CN103004464A - Novel strain of hypsizigus marmoreus - Google Patents
Novel strain of hypsizigus marmoreus Download PDFInfo
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Abstract
The invention provides a novel strain Finc-B-1 of hypsizigus marmoreus. The collection number of the novel strain Finc-B-1 of the hypsizigus marmoreus is China CCTCCNO: M2012308, wherein CCTCCNO refers to center for type culture collection number. According to the novel strain Finc-B-1 provided by the invention, the cultivation period is effectively shortened, and the production cost is reduced. Moreover, the strain has the excellent characteristics that the per unit yield is high, the fruit body appearance is attractive and the length for preservation is long, is free of bitter and can be widely popularized to be planted.
Description
[technical field]
The present invention relates to the edible mushroom technical field, more precisely, related to the new bacterial strain of a kind of true Ji mushroom.
[background technology]
True Ji mushroom (Latin title Hypsizygus marmoreus) is subordinate to Basidiomycotina, Hymenomycetes, Agaricales, white mushroom section, belongs to from gill fungus family, beautiful gill fungus, because it has unique crab delicate flavour, so claim that again it is hypsizygus marmoreus.Initial artificial cultivation started from Japan before and after 1978, its delicious flavour, sense of food are tender and crisp, nutritious, and its flavor is brighter than flat mushroom, and its meat is thick than sliding mushroom, and its matter is more tough than mushroom, and mouthfeel is excellent, and its dry product also thick flavor is excessive.True Ji mushroom contains enriches vitamin and 17 seed amino acids, the β that extracts in the fruit body-1, and the 3-D glucan has very high antitumor activity, is a kind of health food low in calories, low-fat.
True Ji mushroom is from cultivating beginning, and what substantially walk is exactly the route that batch production is produced.Since factory culture have produce continuously in the anniversary, large-scale production, mostly realize semi-mechanization, mechanization so that intelligent, growing environment is controlled better, yield and quality has had good assurance.As everyone knows, bacterial classification plays vital effect to the factory culture of edible mushroom.In the market true Ji's mushroom strains have a variety of, but the main better bacterial classification that uses has HOKTO-18, Baudot A-264 and TNN-11 etc.HOKTO-18 is a kind of bacterial classification that has Japanese Big Dipper company of Co., Ltd. (HOKTO) to cultivate, although this bacterial classification is more successful in the plantation of Japanese HOKTO company, but find in the actual planting process of other companies that there is following defective in it: knurl lid mushroom appears in (1) easily, causes yields to decline to a great extent; (2) per unit area yield is lower, and economic benefit is relatively poor; This adaptability that reflects to a certain extent this bacterial classification is relatively poor, is not suitable for spread.Baudot A-264 plants extensively at home, although it possesses the advantage that cultivation cycle is short, the cultivation cycle is short, most consumers reflect that this bacterial strain taste is relatively more bitter, and mouthfeel is relatively poor, and consumer's acceptance is lower.TNN-11 possesses the fruit body shapeliness, without the advantage of bitter taste, but its cultivation and cultivation cycle are long, and the planting environment parameter had higher requirement such as light intensity, ventilation, humidity etc., this brings higher production cost to the producer undoubtedly, thereby has significantly slackened its competitiveness on market.
[summary of the invention]
The technical problem that the present invention solves provides the new bacterial strain Finc-B-1 of a kind of true Ji mushroom, this bacterial strain have cultivate and the cultivation cycle is short, per unit area yield is high, the fruit body outward appearance is good, without characteristics such as bitter taste, long fresh-keeping periods.
The present invention is achieved by the following technical solutions, the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention is by TNN-11 and the only method acquisition of crossbreeding of Baudot A-264, this bacterial strain has been preserved in Chinese Typical Representative culture collection center on August 13rd, 2012, the address: Wuhan, China city Wuhan University, its preserving number is CCTCC M 2012308.
The new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention is characterised in that:
1, morphological feature
Cap: during young mushroom, whole mushroom cap surface is covered with unique circular decorative pattern, color dark brown, the even droplet that distributes, and along with the growth of mushroom lid, the ratio with decorative pattern zone is dwindled, color shoals, and final only have decorative pattern at the top of mushroom lid; When ripe, mushroom lid diameter 1cm～2cm, mushroom lid section circle mountain type, the dark-grey yellowish-brown of mushroom lid middle part color, mushroom lid edge color Huang is dark brown, meat is thick, meat is hard, amount of speckle is medium, big or small medium, the speckle distribution mushroom lid central authorities of speckle, speckle are clear, the mushroom lid is without be full of cracks.Lamella: yellow-white, lamella normal alignment, fabric width is general, density is general.Stem: height 5cm～7cm, stem diameter 0.8cm～1cm, give birth to centered by cap and the stem connected mode, in thick, white, without hair, the ratio of mushroom lid diameter and stem length is medium, meat is hard.Mycelia pure white dense, aerial hyphae is flourishing, average 80 of fruit body all living creatures's type, number of productive tiller.
2, biological property
Nutrient component source: corncob 40～50%(percentage by weight, lower with), weed tree sawdust 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, wherein, water content 64～66%.
21～25 ℃ of Mycelium culture temperature, humidity 75%, CO
2Concentration 2500～4000ppm, cultivation cycle 80～85 days.
Suitable fruiting temperature is 13～15 ℃, humidity 90～95%, CO
2Concentration 1000～2000ppm, front 1～7 day illumination intensity 50～100Lux, rear 8～20 days illumination intensity 200～300Lux, every day, clearance-type was opened layer bracket lamp stimulation fruiting, 20～23 days cultivation cycles.
Compared with prior art, the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention has shortened cultivation and cultivation cycle effectively, has reduced production cost; And this bacterial strain per unit area yield is high, the fruit body outward appearance is good, and without bitter taste, long fresh-keeping period has the good characteristics of spread plantation.
[description of drawings]
Fig. 1 is the curve map that Finc-B-1, TNN-11 and Baudot A-264 output change with cell age.
Fig. 2 be primer I TS1 and ITS4 to the pcr amplification electrophoresis pattern of 6 bacterial strains, wherein 2 is bacterial strain Baudot A-264,3 is Finc-B-1,4 is TNN-11, the 1,5, the 6th, other true Ji's mushroom bacterial strain of collecting.
Fig. 3 is the ITS phylogenetic tree of Finc-B-1, wherein 3 represents Finc-B-1.
Fig. 4 be primer S29 to the pcr amplification electrophoresis pattern of 6 bacterial strains, same Fig. 2 of implication of numbering 1-6 representative among the figure.
Fig. 5 be primer S37 to the pcr amplification electrophoresis pattern of 6 bacterial strains, same Fig. 2 of implication of numbering 1-6 representative among the figure.
Fig. 6 be primer S45 to the pcr amplification electrophoresis pattern of 6 bacterial strains, same Fig. 2 of implication of numbering 1-6 representative among the figure.
Fig. 7 is the pcr amplification electrophoresis pattern of 121 pairs of 6 bacterial strains of primer S, same Fig. 2 of implication of numbering 1-6 representative among the figure.
[embodiment]
The invention will be further described below by specific embodiment.
Present embodiment has illustrated the breeding process of the new bacterial strain Finc-B-1 of true Ji mushroom.Concrete steps are as follows:
1) Selection parent bacterial strain: choosing TNN-11 and Baudot A-264 is hybrid strain, and these two parents' fruit body can buy in market, in Academy of Agricultural Sciences, Shanghai City DSMZ preservation is arranged also.
2) preparation medium
PDA medium: every 1L medium potato 200g, glucose 20g, agar 20g.
Pedigree seed culture medium: wood chip 55%-70%, rice bran 15%-20%, wheat bran 5%-10%, corn flour 8-35% are as cultivating raw material.Keep the skin wet, make the water content in the medium reach 60～62%.
Cultivated species medium: corncob 40～50%, soybean skin 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, water content 64～66%.
3) obtain spore: the parent through the activation of test tube kind, turn the triangular flask kind, turn original seed, turn cultivated species, cultivation, mycelium stimulation, cultivation fruiting, after parachute-opening, adopt the outstanding method of hook to receive spore, place 17 ℃ of environment kinds and namely obtained spore in 2 days.
4) spore coating: obtain spore suspension with sterile water dilution spore, addition by sterile water is regulated spore concentration, observe at 10 * 40 microscopicallies, when there is 3-4 spore in each visual field, it is desired concn, 1 spore suspension of every plate is coated on the PDA medium equably with spreader.
5) obtain Monospore mycelium: cultivated about 10 days, and at sparse position picking monospore bacterium colony, forwarded on the slant medium for 21 ℃.
6) uninucleate hyphae microscopy: in 10 * 40 fields of microscope, seeking 3～5 visuals field in different zones, then is uninucleate hyphae without clamp connection.
7) pairing hybridization: it is fast to select to send out bacterium speed, the dense pure white uninucleate hyphae pairing of mycelia.
8) heterozygote microscopy: in 10 * 40 fields of microscope, seeking 3～5 visuals field in different zones, if clamp connection is arranged, then is heterozygote.
9) heterozygote picking: forward on the test tube slant medium the regional picking mycelia of hybridization.
10) experiment in cultivation: the heterozygote that obtains is carried out experiment in cultivation and screening according to goal of the invention.
11) hybridization (heterozygote called after " TN4 ") by TNN-11 and Baudot A-264, obtain 400 heterozygotes, from 400 heterozygotes screening and culturing and the cultivation cycle is short, per unit area yield is high, the fruit body shapeliness, without the purpose bacterial strain of bitter taste etc., be numbered " TN4-52 ", called after " Finc-B-1 ".
The new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention is preserved in Chinese Typical Representative culture collection center on August 13rd, 2012, and deposit number is CCTCC M 2012308, and bacterial strain is survived after testing, and classification position is that beautiful gill fungus belongs to true Ji mushroom.
The test of embodiment 2 cell ages
Present embodiment illustrates advantage and the characteristics of the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention by the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention and existing TNN-11, Baudot A-264 are carried out the contrast of cultivation cycle, cultivation cycle and per unit area yield.
The cultivated species medium that present embodiment adopts is:
Corncob 40～50%, weed tree sawdust 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, wherein, water content 64～66%.
Take TNN-11 as contrast, 6 cell ages were set from 70 days to 100 days, per 5 days is a cell age, 64 bottles of every cell ages.
Cultivate and the planting environment condition:
Cultivation temperature is 21～25 ℃, humidity 75%, CO
2Concentration 2500～4000ppm.The cultivating chamber Temperature Setting is 13～15 ℃, humidity 90～95%, CO
2Concentration 1000～2000ppm, front 1～7 day illumination intensity 50～100Lux, rear 8～20 days illumination intensity 200～300Lux, every day, clearance-type was opened layer bracket lamp.
See also Fig. 1 and table 1, the best cell age of contrast TNN-11, Baudot A-264 and Finc-B-1 is respectively 95 days, 85 days and 85 days, because they obtain respectively the highest per unit area yield in these two cell ages.Simultaneously when their obtained the highest per unit area yield, the average cultivation cycle of TNN-11, Baudot A-264 and each bacterial strain of Finc-B-1 was respectively 24 days, 22 days and 21.5 days.When cell age was extended to 95 days from 75 days, the cultivation cycle of TNN-11 was approximately 24 days, and the cultivation cycle of Finc-B-1 reduced to 20 days from 23 days.In 75 days to 95 days cell age scope, the per unit area yield of each cell age of Finc-B-1 is higher than TNN-11 and Baudot A-264.This shows that under the identical cultivation condition, compare parent strain TNN-11 and Baudot A-264, Finc-B-1 sporophore growth speed has clear superiority.
The test of table 1 cell age
Uniformity and stability when present embodiment is planted in different culture medium prescription A, B Finc-B-1 are tested.Wherein,
Culture medium A is:
Corncob 40～50%, weed tree sawdust 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, wherein water content 64～66%.
Medium B is:
Corncob 20～30%, weed tree sawdust 20～30%, soybean skin 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, wherein water content 64～66%.
The culture bottle volume is: 850cc.
Take TNN-11 as contrast, the cultivation effect of test Finc-B-1 bacterial strain.6 small-scale cultivations of this experimental scheme (300 bottles/time) are numbered test 1,2,3,4,5 and 6, and wherein, test 1,2 and 3 used medium prescriptions are prescription A, and test 4,5 and 6 used medium prescriptions are prescription B.The parameter of cultivation and cultivation condition is identical with embodiment 2, does not repeat them here.
Table 2 uniformity and stability test
As known from Table 2, in prescription A medium, the Finc-B-1 culture period is about 85 days, and cultivation is stabilized in 22 days, per unit area yield 〉=177g/ bottle; In prescription B medium, the Finc-B-1 culture period is about 76 days, and cultivation is stabilized in 22 days, and per unit area yield reaches the 200g/ bottle.It is stable that the cultivation cycle of Finc-B-1, cultivation cycle, per unit area yield all keep in different medium, show that thus it has good stability and repeated, and the fruit body shape difference is little between bottle and the bottle, and uniformity is strong.
Principal item TNN-11, packs with 155～165g/ rapidly after Finc-B-1 and TNN-11 gather as contrast on the market, each 20 bag, in freezer (3 ℃) preservation 13 days, then take out and be placed under 22 ℃ of room temperature conditions record Fruitbody situation of change.
Result of the test sees also table 3, Finc-B-1 after gathering 22 days still intact, and TNN-11 gather after the 22nd day existing serious rotten and lose the mushroom of commercial value, so Finc-B-1 has preferably freshness date than principal item TNN-11 on the market.
Table 3 freshness date
Annotate: zero is normal, and △ has one or two root entity to begin to go bad, * have no commercial value, rotten.
Choose respectively 500g of fresh Finc-B-1, TNN-11 and Baudot A-264 fruit body, fry, the room temperature cooling is randomly drawed 100 people and is carried out blind test with regard to Finc-B-1 and two bitter taste and fragility that contrast between the parent, and test result is referring to table 4.
As known from Table 4, in 100 persons of foretasting, 89% think the not bitter or little hardship only of Finc-B-1,70% think is more crisp.The gustation test statistics is the result show, Finc-B-1 has happy people's taste, more is subjected to consumer's favor.
Table 4 gustation test
The morphological feature of Finc-B-1 sees also table 5.
Cap: during young mushroom, whole mushroom cap surface is covered with unique circular decorative pattern, color dark brown, the even droplet that distributes, and along with the growth of mushroom lid, the ratio with decorative pattern zone is dwindled, color shoals, and final only have decorative pattern at the top of mushroom lid; When ripe, mushroom lid diameter 1cm～2cm, mushroom lid section circle mountain type, the dark-grey yellowish-brown of mushroom lid middle part color, mushroom lid edge color Huang is dark brown, meat is thick, meat is hard, amount of speckle is medium, big or small medium, the speckle distribution mushroom lid central authorities of speckle, speckle are clear, the mushroom lid is without be full of cracks.Lamella: yellow-white, lamella normal alignment, fabric width is general, density is general.Stem: height 5cm～7cm, stem diameter 0.8cm～1cm, give birth to centered by cap and the stem connected mode, in thick, white, without hair, the ratio of mushroom lid diameter and stem length is medium, meat is hard.Mycelia pure white dense, aerial hyphae is flourishing, average 80 of fruit body all living creatures's type, number of productive tiller.
Table 5Finc-B-1 morphological feature
Annotate: represents that Finc-B-1 meets this characteristic
The biological property of Finc-B-1:
The nutrient component source: corncob 40～50%, weed tree sawdust 20～30%, rice bran 10～15%, wheat bran 15～20%, corn flour 5%, wherein, water content 64～66%.21～25 ℃ of Mycelium culture temperature, humidity 75%, CO
2Concentration 2500～4000ppm, cultivation cycle 80～85 days.Suitable fruiting temperature is 13～15 ℃, humidity 90～95%, CO
2Concentration 1000～2000ppm, front 1～7 day illumination intensity 50～100Lux, rear 8～20 days illumination intensity 200～300Lux, every day, clearance-type was opened layer bracket lamp stimulation fruiting, 20～23 days cultivation cycles.
Embodiment 7 antagonistic effects
Present embodiment is take the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention and parent TNN-11 thereof and Baudot A-264 as test material, and used medium is the PDA medium.
Test arranges two groups of contrasts, and every dull and stereotyped 3 vaccinations become 1 straight line, vaccination 2cm apart between same kind, and 24 ℃ of constant temperature culture 26 days, cultivation results is as non-antagonism reference; Vaccination is at a distance of 2cm between different cultivars, 24 ℃ of constant temperature culture 26 days, and cultivation results is as the antagonism reference.TNN-11, Finc-B-1 and Baudot A-264, from left to right straight line is inoculated on the PDA medium successively, with vaccination at a distance of 2cm, 24 ℃ of constant temperature culture 26 days, the result contrasts respectively between definite Finc-B-1 and the parent strain whether antagonism is arranged with two groups of contrasts.
The antagonistic effect result shows: TNN-11 and Baudot A-264 self face-off is cultivated does not have antagonism, the handing-over of culture dish front bacterium colony, and culture dish back side bacterium colony joins into white line.TNN-11 and Baudot A-264 have obvious antagonism each other, and the positive two bacterium colonies handing-over of culture dish, culture dish back side bacterium colony join into yellow line.The culture dish front bacterium colony handing-over that Finc-B-1 and parent TNN-11 and Baudot A-264 face-off are cultivated, culture dish back side bacterium colony joins into yellow line, the mutual antagonism of this phenomenon and TNN-11 and Baudot A-264 is identical, and it is different with Baudot A-264 self face-off cultivation phenomenon from TNN-11, illustrating between Finc-B-1 and two parent strains has antagonism, and Finc-B-1 is a new bacterial strain of true Ji mushroom.
Table 6 antagonistic effect
| Kind | TNN-11 | Baudot A-264 | Finc-B-1 |
| TNN-11 | - | + | + |
| Baudot-A264 | + | - | + |
The embodiment 8ITS evaluation of checking order
ITS order-checking flow process entrusts Sangon Biotech's (hereinafter to be referred as giving birth to the worker) to carry out, and comprises following process:
Genomic extraction: extract according to giving birth to worker SK1375 fungal genomic DNA extraction agent box specification.Resulting dna solution is placed-20 ℃, for subsequent use.
Pcr amplification and Establishing (25 μ l): genomic templates 1 μ l, upstream primer (10 μ M) 0.5 μ l, downstream primer (10 μ M) 0.5 μ l, dNTP mix (10mM each) 0.5 μ l, 10 * Taq reaction buffer, 2.5 μ l, Taq (5u/ μ l) 0.2 μ l adds water to 25 μ l.PCR program: 94 ℃ of 5mim of denaturation; 94 ℃ of 30s, 55 ℃ of 35s, 72 ℃ of 1min, 35 circulations, 72 ℃ are extended 8min.The PCR product that amplification obtains is by agarose gel electrophoresis, and its electrophoresis result is seen Fig. 2.
The DNA Ago-Gel reclaims glue purification and conventional method is adopted in order-checking.
This test strain Finc-B-1ITS sequence sequencing result is seen sequence table, checks in from GenBank, and the ITS sequence similarity that Finc-B-1 bacterial strain ITS sequence and beautiful gill fungus belong to true Ji mushroom is the highest.And from GenBank, check in the ITS sequence that belongs to the close kind of true Ji mushroom with beautiful gill fungus, and use the phylogenetic tree that MEGA4.0 constructs strains tested, see also Fig. 3.
Embodiment 9RAPD analyzes
Present embodiment is with the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention and parent TNN-11 and Baudot-A264, and other 3 true Ji's mushroom bacterial strains (what choose in the present invention is brown crab-flavor mushroom) of collecting from market are analytic target, utilize the RAPD analytical technology, further identify the hereditary capacity of the new bacterial strain of the true Ji mushroom of the present invention.DNA extraction in the present embodiment, PCR system, agarose gel electrophoresis system are identical with embodiment 8, do not repeat them here.
See also table 7, during RAPD analyzes, filter out 5 true Ji's mushroom strains that 4 primers can be distinguished clearly Finc-B-1 and collect from the market from 20 random primers, these 4 random primers are respectively S29, S37, S45 and S 121.
Table 7 random primer
| Numbering | The primer numbering | Sequence | Numbering | The | Sequence | ||
| 1 | | TGCTCTGCCC | 11 | | GTGTGCCCCA | ||
| 2 | S23 | AGTCAGCCAC | 12 | | CTCTGGAGAC | ||
| 3 | S29 | GGGTAACGCC | 13 | | GTCCCGACGA | ||
| 4 | S31 | CAATCGCCGT | 14 | | ACTTCGCCAC | ||
| 5 | S37 | GACCGCTTGT | 15 | | ACTGGGACTC | ||
| 6 | S38 | AGGTGACCGT | 16 | S99 | GTCAGGGCAA | ||
| 7 | S42 | GGACCCAACC | 17 | S121 | ACGGATCCTG | ||
| 8 | S43 | GTCGCCGTCA | 18 | S122 | GAGGATCCCT | ||
| 9 | S45 | TGAGCGGACA | 19 | S265 | GGCGGATAAG | ||
| 10 | S47 | TTGGCACGGG | 20 | S1327 | ACGCGACAGA |
The result that RAPD analyzes is as follows:
1) the corresponding collection of illustrative plates of primer S29 is Fig. 4, in its analysis diagram, the position at a place Finc-B-1 of the present invention has the DNA band, bacterial strain 1,2 is without band, position Finc-B-1 that Finc-B-1 of the present invention is different from bacterial strain 1 and 2, b place is described without the DNA band, and bacterial strain 4,5,6 there is the DNA band, illustrate that Finc-B-1 of the present invention is different from bacterial strain 4,5 and 6, so other true Ji mushroom kind that primer S29 can distinguish Finc-B-1 of the present invention and collect.
2) the corresponding collection of illustrative plates of primer S37 is Fig. 5, in its analysis diagram, the position at c place Finc-B-1 of the present invention is without the DNA band, and bacterial strain 1,2 has band, and the position Finc-B-1 at e place has the DNA band, bacterial strain 1,2 is without band, position Finc-B-1 of the present invention that Finc-B-1 of the present invention is different from bacterial strain 1 and 2, d place is described without the DNA band, and bacterial strain 4,5,6 there is the DNA band, primer S37 illustrates that Finc-B-1 of the present invention is different from bacterial strain 4,5 and 6, so can distinguish Finc-B-1 and other bacterial classification.
3) the corresponding collection of illustrative plates of primer S45 is Fig. 6, in its analysis diagram, the position at f place Finc-B-1 of the present invention has the DNA band, bacterial strain 1,2 and 4 is without band, illustrate Finc-B-1 of the present invention be different from bacterial strain 1,2 and the position Finc-B-1 of the present invention at 4, g place the DNA band is arranged, and bacterial strain 5,6 is without the DNA band, primer S45 illustrates that Finc-B-1 of the present invention is different from bacterial strain 5,6, so can distinguish Finc-B-1 of the present invention and other kind.
4) primer S 121 corresponding collection of illustrative plates are Fig. 7, in its analysis diagram, the position at h place Finc-B-1 of the present invention has the DNA band, bacterial strain 1,2 and 4 is without band, illustrate Finc-B-1 of the present invention be different from bacterial strain 1,2 and the position Finc-B-1 of the present invention at 4, i place without the DNA band, and bacterial strain 5,6 has the DNA band, primer S121 illustrates that Finc-B-1 of the present invention is different from bacterial strain 5,6, so can distinguish Finc-B-1 of the present invention and other bacterial classifications.
Above analysis result shows, the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention is similar with other 5 to exist obvious hereditary difference for the examination bacterial classification, random primer S29, S37, S45 and S 121 all can reflect the new bacterial strain Finc-B-1 of the true Ji mushroom of the present invention specific band, and it is a kind of brand-new new bacterial strain of true Ji mushroom that this result further specifies Finc-B-1 of the present invention.
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.
Claims (1)
- A true Ji mushroom ( Hypsizygus marmoreus) new bacterial strain Finc-B-1, its deposit number is CCTCC NO:M 2012308.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154340A (en) * | 2015-08-21 | 2015-12-16 | 山东晨阳菌业有限公司 | Breeding method of hypsizigus marmoreus strain |
| CN105165635A (en) * | 2015-10-22 | 2015-12-23 | 青岛丰科生物科技有限公司 | Hypsizygus marmoreus strain and application thereof |
| CN106434381A (en) * | 2016-10-31 | 2017-02-22 | 上海丰科生物科技股份有限公司 | Pure white hypsizigus marmoreus strain, and molecular marker, specific primer pair and application thereof |
| CN110184201A (en) * | 2019-06-20 | 2019-08-30 | 福建农林大学 | A kind of true pleurotus cornucopiae bacterial strain and its selection |
| WO2022170960A1 (en) * | 2021-02-10 | 2022-08-18 | 上海丰科生物科技股份有限公司 | Hypsizigus marmoreus strain, spore, fruiting body and use thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105802862B (en) * | 2016-05-31 | 2019-01-25 | 上海丰科生物科技股份有限公司 | True pleurotus cornucopiae bacterial strain and its culture, cultural method and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007037455A (en) * | 2005-08-03 | 2007-02-15 | Asahimatsu Shokuhin Kk | Strain having ability to form white pleurotus eryngii and white pleurotus eryngii obtained by culturing the strain |
| JP2007053928A (en) * | 2005-08-23 | 2007-03-08 | Kinokkusu:Kk | New fungal strain and method for culturing new fruit body |
| CN101292603A (en) * | 2007-04-29 | 2008-10-29 | 上海丰科生物科技股份有限公司 | Seed source propagate propagation expanding method and protospecies production method for white crab flavour mushroom |
| CN101317528A (en) * | 2007-06-08 | 2008-12-10 | 上海丰科生物科技股份有限公司 | Propagation method for seed source of beech mushroom |
| CN102047839A (en) * | 2009-11-06 | 2011-05-11 | 上海丰科生物科技股份有限公司 | Hypsizigus marmoreus mon-mon cross-breeding method |
| CN102204476A (en) * | 2010-03-31 | 2011-10-05 | 宝生物工程株式会社 | Hypsizigus marmoreus bacterial strain and production method of hypsizigus marmoreus fruit body |
-
2012
- 2012-11-12 CN CN201210452001.0A patent/CN103004464B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007037455A (en) * | 2005-08-03 | 2007-02-15 | Asahimatsu Shokuhin Kk | Strain having ability to form white pleurotus eryngii and white pleurotus eryngii obtained by culturing the strain |
| JP2007053928A (en) * | 2005-08-23 | 2007-03-08 | Kinokkusu:Kk | New fungal strain and method for culturing new fruit body |
| CN101292603A (en) * | 2007-04-29 | 2008-10-29 | 上海丰科生物科技股份有限公司 | Seed source propagate propagation expanding method and protospecies production method for white crab flavour mushroom |
| CN101317528A (en) * | 2007-06-08 | 2008-12-10 | 上海丰科生物科技股份有限公司 | Propagation method for seed source of beech mushroom |
| CN102047839A (en) * | 2009-11-06 | 2011-05-11 | 上海丰科生物科技股份有限公司 | Hypsizigus marmoreus mon-mon cross-breeding method |
| CN102204476A (en) * | 2010-03-31 | 2011-10-05 | 宝生物工程株式会社 | Hypsizigus marmoreus bacterial strain and production method of hypsizigus marmoreus fruit body |
| KR20110109959A (en) * | 2010-03-31 | 2011-10-06 | 다카라 바이오 가부시키가이샤 | Hypsizigus marmoreus strain and method for manufacturing of fruiting body of hypsizigus marmoreus |
Non-Patent Citations (1)
| Title |
|---|
| 刘明广等: "真姬菇杂交菌株H11的选育", 《食用菌》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154340A (en) * | 2015-08-21 | 2015-12-16 | 山东晨阳菌业有限公司 | Breeding method of hypsizigus marmoreus strain |
| CN105154340B (en) * | 2015-08-21 | 2018-11-13 | 山东晨阳菌业有限公司 | A kind of selection of true pleurotus cornucopiae strain |
| CN105165635A (en) * | 2015-10-22 | 2015-12-23 | 青岛丰科生物科技有限公司 | Hypsizygus marmoreus strain and application thereof |
| CN105165635B (en) * | 2015-10-22 | 2017-05-24 | 青岛丰科生物科技有限公司 | Hypsizygus marmoreus strain and application thereof |
| CN106434381A (en) * | 2016-10-31 | 2017-02-22 | 上海丰科生物科技股份有限公司 | Pure white hypsizigus marmoreus strain, and molecular marker, specific primer pair and application thereof |
| CN106434381B (en) * | 2016-10-31 | 2018-12-21 | 上海丰科生物科技股份有限公司 | Pure white true pleurotus cornucopiae bacterial strain and its molecular labeling, specific primer to and application |
| CN110184201A (en) * | 2019-06-20 | 2019-08-30 | 福建农林大学 | A kind of true pleurotus cornucopiae bacterial strain and its selection |
| WO2022170960A1 (en) * | 2021-02-10 | 2022-08-18 | 上海丰科生物科技股份有限公司 | Hypsizigus marmoreus strain, spore, fruiting body and use thereof |
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