of whole strain diameter weighs 1~10kg.Bacterium
Lid 2~5cm of diameter, bacterium leaf 2~6mm of thickness, stem is mostly level-one branch, and second branch is few, and for mushroom type shaped like chrysanthemum, mushroom color is white
Color.The kind belongs to middle warm type kind, and misshapen mushroom is few, and change of tide is fast, and fiber is few, and commodity is high, referring to attached drawing 9,10.
Claims (3)
1. a kind of white chestnut mushroom kind, is named as and moves white M-520, in chestnut mushroom Qianxi on the basis of No. 1 bacterial strain, using ARTP mutagenesis
Technology breeding forms, and the bacterial strain of the kind is stepped in the preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center
Charging to volume number is:CGMCC No.15876.
2. white chestnut mushroom kind according to claim 1, soil covering culture 16~50cm of whole strain diameter weigh 1~10kg;Cap
2~5cm of diameter, bacterium leaf 2~6mm of thickness, for mushroom type shaped like chrysanthemum, mushroom color is white.
3. the selection of white chestnut mushroom kind described in a kind of claim 1, using ARTP induced-mutation technique breeding, feature exists
In:10 μ L chestnut mushroom protoplast suspension slide glasses are put into ARTP mutagenesis instrument fixed bit to postpone, are 8 SLM, mutagenesis in throughput
The processing time is set as under the conditions of 30-40 s, can get mutant.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810768050.2A CN108823107A (en) | 2018-07-13 | 2018-07-13 | A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810768050.2A CN108823107A (en) | 2018-07-13 | 2018-07-13 | A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108823107A true CN108823107A (en) | 2018-11-16 |
Family
ID=64137304
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810768050.2A Withdrawn CN108823107A (en) | 2018-07-13 | 2018-07-13 | A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108823107A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113174339A (en) * | 2021-06-10 | 2021-07-27 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Oomycopora ovales high-yield strain HMGIM-T43 and breeding method thereof |
CN116640673A (en) * | 2023-07-07 | 2023-08-25 | 上海市农业科学院 | Low-temperature-resistant straw mushroom strain and preparation method thereof |
Citations (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH1175592A (en) * | 1997-09-10 | 1999-03-23 | Godo Shiyusei Kk | Whitened grifola frondosa and its production |
JP2000167532A (en) * | 1998-12-03 | 2000-06-20 | Yukiguni Maitake Co Ltd | Decomposition of harmful environmental pollutant using edible mushroom |
JP2001197884A (en) * | 2000-01-18 | 2001-07-24 | Sakamoto Bio:Kk | Harmful substance removing agent containing decaying fungi and its waste fungus bed |
CN1398979A (en) * | 2002-08-16 | 2003-02-26 | 维京仲华(上海)生物医药科技有限公司 | Fermentation process of ash tree flower and production process of its polyglycopeptide |
KR100511182B1 (en) * | 2003-04-18 | 2005-08-30 | 농업회사법인 하나바이오텍(주) | The cultivating method of white Maitake (Grifola Frondosa) as short term and high yield |
JP2006232751A (en) * | 2005-02-25 | 2006-09-07 | Gunma Prefecture | Endothelial cell damage inhibitor and its use |
US20080213850A1 (en) * | 2005-02-28 | 2008-09-04 | Yukiguni Maitake Co., Ltd. | Pretreatment of Waste Mushroom Bed and Method of Converting the Same to Yield Sugars and Ethanol |
CN102690137A (en) * | 2012-06-15 | 2012-09-26 | 福建农林大学 | Substrate and method for culturing grifola frondosa (fr.) S.F.Gray by utilizing weeds |
CN103081720A (en) * | 2012-12-21 | 2013-05-08 | 孙思伦 | Isolated culture and cultivation method of white wild Grifola frondosa |
CN104757252A (en) * | 2015-04-22 | 2015-07-08 | 福建农林大学 | Preparation method of maitake protein zymolyte having antioxidant activity |
USPP25856P3 (en) * | 2012-10-18 | 2015-09-01 | Hokuto Corporation | Maitake mushroom named ‘Grifon-7’ |
CN105733956A (en) * | 2016-02-18 | 2016-07-06 | 天津泰创生物科技有限公司 | Ganoderma capense (Lloyd) Teng glycopeptide production fungi and application thereof |
JP2017113698A (en) * | 2015-12-24 | 2017-06-29 | エンザイム株式会社 | Biological treatment method for paper pulp waste water |
CN107509530A (en) * | 2017-09-19 | 2017-12-26 | 迁西县林中宝生物科技有限公司 | A kind of big bacteria stick method that earthing fixed point fruiting does not cultivate chestnut mushroom |
US20180000013A1 (en) * | 2016-02-26 | 2018-01-04 | Functional Fungi, Llc | Enhanced Mycelium Growth Medium and Method |
CN107779406A (en) * | 2017-09-24 | 2018-03-09 | 吉林农业大学 | Grifola frondosus facilityization cultivates new varieties and its liquid fermentation strain preparation method |
CN108048335A (en) * | 2017-12-25 | 2018-05-18 | 内蒙古自治区农牧业科学院 | Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection |
CN110396476A (en) * | 2019-07-05 | 2019-11-01 | 江苏大学 | One plant of Grifola frondosa strain obtained by ultraviolet mutagenesis |
-
2018
- 2018-07-13 CN CN201810768050.2A patent/CN108823107A/en not_active Withdrawn
Patent Citations (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH1175592A (en) * | 1997-09-10 | 1999-03-23 | Godo Shiyusei Kk | Whitened grifola frondosa and its production |
JP2000167532A (en) * | 1998-12-03 | 2000-06-20 | Yukiguni Maitake Co Ltd | Decomposition of harmful environmental pollutant using edible mushroom |
JP2001197884A (en) * | 2000-01-18 | 2001-07-24 | Sakamoto Bio:Kk | Harmful substance removing agent containing decaying fungi and its waste fungus bed |
CN1398979A (en) * | 2002-08-16 | 2003-02-26 | 维京仲华(上海)生物医药科技有限公司 | Fermentation process of ash tree flower and production process of its polyglycopeptide |
KR100511182B1 (en) * | 2003-04-18 | 2005-08-30 | 농업회사법인 하나바이오텍(주) | The cultivating method of white Maitake (Grifola Frondosa) as short term and high yield |
JP2006232751A (en) * | 2005-02-25 | 2006-09-07 | Gunma Prefecture | Endothelial cell damage inhibitor and its use |
US20080213850A1 (en) * | 2005-02-28 | 2008-09-04 | Yukiguni Maitake Co., Ltd. | Pretreatment of Waste Mushroom Bed and Method of Converting the Same to Yield Sugars and Ethanol |
CN102690137A (en) * | 2012-06-15 | 2012-09-26 | 福建农林大学 | Substrate and method for culturing grifola frondosa (fr.) S.F.Gray by utilizing weeds |
USPP25856P3 (en) * | 2012-10-18 | 2015-09-01 | Hokuto Corporation | Maitake mushroom named ‘Grifon-7’ |
CN103081720A (en) * | 2012-12-21 | 2013-05-08 | 孙思伦 | Isolated culture and cultivation method of white wild Grifola frondosa |
CN104757252A (en) * | 2015-04-22 | 2015-07-08 | 福建农林大学 | Preparation method of maitake protein zymolyte having antioxidant activity |
JP2017113698A (en) * | 2015-12-24 | 2017-06-29 | エンザイム株式会社 | Biological treatment method for paper pulp waste water |
CN105733956A (en) * | 2016-02-18 | 2016-07-06 | 天津泰创生物科技有限公司 | Ganoderma capense (Lloyd) Teng glycopeptide production fungi and application thereof |
US20180000013A1 (en) * | 2016-02-26 | 2018-01-04 | Functional Fungi, Llc | Enhanced Mycelium Growth Medium and Method |
US10609873B2 (en) * | 2016-02-26 | 2020-04-07 | Ancient Brands, Llc | Enhanced mycelium growth medium and method |
CN107509530A (en) * | 2017-09-19 | 2017-12-26 | 迁西县林中宝生物科技有限公司 | A kind of big bacteria stick method that earthing fixed point fruiting does not cultivate chestnut mushroom |
CN107779406A (en) * | 2017-09-24 | 2018-03-09 | 吉林农业大学 | Grifola frondosus facilityization cultivates new varieties and its liquid fermentation strain preparation method |
CN108048335A (en) * | 2017-12-25 | 2018-05-18 | 内蒙古自治区农牧业科学院 | Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection |
CN110396476A (en) * | 2019-07-05 | 2019-11-01 | 江苏大学 | One plant of Grifola frondosa strain obtained by ultraviolet mutagenesis |
Non-Patent Citations (11)
Title |
---|
CHEN, HSIAN-LING等: "Phenolic profiles and antioxidant abilities of various extracts from white maitake mushrooms (Grifolafrondosa)", 《FASEB JOURNAL》 * |
GARGANO, MARIA LETIZIA等: "Ecology, phylogeny, and potential nutritional and medicinal value of a rare white "maitake" collected in a Mediterranean forest", 《DIVERSITY-BASEL》 * |
孙艳颖: "香菇、滑子菇新品种选育研究", 《中国优秀硕士学位论文全文数据库(电子期刊)农业科技辑》 * |
宋鹤臣: "白色灰树花简介 ", 《特种经济动植物》 * |
张绪璋: "白色灰树花GF-8及栽培要点 ", 《食用菌》 * |
张绪璋: "稀有珍贵真菌――白色灰树花 ", 《北京农业》 * |
张绪璋等: "纯白灰树花的营养成分分析初报 ", 《福建农业大学学报》 * |
李怡彬等: "灰树花与白灰树花子实体蛋白质营养评价", 《中国农学通报》 * |
陈石良等: "深层发酵灰树花菌株的诱变筛选", 《食用菌》 * |
鲍正光: "白色灰树花及栽培技术 ", 《北京农业》 * |
鲍正光: "白色灰树花栽培技术 ", 《农村实用技术》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113174339A (en) * | 2021-06-10 | 2021-07-27 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Oomycopora ovales high-yield strain HMGIM-T43 and breeding method thereof |
CN113174339B (en) * | 2021-06-10 | 2022-08-02 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Oomycopora ovales high-yield strain HMGIM-T43 and breeding method thereof |
CN116640673A (en) * | 2023-07-07 | 2023-08-25 | 上海市农业科学院 | Low-temperature-resistant straw mushroom strain and preparation method thereof |
CN116640673B (en) * | 2023-07-07 | 2024-03-26 | 上海市农业科学院 | Low-temperature-resistant straw mushroom strain and preparation method thereof |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108048334B (en) | Establishment method of symbiotic system of gloeosporium fungi for promoting germination of cymbidium and cattleya seeds | |
CN1704469A (en) | Method for producing Chinese aweto fungus | |
Brown | Soil bacteriostasis limitation in growth of soil and rhizosphere bacteria | |
CN111876333B (en) | Lentinus edodes strain XG-3 capable of producing polysaccharide at high yield and application thereof | |
CN103710271A (en) | Morchella esculenta bacterial strain and culture method thereof | |
CN108865895A (en) | Paecilomyces hepiali chen ZJB18001 and its application | |
CN102119631A (en) | Grifola frondosa strain for producing polysaccharide with composite raw material of rice bran and wheat bran | |
CN101558766B (en) | Trichoderma solid granules for preventing and controlling tobacco soil-borne fungus diseases and preparation method thereof | |
CN108823107A (en) | A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding | |
CN105331548A (en) | Lepista nuda strain and liquid culture and preparation method thereof | |
CN111394507A (en) | Biocontrol fungus identification method for betel nut taro soft rot disease | |
CN107557313A (en) | A kind of composite conditioner for being passivated farmland cadmium pollution and promoting increasing production of rice | |
CN102676444B (en) | Culture medium for promoting growth of bacillus and application thereof | |
CN106591173A (en) | Bacillus flexus HL-37 capable of activating soil heavy metal cadmium, and applications thereof | |
CN107099465B (en) | Growth-promoting bacterial strain and its microorganism seedling medium for promoting cuttage tea shoot to take root | |
CN115261233B (en) | Biocontrol fungus for stem rot of saffron crocus and application thereof | |
CN110452821B (en) | Rhizosphere fungus capable of promoting development of adventitious roots and secondary roots of nursery stocks and application thereof | |
CN114766285A (en) | Ganoderma leucocontextum strain L4495 and cultivation method and application thereof | |
CN112746037B (en) | Streptomyces castochromogenes strain CPAT-W03 and application thereof | |
CN107699531A (en) | A kind of hippophae plant Frankia isolated culture method | |
CN108570442B (en) | Method for rapidly inducing spore production of anthrax | |
CN103114125A (en) | Indoor screening method of disease-resistant variety of rainbow conk | |
CN113046249A (en) | Verticillium lecanii LL-01 and biocontrol application thereof | |
CN110172411A (en) | A kind of scab shape Xylaria strain ZJ1811 and its cultural method and application | |
CN111411046A (en) | Dark color endophytic fungus agent and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20181116 |