CN105483019B - A kind of method of lucidum strain purification and rejuvenation - Google Patents
A kind of method of lucidum strain purification and rejuvenation Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 22
- 230000003716 rejuvenation Effects 0.000 title claims abstract description 22
- 238000000746 purification Methods 0.000 title claims abstract description 12
- 240000008397 Ganoderma lucidum Species 0.000 claims abstract description 57
- 235000001637 Ganoderma lucidum Nutrition 0.000 claims abstract description 57
- 238000004519 manufacturing process Methods 0.000 claims abstract description 46
- 238000011031 large-scale manufacturing process Methods 0.000 claims abstract description 24
- 241000894007 species Species 0.000 claims abstract description 15
- 238000000926 separation method Methods 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 8
- 230000008859 change Effects 0.000 claims abstract description 5
- 238000012258 culturing Methods 0.000 claims abstract description 4
- 238000012216 screening Methods 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims description 86
- 239000001963 growth medium Substances 0.000 claims description 41
- 244000062793 Sorghum vulgare Species 0.000 claims description 27
- 235000019713 millet Nutrition 0.000 claims description 27
- 238000012360 testing method Methods 0.000 claims description 24
- 238000004321 preservation Methods 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 239000002023 wood Substances 0.000 claims description 21
- 239000012092 media component Substances 0.000 claims description 20
- 235000015097 nutrients Nutrition 0.000 claims description 20
- 235000007164 Oryza sativa Nutrition 0.000 claims description 17
- 235000009566 rice Nutrition 0.000 claims description 17
- 235000015099 wheat brans Nutrition 0.000 claims description 17
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 13
- 241000894006 Bacteria Species 0.000 claims description 12
- 244000068988 Glycine max Species 0.000 claims description 12
- 235000010469 Glycine max Nutrition 0.000 claims description 12
- 238000004458 analytical method Methods 0.000 claims description 12
- 229910052602 gypsum Inorganic materials 0.000 claims description 12
- 239000010440 gypsum Substances 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 8
- 230000035784 germination Effects 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 4
- 238000012549 training Methods 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 3
- -1 Vb10.03~0.07 part Substances 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000003501 co-culture Methods 0.000 claims description 2
- 241000219492 Quercus Species 0.000 claims 7
- 240000007594 Oryza sativa Species 0.000 claims 3
- 238000006243 chemical reaction Methods 0.000 abstract description 9
- 241000233866 Fungi Species 0.000 abstract description 3
- 230000007850 degeneration Effects 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- 230000009467 reduction Effects 0.000 abstract description 3
- 238000009395 breeding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract description 2
- 241000209094 Oryza Species 0.000 description 14
- 241000207961 Sesamum Species 0.000 description 11
- 235000003434 Sesamum indicum Nutrition 0.000 description 11
- 238000005516 engineering process Methods 0.000 description 9
- 239000000463 material Substances 0.000 description 6
- 230000009466 transformation Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 241000222336 Ganoderma Species 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
The present invention relates to microorganism fungus kind breeding technique field, especially a kind of method of lucidum strain purification and rejuvenation.The specific healthy and strong ganoderma lucidum individual tissue separation and Extraction parent species using within three generations and three generations, take into full account that lucidum strain is cultivating change of the different phase of production to nutritional need, utilize different culture mediums, progressively carry out the first screening and culturing of strain, second of culture, third time is cultivated, 4th culture, extend Primary spawn, expand reincarnation production kind culture, and the large-scale production application in workshop, effectively solve the problems, such as lucidum strain spawn degeneration caused by Long Term Passages culture, improve the biological conversion ratio of lucidum strain production, reduction pollution scrappage, strain guarantee is provided for the scale sustainability production of ganoderma lucidum.
Description
Technical field
The present invention relates to microorganism fungus kind breeding technique field, especially a kind of method of lucidum strain purification and rejuvenation.
Technical background
Ganoderma lucidum is a kind of important medicinal fungi cultivated extensively, and production equally has degeneration with strain with other microorganisms
Problem.At present, China ganoderma lucidum respectively cultivates the first generation parent species of base production and is substantially introduction, the one-level parent species kind of introduction
Property it is excellent, after repeatedly switching original seed, production kind when, the component of culture medium is all by 80%~85% wood chip, 15%~20%
Wheat bran or rice bran are constituted, and do not take into full account that lucidum strain is cultivating change of the different phase of production to nutritional need, two
After generation, ganoderma lucidum grows continuous weak tendency, and resistance is poor, ultimately results in the planting household underproduction or even total crop failure.
At present, shown as when glossy ganoderma cultivating is produced：
The first generation (year)：Mycelia material feeding quickly, culture medium mycelia pure white, growth cycle length sturdy all the time, yield high-quality
High, biological conversion ratio can >=5.3%, and resistance is very strong, even in the simple and crude inoculation bacteria condition ring of common planting household
Inadequate cleaning ambient near border, or village people and animals life cultivates place, and it is zero or few that can also reach pollution scrappage, pollution
Rate≤1%；
The second generation (year)：The parent species of introduction are through succeeding preservation, or tissue separation, and mycelia material feeding is fast or very fast, culture medium
Relatively sturdy pure white, growth cycle is longer, yield high-quality is good, biological conversion ratio can reach 5%, resistance all the time for mycelial growth
Relatively strong, pollution rate is less than≤3%；
The third generation (year)：The parent species of introduction again succeeding preservation or tissue separation, mycelia material feeding is slack-off, culture medium mycelia life
Long not sturdy enough, pure white duration is shortened, growth and maturity cycle time, yield and quality are relatively good, biological conversion ratio 4.5
~4.9%, resistance is weaker, production is planted and culture medium bacterium wood pollution scrappage is carried on the back up to 5~10%, about 5% or so fructification
Face has downright bad patch；
Forth generation, the 5th generation (year) and after：Parent species is succeeding preservations again or organize separation, and mycelia material feeding is slow, training
Support that base mycelial growth is thin and delicate, mycelia more than 40 ages in days normally becomes brown patch, growth and maturity cycle and greatly shortened, from original
Plant, production kind to culture medium bacterium wood usually occurs --- culture base-material surface still has 1/3,1/4 or even 1/2 area mycelia slow
Fail to have position late, and vaccination just grows fructification.It is most once the culture medium bacterium wood pollution scrappage after de- bag earthing
Up to more than 30%, biological conversion ratio is reduced to less than 4.3%, is often no lack of total crop failure.And can only typically be relied on whenever now
" new " parent species are introduced again, to improve biological conversion ratio, the reduction pollution scrappage of ganoderma lucidum production, add production cost.
The content of the invention
For above-mentioned prior art, it is an object of the invention to overcome in the prior art lucidum strain through multiple squamous subculture
The problem of biological conversion ratio is low afterwards, pollution scrappage is significantly increased, height is still stablized so as to provide after a kind of ganoderma lucidum squamous subculture
Production, strong stress resistance, the method for the lucidum strain purification and rejuvenation of bacterium rod biological transformation ratio >=5%.
To achieve these goals, the present invention provides following technical scheme, a kind of method of lucidum strain purification and rejuvenation, bag
Include following steps：
(1) the first screening and culturing of strain：Using the healthy and strong ganoderma lucidum individual tissue separation and Extraction within three generations and three generations
Parent species, coordinate potato as culture medium using agar or millet, regular bevel culture 10~15 days, then be placed in ultralow temperature -80~-
50 DEG C of preservations；
(2) second of culture：The lucidum strain of selecting step (1) culture, using millet powder and robur leaf juice as culture medium, often
Advise inclined-plane culture 10~15 days, then carry out ultralow temperature preservation；
(3) third time is cultivated：The lucidum strain of selecting step (2) culture, using millet powder and oak leaf powder as culture medium, often
Advise inclined-plane culture 10~15 days, then carry out ultralow temperature preservation；
(4) the 4th cultures：The lucidum strain of selecting step (3) culture, using oak leaf powder, Vb1 and water as culture medium, often
Inclined-plane culture is advised, stops culture when the full test tube of mycelial growth, then carry out ultralow temperature preservation；
(5) original seed is extended：Selecting step (4) culture lucidum strain, using the mixture of robur wood chip, wheat bran or rice bran as
Culture medium, using wide-mouth bottle, tampon cellar culture 18~25 days, it was four layers of hospital gauze sealing cover bottleneck then to change tampon, after
Continuous culture 10~20 days and preservation；
(6) reincarnation production kind is expanded：The lucidum strain of selecting step (5) culture, with the mixture of robur wood chip, wheat bran or rice bran
For culture medium, through steam sterilizing, splitting, inoculation, and add bundle outer bag hood guard, cellar culture, as 8~10cm of inoculation hole bacterium germination diameter
Overlay film removes outer hood guard bag afterwards, continues cellar culture one and co-cultures 30~40 days and preservation；
(7) application production：The lucidum strain that step (6) is cultivated is used for workshop large-scale production.
Further, second of culture of the step (2) nutrient media components mass ratio used is millet powder：Robur leaf juice
=(2.6~3.5):L, preferably millet powder：Robur leaf juice=3:l.
Further, it is millet powder that step (3) third time, which cultivates nutrient media components mass ratio used,：Oak leaf powder
=(0.5~1.5):L, preferably millet powder：Oak leaf powder=1:l.
Further, the mass component of the 4th culture of the step (4) culture medium used is：Oak leaf powder 800~
1200 parts, Vb10.03~0.07 part, 1300~1700 parts of water, be preferably：1000 parts of oak leaf powder, Vb10.05 parts, water 1500
Part.
Further, bacterium is worked as using test tube, tampon regular bevel culture in the step (1), step (2), step (3)
Silk stops culture from when being 3~7mm with a distance from test tube bottom.
Further, using wide-mouth bottle, tampon cellar culture in the step (5), when culture bacterium germination is into wide-mouth bottle one
It is four layers of hospital gauze sealing cover bottleneck that tampon is changed when half, continues to cultivate, and when mycelia hair is full in wide-mouth bottle, stops cultivating and protecting
Hide.
Further, step (5) the extension original seed, step (6) expand the mass component of the culture medium used in reincarnation production kind
For：220~280 parts of 700~750 parts of robur wood chip, wheat bran or rice bran, 15~25 parts of analysis for soybean powder, 8~13 parts of gypsum, water 1100
~1300 parts.
The beneficial effects of the present invention are：The method for the lucidum strain purification and rejuvenation that the present invention is provided, takes into full account ganoderma lucidum
Strain is cultivating change of the different phase of production to nutritional need, using different culture mediums, can effectively solve Ganoderma Lucidum
Plant the spawn degeneration problem caused by Long Term Passages culture；Increased by being arrived in the training of the first sieve of strain between the big productive culture in workshop
Time middle transition culture is added, robur leaf juice, oak leaf powder, robur wood chip is added respectively in multiple transition culture medium
Deng realizing being changed stepwise for culture medium, the wooden media environment of robur section for making strain adapt to produce greatly gradually improves lucidum strain
Biological conversion ratio, the reduction pollution scrappage of production, strain guarantee is provided for the scale sustainability production of ganoderma lucidum.
Embodiment
Embodiment to the present invention is explained as follows：
A kind of method of lucidum strain purification and rejuvenation, comprises the following steps：
(1) the first screening and culturing of strain：In annual July, using the healthy and strong ganoderma lucidum individual within three generations and three generations
Separation and Extraction parent species are organized, coordinates potato as culture medium using agar or millet, uses test tube, silica gel plug regular bevel culture
10~15 days, culture is stopped when Mycelium Growth of Ganoderma lucidum is to from 3~7mm of test tube bottom；Again by test tube be placed in ultralow temperature -80~-
50 DEG C of preservations to October, ultralow temperature preservation makes mycelia in a dormant state, meets production turnover needs.
(2) second of culture：The lucidum strain test tube of optional step (1) culture, takes apart from 3~10mm of test tube bottom, spirit
The mycelia block of the diameter 2mm sizes at the dense regular place of sesame mycelia together with culture medium, be placed in it is new using millet powder and robur leaf juice as
In the middle part of the test tube of culture medium, using tampon regular bevel culture 10~15 days, when Mycelium Growth of Ganoderma lucidum to from test tube bottom 3~
Stop culture during 7mm, then test tube is placed in ultralow temperature preservation.This stylish nutrient media components is millet powder：Robur leaf juice=
(2.6~3.5):L, nutrient media components is preferably millet powder：Robur leaf juice=3:l.
(3) third time is cultivated：The lucidum strain test tube of optional step (2) culture, takes apart from 3~10mm of test tube bottom, spirit
The mycelia block of the diameter 2mm sizes at the dense regular place of sesame mycelia together with culture medium, be placed in it is new using millet powder and oak leaf powder as
In the middle part of the test tube of culture medium, using tampon regular bevel culture 10~15 days, when Mycelium Growth of Ganoderma lucidum to from test tube bottom 3~
Stop culture during 7mm, then test tube is placed in ultralow temperature preservation.This stylish nutrient media components is millet powder：Oak leaf powder=
(0.5~1.5):L, the preferred millet powder of nutrient media components：Oak leaf powder=1:l.
(4) the 4th cultures：Optional step (3) third time culture lucidum strain test tube, take apart from test tube bottom 3~
The mycelia block of the diameter 2mm sizes at the dense regular place of 10mm, Ganoderma lucidum mycelium is placed in new with oak leaf powder, Vb1 together with culture medium
With test tube middle part of the water for culture medium, using tampon regular bevel culture 13~17 days, stop when Ganoderma lucidum mycelium covers with test tube
Culture, then test tube is placed in ultralow temperature preservation.This stylish culture medium mass component be 800~1200 parts of oak leaf powder,
1300~1700 parts of Vb10.03~0.07 part, water；Nutrient media components is preferred：1000 parts of oak leaf powder, Vb10.05 parts, water 1500
Part.
(5) original seed is extended：With robur wood chip, wheat bran or rice bran, analysis for soybean powder, gypsum etc. for culture medium, wherein culture matrix
Amount component is 220~280 parts of 700~750 parts of robur wood chip, wheat bran or rice bran, 15~25 parts of analysis for soybean powder, 8~13 parts of gypsum, water
1100~1300 parts, the lucidum strain of the 4th culture of inoculation step (4), uses tampon cellar culture 18~25 in wide-mouth bottle
My god, tampon is changed when the half of culture bacterium germination to wide-mouth bottle and continues to cultivate 10~20 days for four layers of hospital gauze sealing cover bottleneck, when
Stop culture and preservation when mycelia hair is full in wide-mouth bottle.
(6) reincarnation production kind is expanded：With robur wood chip, wheat bran or rice bran, analysis for soybean powder, gypsum etc. for culture medium, wherein culture medium
Mass component be 220~280 parts of 700~750 parts of robur wood chip, wheat bran or rice bran, 15~25 parts of analysis for soybean powder, 8~13 parts of gypsum,
1100~1300 parts of water, through steam sterilizing, splitting, every bag of 1.5~1.9kg, the ganoderma lucidum cultivated in the hole inoculation step (5) of bag body five
Strain, and add bundle outer bag hood guard, cellar culture；Overlay film removes outer hood guard bag after inoculation hole bacterium germination 8~10cm of diameter, continues conventional
Culture, stops culture and preservation when bag material mycelia hair is full, wherein culture period is 30~40 days altogether.
(7) application production：The lucidum strain that step (6) is cultivated is used for workshop large-scale production.
Embodiment 1：
Number red sesame lucidum variety of Shanghai agriculture is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, separation and Extraction parent species,
The purification of lucidum strain, rejuvenation are carried out through above-mentioned steps and applied to workshop large-scale production.Wherein, the culture of second of culture
Base component is millet powder：Robur leaf juice=2.6:l；The nutrient media components of third time culture is millet powder：Oak leaf powder=0.5:
l；4th time culture culture medium mass component be：800 parts of oak leaf powder consumption, 0.03 part of Vb1 consumptions, 1300 parts of water consumption；
Primary spawn is extended, nutrient media components is calculated according to the mass fraction, 280 parts of 700 parts of robur wood chip consumption, wheat bran or rice bran consumption,
15 parts of analysis for soybean powder consumption, 8 parts of gypsum consumption, 1100 parts of water consumption；Expand reincarnation production kind culture, nutrient media components is according to the mass fraction
Calculate, 220 parts of 750 parts of robur wood chip consumption, wheat bran or rice bran consumption, 20 parts of analysis for soybean powder consumption, 10 parts of gypsum consumption, water consumption
1100 parts；The lucidum strain of previous step culture is used for workshop large-scale production.Repeat the above steps, produce three batches, often
The large-scale production of batch workshop uses bavin Duan Mu (moisture content 45% or so) 100kg, and the accumulative ganoderma lucidum produced and conidia powder are total
Measure and reach 14.55kg for 15.45kg, pollution learies；And control group, routinely squamous subculture progress workshop large-scale production, life
The ganoderma lucidum of output and conidia powder total amount are 11.85kg, pollute learies up to 100.95kg.
Embodiment 2：
Number red sesame lucidum variety of Shanghai agriculture is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, separation and Extraction parent species,
The purification of lucidum strain, rejuvenation are carried out through above-mentioned steps and applied to workshop large-scale production.Wherein, the culture of second of culture
Base component is millet powder：Robur leaf juice=3.5:l；The nutrient media components of third time culture is millet powder：Oak leaf powder=1.5:
l；4th culture, culture medium mass component be oak leaf powder consumption be 1200 parts, Vb1 consumptions be 0.07 part, water consumption be
1700 parts；Primary spawn is extended, nutrient media components is calculated according to the mass fraction, 750 parts of robur wood chip consumption, wheat bran or rice bran consumption
220 parts, 18 parts of analysis for soybean powder consumption, 11 parts of gypsum consumption, 1250 parts of water consumption；Expand reincarnation production kind culture, wherein nutrient media components
Calculate according to the mass fraction, 250 parts of 720 parts of robur wood chip consumption, wheat bran or rice bran consumption, 22 parts of analysis for soybean powder consumption, gypsum consumption
10 parts, 1300 parts of water consumption；The lucidum strain of previous step culture is used for workshop large-scale production.Repeat the above steps, it is raw
Three batches are produced, bavin Duan Mu (moisture content 45% or so) 100kg, the accumulative ganoderma lucidum produced are used per the large-scale production of batch workshop
And conidia powder total amount is 15.66kg, pollutes learies up to 14.16kg；And control group, routinely squamous subculture carry out workshop scale
Metaplasia is produced, and the ganoderma lucidum produced and conidia powder total amount are 12.24kg, pollute learies up to 96.51kg.
Embodiment 3：
Number red sesame lucidum variety of Shanghai agriculture is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, separation and Extraction parent species,
The purification of lucidum strain, rejuvenation are carried out through above-mentioned steps and applied to workshop large-scale production.Wherein, the culture of second of culture
Base component is millet powder：Robur leaf juice=3:l；The nutrient media components of third time culture is millet powder：Oak leaf powder=1:l；The
Four cultures, culture medium mass component be oak leaf powder consumption be 1000 parts, Vb1 consumptions be 0.05 part, water consumption be 1500 parts；
Primary spawn is extended, nutrient media components is calculated according to the mass fraction, 250 parts of 720 parts of robur wood chip consumption, wheat bran or rice bran consumption,
20 parts of analysis for soybean powder consumption, 10 parts of gypsum consumption, 1200 parts of water consumption；Expand reincarnation production kind culture, nutrient media components presses mass fraction
Calculate, 270 parts of 700 parts of robur wood chip consumption, wheat bran or rice bran consumption, 21 parts of analysis for soybean powder consumption, 12 parts of gypsum consumption, water consumption
1280 parts；The lucidum strain of previous step culture is used for workshop large-scale production.Repeat the above steps, produce three batches, often
The large-scale production of batch workshop uses bavin Duan Mu (moisture content 45% or so) 100kg, and the accumulative ganoderma lucidum produced and conidia powder are total
Measure and reach 13.80kg for 15.93kg, pollution learies；And control group, routinely squamous subculture progress workshop large-scale production, life
The ganoderma lucidum of output and conidia powder total amount are 12.48kg, pollute learies up to 88.62kg.
Embodiment 4：
Number lucidum variety of Mount Huang spore sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 1
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 16.80kg, pollution learies
Up to 12.33kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
21.87kg is reached for 13.95kg, pollution learies.
Embodiment 5：
Number lucidum variety of Mount Huang spore sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 2
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 17.28kg, pollution learies
Up to 11.61kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
20.49kg is reached for 14.16kg, pollution learies.
Embodiment 6：
Number lucidum variety of Mount Huang spore sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 3
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 17.67kg, pollution learies
Up to 9.60kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount are
14.49kg, pollution learies reach 18.72kg.
Embodiment 7：
Number lucidum variety of Mount Huang purple sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, keeps same with example 1
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 15.60kg, pollution learies
Up to 12.78kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
103.68kg is reached for 12.31kg, pollution learies.
Embodiment 8：
Number lucidum variety of Mount Huang purple sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, keeps same with example 2
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 15.18kg, pollution learies
Up to 12.57kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
101.13kg is reached for 11.82kg, pollution learies.
Embodiment 9：
Number lucidum variety of Mount Huang purple sesame is chosen, is organized using the healthy and strong ganoderma lucidum individual of the third generation, keeps same with example 3
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 16.29kg, pollution learies
Up to 12.12kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
96.72kg is reached for 12.66kg, pollution learies.
Embodiment 10：
Number lucidum variety of Mount Huang rainbow conk is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 1
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 16.77kg, pollution learies
Up to 11.16kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
23.79kg is reached for 14.04kg, pollution learies.
Embodiment 11：
Number lucidum variety of Mount Huang rainbow conk is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 2
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 17.12kg, pollution learies
Up to 10.02kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount
21.51kg is reached for 14.19kg, pollution learies.
Embodiment 12：
Number lucidum variety of Mount Huang rainbow conk is chosen, is organized using the healthy and strong ganoderma lucidum individual of the second generation, keeps same with example 3
The production technology of sample, the batch of duplication of production three, the accumulative ganoderma lucidum produced and conidia powder total amount are 17.58kg, pollution learies
Up to 8.88kg；And control group, routinely squamous subculture progress workshop large-scale production, the ganoderma lucidum produced and conidia powder total amount are
14.55kg, pollution learies reach 19.08kg.
Every data statistics that examples detailed above is drawn is calculated as follows shown in table：
Wherein, biological transformation ratio=ganoderma lucidum and spore powder yield/wooden consumption * 100% of bavin section；
Total amount/wooden consumption * 100% of bavin section is scrapped in pollution scrappage=pollution.
The present invention is can be seen that by upper table data to organize by using the healthy and strong ganoderma lucidum individual within three generations and three generations
Parent species are repeatedly purified, rejuvenation culture by separation and Extraction parent species by different culture medium and training method, so that life is made
The low high-quality lucidum strain of thing high conversion rate, pollution scrappage.Compared to strain conventional treatment method, strain of the invention exists
After squamous subculture, yield is higher, more stable, strong stress resistance, its biological transformation ratio >=5% produced, pollution scrappage≤5%.
It is above-mentioned that invention is exemplarily described, it is clear that the present invention, which is implemented, to be not subject to the restrictions described above, only
This insubstantial for employing inventive concept and technical scheme of the present invention progress is improved, or the not improved design by invention
Other occasions are directly applied to technical scheme, within the scope of the present invention.
Claims (10)
1. a kind of method of lucidum strain purification and rejuvenation, it is characterised in that comprise the following steps：
(1) the first screening and culturing of strain：Using the healthy and strong ganoderma lucidum individual tissue separation and Extraction parent species within three generations and three generations,
Potato is coordinated as culture medium using agar or millet, regular bevel culture 10~15 days, then it is placed in ultralow temperature -80~-50 DEG C guarantor
Hide；
(2) second of culture：The lucidum strain of selecting step (1) culture, with millet powder and robur (Quercus Linn) leaf juice
For culture medium, regular bevel culture 10~15 days, then carry out ultralow temperature preservation；
(3) third time is cultivated：The lucidum strain of selecting step (2) culture is conventional oblique using millet powder and oak leaf powder as culture medium
Face is cultivated 10~15 days, then carries out ultralow temperature preservation；
(4) the 4th cultures：The lucidum strain of selecting step (3) culture is conventional oblique using oak leaf powder, Vb1 and water as culture medium
Face is cultivated, and stops culture when the full test tube of mycelial growth, then carry out ultralow temperature preservation；
(5) original seed is extended：The lucidum strain of selecting step (4) culture, using the mixture of wood chip containing robur and wheat bran or rice bran as
Culture medium, using wide-mouth bottle, tampon cellar culture 18~25 days, it was four layers of hospital gauze sealing cover bottleneck then to change tampon, after
Continuous culture 10~20 days and preservation；
(6) reincarnation production kind is expanded：The lucidum strain of selecting step (5) culture, with wood chip containing robur and the mixture of wheat bran or rice bran
For culture medium, through steam sterilizing, splitting, inoculation, and add bundle outer bag hood guard, cellar culture, as 8~10cm of inoculation hole bacterium germination diameter
Overlay film removes outer hood guard bag afterwards, continues cellar culture, and one co-cultures 30~40 days and preservation；
(7) application production：The lucidum strain that step (6) is cultivated is used for workshop large-scale production.
2. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that the step (2) second
Nutrient media components mass ratio used in secondary culture is millet powder：Robur leaf juice=(2.6~3.5):l.
3. a kind of method that lucidum strain is purificated and rejuvenated according to claim 2, it is characterised in that the step (2) second
Nutrient media components mass ratio used in secondary culture is millet powder：Robur leaf juice=3:l.
4. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that the step (3) the 3rd
Nutrient media components mass ratio used in secondary culture is millet powder：Oak leaf powder=(0.5~1.5):l.
5. a kind of method that lucidum strain is purificated and rejuvenated according to claim 4, it is characterised in that the step (3) the 3rd
Nutrient media components mass ratio used in secondary culture is millet powder：Oak leaf powder=1:l.
6. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that the step (4) the 4th
The mass component of culture medium used in secondary culture is：800~1200 parts of oak leaf powder, Vb10.03~0.07 part, water 1300~
1700 parts.
7. a kind of method that lucidum strain is purificated and rejuvenated according to claim 6, it is characterised in that the step (4) the 4th
The mass component of culture medium used in secondary culture is：1000 parts of oak leaf powder, 0.05 part of Vb1,1500 parts of water.
8. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that the step (1), step
(2), using test tube, tampon regular bevel culture in step (3), when mycelia is from stopping training when being 3~7mm with a distance from test tube bottom
Support.
9. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that make in the step (5)
With wide-mouth bottle, tampon cellar culture, it is four layers of hospital gauze sealing cover bottleneck that tampon is changed when cultivating bacterium germination half into wide-mouth bottle,
Continue to cultivate, when mycelia hair is full in wide-mouth bottle, stop culture and preservation.
10. a kind of method that lucidum strain is purificated and rejuvenated according to claim 1, it is characterised in that：Step (5) extension
The mass component of culture medium that original seed, step (6) expand used in reincarnation production kind is：700~750 parts of robur wood chip, wheat bran or rice bran
220~280 parts, 15~25 parts of analysis for soybean powder, 8~13 parts of gypsum, 1100~1300 parts of water.
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Non-Patent Citations (3)
| Title |
|---|
| 初洋等.野生灵芝的驯化栽培技术.《北方园艺》.2010,(第2010年17期),摘要，第217页左栏第3段至218页左栏第5段，第218页右栏第2段. * |
| 卢巧英等.盆景灵芝不同杂交菌株在不同培养基上菌丝生长情况研究初报.《农业与技术》.2014,第34卷(第1期),参见第26页左栏表1. * |
| 闫茂华.栎叶、废弃茶叶栽培灵芝试验.《食用菌》.2000,(第2000年2期),第23页左栏第1段-第2段. * |
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