CN102668993B - New bacterial strain of pholiota squarrosoides and sporocarp culture method thereof - Google Patents
New bacterial strain of pholiota squarrosoides and sporocarp culture method thereof Download PDFInfo
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Abstract
The invention discloses a new bacterial strain of pholiota squarrosoides and a sporocarp culture method thereof. The pholiota squarrosoides provided by the invention is specifically pholiota squarrosoides HS4 which has a preservation number of CGMCC No.6062 in the China General Microbiological Culture Collection Center (CGMCC). The hypha of the pholiota sguarrosoides HS4 with the preservation number of CGMCC No.6062 provided by the invention has high growth speed and short sackful time (which is advanced by 10d to 12d compared with that of pholiota adipose No.1), and can be taken as a hybridized parent to breed a new strain with high character hyphae breeding speed, short period and high yield. In addition, the pholiota squarrosoides HS4 with the preservation number of CGMCC No.6062 has high yield, and the biology efficiency of the first two tides of mushrooms reaches 51.29% which is higher than the biology efficiency of 42.67% of the pholiota adipose No.1.
Description
Technical field
The present invention relates to a kind of New bacterial strain of pholiota squarrosoides and its sporocarp culture method.
Background technology
The yellow umbrella of squarrosoides umbrella scientific name Pholiota squarrosoides (Peck) Sacc. alias point squama, sharp squama handle mushroom, thorn mushroom.Belong to mycota, Basidiomycota, Hymenomycetes, Agaricales, Strophariaceae, ring rust umbrella category on taxology.Squarrosoides destroying angel lovely luster is covered with pyramidal scale in golden yellow on cap stem.The Pseudomonas is in middle low temperature type edible fungus, and summer and autumn largely stub and generation on wood in woods, yield is higher；Its fructification is searched for food by people for many years rich in protein, carbohydrate, vitamin and multi mineral prime element, delicious flavour, unique flavor.Squarrosoides umbrella belongs to Strophariaceae, ring rust umbrella category with yellow umbrella, and morphological feature is more close；Mating reaction can occur for the single-ascospore strain of the two, therefore, can be used as the excellent new strains of hybrid strain seed selection.At present, the document on squarrosoides umbrella is less, the report for not yet having domesticating and cultivating.
Squarrosoides destroying angel is more medium big.Bacteria cover diameter 3-12cm, hemispherical is last flat to flat hemispherical, sometimes slightly convex, the dry tack free in middle part, and yellowish-brown or drabon colour band pink, the pyramidal color of scale are deeper, and middle part is more, easy to fall off, involute when lid edge is young, often adheres to collarium（Velum）Relic.Band milk yellow, pleat edge serrulate, Length discrepancy after bacterial context white.Stem is closely cylindrical, long 3-12cm, thick 0.8-1.5cm.Expanded to base portion, it is more than collarium white, and there is the graininess scale of similar lid color below, it is easy to fall off, it is internal soft to becoming hollow.Collarium film quality, above white, below band brown, it is frangible it is broken disappear.Spore print white.Spore is colourless, false starch reaction, smooth, ellipse,（7.3-8.1）μm×（2.3-3）μm.Utricule is bar-shaped, colourless or light brown,（20-50）μm×（8-12）μm.Squarrosoides umbrella summer and autumn is more in dragon spruce, fir, Pinuns densata, Korean pine and the scattered or all living creatures in mixed forest ground.
The content of the invention
The invention provides a kind of New bacterial strain of pholiota squarrosoides and its sporocarp culture method.
The New bacterial strain of pholiota squarrosoides that the present invention is provided is picked up from the persimmon tree of ChangPing, Beijing City, and squarrosoides umbrella is accredited as through Microbe Inst., Chinese Academy of Sciences fourth of the twelve Earthly Branches Mr. morning mist, and organizes separation to obtain original strain, is squarrosoides umbrella by the Strain Designation（Pholiota squarrosoides）HS4, the bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 26th, 2012（Abbreviation CGMCC, address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3）, deposit number is CGMCC No.6062.
The squarrosoides umbrella（Pholiota squarrosoides）Applications of the HS4CGMCC No.6062 in being cross-breeding as parent falls within protection scope of the present invention.
Cultivate the squarrosoides umbrella（Pholiota squarrosoides）The fructification that HS4CGMCC No.6062 are obtained falls within protection scope of the present invention.
Cultivate the squarrosoides umbrella（Pholiota squarrosoides）The mycelium that HS4CGMCC No.6062 are obtained falls within protection scope of the present invention.
The squarrosoides umbrella provided by the present invention（Pholiota squarrosoides）The cultural method of HS4CGMCC No.6062 fructifications, it may include following steps：
（1）Inoculation and bacterium germination：By the squarrosoides umbrella（Pholiota squarrosoides）HS4 CGMCC No.6062 cultigen access culture medium for cultivating, is 23 DEG C -25 DEG C in environment temperature, relative air humidity is 50%-60%, and well-ventilated, lucifuge culture 24-26d, mycelia covers with culture medium；
（2）Low temperature stimulation：3-5d is handled under the conditions of being transferred to -5 DEG C -5 DEG C；
（3）Fruiting：Environment temperature is transferred to for 15 DEG C -20 DEG C, relative air humidity 80%-90%, illumination 300-600Lux, well-ventilated, gas concentration lwevel between 2000-2300ppm under conditions of culture 20-22d formed to fruit body primordium, continue to cultivate；
（4）Harvesting：After 6-8d, fructification stem length to 7-9cm, cap is not yet fully deployed, you can harvesting.
In above-mentioned cultural method, the culture medium for cultivating is specifically mixed by cotton seed hulls, wood chip, wheat bran, corn flour, gypsum, lime and water；Wherein, the percent mass proportioning of the cotton seed hulls, the wood chip, the wheat bran, the corn flour, the gypsum and the lime is 60%：18%：15%：5%：1%：1%；Weight/mass percentage composition of the water in the culture medium for cultivating is 60%-65%.
In above-mentioned cultural method, the cultigen can be obtained by the following method：
（1）It is prepared by parent species：By the squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 are inoculated on mother culture media, 24 DEG C of -28 DEG C of lucifuge culture 7-10d, and obtained mycelium is as parent species；If gained parent species are used not in time, 4 DEG C can be stored in.
（2）It is prepared by original seed：By step（1）The parent species of gained are transferred on pedigree seed culture medium, and cultivation temperature is 23 DEG C -25 DEG C, and relative air humidity is 50%-60%, and container is covered with well-ventilated, lucifuge culture to mycelia, obtains the original seed.Original seed general room temperature storage period, is no more than 15d, and low-temp storage is no more than 20d, and expired strain can not be used.
（3）It is prepared by cultigen：By step（2）The original seed of gained is transferred on culture medium for cultivating, environment temperature be 23 DEG C -25 DEG C, relative air humidity be that culture 24-26d mycelia covers with bacterium bag under conditions of 50%-60%, well-ventilated, lucifuge, obtain the cultigen.
The squarrosoides umbrella（Pholiota squarrosoides）Application of the HS4CGMCC No.6062 fructification in food processing falls within protection scope of the present invention.
The squarrosoides umbrella（Pholiota squarrosoides）Thick flavor after HS4CGMCC No.6062 fructification is dry, quality is crisp when eating.
Squarrosoides umbrella provided by the present invention（Pholiota squarrosoides）HS4CGMCC No.6062 mycelial growth rate is fast, and the mycelia purseful time is short, it may be considered that as hybrid strain, the new strains that Breeding Traits mycelia speed is fast, the cycle is short, yield is high.In addition, squarrosoides umbrella（Pholiota squarrosoides）HS4 CGMCC No.6062 yield is high, and the biological efficiency of first two damp mushroom is up to 51.29%, more than existing yellow umbrella（Pholiota adipose）
The 42.67% of No. 1 cultivar.
Preservation explanation
It is recommended that Classification And Nomenclature：Squarrosoides umbrella
Latin name：（Pholiota squarrosoides）
Join the biomaterial of Ju：HS4
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 April 26th, 2012
Collection is registered on the books numbering：CGMCC No.6062
Brief description of the drawings
Fig. 1 is squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 sporophore shape.
Fig. 2 is squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 spore shape.
Fig. 3 is squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 hypha form, arrow meaning is the clamp connection of mycelia.
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
Yellow umbrella（Pholiota adipose）The such as No. 1 Meng Junlong, Liang Zhiying, Liu Jingyu Huang umbrella high yield and quality cultivation model study edible fungi of china .2006（25）：27-30.
The formula of various culture mediums is as follows used by following embodiments：
（1）Mother culture media：Potato 200g, glucose 20g, peptone 2g, MgSO40.2g、KH2PO40.25g、K2HPO40.25g, VB10.5mg, agar powder 15g, water 1000mL, pH7.0.
（2）Pedigree seed culture medium：Mixed by wood chip, wheat bran, gypsum, lime and water；Wherein, the percent mass proportioning of the wood chip, the wheat bran, the gypsum and the lime is 78%：20%：1%：1%；Weight/mass percentage composition of the water in the culture medium for cultivating is 60%-65%.
（3）Culture medium for cultivating：Mixed by cotton seed hulls, wood chip, wheat bran, corn flour, gypsum, lime and water；Wherein, the percent mass proportioning of the cotton seed hulls, the wood chip, the wheat bran, the corn flour, the gypsum and the lime is 60%：18%：15%：5%：1%：1%；Weight/mass percentage composition of the water in the culture medium for cultivating is 60%-65%.
Embodiment 1, squarrosoides umbrella（Pholiota squarrosoides）HS4 separation and identification
Bacterial strain of the present invention（It is denoted as HS4）Wild fructification pick up from the persimmon tree of ChangPing, Beijing City, be accredited as squarrosoides umbrella through Microbe Inst., Chinese Academy of Sciences fourth of the twelve Earthly Branches Mr. morning mist（Pholiota squarrosoides）.Further, by the observation to its fructification and mycelium morphology feature, and genetic interval sequence（ITS）Sequencing, judges the biological classification name of the bacterial strain.
First, morphological feature
1st, sporophore shape feature
Bacterial strain HS4 fructifications of the present invention are general medium big（Fig. 1）.Bacteria cover diameter 3-12cm, hemispherical is last flat to flat hemispherical, sometimes slightly convex, the dry tack free in middle part, yellowish-brown or drabon colour band pink, and scale is pyramidal, thorn-like, and color is deeper, and middle part is more, easy to fall off, involute when lid edge is young, often adheres to collarium（Velum）Relic.Band milk yellow after bacterial context white.Lamella is slightly close, growing straight, near-white, afterwards in shallow dark brown.Stem is closely cylindrical, long 4.5-8cm, thick 0.7-1.0cm, cellulosic, and interior real color is similar to cap, and the part of bottom 2/3 is covered with the scale of shallow rotten leaf color, collarium above near-white, no scale.Collarium is easy to fall off.Spore print is in rust brown.Spore is oval or subsphaeroidal, diameter（7.2-8.4）μm×（4.1-5.0）μm（Fig. 2）.Under an optical microscope, mycelia has clamp connection（Fig. 3）.Thick flavor after the bacterial strain fructification is dry, quality is crisp when eating.
2nd, the whole growth phase mycelia of mycelium morphology feature bacterial strain HS4 of the present invention is white, fine and close.
By above-mentioned Morphological Identification, bacterial strain HS4 of the present invention is tentatively judged as squarrosoides umbrella（Pholiota squarrosoides）.
2nd, rDNA-ITS is sequenced
RDNA-ITS sequencings are carried out to isolated mycelium from the fructification of collection using molecular biology method, its sequencing result is as shown in sequence 1 in sequence table.By sequence 1 and Genbank No.：FJ810180（Yellow umbrella（Pholiota adipose）DNAMAN shearing comparison processing is carried out, finds its homology up to 98.83%；Proposed in the research for the ectotrophic mycorrhiza group progress Molecular Identification that Renske etc. is distributed in soil layer, the rDNA ITS sequences for examination are compared by carrying out BLAST retrievals with NCBI GenBank databases, sequence similarity >=99%, it is possible to authenticate be mutually of the same race；Sequence similarity is 95%~99%, it is possible to authenticate be same genus；Sequence similarity≤95%, it is possible to authenticate be mutually equal（Renske L, Paula L, Thom W K, et al.Molecular identification of ectomycorrhizal mycelium in soil horizons.Applied and environmental microbiology, 2003,69 (1):327-333.）.Therefore, bacterial strain of the present invention and yellow umbrella be can conclude that（Pholiota adipose）Belong to ring rust umbrella category.
In view of above-mentioned morphological feature and rDNA-ITS sequencing identification results, mycota, Basidiomycota, Hymenomycetes, Agaricales, Strophariaceae, the squarrosoides umbrella of ring rust umbrella category are accredited as by bacterial strain HS4 of the present invention（Pholiota squarrosoides）.The bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 26th, 2012（Abbreviation CGMCC, address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3）, deposit number is CGMCCNo.6062.
Embodiment 2, squarrosoides umbrella（Pholiota squarrosoides）The artificial cultivation of HS4CGMCC No.6062 fructifications
First, the preparation of strain
（1）It is prepared by parent species：The squarrosoides umbrella that embodiment 1 is obtained（Pholiota squarrosoides）HS4CGMCC No.6062 are inoculated on mother culture media, 24 DEG C of -28 DEG C of lucifuge culture 7-10d, and obtained mycelium is as parent species；If gained parent species are used not in time, 4 DEG C can be stored in.
（2）It is prepared by original seed：By step（1）The parent species of gained are transferred on pedigree seed culture medium, and cultivation temperature is 23 DEG C -25 DEG C, and relative air humidity is 50%-60%, and well-ventilated, lucifuge culture to mycelia covers with container, obtains original seed.Original seed general room temperature storage period, is no more than 15d, and low-temp storage is no more than 20d, and expired strain can not be used.
（3）It is prepared by cultigen：By step（2）The original seed of gained is transferred on culture medium for cultivating, environment temperature be 23 DEG C -25 DEG C, relative air humidity be that culture 24-26d mycelia covers with bacterium bag under conditions of 50%-60%, well-ventilated, lucifuge, obtain cultigen.
2nd, cultural method
Squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 cultivation flow is specific as follows：Cultivate matrix manufacturing → cooling inoculation → bacterium germination → low temperature stimulation → fruiting → harvesting
（1）Cultivate matrix manufacturing and pack
Formula according to culture medium for cultivating above, which is prepared, obtains culture medium for cultivating, and it is conventionally prepared into bacterium bag, and the bacterium bag specification used is 17cm × 33cm × 0.04cm high pressure resistant polybag, per packed siccative（Other components in culture medium for cultivating in addition to water）0.40kg.Through autoclaving, bacterium bag to be seeded is obtained after cooling.
（2）Inoculation and bacterium germination：The squarrosoides umbrella that will be obtained by above-mentioned steps one（Pholiota squarrosoides）HS4CGMCC No.6062 cultigen uniformly accesses step（1）The bacterium bag of preparation, every bag of inoculum concentration is 3.14 × 10-5m3.In environment temperature it is 23 DEG C -25 DEG C after inoculation, relative air humidity is 50%-60%, well-ventilated, is cultivated under the conditions of lucifuge, and mycelia can cover with culture medium after 24-26d；
（2）Low temperature stimulation：3-5d is handled under the conditions of being transferred to -5 DEG C -5 DEG C；
（3）Fruiting：Environment temperature is transferred to for 15 DEG C -20 DEG C, relative air humidity 80%-90%, illumination 300-600Lux, well-ventilated, gas concentration lwevel between 2000-2300ppm under conditions of culture 20-22d formed to fruit body primordium, continue to cultivate；
（4）Harvesting：After 6-8d, fructification stem length to 7-9cm, cap is not yet fully deployed, you can the first damp mushroom of harvesting.Mycelium stimulation is carried out after harvesting, into mycelia convalescence, recovers aforesaid operations again after mycelia is replied and carries out fruiting, the second damp mushroom of harvesting can typically adopt 3-4 damp mushrooms.
Embodiment 3, squarrosoides umbrella（Pholiota squarrosoides）The detection of HS4CGMCC No.6062 mycelial growth rates, yield and nutritional ingredient
The present embodiment is with yellow umbrella（Pholiota adipose）No. 1 is control（Currently without the report cultivated on squarrosoides umbrella, control can only be used as from the yellow umbrella of its sibling species）, detect squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 mycelial growth rate, yield and nutritional ingredient.
First, the measure of mycelial growth rate
1st, experiment material
Squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 and yellow umbrella（Pholiota adipose）Parent species, original seed and the cultigen of No. 1 bacterial strain, the step one of its preparation method be the same as Example 2.
2nd, experimental method
Parent species Growth rate is determined：The parent species of two kinds of bacterial strains are inoculated in respectively after the center of Mother culture primary surface, culture 2-3d, record mycelia position（A）, record mycelia position after 3-5d（B）, measure the distance between A and B with ruler, divided by the number of days between two positions is the per day speed of growth of mycelia, five repetitions of Setup Experiments, results averaged.
Cultigen Growth rate is determined：The original seed of two bacterial strains is inoculated in the cultivating bag center equipped with culture medium for cultivating, makes to be uniformly distributed, after after mycelium germination 3-5d, mycelia position is recorded（A）, record mycelia position after 5-7d（B）, measure the distance between A and B with ruler, divided by the number of days between two positions is the per day speed of growth of mycelia, five repetitions of Setup Experiments, results averaged.
3rd, experimental result
Measurement result to parent species mycelial growth rate is as shown in table 2, squarrosoides umbrella（Pholiota squarrosoides）The mycelial growth rate of HS4CGMCC No.6062 parent species is up to 6.85 ± 0.15mm/d, far above the yellow umbrella of control（Pholiota adipose）4.33 ± 0.05mm/d of mycelial growth rate of No. 1 parent species.
The measurement result of the parent species mycelial growth rate of table 2（Unit：mm/d）
| Bacterial strain | Repeat 1 | Repeat 2 | Repeat 3 | Repeat 4 | Repeat 5 | Mean+SD |
| HS4 | 7.00 | 6.65 | 6.75 | 6.85 | 7.00 | 6.85±0.15 |
| No. 1 | 4.30 | 4.40 | 4.30 | 4.30 | 4.35 | 4.33±0.05 |
Measurement result to cultigen mycelial growth rate is as shown in table 3, squarrosoides umbrella（Pholiota squarrosoides）The mycelial growth rate of HS4CGMCC No.6062 cultigens is up to 3.02 ± 0.01mm/d, higher than the yellow umbrella of control（Pholiota adipose）2.51 ± 0.02mm/d of mycelial growth rate of No. 1 cultigen.
The measurement result of the cultigen mycelial growth rate of table 3（Unit：mm/d）
| Bacterial strain | Repeat 1 | Repeat 2 | Repeat 3 | Repeat 4 | Repeat 5 | Mean+SD mm/d |
| HS4 | 3.01 | 3.03 | 3.03 | 3.04 | 2.99 | 3.02±0.02 |
| No. 1 | 2.50 | 2.53 | 2.51 | 2.52 | 2.49 | 2.51±0.02 |
2nd, the measure of yield
By the squarrosoides umbrella of the present invention（Pholiota squarrosoides）HS4CGMCC No.6062 are with being used as the yellow umbrella compareed（Pholiota adipose）No. 1 is cultivated according to the cultural method of embodiment 2 respectively, harvests first two damp mushroom, its cultivation period and yield are compared.Three repetitions of Setup Experiments, it is each to repeat to cultivate 150 bacterium bags, results averaged.
1st, cultivation period
Under cultivation condition described in embodiment 2, squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCCNo.6062 bacteria developing period（From the time for being inoculated into mycelia and covering with bacterium bag）For 24-26d（Umbrella more yellow than control（Pholiota adipose）No. 1 shifts to an earlier date 10-12d）, low temperature stimulation 3-5d, fruit body primordium formation phase 20-22d under Cultivation condition, fruiting body differentiation to calculate to 6-8d is harvested and need 53-61d altogether from being seeded to the first damp mushroom of harvesting.Mushroom tide interval 14-16d, the second damp mushroom fruiting body differentiation is to harvesting 8-10d.The whole production cycle for going out two damp mushrooms is 75-87d.
Under cultivation condition described in embodiment 2, yellow umbrella（Pholiota adipose）No. 1 bacteria developing period is 34-38d, low temperature stimulation 3-5d, fruit body primordium formation phase 10-12d under Cultivation condition, and fruiting body differentiation to calculate to 4-6d is harvested and need 51-61d altogether from being seeded to the first damp mushroom of harvesting.Mushroom tide interval 12-15d, the second damp mushroom fruiting body differentiation is to harvesting 5-8d.The whole production cycle for going out two damp mushrooms is 68-84d.
2nd, yield
Yield is represented with " biological efficiency ".The biological efficiency refers to edible mushroom fresh weight and compost dry weight used（Other components in culture medium for cultivating in addition to water）The ratio between, conventional percentage is represented.Such as dry composts of 100kg produce 80kg fresh food bacterium, then the biological efficiency of this edible mushroom is 80%.
Under cultivation condition described in embodiment 2, squarrosoides umbrella（Pholiota squarrosoides）The biological efficiency of first two damp mushrooms of HS4CGMCCNo.6062 is up to 51.29 ± 1.68%, higher than the yellow umbrella as control（Pholiota adipose）The biological efficiency 42.67 ± 1.35% of the damp mushroom of 1 number two, refers to table 4.
The yield of first two damp mushroom of table 4（Biological efficiency）Statistics（Unit：%）
| Bacterial strain | Repeat 1 | Repeat 2 | Repeat 3 | Mean+SD |
| HS4 | 52.87 | 51.49 | 49.52 | 51.29±1.68 |
| No. 1 | 41.52 | 42.33 | 44.15 | 42.67±1.35 |
3rd, the analysis of nutritional ingredient
The squarrosoides umbrella harvested from step 2（Pholiota squarrosoides）HS4CGMCC No.6062 and the yellow umbrella as control（Pholiota adipose）500g is each randomly selected in first two damp mushroom of No. 1 and carries out nutritional ingredient（Amino acid content and vitamin content）Analysis, determined by Pu Ni test centers.Amino acid（18 kinds of amino acid in table 5）In in addition to tryptophan, the measure of amino acid is with reference to GB/T 5009.124-2003 in reference food；Thiamine（Vitamin B1）Measure with reference to GB/T 5009.84-2003；Riboflavin（Vitamin B2）Measure with reference to GB/T 5009.85-2003；The measure of vitamin B6 is with reference to GB/T 5009.154-2003；Nicotinic acid and niacinamide（Vitamin B3, PP）Measure with reference to GB 5413.15-2010.3 repetitions of Setup Experiments, results averaged.
Its amino acid and vitamin content as shown in table 5 and table 6, squarrosoides umbrella（Pholiota squarrosoides）Amino acid content is less than the yellow umbrella as control in HS4CGMCC No.6062 fructification（Pholiota adipose）No. 1, this is probably because the two bacterial strains belong to different biological species；And on vitamin level, its vitamin B2 content is yellow umbrella（P.adipose）7 times of No. 1, vitamin B3 is slightly below yellow umbrella（P.adipose）No. 1, vitamin B1 and B6 contents are more or less the same.In summary analyze, squarrosoides umbrella（Pholiota squarrosoides）The nutritional ingredient of HS4CGMCC No.6062 bacterial strains is slightly poorer than the yellow umbrella of control strain（P.adipose）No. 1, but in view of its mycelial growth rate is very fast and yield is higher, thus can as yellow umbrella production bacterial strain hybrid strain, seed selection has the new strains of the two merit concurrently.
The squarrosoides umbrella of table 5（Pholiota squarrosoides）HS4CGMCC No.6062 and yellow umbrella（Pholiota adipose）18 amino acid contents in 1 work song entity compare（Unit：g/100g）
| Amino acid | No. 1 | HS4 | Amino acid | No. 1 | HS4 |
| Asparatate | 1.47±0.01 | 0.85±0.00 | Methionine | 1.04±0.02 | 1.08±0.03 |
| Threonine | 0.76±0.01 | 0.47±0.00 | Isoleucine | 0.74±0.01 | 0.50±0.01 |
| Serine | 0.78±0.01 | 0.48±0.00 | Leucine | 1.20±0.01 | 0.80±0.02 |
| Glutamic acid | 3.23±0.05 | 1.54±0.00 | Tyrosine | 0.40±0.00 | 0.24±0.00 |
| Proline | 0.71±0.03 | 0.39±0.02 | Phenylalanine | 0.81±0.01 | 0.58±0.02 |
| Glycine | 0.67±0.01 | 0.40±0.00 | Lysine | 0.83±0.01 | 0.50±0.01 |
| Alanine | 0.89±0.01 | 0.54±0.00 | Histidine | 0.33±0.00 | 0.20±0.01 |
| Cystine | 0.14±0.01 | 0.09±0.01 | Tryptophan | 0.20±0.00 | 0.10±0.01 |
| Valine | 0.71±0.00 | 0.42±0.01 | Arginine | 1.11±0.02 | 0.44±0.00 |
Note：Unit g/100g, refers to that every 100g does the grams for containing corresponding amino acid in fructification.
The squarrosoides umbrella of table 6（Pholiota squarrosoides）HS4CGMCC No.6062 and yellow umbrella（Pholiota adipose）Vitamin content in 1 work song entity compares（Unit：mg/100g）
| Vitamin | No. 1 | HS4 |
| Vitamin B2 | 0.30±0.01 | 2.10±0.00 |
| Vitamin B1 | 0.21±0.00 | 0.30±0.00 |
| Vitamin B6 | 0.37±0.01 | 0.28±0.00 |
| Vitamin B3 | 75.00±2.00 | 50.75±0.75 |
Note：Unit mg/100g, refers to that every 100g does the milligram number for containing corresponding vitamin in fructification.
Summary result, squarrosoides umbrella provided by the present invention（Pholiota squarrosoides）HS4CGMCC No.6062 mycelial growth rates are very fast, and yield is high, rich in nutrition content, can as Huang umbrella strain breeding thereof Parents.
Claims (2)
1. the cultural method of fructification, comprises the following steps：
（1）Inoculation and bacterium germination：By squarrosoides umbrella（Pholiota squarrosoides）HS4CGMCC No.6062 cultigen access culture medium for cultivating, is 23 DEG C -25 DEG C in environment temperature, relative air humidity is to be cultivated under the conditions of 50%-60%, lucifuge, and culture medium is covered with to mycelia；
（2）Low temperature stimulation：After mycelia covers with culture medium, 3-5d is handled under the conditions of being transferred to -5 DEG C -5 DEG C；
（3）Fruiting：Environment temperature is transferred to for 15 DEG C -20 DEG C, relative air humidity 80%-90%, illumination 300-600Lux, gas concentration lwevel between 2000-2300ppm under conditions of culture obtain fructification.
2. according to the method described in claim 1, it is characterised in that：The culture medium for cultivating is mixed by cotton seed hulls, wood chip, wheat bran, corn flour, gypsum, lime and water；Wherein, the percent mass proportioning of the cotton seed hulls, the wood chip, the wheat bran, the corn flour, the gypsum and the lime is 60%：18%：15%：5%：1%：1%；Weight/mass percentage composition of the water in the culture medium for cultivating is 60%-65%.
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| CN1875682A (en) * | 2006-07-04 | 2006-12-13 | 彭俊 | Stropharia rugoso-annulata plantation method |
| CN102172168A (en) * | 2010-12-08 | 2011-09-07 | 中国科学院新疆理化技术研究所 | Method for separating, domesticating and cultivating wild Pholiota squarrosa strain |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1875682A (en) * | 2006-07-04 | 2006-12-13 | 彭俊 | Stropharia rugoso-annulata plantation method |
| CN102172168A (en) * | 2010-12-08 | 2011-09-07 | 中国科学院新疆理化技术研究所 | Method for separating, domesticating and cultivating wild Pholiota squarrosa strain |
Non-Patent Citations (2)
| Title |
|---|
| Analysis of Cultural Characteristics and Phylogenic Relationships of Collected Strains of Pholiota species;Yong-Hyun Cho et，al.;《Mycobiology》;20031231;第31卷(第4期);全文 * |
| 尖鳞伞特性及驯化栽培研究;常冀等;《食用菌》;20001130(第6期);第13页右栏第1段，和1.1菌种制作"部分，第14页表1和右栏16-20行；第13页倒数第6行-第14页第2行 * |
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