CN107493979A - Cordyceps militaris ascospore cross breeding method based on Protocols in Molecular Biology - Google Patents
Cordyceps militaris ascospore cross breeding method based on Protocols in Molecular Biology Download PDFInfo
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Abstract
The present invention relates to a kind of Cordyceps militaris ascospore cross breeding method based on Protocols in Molecular Biology, specifically include：Induced in gnotobasis and separate Cordyceps militaris monospore and cultivate and form bacterium colony；The bacterium colony formed using round pcr to the monospore of separation is carried out mating type gene MAT1 11, MAT1 12 and MAT1 21 and detected；The bacterium colony for not containing MAT1 21 monospore but containing mating type gene MAT1 11 and MAT1 12 and being formed is chosen, the bacterium colony that the monospore with not containing MAT1 11 and MAT1 12 but containing mating type gene MAT1 21 is formed takes appropriately sized mycelium to be placed in nutrient solution and carries out light culture respectively；Bacterium solution after the combination culture of debita spissitudo is inoculated into Cordyceps militaris synthetic medium, after cultivating a period of time, the fruiting bodies of cordyceps militaris of new varieties is grown on synthetic medium.The Cordyceps militaris new varieties that this method is cultivated are easier to obtain preferable yield and fructification activity substance content, the fruiting body yield and activity substance content turned out by counting various combination, can obtain can stably produce the combination of high-quality fruiting bodies of cordyceps militaris, this method cycle is shorter, workload is smaller, is a kind of efficient Cordyceps militaris breeding method.
Description
Technical field
The present invention relates to biological technical field, and in particular to a kind of Cordyceps militaris ascospore based on Protocols in Molecular Biology
Cross breeding method.
Background technology
In the sexual reproduction of fungi, mating type can illustrate hereditary control situation and the zoogamy development of fungi
Process, therefore, mating type gene plays decisive role, and the affine performance of property to the sexual control of fungi and genetic evolution
Type genetic determinant.The mating system of fungi known is divided into P.drechsleri and the major class of heterothallism two.P.drechsleri is a kind of
Autologous zoogamy mode that can be pregnant, heterothallism are a kind of autologous infertile zoogamy modes.Heterothallism is according to control
The incompatibility factor of mating type is a pair or two pairs, is divided into two classes：The different ancestor of two polarity with a pair of incompatibility factors
Coordinate (bipolar heterot hallism) and the quadripolarity heterothallism with two pairs of incompatibility factors
(tetrapolar heterot hallism)。
Cordyceps militaris is two polarity heterothallic fungus.Two kinds of different mating types of Cordyceps militaris are by single mating site
What MAT1 was determined.One of them is MAT-alpha（MAT1-1）, it contains the ORFs of a coding for alpha box albumen, this
Contain two mating type genes in site：MAT1-1-1 and MAT1-1-2.Another is MAT-HMG (MAT1-2), and it contains one
A kind of ORFs of regulatory protein is encoded, and this regulatory protein contains HMG（High Mobility Group
Proteins）DNA calmodulin binding domain CaMs on albumen.MAT-HMG includes a mating type gene：MAT1-2-1.
Heterothallic fungi breeding, Main Means are according to the characteristic of fungi zoogamy, implement crossbreeding.Substantially
Method is exactly that the monospore of two different double-core bacterial strains of genotype is separated, and is matched two-by-two.If the two monospore
To belong to different sexes, i.e., can be affine after pairing containing the different incompatibility factors.Each of which is sprouted
Into uninucleate hyphae can mate to form the dicaryon with knot physical capabilities.Many different hybridization dicaryons are gone out
Grass experiment, desired quality strains can be filtered out according to careless result of the test is gone out.
PCR（PCR）It is a kind of Protocols in Molecular Biology for being used to amplify the specific DNA fragmentation of amplification.Profit
With after PCR amplifications Cordyceps militaris DNA, again by electrophoretic techniques, the mesh to the detection of Cordyceps militaris monospore mating type gene can be reached
, then pointedly select the monospore for belonging to different sexes to be matched two-by-two, greatly improve the efficiency of crossbreeding.
Currently, cultivating Cordyceps militaris, it obtains the mode of strain mainly using vegetative propagation technique, i.e., directly from
Mycelial technology is separated on Cordyceps militaris tissue block, its strain inhereditary feature is very single, it is impossible to which multidirectional selection is more preferably excellent
Strain, although the technology can keep the outward appearance condition of fruiting bodies of cordyceps militaris, with the degeneration of strain, the content of its active material
Decline clearly, therefore its active material of most fruiting bodies of cordyceps militaris is extremely low in the market, does not reach as medicinal
Purpose.
And after being identified by Protocols in Molecular Biology Cordyceps militaris ascospore mating type, then by different genes mating type
Monospore carries out sexual hybridization, make use of biological gene restructuring and the characteristic separated so that its strain has genetic diversity, can
It is higher and the strain of more active materials can be enriched with to filter out fruiting bodies of cordyceps militaris yield.
Now report discloses a variety of Cordyceps militaris breeding modes, and based on the Cordyceps militaris ascus spore of Protocols in Molecular Biology auxiliary
Sub- crossbreeding mode there is no report.
The content of the invention
The present invention relates to a kind of Cordyceps militaris ascospore cross breeding method based on Protocols in Molecular Biology, solves pupa
The problem of cordyceps species hereditary capacity itself causes fruiting body yield and relatively low activity substance content.This method cycle is shorter,
Workload is smaller, is a kind of efficient Cordyceps militaris breeding method.
The present invention is mainly achieved through the following technical solutions：
Cordyceps militaris spawn used in the present invention, come from the strain library of Changde Yandi Biotechnology Co., Ltd., specific strain
For one kind in megaspore head kind, microspore head kind, tip conidia powder kind；This method comprises the following steps that：
(1) aseptically, fresh, ripe fruiting bodies of cordyceps militaris is suspended from above water agar, avoids fructification
Contacted with culture medium；
(2) transfer them in biochemical cultivation case, 21 DEG C of lucifuge cultures 4-5 days, ascospore is launched naturally and arrive PDA culture medium
On；
(3) picking only has the bacterium colony of single ascospore under Stereo microscope, adds 2ml sterilized water, stirs,
Spore suspension is prepared into, takes 200 microlitres of spore suspension, is coated with PDA plate culture medium, is positioned over 21 DEG C of biochemical cultivation cases
Middle lucifuge culture；
(4) after cultivating 3-4 days, macroscopic ascospore single bacterium colony is chosen on superclean bench, new PDA is transferred to and puts down
On plate culture medium, single bacterium colony is only cultivated on each PDA plate culture medium；
(5) after cultivating 5 days, the single ascospore bacterium colony of purifying is taken to extract its DNA, the son using PCR amplification techniques to extraction
Cystospore single bacterium colony DNA carries out mating type gene MAT1-1-1, MAT1-1-2 and MAT1-2-1 detection；
(6) nutrient solution is configured：By glucose 2%, peptone 0.5%, beef extract 0.5%, epsom salt 0.1%, potassium dihydrogen phosphate
0.1%th, vitamin b1 0.001%, vitamin b12 0.001%, water 96.798% form, and are well mixed；
(7) by step（6）Middle nutrient solution is dispensed into triangular flask, autoclave sterilization 20~30 minutes, and taking-up is cooled to normal temperature
It is standby；
(8) selecting step（5）The list for not containing MAT1-2-1 but containing mating type gene MAT1-1-1 and MAT1-1-2 of detection
The bacterium colony of sporogenesis, the monospore with not containing MAT1-1-1 and MAT1-1-2 but containing mating type gene MAT1-2-1 are formed
Bacterium colony take the mycelium block of 2~5% sizes to be inoculated in step respectively（7）In the nutrient solution made, it is placed in concussion shaking table, if
Put 19 DEG C, lucifuge, rotating speed 150rpm Shaking cultures 4-5 days；
(9) by step（8）Cultured mycelium is inoculated on synthetic medium, be transferred to 17~25 DEG C of temperature, humidity 65~
In the environment of 85%, light culture 5~7 days, treat that media surface grows a large amount of white hyphas；
(10) under conditions of step (9), 200~2000lux of intensity of illumination is set, carries out seeing light annesl, generated by former base,
Go out flower bud growth, go out grass growth three phases, treat that cordyceps militaris sporocarp length harvests to 4~5cm height, then by fructification.
The present invention has following 3 big advantages：
（1）The present invention has genetic diversity using the restructuring and stalling characteristic, its strain of gene in biological sexual reproduction process,
Therefore developing new product variety can be used for；
（2）The present invention is detected by gene mating type, and the ascospore using different mating types is hybridized, and improves Cordyceps militaris
The efficiency of breeding of new variety, the crossbreeding cycle is shortened, reduces workload；
（3）The fruiting bodies of cordyceps militaris cultivated by present invention screening, its yield and active material substantially increase, and improve economy
Benefit and medical value.
Embodiment
To make the object, technical solutions and advantages of the present invention of greater clarity, with reference to embodiment, to this
Invention is further described.It should be understood that these descriptions are merely illustrative, and it is not intended to limit the scope of the present invention.
Embodiment 1：
(1) aseptically, by fresh, ripe fruiting bodies of cordyceps militaris（The fructification comes from Changde Yan Di, also known as Shen Nong, a legendary ruler's biotechnology
The megaspore head kind that Co., Ltd's strain library nurtures）It is suspended from above water agar, avoids fructification from being connect with culture medium
Touch；
(2) transfer them in biochemical cultivation case, 21 DEG C of lucifuge cultures 4-5 days, ascospore is launched naturally and arrive PDA culture medium
On；
(3) picking only has the bacterium colony of single ascospore under Stereo microscope, adds 2ml sterilized water, stirs, system
It is standby into spore suspension, take 200 microlitres of spore suspension, be coated with PDA plate culture medium, be positioned in 21 DEG C of biochemical cultivation cases
Lucifuge culture；
(4) after cultivating 3-4 days, macroscopic ascospore single bacterium colony is chosen on superclean bench, new PDA is transferred to and puts down
On plate culture medium, single bacterium colony is only cultivated on each PDA plate culture medium；
(5) after cultivating 5 days, the single ascospore bacterium colony of purifying is taken to extract its DNA, the son using PCR amplification techniques to extraction
Cystospore single bacterium colony DNA carries out mating type gene MAT1-1-1, MAT1-1-2 and MAT1-2-1 detection；
(6) nutrient solution is configured：By glucose 2%, peptone 0.5%, beef extract 0.5%, epsom salt 0.1%, potassium dihydrogen phosphate
0.1%th, vitamin b1 0.001%, vitamin b12 0.001%, water 96.798% form, and are well mixed；
(7) by step（6）Middle nutrient solution is dispensed into triangular flask, autoclave sterilization 20~30 minutes, and taking-up is cooled to normal temperature
It is standby；
(8) selecting step（5）The list for not containing MAT1-2-1 but containing mating type gene MAT1-1-1 and MAT1-1-2 of detection
The bacterium colony of sporogenesis, the monospore with not containing MAT1-1-1 and MAT1-1-2 but containing mating type gene MAT1-2-1 are formed
Bacterium colony take the mycelium block of 2~5% sizes to be inoculated in step respectively（7）In the nutrient solution made, it is placed in concussion shaking table, if
Put 19 DEG C, lucifuge, rotating speed 150rpm Shaking cultures 4-5 days；
(9) by step（8）Cultured mycelium is inoculated on synthetic medium, be transferred to 17~25 DEG C of temperature, humidity 65~
In the environment of 85%, light culture 5~7 days, treat that media surface grows a large amount of white hyphas；
(10) under conditions of step (9), 200~2000lux of intensity of illumination is set, carries out seeing light annesl, generated by former base,
Go out flower bud growth, go out grass growth three phases, treat that cordyceps militaris sporocarp length harvests to 4~5cm height, then by fructification.
Embodiment 2：
(1) aseptically, by fresh, ripe fruiting bodies of cordyceps militaris（The fructification comes from Changde Yan Di, also known as Shen Nong, a legendary ruler's biotechnology
The microspore head kind that Co., Ltd's strain library nurtures）It is suspended from above water agar, avoids fructification from being connect with culture medium
Touch；
(2) transfer them in biochemical cultivation case, 21 DEG C of lucifuge cultures 4-5 days, ascospore is launched naturally and arrive PDA culture medium
On；
(3) picking only has the bacterium colony of single ascospore under Stereo microscope, adds 2ml sterilized water, stirs,
Spore suspension is prepared into, takes 200 microlitres of spore suspension, is coated with PDA plate culture medium, is positioned over 21 DEG C of biochemical cultivation cases
Middle lucifuge culture；
(4) after cultivating 3-4 days, macroscopic ascospore single bacterium colony is chosen on superclean bench, new PDA is transferred to and puts down
On plate culture medium, single bacterium colony is only cultivated on each PDA plate culture medium；
(5) after cultivating 5 days, the single ascospore bacterium colony of purifying is taken to extract its DNA, the son using PCR amplification techniques to extraction
Cystospore single bacterium colony DNA carries out mating type gene MAT1-1-1, MAT1-1-2 and MAT1-2-1 detection；
(6) nutrient solution is configured：By glucose 2%, peptone 0.5%, beef extract 0.5%, epsom salt 0.1%, potassium dihydrogen phosphate
0.1%th, vitamin b1 0.001%, vitamin b12 0.001%, water 96.798% form, and are well mixed；
(7) by step（6）Middle nutrient solution is dispensed into triangular flask, autoclave sterilization 20~30 minutes, and taking-up is cooled to normal temperature
It is standby；
(8) selecting step（5）The list for not containing MAT1-2-1 but containing mating type gene MAT1-1-1 and MAT1-1-2 of detection
The bacterium colony of sporogenesis, the monospore with not containing MAT1-1-1 and MAT1-1-2 but containing mating type gene MAT1-2-1 are formed
Bacterium colony take the mycelium block of 2~5% sizes to be inoculated in step respectively（7）In the nutrient solution made, it is placed in concussion shaking table, if
Put 19 DEG C, lucifuge, rotating speed 150rpm Shaking cultures 4-5 days；
(9) by step（8）Cultured mycelium is inoculated on synthetic medium, be transferred to 17~25 DEG C of temperature, humidity 65~
In the environment of 85%, light culture 5~7 days, treat that media surface grows a large amount of white hyphas；
(10) under conditions of step (9), 200~2000lux of intensity of illumination is set, carries out seeing light annesl, generated by former base,
Go out flower bud growth, go out grass growth three phases, treat that cordyceps militaris sporocarp length harvests to 4~5cm height, then by fructification.
Embodiment 3：
(1) aseptically, by fresh, ripe fruiting bodies of cordyceps militaris（The fructification comes from Changde Yan Di, also known as Shen Nong, a legendary ruler's biotechnology
The tip conidia powder kind that Co., Ltd's strain library nurtures）It is suspended from above water agar, avoids fructification and culture medium
Contact；
(2) transfer them in biochemical cultivation case, 21 DEG C of lucifuge cultures 4-5 days, ascospore is launched naturally and arrive PDA culture medium
On；
(3) picking only has the bacterium colony of single ascospore under Stereo microscope, adds 2ml sterilized water, stirs,
Spore suspension is prepared into, takes 200 microlitres of spore suspension, is coated with PDA plate culture medium, is positioned over 21 DEG C of biochemical cultivation cases
Middle lucifuge culture；
(4) after cultivating 3-4 days, macroscopic ascospore single bacterium colony is chosen on superclean bench, new PDA is transferred to and puts down
On plate culture medium, single bacterium colony is only cultivated on each PDA plate culture medium；
(5) after cultivating 5 days, the single ascospore bacterium colony of purifying is taken to extract its DNA, the son using PCR amplification techniques to extraction
Cystospore single bacterium colony DNA carries out mating type gene MAT1-1-1, MAT1-1-2 and MAT1-2-1 detection；
(6) nutrient solution is configured：By glucose 2%, peptone 0.5%, beef extract 0.5%, epsom salt 0.1%, potassium dihydrogen phosphate
0.1%th, vitamin b1 0.001%, vitamin b12 0.001%, water 96.798% form, and are well mixed；
(7) by step（6）Middle nutrient solution is dispensed into triangular flask, autoclave sterilization 20~30 minutes, and taking-up is cooled to normal temperature
It is standby；
(8) selecting step（5）The list for not containing MAT1-2-1 but containing mating type gene MAT1-1-1 and MAT1-1-2 of detection
The bacterium colony of sporogenesis, the monospore with not containing MAT1-1-1 and MAT1-1-2 but containing mating type gene MAT1-2-1 are formed
Bacterium colony take the mycelium block of 2~5% sizes to be inoculated in step respectively（7）In the nutrient solution made, it is placed in concussion shaking table, if
Put 19 DEG C, lucifuge, rotating speed 150rpm Shaking cultures 4-5 days；
(9) by step（8）Cultured mycelium is inoculated on synthetic medium, be transferred to 17~25 DEG C of temperature, humidity 65~
In the environment of 85%, light culture 5~7 days, treat that media surface grows a large amount of white hyphas；
(10) under conditions of step (9), 200~2000lux of intensity of illumination is set, carries out seeing light annesl, generated by former base,
Go out flower bud growth, go out grass growth three phases, treat that cordyceps militaris sporocarp length harvests to 4~5cm height, then by fructification.
Table 1 below be this patent embodiment 1 using mycelium tangle fusion hybridizing method obtain megaspore head Hybrid with
Two parents（Non-hybridized megaspore head male parent, megaspore are maternal）The data contrasted respectively.
Table 1：
Table 2 below be this patent embodiment 2 using mycelium tangle fusion hybridizing method obtain microspore head Hybrid with
Two parents（Non-hybridized microspore head male parent, microspore are maternal）The data contrasted respectively.
Table 2：
Table 3 below be this patent embodiment 3 using mycelium tangle fusion hybridizing method obtain megaspore head Hybrid with
Two parents（Non-hybridized megaspore head male parent, megaspore are maternal）The data contrasted respectively.
Table 3
It can be seen from upper table 1, table 2, the result of table 3, fruiting bodies of cordyceps militaris its yield that the present invention nurtures is higher, reduces list
Position cost；Cordycepin, adenosine content are higher simultaneously, and its medical value is also very high.
Although embodiments of the present invention are described in detail, it should be understood that, without departing from the present invention's
In the case of spirit and scope, embodiments of the present invention can be made with various changes, replacement and change.
Claims (1)
1. the Cordyceps militaris ascospore cross breeding method based on Protocols in Molecular Biology, it is characterised in that comprise the following steps：
（1）Aseptically, fresh, ripe fruiting bodies of cordyceps militaris is suspended from above water agar, avoids fructification
Contacted with culture medium；The fruiting bodies of cordyceps militaris is by megaspore head product in the strain library of Changde Yandi Biotechnology Co., Ltd.
What a kind of in kind, microspore head kind, tip conidia powder kind nurtured；
（2）Transfer them in biochemical cultivation case, 21 DEG C of lucifuge cultures 4-5 days, ascospore is launched naturally and arrive PDA culture medium
On；
（3）Picking only has the bacterium colony of single ascospore under Stereo microscope, adds 2ml sterilized water, stirs,
Spore suspension is prepared into, takes 200 microlitres of spore suspension, is coated with PDA plate culture medium, is positioned over 21 DEG C of biochemical cultivation cases
Middle lucifuge culture；
(4) after cultivating 3-4 days, macroscopic ascospore single bacterium colony is chosen on superclean bench, new PDA is transferred to and puts down
On plate culture medium, single bacterium colony is only cultivated on each PDA plate culture medium；
(5) after cultivating 5 days, the single ascospore bacterium colony of purifying is taken to extract its DNA, the son using PCR amplification techniques to extraction
Cystospore single bacterium colony DNA carries out mating type gene MAT1-1-1, MAT1-1-2 and MAT1-2-1 detection；
(6) nutrient solution is configured：By glucose 2%, peptone 0.5%, beef extract 0.5%, epsom salt 0.1%, potassium dihydrogen phosphate
0.1%th, vitamin b1 0.001%, vitamin b12 0.001%, water 96.798% form, and are well mixed；
(7) by step（6）Middle nutrient solution is dispensed into triangular flask, autoclave sterilization 20~30 minutes, and taking-up is cooled to normal temperature
It is standby；
(8) selecting step（5）The list for not containing MAT1-2-1 but containing mating type gene MAT1-1-1 and MAT1-1-2 of detection
The bacterium colony of sporogenesis, the monospore with not containing MAT1-1-1 and MAT1-1-2 but containing mating type gene MAT1-2-1 are formed
Bacterium colony take the mycelium block of 2~5% sizes to be inoculated in step respectively（7）In the nutrient solution made, it is placed in concussion shaking table, if
Put 19 DEG C, lucifuge, rotating speed 150rpm Shaking cultures 4-5 days；
(9) by step（8）Cultured mycelium is inoculated on synthetic medium, be transferred to 17~25 DEG C of temperature, humidity 65~
In the environment of 85%, light culture 5~7 days, treat that media surface grows a large amount of white hyphas；
(10) under conditions of step (9), 200~2000lux of intensity of illumination is set, carries out seeing light annesl, generated by former base,
Go out flower bud growth, go out grass growth three phases, treat that cordyceps militaris sporocarp length harvests to 4~5cm height, then by fructification.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122049A (en) * | 2018-08-03 | 2019-01-04 | 内蒙古农业大学 | A kind of more spore separators of Cordyceps militaris and method |
| CN111527987A (en) * | 2020-06-10 | 2020-08-14 | 上海市农业科学院 | Method for improving yield of cordyceps militaris sporocarp by two-step inoculation |
| CN117063780A (en) * | 2023-10-17 | 2023-11-17 | 四川朕源生物科技有限公司 | Cordyceps sinensis ascospore harvesting culture medium and harvesting method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004033061A (en) * | 2002-07-01 | 2004-02-05 | Tohaku Nosan Kigyo Kumiai | Artificial cultivation method for cordyceps spp |
| CN1995321A (en) * | 2006-12-29 | 2007-07-11 | 上海市农业科学院 | Northern aweto strain and its selection breeding method |
| CN103347997A (en) * | 2010-02-04 | 2013-10-09 | 帝斯曼知识产权资产管理有限公司 | Process for preparing filamentous fungal strains having a sexual cycle and a process for preparing sexually crossed filamentous fungal strains |
| CN103609331A (en) * | 2013-11-18 | 2014-03-05 | 安徽农业大学 | Method for rejuvenating cordyceps militaris strains and increasing sporocarp yield by mating type |
| CN106434879A (en) * | 2016-08-29 | 2017-02-22 | 沈阳农业大学 | Method for quickly and efficiently detecting different strain mating types of cordyceps militaris |
| CN106591213A (en) * | 2017-02-09 | 2017-04-26 | 山东省蚕业研究所 | Method for rapidly separating and cultivating single spores from cordyceps militaris fruiting bodies |
| CN107173051A (en) * | 2017-05-20 | 2017-09-19 | 天津市东方中滨农业科技有限公司 | A kind of method and its application of many spore crossbreeding of Cordyceps militaris |
-
2017
- 2017-10-16 CN CN201710956892.6A patent/CN107493979A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004033061A (en) * | 2002-07-01 | 2004-02-05 | Tohaku Nosan Kigyo Kumiai | Artificial cultivation method for cordyceps spp |
| CN1995321A (en) * | 2006-12-29 | 2007-07-11 | 上海市农业科学院 | Northern aweto strain and its selection breeding method |
| CN103347997A (en) * | 2010-02-04 | 2013-10-09 | 帝斯曼知识产权资产管理有限公司 | Process for preparing filamentous fungal strains having a sexual cycle and a process for preparing sexually crossed filamentous fungal strains |
| CN103609331A (en) * | 2013-11-18 | 2014-03-05 | 安徽农业大学 | Method for rejuvenating cordyceps militaris strains and increasing sporocarp yield by mating type |
| CN106434879A (en) * | 2016-08-29 | 2017-02-22 | 沈阳农业大学 | Method for quickly and efficiently detecting different strain mating types of cordyceps militaris |
| CN106591213A (en) * | 2017-02-09 | 2017-04-26 | 山东省蚕业研究所 | Method for rapidly separating and cultivating single spores from cordyceps militaris fruiting bodies |
| CN107173051A (en) * | 2017-05-20 | 2017-09-19 | 天津市东方中滨农业科技有限公司 | A kind of method and its application of many spore crossbreeding of Cordyceps militaris |
Non-Patent Citations (2)
| Title |
|---|
| 张磊等: "不同分离方法获得的冬虫夏草无性型菌株的交配型", 《安徽农业大学学报》 * |
| 鲍大鹏: "运用交配型基因分子标记辅助蛹虫草亲和配对", 《核农学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122049A (en) * | 2018-08-03 | 2019-01-04 | 内蒙古农业大学 | A kind of more spore separators of Cordyceps militaris and method |
| CN111527987A (en) * | 2020-06-10 | 2020-08-14 | 上海市农业科学院 | Method for improving yield of cordyceps militaris sporocarp by two-step inoculation |
| CN111527987B (en) * | 2020-06-10 | 2021-11-26 | 上海市农业科学院 | Method for improving yield of cordyceps militaris sporocarp by two-step inoculation |
| CN117063780A (en) * | 2023-10-17 | 2023-11-17 | 四川朕源生物科技有限公司 | Cordyceps sinensis ascospore harvesting culture medium and harvesting method thereof |
| CN117063780B (en) * | 2023-10-17 | 2023-12-26 | 四川朕源生物科技有限公司 | Cordyceps sinensis ascospore harvesting culture medium and harvesting method thereof |
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