CN103004464A - Novel strain of hypsizigus marmoreus - Google Patents
Novel strain of hypsizigus marmoreus Download PDFInfo
- Publication number
- CN103004464A CN103004464A CN2012104520010A CN201210452001A CN103004464A CN 103004464 A CN103004464 A CN 103004464A CN 2012104520010 A CN2012104520010 A CN 2012104520010A CN 201210452001 A CN201210452001 A CN 201210452001A CN 103004464 A CN103004464 A CN 103004464A
- Authority
- CN
- China
- Prior art keywords
- finc
- mushroom
- bacterial strain
- days
- present
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 230000001580 bacterial effect Effects 0.000 claims description 61
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 60
- 241001534815 Hypsizygus marmoreus Species 0.000 claims description 3
- 235000013399 edible fruits Nutrition 0.000 abstract description 11
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 239000002609 medium Substances 0.000 description 24
- 238000012360 testing method Methods 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 241000894006 Bacteria Species 0.000 description 8
- 241000209094 Oryza Species 0.000 description 7
- 235000007164 Oryza sativa Nutrition 0.000 description 7
- 240000008042 Zea mays Species 0.000 description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 7
- 230000008485 antagonism Effects 0.000 description 7
- 235000005822 corn Nutrition 0.000 description 7
- 235000013312 flour Nutrition 0.000 description 7
- 235000013372 meat Nutrition 0.000 description 7
- 235000009566 rice Nutrition 0.000 description 7
- 235000015099 wheat brans Nutrition 0.000 description 7
- 241000196324 Embryophyta Species 0.000 description 6
- 238000012408 PCR amplification Methods 0.000 description 6
- 238000001962 electrophoresis Methods 0.000 description 6
- 238000005286 illumination Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 241000233866 Fungi Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 235000019658 bitter taste Nutrition 0.000 description 5
- 241000446313 Lamella Species 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000000796 flavoring agent Substances 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 230000000877 morphologic effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000002255 vaccination Methods 0.000 description 4
- 230000003042 antagnostic effect Effects 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000014860 sensory perception of taste Effects 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 230000000007 visual effect Effects 0.000 description 3
- 238000007400 DNA extraction Methods 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000004744 fabric Substances 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000032696 parturition Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 235000019640 taste Nutrition 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000222485 Agaricales Species 0.000 description 1
- 241000222382 Agaricomycotina Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 230000037237 body shape Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- DMBHHRLKUKUOEG-UHFFFAOYSA-N diphenylamine Chemical compound C=1C=CC=CC=1NC1=CC=CC=C1 DMBHHRLKUKUOEG-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000019590 thick flavour Nutrition 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Images
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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210452001.0A CN103004464B (en) | 2012-11-12 | 2012-11-12 | Novel strain of hypsizigus marmoreus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210452001.0A CN103004464B (en) | 2012-11-12 | 2012-11-12 | Novel strain of hypsizigus marmoreus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103004464A true CN103004464A (en) | 2013-04-03 |
| CN103004464B CN103004464B (en) | 2014-07-09 |
Family
ID=47953995
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210452001.0A Active CN103004464B (en) | 2012-11-12 | 2012-11-12 | Novel strain of hypsizigus marmoreus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103004464B (en) |
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 |
|---|---|---|---|---|
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103004464B (en) | 2014-07-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103004464B (en) | Novel strain of hypsizigus marmoreus | |
| CN105165635B (en) | Hypsizygus marmoreus strain and application thereof | |
| CN106676016B (en) | The true pleurotus cornucopiae bacterial strain of the brown of high yield glucan and its cultural method and application | |
| CN105802862B (en) | True pleurotus cornucopiae bacterial strain and its culture, cultural method and application | |
| CN103210849B (en) | Lyophyllum decastes new bacterial strain | |
| CN102668885B (en) | Pholiota adipose new strain and method for cultivating fruiting body of pholiota adiposa new strain | |
| CN107881118B (en) | Light yellow flammulina velutipes JK05 strain and application thereof | |
| CN103202175A (en) | Novel strain of lyophyllum decastes | |
| CN103477993B (en) | Pure white new hypsizigus marmoreus bacterial strain | |
| CN107227263A (en) | White gold needle mushroom JK01 bacterial strains and its application | |
| CN103503780B (en) | New pure white hypsizigus marmoreus strain | |
| CN106434382B (en) | Pure white true pleurotus cornucopiae bacterial strain and its molecular labeling, specific primer to and application | |
| CN114250152A (en) | Six-sister morchella Guizhou MS630 and application thereof | |
| CN107557307A (en) | A kind of lab screening method of high temperature resistant cloud ear bacterial strain | |
| CN103030459A (en) | Edible fungus mother culture medium and mother culture preparation method | |
| CN105230342B (en) | Lyophyllum decastes bacterial strain and its cultural method and application | |
| CN104620855A (en) | Pleurotus spodotecus strain, and culture medium, culture method and application of pleurotus spodotecus strain and a culture medium | |
| CN103503779B (en) | New pure white hypsizigus marmoreus strain | |
| CN107641600A (en) | Suitable for the flat mushroom JK02 bacterial strains and its cultural method of low temperature fruiting and application | |
| CN106434381B (en) | Pure white true pleurotus cornucopiae bacterial strain and its molecular labeling, specific primer to and application | |
| CN115074259B (en) | Alkali-resistant lentinus edodes strain golden fragrance 194 and application thereof | |
| CN109220514B (en) | Separation and artificial domestication cultivation method of new wild edible fungi | |
| CN107926478A (en) | A kind of bacterial strain of suitable factory culture white gold needle mushroom and its application | |
| Sindhu et al. | Cropping duration and non-rhizomorphic mycelial phenotype of Pleurotus djamor woody1 co-segregate in the hybrid progenies | |
| CN115093976B (en) | White beech mushroom ZHSjg-byg and cultivation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: QINGDAO FENGKE BIOTECHNOLOGY CO., LTD. Free format text: FORMER OWNER: SHANGHAI FINC BIO-TECH INC. Effective date: 20140729 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 201401 FENGXIAN, SHANGHAI TO: 266221 QINGDAO, SHANDONG PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20140729 Address after: Three road 266221 Jimo City Tianshan Daxin Town, Shandong province Qingdao City No. 68 Patentee after: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Address before: 201401 Shanghai City, Fengxian District modern agriculture park Gao Feng Road, No. 968 Patentee before: Shanghai Finc Bio-tech Inc. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Hypsizigus marmoreus bacterial strain and production method of hypsizigus marmoreus fruit body Effective date of registration: 20150727 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2015990000599 |
|
| PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20160808 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2015990000599 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Hypsizigus marmoreus bacterial strain and production method of hypsizigus marmoreus fruit body Effective date of registration: 20160808 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2016990000684 |
|
| PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20171012 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2016990000684 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Hypsizigus marmoreus bacterial strain and production method of hypsizigus marmoreus fruit body Effective date of registration: 20171017 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2017370010067 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20181113 Granted publication date: 20140709 Pledgee: Qingdao high technology financing Company limited by guarantee Pledgor: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Registration number: 2017370010067 |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20181128 Address after: 066607 South of Changhuang Highway and East of Planned Coastal Expressway in Dapuhe Town, Changli County, Qinhuangdao City, Hebei Province Co-patentee after: Shanghai Finc Bio-tech Inc. Patentee after: HEBEI FENGKE BIOTECHNOLOGY CO.,LTD. Co-patentee after: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. Address before: 266221 68 Tianshan Third Road, Daxin Town, Jimo City, Qingdao City, Shandong Province Patentee before: QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Agaricus martensii strain Effective date of registration: 20230222 Granted publication date: 20140709 Pledgee: Industrial Bank Co.,Ltd. Shanghai Hongkou sub branch Pledgor: Shanghai Finc Bio-tech Inc.|QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd.|HEBEI FENGKE BIOTECHNOLOGY CO.,LTD. Registration number: Y2023310000040 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20140709 Pledgee: Industrial Bank Co.,Ltd. Shanghai Hongkou sub branch Pledgor: Shanghai Finc Bio-tech Inc.|QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd.|HEBEI FENGKE BIOTECHNOLOGY CO.,LTD. Registration number: Y2023310000040 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Zhenji mushroom strain Granted publication date: 20140709 Pledgee: Industrial Bank Co.,Ltd. Shanghai Hongkou sub branch Pledgor: Shanghai Finc Bio-tech Inc.|QINGDAO FENGKE BIOTECHNOLOGY Co.,Ltd.|HEBEI FENGKE BIOTECHNOLOGY CO.,LTD. Registration number: Y2024310000173 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |