CN117487672A - Yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation - Google Patents
Yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation Download PDFInfo
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- CN117487672A CN117487672A CN202311842136.2A CN202311842136A CN117487672A CN 117487672 A CN117487672 A CN 117487672A CN 202311842136 A CN202311842136 A CN 202311842136A CN 117487672 A CN117487672 A CN 117487672A
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- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims abstract description 5
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- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 1
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Abstract
A yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation belongs to the field of edible fungus breeding, and is preserved in China general microbiological culture Collection center (CGMCC) No.40892, and is JSJ-Spar16-1, classified and named Sparassis crispaSparassis sp.ITS sequence is shown in SEQ ID NO:1 shows that the strain is She Xiu cocci guangdong through ITS sequence comparisonSparassis latifolia). The strain has characteristic DNA sequence fragments which are different from other strains and are shown in SEQ ID NO: 2. The strain has the advantages of fast hypha growth, strong antibacterial capability, strong primordium, high fruiting rate of 90% or more, high biological conversion rate of 68.70% or more, a growth and development period of only 94-97 days, yellow or pale yellow color, good commodity characters, and good application prospects in the aspects of breeding, industrialized production and cultivation and the like.
Description
Technical Field
The invention belongs to the field of edible fungus breeding, and particularly relates to a yellow or pale yellow Sparassis crispa strain JSJ-Spar16-1 suitable for artificial cultivation.
Background
She Xiu cocci (Pediococcus furiosus)Sparassis latifolia) Belonging to Basidiomycota（Basidiomycota）Agaricus class（Agaricomycetes）The order of Polyporus（Polyporales）Sparassis Crispa (Sparassis)（Sparassidaceae）Genus SparassisSparassis). Sparassis crispa is a rare edible and medicinal fungus, contains very rich beta-glucan, and is the edible fungus with the highest beta-glucan content in the currently known edible fungus. At present, the researches on Sparassis crispa are mainly focused on biological characteristics, glucan, activity researches thereof and the like, but the researches on the genetic breeding of Sparassis crispa, domestication cultivation of new varieties, liquid submerged fermentation technology and the like are still weaker. Only 2 sparassis crispa varieties which are identified or identified in the industry of China are identified as Minxiu No. 1 in 2013 respectively, and the sparassis crispa is obtained by separating and purifying wild sparassis crispa, and the fruiting body is white or milky white, the cultivation period is 120 days, and the biological efficiency is 51.07%; and "Minxiu No. 2" identified in 2021, the fruiting body is pure white to milky white in color, the cultivation period is 106 days, and the yield is 223 g/bag on average. (quoted from https:// www.chinaseed114.com/seed/10/seed_48843.Html and https:// www.putian.gov.cn/zwgk/zdlyxxgk/shgyjsly/tpgjly/gzqk/202302/t 20230213_1799355.Htm, respectively). The problems of single sparassis crispa cultivar and serious homogenization, lack of abundant materials in genetic breeding, insufficient genetic breeding research and the like are one of the most fundamental reasons for restricting the development of sparassis crispa industry. Therefore, the breeding work of the new sparassis crispa variety is urgent.
Disclosure of Invention
The invention aims to provide a yellow or pale yellow sparassis crispa strain suitable for artificial cultivation, which is different from all the existing cultivated strains in morphology and genetics, is superior to the existing cultivated strains in yield, period, resistance and the like, provides rich materials for development of sparassis crispa industry and genetic breeding work, and promotes rapid development of sparassis crispa industry.
The technical scheme adopted by the invention is as follows:
a yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation is preserved in China general microbiological culture Collection center (CGMCC), the preservation unit address is North Xicilu No. 1, 3 in the Korean region of Beijing, the preservation date is 2023, 10 months and 7 days, the preservation number is CGMCC No.40892, the strain is JSJ-Spar16-1, and the strain is named Sparassis crispaSparassis sp.ITS sequence is shown in SEQ ID NO:1 shows that the strain is She Xiu cocci guangdong through ITS sequence comparisonSparassis latifolia) Belonging to Polyporales, sparassis of Sparassis.
The strain has a characteristic DNA sequence which is different from other strains, and the fragment of the characteristic DNA sequence is shown in SEQ ID NO: 2.
The fruiting body shape characteristics of the strain are as follows: the fruiting body is yellow or pale yellow, the edge of the fruiting body is small wavy, and the spore is pale yellow or beige, and the basidiomycete is wide and elliptical; the fruiting body comprises a handle, a plurality of flat branches which are staggered with each other and grow upwards from the handle, and a plurality of irregularly-bent petals formed at the tail ends of the branches, wherein the edges of the petals are in a small wave shape and are similar to a hydrangea flower and meat.
The strain has white mycelium, aerogenic mycelium, quick mycelium growth, good colony morphology consistency, obvious hypha locking combination and even chlamydospores in a culture medium formula of 18-20g/L glucose, 4-6g g/L corn meal, 4-6g g/L glutinous rice flour, 2-4g g/L fish meal peptone, 3-g g/L potassium dihydrogen phosphate, 1.5g g/L magnesium sulfate and 16-g g/L agar.
The invention provides a yellow or pale yellow sparassis crispa strain suitable for artificial cultivation, and the fruiting body of the strain is yellow or pale yellow, which is different from all the existing artificially cultivated white sparassis crispa strains. The yellow or pale yellow sparassis crispa strain suitable for artificial cultivation provided by the invention has characteristic DNA fragments different from other strains. The yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation, the matched cultivation substrate and the cultivation management method provided by the invention enable hyphae of the Sparassis crispa strain to grow fast, have strong antibacterial capability, white and robust primordium, high fruiting rate of 90% or more, high biological conversion rate of 68.70% or more, and short growth and development period of only 94-97 days.
The invention breeds yellow or pale yellow sparassis crispa varieties suitable for artificial cultivation by developing wild resource collection and systematic domestication and breeding, provides rich materials for development of sparassis crispa industry and genetic breeding work, and promotes rapid development of sparassis crispa industry.
Drawings
The yellow or pale yellow sparassis crispa strain suitable for artificial cultivation is preserved in China general microbiological culture Collection center (CGMCC), and the preservation date is 2023, 10 and 7 days, and the preservation number is CGMCC No.40892.
FIG. 1 shows the fruiting body shape of the strain of the invention compared with the fruiting body shape of the control strain, wherein A is the fruiting body shape of the strain of the invention, and B is the fruiting body shape of the control strain;
FIG. 2 shows the growth of the strain of the present invention in the fruiting pouch compared with the growth of the control strain, wherein A is the growth of the strain of the present invention in the fruiting pouch, and B is the growth of the control strain in the fruiting pouch.
FIG. 3 shows fruiting of the strain of the invention in a mushroom house.
FIG. 4 is a gel diagram of DNA molecular fragments specific to the strain of the present invention.
Detailed Description
In order to more clearly describe the technical scheme of the present invention, the technical scheme of the present invention will be clearly and completely described below with reference to the embodiments and the accompanying drawings. It is apparent that these embodiments and figures are only some, but not all, embodiments and figures of the present invention. The invention and its scope include, but are not limited to, embodiments. All modifications which can be made by a person skilled in the art without having to make any inventive effort are within the scope of the invention as claimed.
The percentages of the present invention are mass percentages unless otherwise indicated.
Examples
A yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation is preserved in China general microbiological culture Collection center with preservation number of CGMCC No.40892, and is JSJ-Spar16-1 named Sparassis crispaSparassis sp.。
The strain parent of the invention is obtained from wild sparassis crispa fruiting bodies of old junshan town in the university of Yunnan province in 9 months of 2022, and pure culture mycelium is obtained through tissue separation, purification and identification.
The strain is She Xiu coccus guangdongSparassis latifolia) Belonging to Polyporales, sparassis of Sparassis. ITS ITS sequence is shown in SEQ ID NO: 1.
TGGGCTTGCGGAGTCGAGCGAGGTTGTAGCTGGCCTTCTCGGAGGCATCGTGCACGCCCTGCCCGTCCCATATCATACCTGTGAACTTTTTGGTAGGCGGGTTTGTGTCGGCCTCGAAAGGGGTCGGCCGGCCCTCCGGCCGTCTTTATATACACACCATACGAGTCTTTAGAATGTTTGTGCGTCTCGACGCATCTTATATATAACTTTCAGCGACGGATCTCTTGGCTCTCGCATCGATGAAGAACGCAGCGAAACGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACCTTGCGCTCCTCGGTATTCCGAGGAGCATGCCTGTTTGAGTGTCATGAAATTATCAACCCCTCCTCCTTCATCGGTGGTGGGGCTTGGACTTGGAGGCTTTGCGGGCTTTTAACGAGTCGGCTCCTCTCAAATGCATTAGCTCGAACCCCTGCGGATCGGCCGTCGGTGTGATATAATGTCTACGTCGTGGTCGTGAGCGTCGGATCGGCTTCTAATGGTCCCCTTTCGGAGGCGGAATTTGAACTTGTGACCTCAAATCAGGTAGGACTACCCGCTGAACTTAAGCATATCAATAAGCGGAGGAA（SEQ ID NO：1）
The strain has a characteristic DNA sequence which is different from other strains, and the fragment of the characteristic DNA sequence is shown in SEQ ID NO: 2.
TGCGGAGAGAGAGAGAGAGCGTCGCGTCGAATGCTGTCGACGTGGGACCGAAGACTTCCTTCTCCTCTGGACTAACTTTGAAGGTTGACTCCAGTCTCTTCGGAAAACCATCTACTTTGCTACCTTTCGATAATCCCGGCTCTGAGACGAAGTCGGACCCACGAGAGAGTTCGCCGTTCGGGATCCCTTCTCTCTCTCTCTCAGCA（SEQ ID NO：2）
The cultivation method of the yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation comprises the following steps:
(1) Preparing strains:
a. mother culture
The mother culture adopts F1 mother culture medium, the formula of which is glucose 18-20g, corn flour 4-6g, glutinous rice flour 4-6g, fish meal peptone 2-4g, potassium dihydrogen phosphate 3g, magnesium sulfate 1.5g, agar 16g, and water is used for constant volume to 1 liter. The formula range is that the gradient experiment of each component in the formula proves that the strain has better culture effect, the mycelium of the strain in the formula of the F1 mother culture medium is white, the mycelium grows fast, the colony morphology consistency is good, the hypha lock-like combination is obvious, and chlamydospores are even.
The mother culture medium adopted in the embodiment comprises 20g of glucose, 5g of corn flour, 5g of glutinous rice flour, 3g of fish meal peptone, 3g of monopotassium phosphate, 1.5g of magnesium sulfate and 16g of agar, and is prepared by water for constant volume to 1 liter, sterilizing, packaging and cooling. Inoculating purified Sparassis crispa mycelium into the culture medium, and culturing in dark at 24+ -1deg.C.
b. Stock culture
The stock culture formula comprises 72% of pine sawdust, 9% of sweet potato powder, 12% of sorghum powder, 6% of corn powder, 1% of fish meal peptone and 64% of water content. Preparing a culture medium according to the formula, filling the culture medium into a 350 milliliter wide-mouth tissue culture bottle after preparing, sterilizing at high temperature and high pressure, and cooling for later use. And (3) cutting the mother seeds prepared in the step (a) into small pieces, inoculating the small pieces to a stock culture medium, and uniformly stirring by using a sterile tool. Dark culturing at 24+ -1deg.C for 20-25 days.
c. Cultivation seed culture (solid strain or liquid strain)
The solid strain culture medium comprises 72% of pine sawdust, 9% of sweet potato powder, 12% of sorghum powder, 6% of corn powder, 1% of fish meal peptone and 64% of water content of the culture medium. Preparing a culture medium according to the formula, filling the culture medium into a 350 milliliter wide-mouth tissue culture bottle after preparing, sterilizing at high temperature and high pressure, and cooling for later use. Inoculating the stock seeds prepared in the step b into a culture medium of cultivated species, and uniformly mixing and stirring. Dark culturing at 24+ -1deg.C for 20-30 days.
The liquid strain culturing method comprises the following steps:
the mother seeds or the stock seeds are used for preparing liquid seed liquid for fermentation, and the preparation method of the liquid seed liquid comprises the steps of crushing the stock seeds and using a sterile conical bottle according to strains: sterile water was 1: mixing the above materials at a ratio of 2-3 to obtain seed solution. The formula of the fermentation tank for preparing the liquid strain is as follows: 18-20g/L of glucose, 4-6g/L of corn meal, 4-6g/L of glutinous rice flour, 2-4g/L of fish meal peptone, 3g/L of monopotassium phosphate and 1.5g/L of magnesium sulfate. The formula range of the liquid strain fermentation tank is that the liquid strain fermentation tank has better culture effect after the gradient experiment of each component in the formula is carried out. The fermentation tank formula of the embodiment comprises 20g/L glucose, 5g/L corn meal, 5g/L glutinous rice flour, 3g/L fish meal peptone, 3g/L potassium dihydrogen phosphate and 1.5g/L magnesium sulfate.
The preparation method comprises the steps of dissolving the formula in water according to a certain proportion, pouring the mixture into a fermentation tank, sterilizing, cooling, inoculating the prepared seed liquid into the fermentation tank according to an inoculum size of 1-2%, and culturing at constant temperature of 24+/-1 ℃ for 7-9 days. This example was incubated for 8 days.
(2) Preparing a cultivated fruiting bag:
the fruiting bag comprises the following formula: 62-68% of whole pine sawdust, 25-35% of carbon-nitrogen source compound and 10-15% of soil; the carbon-nitrogen source compound is sweet potato powder: rice flour and/or tapioca flour and/or corn flour: the mass ratio of the peptone is 7-10:4-6:1-2, the fruiting content of which is 63-65% of water and the pH value of which is 5-6. The numerical range of the fruiting bag formula is a better formula determined after gradient experiments of all components in the formula are carried out. The whole pine sawdust is adopted in the embodiment, the carbon-nitrogen source compound is 30% and the soil is 10%. Fruiting contained approximately 64% water, pH5.5.
(3) Cultivation and management: culturing the cultured strain in fruiting bag, controlling the culture temperature at 23-25deg.C in mycelium culture stage, and culturing in dark for 40-45 days, wherein the carbon dioxide concentration in the culture chamber is less than or equal to 3000ppm, and the carbon dioxide concentration in the fungus bag is less than or equal to 3%; controlling the induction temperature at 16-19 ℃ in the primordial induction stage, inducing for 20-25 days, illuminating for 500-2000Lx, irradiating for more than or equal to 11 hours every day, inducing the indoor carbon dioxide concentration to be less than or equal to 2000ppm, the carbon dioxide concentration in the bag to be less than or equal to 2%, and the air humidity to be 70% -80%; controlling the temperature at 18-20deg.C during fruiting body development stage, culturing for 15-20 days, illuminating for 500-2000Lx, illuminating for more than or equal to 11 hr every day, wherein indoor carbon dioxide concentration is less than or equal to 1000ppm, carbon dioxide concentration in bag inner side wall is less than or equal to 1%, and air humidity is more than or equal to 95%; the whole growth and development period is 94-97 days. The mycelium grows fast, the antibacterial capability is strong, the primordium is white and strong, the fruiting rate is more than 90%, and the biological conversion rate is more than 68.70%. The above-mentioned technical parameter ranges are preferred technical parameters determined after the gradient experiment is performed.
In the embodiment, the culture temperature is controlled to be 24 ℃ in the mycelium culture stage, the culturing is performed in a dark state for 42 days, the carbon dioxide concentration in a culture chamber is not more than 2800ppm, and the carbon dioxide concentration in a fungus bag is not more than 2%. Controlling the induction temperature at 18 ℃ in the primordial induction stage, inducing for 22 days, illuminating 500-2000Lx, illuminating for not less than 12 hours every day, inducing the indoor carbon dioxide concentration to be not more than 1800ppm, the carbon dioxide concentration in the bag to be not more than 2%, and the air humidity to be about 75%. The temperature is controlled at 18-20deg.C at fruiting body development stage, culturing for 18 days, illuminating for 500-2000Lx, illuminating for not less than 12 hr every day, indoor carbon dioxide concentration not exceeding 1000ppm, carbon dioxide concentration not exceeding 1% at bag inner side wall, and air humidity not less than 95%. The whole growth and development period is 95 days. The mycelium grows fast, the capability of resisting bacteria is strong, the primordium is white and strong, the fruiting rate reaches 90%, and the biological conversion rate reaches 68.70%.
Fruiting management is carried out on the pure culture of Sparassis crispa obtained through first generation separation according to the method to obtain mature fruiting bodies, then tissue separation or basidiospore separation is carried out on the mature fruiting bodies to obtain pure culture, fruiting management is carried out on the pure culture to obtain mature fruiting bodies, indoor artificial cultivation and domestication are circularly and repeatedly carried out for 7 rounds, and finally the strain with stable properties and yield, namely the strain JSJ-Spar16-1 is obtained. Table 1 counts the results of 7 runs:
table 17 results of the test for fruiting by means of acclimatization
Further, the stable strain obtained by the 7-wheel train domestication was subjected to fruiting verification for 3 more times, 100 bags were repeated each time, and fruiting conditions and fruiting body properties were recorded, as shown in table 2 and fig. 1.
TABLE 2 statistics of production traits of the strain JSJ-Spar16-1 of the invention
As can be seen from FIG. 1, the fruiting body of the strain of the invention is yellow or pale yellow, the edge of the fruiting body is slightly wavy, and the spore print is pale yellow, which is obviously different from the white or milky white Sparassis crispa strain cultivated artificially at present. The fruiting body shape of the strain is characterized as follows:
the fruiting body is yellow or pale yellow, the edge of fruiting body is small wavy, and the spore is pale yellow or beige, and the basidiomycete is wide and elliptical. The fruiting body comprises a handle, a plurality of flat branches which are staggered with each other and grow upwards from the handle, and a plurality of irregularly-bent petals formed at the tail ends of the branches, wherein the edges of the petals are in a small wave shape and are similar to a hydrangea flower and meat.
The strain has fast hypha growth, strong antibacterial capability, white and robust primordium, fruiting rate of 90%, biological conversion rate of 68.70% and growth and development period of 94-97 days.
Construction of sparassis crispa SCAR molecular markers:
and analyzing the genetic diversity of Sparassis crispa based on ISSR markers and constructing molecular markers. Using 100 ISSR random primers published by Columbia university (see reference: zhengqing. Application of DNA molecular marker technology in plant research [ M ]. Beijing: chemical industry Press, 2005.), after multiple rounds of screening, specific primers were obtained for amplifying a DNA fragment specific to the JSJ-Spar16-1 strain, the DNA fragment being 206bp in size, and deleting the fragment in other strains, see FIG. 4. In FIG. 4, M is Mark, CK is a blank, lanes 1 to 8 are strains according to the present invention, and lanes 9 to 12 are control strains. As can be seen, the strains of the invention have characteristic DNA fragments which are distinguished from the control strain. The characteristic DNA sequence fragment of the strain is shown as SEQ ID NO: 2.
Claims (4)
1. A yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.40892, and is JSJ-Spar16-1, classified and named Sparassis crispaSparassis sp.ITS sequence is shown in SEQ ID NO:1, the strain is a Guangdong She Xiu coccus belonging to the genus Sparassis of the order Polyporaceae by ITS sequence alignment.
2. A strain of sparassis crispa suitable for artificial cultivation as claimed in claim 1, said strain having a characteristic DNA sequence fragment as set forth in SEQ ID NO: 2.
3. A strain of sparassis crispa suitable for artificial cultivation as defined in claim 1, the fruiting body of said strain being characterised by: the fruiting body is yellow or pale yellow, the edge of the fruiting body is small wavy, and the spore is pale yellow or beige, and the basidiomycete is wide and elliptical; the fruiting body comprises a handle, a plurality of flat branches which are staggered with each other and grow upwards from the handle, and a plurality of irregularly-bent petals formed at the tail ends of the branches, wherein the edges of the petals are in a shape of small waves, a embroidered ball, and the meat quality.
4. A yellow or pale yellow Sparassis crispa strain suitable for artificial cultivation as claimed in claim 1, wherein the strain has white mycelium, aerial mycelium and even chlamydospores in a culture medium formula of 18-20g/L glucose, 4-6g g/L corn flour, 4-6g g/L glutinous rice flour, 2-4g g/L fish meal peptone, 3-g g/L potassium dihydrogen phosphate, 1.5g g/L magnesium sulfate and 16-g g/L agar.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102863271A (en) * | 2012-09-27 | 2013-01-09 | 中国科学院微生物研究所 | Synthetic medium of broad-leave Sparassis crispa |
| CN104429604A (en) * | 2014-12-04 | 2015-03-25 | 福建省农业科学院食用菌研究所 | Sparassis crispa liquid strain culture medium and culture method |
| KR20150073504A (en) * | 2013-12-23 | 2015-07-01 | 전남대학교산학협력단 | A Novel Sparassis latifolia strain |
| CN104928189A (en) * | 2015-05-29 | 2015-09-23 | 吉林大学 | High-yield sparassis crispa mutant strain and culture method |
| CN110777135A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Fermentation method for high yield of β -glucan based on sparassis crispa |
| CN112592835A (en) * | 2020-12-11 | 2021-04-02 | 云南菌视界生物科技有限公司 | Tremella aurantialba strain suitable for industrial cultivation |
| WO2022195581A1 (en) * | 2021-03-16 | 2022-09-22 | Cannabotech Ltd. | Novel mushroom strains, extracts thereof and compositions comprising them |
| CN115226571A (en) * | 2022-07-25 | 2022-10-25 | 江苏品品鲜生物科技股份有限公司 | Sparassis crispa liquid strain culture medium and culture method thereof |
| CN116042896A (en) * | 2022-12-09 | 2023-05-02 | 福建省农业科学院食用菌研究所 | Indel molecular marker for identifying SP-A strain of Pediococcus guangdalina She Xiu and application |
-
2023
- 2023-12-29 CN CN202311842136.2A patent/CN117487672B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102863271A (en) * | 2012-09-27 | 2013-01-09 | 中国科学院微生物研究所 | Synthetic medium of broad-leave Sparassis crispa |
| KR20150073504A (en) * | 2013-12-23 | 2015-07-01 | 전남대학교산학협력단 | A Novel Sparassis latifolia strain |
| CN104429604A (en) * | 2014-12-04 | 2015-03-25 | 福建省农业科学院食用菌研究所 | Sparassis crispa liquid strain culture medium and culture method |
| CN104928189A (en) * | 2015-05-29 | 2015-09-23 | 吉林大学 | High-yield sparassis crispa mutant strain and culture method |
| CN110777135A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Fermentation method for high yield of β -glucan based on sparassis crispa |
| CN112592835A (en) * | 2020-12-11 | 2021-04-02 | 云南菌视界生物科技有限公司 | Tremella aurantialba strain suitable for industrial cultivation |
| WO2022195581A1 (en) * | 2021-03-16 | 2022-09-22 | Cannabotech Ltd. | Novel mushroom strains, extracts thereof and compositions comprising them |
| CN115226571A (en) * | 2022-07-25 | 2022-10-25 | 江苏品品鲜生物科技股份有限公司 | Sparassis crispa liquid strain culture medium and culture method thereof |
| CN116042896A (en) * | 2022-12-09 | 2023-05-02 | 福建省农业科学院食用菌研究所 | Indel molecular marker for identifying SP-A strain of Pediococcus guangdalina She Xiu and application |
Non-Patent Citations (2)
| Title |
|---|
| LEE 等: "Noble strain of Sparassis latifolia produces high content of b-glucan", ASIAN PACIFIC JOURNAL OF TROPICAL BIOMEDICINE, vol. 5, no. 8, 17 July 2015 (2015-07-17), pages 629 - 635 * |
| 杨驰 等: "基于ITS 序列和BOX-PCR 鉴定广叶绣球菌菌株", 中国食用菌, vol. 35, no. 5, 15 September 2016 (2016-09-15), pages 37 - 39 * |
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