CN103202175A - Novel strain of lyophyllum decastes - Google Patents
Novel strain of lyophyllum decastes Download PDFInfo
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- CN103202175A CN103202175A CN201310050085XA CN201310050085A CN103202175A CN 103202175 A CN103202175 A CN 103202175A CN 201310050085X A CN201310050085X A CN 201310050085XA CN 201310050085 A CN201310050085 A CN 201310050085A CN 103202175 A CN103202175 A CN 103202175A
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Abstract
The invention relates to a novel strain Finc-LD-2 of lyophyllum decastes. The preservation number of the novel strain is China center for type culture collection number (CCTCCNO): M2013006. The Finc-LD-2 stain has the advantages of being short in cultivation and planting cycle, strong in infectious microbe resistance, free of earthing in a planting step, concentrated in collection period and suitable for factorization population and planting.
Description
[technical field]
The present invention relates to the edible mushroom technical field, or rather, relate to the new bacterial strain of a kind of Lyophyllum decastes.
[background technology]
Lyophyllum decastes (Latin title Lyophyllum decastes), be subordinate to Basidiomycotina, Hymenomycetes, Agaricales, white mushroom section, belong to from the pleat umbrella, be to eat the good wild edible fungus that Mycota is competitively studied cultivation both at home and abroad, the Lyophyllum decastes bacterial context is plump fine and smooth, A sweety scent assails the nostrils, the crisp cunning of mouthfeel, delicious flavour, nutritious, and to the repair of human body cell with having demonstrated great superiority aspect treatment cardiovascular disease, anti-ageing the waiting for a long time, be a kind of famous and precious edible mushroom that eats the medicine dual-purpose.Since 2006, Lyophyllum decastes began a small amount of artificial cultivation and achieving success in Japan.
[summary of the invention]
Characteristics such as the technical problem that the present invention solves provides the new bacterial strain Finc-LD-2 of a kind of Lyophyllum decastes, and this bacterial strain has cultivation and cultivation cycle weak point, and anti-assorted bacterium ability is strong, and the cultivation link does not need earthing, and picking time is concentrated.
The present invention is achieved by the following technical solutions, and the new bacterial strain Finc-LD-2 of Lyophyllum decastes of the present invention is by gathering the wild resource in Yunnan, through domesticating and cultivating, obtaining through screening system again.This bacterial strain has been preserved in Chinese typical culture collection center, address on January 8th, 2013: Chinese Wuhan City Wuhan University, and its preserving number is CCTCC NO:M2013006, its biology specific name is Lyophyllum decastes (Lyophyllum decastes).
The new bacterial strain Finc-LD-2 of Lyophyllum decastes of the present invention is characterised in that:
1, morphological feature
The mycelia of Finc-LD-2: aerial hyphae is dense; Cap: diameter is 3 ± 2cm, and it is light yellow that color is, rounded, upper flat, and the cap periphery is the strip ring grain, and Gai Yuan has small sawtooth, the no scale in cap surface, quality is tender and crisp, frangible; Lamella: light yellow, the lamella section is crescent; Stem: length is 6～15cm, stem top is pale yellow, and the bottom is pure white, near the stem interior solid consolidation at cap place, away from cap place stem hollow, there are white aerial hyphae in stem and cap binding site, and stem has scale, scale main give birth in the middle and upper part near cap, it is irregular to distribute, stem clusters, and base portion has the joint phenomenon, sees also Fig. 1.
2, biological property
The nutrient component source: wood chip, corncob, rice bran, wheat bran, corn flour, cotton seed hulls, soybean skin, forest humus etc., wherein, water content 64%～66% (percentage by weight, down together).
Cultivate and cultivation condition: the mycelium cultivation temperature is 20～25 ℃, humidity 75%, CO
2Concentration 2000～3500ppm, cultivation cycle 50～70 days.Suitable fruiting temperature is 15～18 ℃, humidity 93%～100%, CO
2Concentration 1000～2000ppm, preceding 1～7 day, intensity of illumination 10～50Lux, 8～27 days illumination intensity 200～300Lux in back, every day, clearance-type was opened layer bracket lamp stimulation fruiting, and the cultivation cycle is about 27 days.
3, genetics characteristic
The genomic DNA of Lyophyllum decastes Finc-LD-2 bacterial strain carries out pcr amplification through universal primer ITS1 and ITS4, obtains the specific DNA segment that size is about 606bp, and this fragment is checked order, and sequence information is referring to sequence table.
The new bacterial strain of Finc-LD-2 of the present invention, have and cultivate and cultivation cycle weak point, be conducive to improve the workshop week transfer efficient that batch production is produced, save production cost, anti-assorted bacterium ability is strong, reduced the production difficulty, the cultivation link does not need earthing, characteristics such as it is concentrated to gather, reduced the manual work intensity of production link widely, and long fresh-keeping period, per unit area yield height, suitable batch production popularizing planting.
[description of drawings]
Fig. 1 is the fruit body outward appearance picture of Lyophyllum decastes Finc-LD-2 bacterial strain of the present invention.
Fig. 2 is Lyophyllum decastes Finc-LD-2 bacterial strain phylogenetic tree of the present invention.
[embodiment]
Below by specific embodiment the present invention is described in further detail.
The domestication of embodiment 1 Finc-LD-2 of the present invention
Present embodiment has illustrated the domestication process of Lyophyllum decastes Finc-LD-2, and concrete steps are as follows:
1) preparation medium
PDA medium: every 1L medium potato 200g, glucose 20g, agar 20g.
2) collection of resources
In September, 2007, in 1800～3000 meters height above sea level areas of Yunnan Binchuan Jizu Shan Mountain, gather wild resource 4 strains, take back the laboratory, organize separation, be inoculated on the PDA medium, obtain to treat 4 of naturalized strains, numbering is respectively FLD-A, FLD-B, FLD-C, FLD-D.
3) strain domestication
It is numerous to use the PDA medium to expand the FLD-B in 4 bacterial strains that obtain, and 22 ℃, humidity 70%, gas concentration lwevel is less than 3000ppm, dark culturing.
To the cultivation of going down to posterity respectively of above-mentioned 4 bacterial strains, numerous 20 of each expansion was inoculated after 12～15 days, chose two the fastest test tube subcultures of a bacterium and expanded numerous, be seeded on the PDA medium, inoculate after 12～15 days, get again and send out two the fastest test tube subcultures of bacterium and expand numerously, repeat 10 circulations, to send out bacterium speed very fast to expanding numerous offspring's test tube, send out the neat homogeneous of bacterium speed, finish preliminary domestication work, and with liquid nitrogen cryopreservation bacterial strain is preserved in academy of agricultural sciences, Shanghai City edible mushroom research institute.
4) factory formula screening
The wheel formula tests screening through how, the female kind of acquisition, cultivated species prescription are:
Mother culture media: wood chip 50%(percentage by weight, down with), rice bran 25%, wheat bran 15% and corn flour 10%, water content is 60%～62%.
The cultivated species medium: wood chip 20%, corncob 15%, forest humus 40%, rice bran 10%, wheat bran 5% and corn flour 10%, water content is 64%～66%.
5) obtain aimed strain: by a series of domestication breeding work, obtain a Lyophyllum decastes bacterial strain, it has cultivates and cultivation cycle weak point, anti-assorted bacterium ability is strong, and the cultivation link does not need earthing, and it is concentrated to gather, characteristics such as per unit area yield height have reached the purpose of domestication breeding strain excellent.This bacterial strain is named as " Finc-LD-2 ".
In being preserved in Chinese typical culture collection center on January 18th, 2013, its preserving number is CCTCC NO:M2013006 to the new bacterial strain Finc-LD-2 of Lyophyllum decastes of the present invention, and bacterial strain is survived after testing, and classification position is for to belong to Lyophyllum decastes from the pleat umbrella.
Embodiment 2 carbon sources are to the influence of Lyophyllum decastes mycelial growth
For trying carbon source: glucose, sucrose and soluble starch.
The basal medium of test carbon source: magnesium sulfate 0.5g, potassium dihydrogen phosphate 1g, peptone 2g, agar 20g add water 1000ml.
In basal medium, add respectively for trying a kind of of carbon source, addition is 20g, pack into behind the mixing in the 250ml triangular flask, every bottle of 150ml puts into 121 ℃ of sterilizations of autoclave 30min, treats to cool off slightly the back and falls dull and stereotyped, the about 20ml medium of every plate, each dull and stereotyped central authorities connects the bacterium piece that a diameter is 5mm, and each processing arranges 5 repetitions, places under 25 ℃ of constant temperatures continuous culture 20 days.Came divided by the interval fate with the difference of twice test number with right-angled intersection method mensuration colony diameter, and be the mycelial growth rate of average every day in per 7 days.
Table 1 carbon source is to the influence of Finc-LD-2 and FLD-B mycelial growth
As shown in Table 1, supply to try carbon source from 3 kinds, Finc-B mycelial growth in the medium that with starch is carbon source is the fastest, and it is centimetre/day that average growth rate reaches 1.74cm/d(, down together), next is sucrose, is glucose at last; And Finc-LD-2 of the present invention average growth rate of mycelia in the medium that with starch and glucose is carbon source is suitable, 1.83cm/d and 1.82cm/d have been reached respectively, than the fast 0.09cm/d of wild strain, it is the slowest to grow in sucrose, and average growth rate is 1.63cm/d.Explanation thus, in the medium of different carbon sources, the average mycelial growth rate of Finc-LD-2 is all fast than wild strain, and starch is best, and glucose takes second place, and the uniformity of growth rate is better, and excursion is little.
Embodiment 3 nitrogenous sources are to the influence of Lyophyllum decastes mycelial growth
For trying nitrogenous source: ammonium sulfate, peptone and wheat bran.
The basal medium of test nitrogenous source: magnesium sulfate 0.5g, potassium dihydrogen phosphate 1g, glucose 20g, agar 20g add water 1000ml.
In the basal medium of measuring the nitrogenous source utilization, add respectively for trying a kind of of nitrogenous source, addition is the absolute mass of 2g(N), pack into behind the mixing in the 250ml triangular flask, every bottled 150ml puts into 121 ℃ of sterilizations of autoclave 30min, treats to cool off slightly the back and falls dull and stereotyped, the about 20ml medium of every plate, each dull and stereotyped central authorities connects the bacterium piece that a diameter is 5mm, and each processing arranges 5 repetitions, places under 25 ℃ of constant temperature culture conditions continuous culture 20 days.Came divided by the interval fate with the difference of twice test number with right-angled intersection method mensuration colony diameter, and be the mycelial growth rate of average every day in per 7 days.
Table 2 nitrogenous source is to the influence of Finc-LD-2 and FLD-B mycelial growth rate
As shown in Table 2, supply to try nitrogenous source from 3 kinds, Finc-B mycelial growth in the medium that with wheat bran is nitrogenous source is the fastest, and average growth rate is 1.83cm/d, is peptone secondly, is ammonium sulfate, only 0.48cm/d at last; And Finc-LD-2 of the present invention average growth rate of mycelia in the medium that with wheat bran is nitrogenous source has reached 1.89cm/d, than the fast 0.06cm/d of wild strain, growth rate is taken second place in peptone, is 1.61cm/d, it is the slowest that ammonium sulfate is that nitrogenous source is then grown, and is 0.73cm/d.Show that thus in the medium of different nitrogen sources, the average mycelial growth rate of Finc-LD-2 is all fast than wild strain, wheat bran is best, and peptone takes second place, and ammonium sulfate is the poorest, and the uniformity of mycelial growth rate is best with peptone.
Embodiment 4 C/N ratios are to the influence of Lyophyllum decastes mycelial growth
Be carbon source with glucose, ammonium sulfate is nitrogenous source, use magnesium sulfate 0.5g, yeast extract 2g, potassium dihydrogen phosphate 1g, agar 20g, add water 1000ml, the preparation C/N ratio is respectively 5 kinds of medium of 20:1,30:1,40:1,50:1 and 60:1, in the 250ml triangular flask of packing into, every bottled 120ml, pH is adjusted into 6.0, puts into 121 ℃ of sterilizations of autoclave 30min, treats to cool off slightly the back and falls dull and stereotyped.The about 20ml medium of every plate is the bacterium piece of 5mm at each dull and stereotyped central authorities inoculation one diameter, and the processing of each C/N ratio arranges 5 repetitions, places under 25 ℃ of constant temperatures continuous culture 20 days.Measured colony diameter with the right-angled intersection method in per 5 days, twice test number difference namely draws the mycelial growth rate of average every day divided by the interval fate, the results are shown in Table 3.
Table 3 C/N ratio is to the influence of the mycelial growth of Finc-LD-2 and FLD-B
As known from Table 3, when C/N ratio increases to 60:1 by 20:1, the mycelial growth rate of Finc-LD-2 and two bacterial strains of FLD-B all presents elder generation and increases the trend that afterwards reduces, when C/N ratio is 40:1, all reach maximum growth rate, Finc-LD-2 is 0.80cm/d, and uniformity is better, and Finc-B is 0.42cm/d.Show that thus Finc-LD-2 bacterial strain of the present invention is cultivated in the medium of different C/N ratios, and is stronger than the adaptive capacity of wild strain FLD-B, it is faster to send out bacterium speed, and the suitableeest C/N ratio of Finc-LD-2 bacterial strain is 40:1.
The influence of the Lyophyllum decastes mycelial growth of embodiment 5pH
Sodium hydroxide solution with 1.0% and 1.0% hydrochloric acid solution are regulated the acid-base value of PDA medium, obtain the pH value and are respectively 6 soda acid gradients of 4.0,5.0,6.0,7.0,8.0 and 9.0.The medium of gained is respectively charged in the 250ml triangular flask, and every bottle of 120ml puts into 121 ℃ of sterilizations of autoclave 30min, and cooling is fallen dull and stereotyped slightly.The dull and stereotyped about 20ml medium of every culture dish, treat to cool off fully afterwards solid plate.During inoculation, with one of the bacterium colony of the aseptic card punch cut-off footpath 5mm of sterilization, be seeded in the dull and stereotyped central authorities of PDA.Same pH condition is done 5 repetitions, places under 25 ℃ of constant temperature culture conditions continuous culture 20 days.Measured colony diameter with the right-angled intersection method in per 5 days, the difference of measuring numerical value twice namely draws the mycelial growth rate of average every day again divided by the interval fate, the results are shown in Table 4.
The influence of the table Finc-LD-2 of 4pH and FLD-B mycelial growth
As shown in Table 4, the optimal pH of Finc-LD-2 and the required medium of FLD-B all is 6.0, and mycelial growth rate is respectively 1.56cm/d and 1.48cm/d, and Finc-LD-2 is faster slightly than FLD-B growth rate.And, in the medium of identical pH condition, to cultivate, the mycelial growth rate of Finc-LD-2 is all slightly fast than FLD-B.
Embodiment 6 morphological features and biological property
The mycelia of morphological feature: Finc-LD-2: aerial hyphae is dense; Cap: diameter is 3 ± 2cm, and yellow of light color is rounded, upper flat, and the cap periphery is the strip ring grain, is circular around the decorative pattern, and Gai Yuan has small sawtooth, the no scale (see figure 1) in cap surface, quality is tender and crisp, frangible; Lamella: light yellow, the lamella section is crescent; Stem: length 6～15cm, top is pale yellow, the bottom is pure white, near cap place stem interior solid consolidation, away from cap place stem hollow, there are white aerial hyphae in stem and cap binding site, stem has scale, scale main give birth in the middle and upper part near cap, it is irregular to distribute, stem clusters, and base portion has the joint phenomenon.
Biological property: nutrient component source: wood chip, corncob, rice bran, wheat bran, corn flour, cotton seed hulls, soybean skin, forest humus etc., wherein, water content 64%～66%.
Cultivate and cultivation condition: the mycelium cultivation temperature is 20～25 ℃, humidity 75%, CO
2Concentration 2000～3500ppm, cultivation cycle 50～70 days.Suitable fruiting temperature is 15～18 ℃, humidity 93～100%, CO
2Concentration 1000～2000ppm, preceding 1～7 day illumination intensity 10～50Lux, 8～27 days the intensity of illumination 200～300Lux in back, every day, clearance-type was opened layer bracket lamp stimulation fruiting, and the cultivation cycle is about 27 days.
Embodiment 7 cultivation comparative trials
Present embodiment passes through from the new bacterial strain Finc-LD-2 of pleat umbrella, with cultivate contrast without habituation of wild bacterial strain FLD-B, investigate the anti-assorted bacterium ability of strains tested, index such as a situation arises, the concentration degree of gathering and the per unit area yield of the original hase (forming fruit body in the future) under the earthing condition not, be applicable to that to embody this bacterial strain the batch production bottle plants advantage and the characteristics of production.
The cultivated species medium that present embodiment adopts is: wood chip 20%, and corncob 15%, forest humus 40%, rice bran 10%, wheat bran 5% and corn flour 10%, water content is 64%～66%.
Cultivation stage pollution rate test is taked to cultivate common best cultivation again and carry out in advance earlier, inoculate 128 bottles, is placed in the high pre-incubation chamber of cleanliness factor cultivation then 15 days, go to common culturing room again, total cultivation cycle 57 days, statistics pollution condition during mycelium stimulation, the result consults table 5.
As known from Table 5, the new bacterial strain Finc-LD-2 of the Lyophyllum decastes pollution rate that obtains by domestication is 0, and the wild strain pollution rate is 3.91%.This shows, compare with wild strain that Finc-LD-2 has stronger anti-assorted bacterium infection ability.
Table 5Finc-LD-2 and FLD-B cultivation stage pollute contrast
Original hase generation number is research object with above-mentioned 128 inoculation bottles in the cultivation process, and under the situation of earthing not, a situation arises sees Table 6 for the original hase of Finc-LD-2 and wild strain FLD-B.As known from Table 6, the new bacterial strain Finc-LD-2 of Lyophyllum decastes by domestication obtains compares with wild strain, and original hase generation ratio is higher, and is stronger for the adaptive capacity of soil covering culture not.
A situation arises contrasts for table 6Finc-LD-2 and FLD-B original hase
Same batch of concentration degree of gathering of statistics is subjects with above-mentioned 128 inoculation bottles when gathering, and result of the test is consulted table 7.As known from Table 7, the new bacterial strain Finc-LD-2 of Lyophyllum decastes that obtains by domestication concentrates on first and gathers, and recovery ratio reaches 75.6%, and the gathering based on the 3rd batch of FLD-B, recovery ratio is 48.9%.This shows, compare with wild strain that the fruit body of Finc-LD-2 cycle of gathering is more concentrated, and concentrate the date of gathering obviously to shift to an earlier date than wild strain.
Table 7Finc-LD-2 and the FLD-B concentration degree of gathering contrasts
The subjects that is used for per unit area yield mensuration is above-mentioned 128 inoculation bottles, as known from Table 8, the new bacterial strain Finc-LD-2 of Lyophyllum decastes by the domestication acquisition, compare with wild strain, the cultivation per unit area yield is higher, under the situation of same test quantity, Finc-LD-2 has obtained higher gross yield, and total amount of increase in production has reached 43.1%.
Per unit area yield contrast after table 8Finc-LD-2 and FLD-B gather
Embodiment 8 fresh-keeping tests
After Finc-LD-2 and FLD-B gather, each 24 bag of picked at random Finc-LD-2 and FLD-B, in freezer (3 ℃～8 ℃) preservation to 55 day, the situation of change of record fruit body quality, result of the test sees Table 9.Wherein, normal: the expression fruit body is fresh fully; Begin to go bad: expression has one and above fruit body to begin mould corruption, but integral body still has commodity value; Rotten fully: two mould corruption of above fruit body, no commodity value.
As shown in Table 9, the new bacterial strain Finc-LD-2 of Lyophyllum decastes by domestication obtains compares with wild strain, and when preservation to 25 day, whole 24 bags are still intact, and FLD-B is in the time of the 15th day, and existing 5 to unwrap the beginning rotten.When continuing preservation to ground in the time of 30 days, 24 bags of FLD-B have all begun rotten or rotten fully, and Finc-LD-2 to the just began rotten or rotten fully for examination 24 bags in the time of 45 days.In the time of the 45th day, all of FLD-B 24 bags are all rotten fully, all lost commercial value, and this moment, Finc-LD-2 still had 15 bags to have commercial value, had reached more than 60%.All 24 bags of Finc-LD-2 are all rotten fully and the preservation time that all lose commercial value is 55 days, prolonged 10 days than FLD-B.Present embodiment shows, the new bacterial strain Finc-LD-2 of Lyophyllum decastes can realize the commodity shelf life more grown, compares with wild strain, and the shelf-life is longer, and the freshness date of Finc-LD-2 fruit body of the present invention has clear superiority.
Table 9Finc-LD-2 and FLD-B cold storing and fresh-keeping test contrast
Embodiment 9 gustations test
Choose fresh Finc-LD-2 and FLD-B, fry, the room temperature cooling is randomly drawed 100 people and is carried out blind test with regard to fragrance, bitter taste and the fragility of Finc-LD-2 and FLD-B, and everyone who participates in investigating fills in the gustation grade form, and standards of grading see Table 10.
Table 10 gustation test standards of grading
This experimental result as known from Table 11, each item rating of Finc-LD-2 is: fragrance 4.5, bitter taste 4.8, fragility 4.3, above-mentioned every appraisal result all is higher than the score of FLD-B, and the new bacterial strain Finc-LD-2 of this explanation Lyophyllum decastes of the present invention has happy people's taste and fragility, more is subjected to consumer's favor.
Table 11Finc-LD-2 and FLD-B gustation comparing result
The embodiment 10ITS evaluation of checking order
Present embodiment student on commission worker biotechnology (Shanghai) limited company (hereinafter to be referred as " giving birth to the worker ") carries out, and may further comprise the steps:
Genomic extraction: extract according to giving birth to worker SK1375 fungal gene group DNA extraction agent box specification.Resulting dna solution is placed-20 ℃ of preservations, standby.
The pcr amplification system is: cumulative volume 25 μ L, genomic templates 1 μ L, fungi rDNA universal primer ITS1 sequence is 5'-TCCGTAGGTGAACCTGCGG-3'(10 μ M) 0.5 μ L, the ITS4 sequence is 5'-TCCTCCGCTTATTGATATGC-3'(10 μ M) 0.5 μ L, dNTP mix (10mM each) 0.5 μ L, 10 * Taq reaction buffer, 2.5 μ L, 5U/ μ L Taq0.2 μ L adds water to 25 μ L.The PCR program: pre-94 ℃ of 5mim of sex change, 94 ℃ of 30s, 55 ℃ of 35s, 72 ℃ of 1min, 35 circulations, 72 ℃ are extended 8min.
Adopt conventional sequence measurement that above-mentioned pcr amplification product is checked order.The ITS sequence sequencing result of this test strain Finc-LD-2 sees also sequence table, and its sequence is the dna fragmentation of 606bp.Check in from GenBank, Finc-LD-2 bacterial strain ITS sequence is the highest with the ITS sequence similarity that belongs to Lyophyllum decastes from the pleat umbrella.Simultaneously, from GenBank, check in and the ITS sequence of close to pleat umbrella symbolic animal of the birth year, planting, use the phylogenetic tree that DNAStar MegAlign software building goes out strains tested, see also accompanying drawing 2.
More than describing is embodiments of the invention only, forgives and can understand, and under the prerequisite that does not depart from the present invention's design, all should be included within the technical conceive of the present invention simple modification of the present invention and replacement.
Sequence table
SEQUENCE LISTING
＜110〉Shanghai Finc Bio-tech Inc.
＜120〉Lyophyllum decastes bacterial strain Finc-LD-2
＜130〉ITS sequence
<160>1
<170>PatentIn version3.3
<210>1
<211>606
<212>DNA
<213>Lyophyllum shimeji
<400>1
tggttgggct gttgctggct cctaggagca tgtgcacgcc tagctccatc tttaccacct 60
gtgcaccttt tgtagacctg gatatctttc gaggaaactc ggtttgagga ctgctgagcg 120
gaaagcccta gcttccttta catttccggt ctatgtcttt acatacccca tattgaatgt 180
aacagaatgt cgtttactgg cctttgtgcc tttaatcaaa tacaactttc agcaacggat 240
ctcttggctc tcgcatcgat gaagaacgca gcgaaatgcg ataagtaatg tgaattgcag 300
aattcagtga atcatcgaat ctttgaacgc acctggcgct ccctggtatt ccggggagca 360
tgtctgtttg agtgtcatta aattctcaac ctttccaact tttgcgagtt tggttaggct 420
tggatgtggg gggtttgcgg gcttcacaga agtcggctcc tctgaaatgc attagtgaaa 480
cctttgttga tcatctcttg gtgtgataat tatctacgcc actagagtgt cagcctcaac 540
tagctgagag gtttttgctt ctaatcgtcc tccaaggaca acttctttac ttttgacctc 600
aaatca
606
Claims (1)
1. new bacterial strain Finc-LD-2 of Lyophyllum decastes, its deposit number is CCTCC NO:M 2013006.
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| CN105230342A (en) * | 2015-10-22 | 2016-01-13 | 上海丰科生物科技股份有限公司 | Lyophyllum decastes strain, cultivation method and application thereof |
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| CN105230342B (en) * | 2015-10-22 | 2018-03-06 | 上海丰科生物科技股份有限公司 | Lyophyllum decastes bacterial strain and its cultural method and application |
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| CN116171797A (en) * | 2023-04-04 | 2023-05-30 | 福建恒绿生物科技有限公司 | Cultivation method of velvet mushrooms |
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