CN107557307A - A kind of lab screening method of high temperature resistant cloud ear bacterial strain - Google Patents
A kind of lab screening method of high temperature resistant cloud ear bacterial strain Download PDFInfo
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Abstract
The invention belongs to bacterial strain screening technical field, discloses a kind of lab screening method of high temperature resistant cloud ear bacterial strain, and bacterial strain cultivates 7d under 6 thermogrades, and each processing sets 3 repetitions；The bacterial strain mycelia disk for activating 5d is inoculated into PDA synthetic mediums center, is placed at 40 DEG C and takes out renewal cultivation after different incubation times；After strains tested activation 5d, picking colony edge mycelia block disk is inoculated in PDA synthesis plating medium center cultures, high-temperature exercise；After strains tested activation culture 8d, for making cultivation strain；After cultivation strain purseful, cultigen is accessed 50d or so is cultivated in fruiting bag culture medium, fruiting bag is put into produce agaric under Cultivation condition respectively.Bacterial strain of the mycelium compared with high temperature resistant and the produce agaric that can normally budding in fruiting high-temperature region is filtered out, obtains relatively resistant to elevated temperatures Germplasms, the excellent cloud ear kind of high temperature resistant for being adapted to Guangxi province cultivation for seed selection from now on lays the foundation.
Description
Technical field
The invention belongs to bacterial strain screening technical field, more particularly to a kind of lab screening method of high temperature resistant cloud ear bacterial strain.
Background technology
Guangxi fungus cultivation has more than 500 years history, is one of 4 big edible mushroom of Guangxi.Due to the black fungus market price in recent years
Stable, economic benefit is obvious, and its cultivated area expands day by day.Most of black fungus kind that Guangxi main is planted at present from the north or
The middle low form kind of the band of Zhejiang Fujian one.Middle low temperature kind is cultivated in Guangxi, runs into warm winter weather, rotten bag easily occurs, tide
Secondary reduction, yield and quality decline.Cloud ear is the distinctive black fungus kind in Guangxi, and auricle sepia is glossy, translucent, enters
The tender crisp and refreshing cunning of mouth, excellent taste, because quality well once ranked front three in national commodity agaric is appraised through comparison, enjoy great prestige at home and abroad.Although cloud
Ear higher warm type kind in belonging in black fungus kind, but influenceed by megathermal climate still larger, therefore cloud ear also mainly collects
In cultivated in North-west Guangxi or osmanthus Mountainous Region, Guangxi most area can not still cultivate.In order to preferably promote Guangxi cloud ear characteristic
The utilization of resource, it is necessary to filter out more resistant to elevated temperatures cloud ear bacterial strain, expand cultural area.
The triage techniques method of edible mushroom high temperature bacterial strain mainly has two ways at present：First, mycelium heat shock method is taken,
After i.e. mycelium cultivates certain time under suitable culture conditions, progress thermostimulation 1-8h under hot conditions is put into, further takes out and puts
Renewal cultivation is carried out at 25 DEG C of normal temperature, measures the speed of growth of mycelia, eliminates the bacterial strain of heat-resisting ability difference；Second, bacterium
The single temperature high temperature resistant cultivation of filament, the mycelia after rigid connection kind is positioned under 40 DEG C of hot conditions and carries out culture 1-24h, then
Taking-up is placed at 25 DEG C of normal temperature and carries out renewal cultivation, measures the speed of growth of mycelia, eliminates the bacterial strain of heat-resisting ability difference.
In summary, the problem of prior art is present be：First, the mycelium of black fungus has certain patience to high temperature,
, can normally restoration ecosystem after mycelium is influenceed by of short duration high temperature.The time that kind is endured hot conditions is longer, degree
The heat-resisting ability of the bigger explanation kind is stronger.It is short, high that incubation time is heat-treated in mycelium high temperature resistant screening technique at present
Warm condition is single, lacks mycelium by low temperature to high temperature acclimation adaptability, stabilization exercise, therefore the high-temperature resistant strain filtered out
Lack representative and stability.2nd, edible mushroom different growing, the tolerance of high temperature is not quite similar.One excellent resistance to height
Warm kind, not only mycelium energy high temperature resistant, fructification also can high temperature resistant fruitings.Edible mushroom high temperature screening varieties technology is main at present
To be screened using mycelium high temperature resistant, the screening of fructification heatproof is less, therefore mycelium high temperature resistant occurs in the kind filtered out, but
Under hot conditions can not normal fruiting the problem of.
The content of the invention
The problem of existing for prior art, the invention provides a kind of lab screening method of high temperature resistant cloud ear bacterial strain.
The present invention is achieved in that a kind of lab screening method of high temperature resistant cloud ear bacterial strain, the high-temperature resistant strain
Lab screening method comprises the following steps：
Step 1, sets 6 thermogrades, and each processing sets 3 repetitions；Activation 5d bacterial strain mycelia disk is inoculated into
PDA synthetic mediums center, is respectively placed in 25 DEG C, 28 DEG C, 31 DEG C, 34 DEG C, 37 DEG C, cultivates 7d in 40 DEG C of incubator, observes
Mycelial growth situation, measure mycelial growth rate；
Step 2, the bacterial strain mycelia disk activated are inoculated into PDA synthesis plating mediums center, are placed in 40 DEG C of incubators
After middle culture respectively 3d, 5d, 7d, 9d, 11d, then remove and be positioned over 25 DEG C of renewal cultivation 7d, each processing sets 3 repetitions, observes
Mycelial growth situation, measure mycelial growth rate；
Step 3, after strains tested activates 5d, picking colony edge mycelia block disk is inoculated in PDA synthesis plating mediums
Center, it is placed at 40 DEG C and cultivates 5d, takes out the renewal cultivation 7d at 28 DEG C；The preferable mycelia block of germination and growth is selected to be inoculated in
On PDA synthetic mediums, 42 DEG C of culture 3d are put into, take out the renewal cultivation 7d at 28 DEG C, then select the preferable bacterium of germination and growth
Silk block is inoculated on PDA synthetic mediums, after being placed in 45 DEG C of culture 5h；Taking-up cultivates 7d at 28 DEG C；Each Temperature Treatment is extensive
After the long 7d of demutation, mycelial growth situation is observed, measures mycelial growth rate.
Step 4：After strains tested activation culture 8d, cultivation strain is made；After cultivation strain purseful, cultigen is accessed
50d or so is cultivated in fruiting bag culture medium, fruiting bag is put into respectively 25 DEG C, 28 DEG C, carry out produce agaric under 31 DEG C of Cultivation conditions, see
Examine the situation of buddingging.
Further, mycelial growth rate (mm/d)=colony radius (mm)/growth number of days (d) in the step 1.
Further, the high temperature resistant cloud ear bacterial strain lab screening method screening bacterial strain TY041, TY026, TY071,
HME2、YE1。
Advantages of the present invention and good effect are：Bacterial strain TY041, TY026, TY071, HME2, YE1, through under different temperatures
Cultural hypha, at 40 DEG C after high-temperature cultivation processing mycelia recover, high-temperature exercise (40 DEG C, 42 DEG C, 45 DEG C), mycelial growth speed
Degree and mycelium growth vigor show relatively stable.The bacterial strain screened restoration ecosystem speed after 40 DEG C of high-temperature cultivations for a long time
Than control strain 916 fast more than 10%.The bacterial strain filtered out by the technical method fruiting temperature of buddingging carries than control strain 916
It is high 6 DEG C.Filter out bacterial strain not only mycelium compared with high temperature resistant, and can in high-temperature region 28-31 DEG C normally budding, obtain it is relatively resistance to
The Germplasms of high temperature, the excellent cloud ear kind of high temperature resistant for being adapted to Guangxi province cultivation for seed selection from now on lay the foundation.
Brief description of the drawings
Fig. 1 is the lab screening method flow diagram of high temperature resistant cloud ear bacterial strain provided in an embodiment of the present invention.
Fig. 2 is that colony growth situation compares signal to different cloud ear bacterial strains provided in an embodiment of the present invention at different temperatures
Figure.
Fig. 3 is different cloud ear bacterial strains provided in an embodiment of the present invention restoration ecosystem speed after different incubation times at 40 DEG C
Schematic diagram.
Fig. 4 is that different cloud ear bacterial strains colony growth diameter of high-temperature exercise at 40 DEG C provided in an embodiment of the present invention compares
Schematic diagram.
Fig. 5 is that different 42 DEG C of high-temperature exercise mycelial growth rates of cloud ear bacterial strain provided in an embodiment of the present invention compare signal
Figure.
Fig. 6 is that different 45 DEG C of high-temperature exercise mycelial growth rates of cloud ear bacterial strain provided in an embodiment of the present invention compare signal
Figure.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
With Guangxi, main black fungus cultivation kind 916 is control at present, to from the wild cloud ear separating obtained 14 in the ground such as Tianlin County, Tiane
Individual bacterial strain is tested, and compares 15 bacterial strains mycelial growth situation at different temperatures by observation；Take exercise under the high temperature conditions
Mycelia recovery situation, filter out the more resistant to elevated temperatures bacterial strain of mycelium；Progress fruiting, which is buddingged, at different temperatures compares, and filters out out
The more resistant to elevated temperatures bacterial strain of mushroom, relatively resistant to elevated temperatures Germplasms are obtained, be adapted to the resistance to of Guangxi province cultivation for seed selection from now on
The excellent cloud ear kind of high temperature lays the foundation.
The application principle of the present invention is explained in detail below in conjunction with the accompanying drawings.
As shown in figure 1, the lab screening method of high temperature resistant cloud ear bacterial strain provided in an embodiment of the present invention comprises the following steps：
S101：6 thermogrades are set, and each processing sets 3 repetitions；Activate 5d bacterial strain mycelia disk (diameter 6mm)
PDA synthetic mediums (culture dish diameter 9cm) center is inoculated into, is respectively placed in 25 DEG C, 28 DEG C, 31 DEG C, 34 DEG C, 37 DEG C, 40 DEG C
Incubator in cultivate 7d, observe mycelial growth situation, measure mycelial growth rate.Mycelial growth rate (mm/d)=bacterium colony half
Footpath (mm)/growth number of days (d)；
S102：The bacterial strain mycelia disk activated is inoculated into PDA synthesis plating mediums center, is placed in 40 DEG C of incubators
After cultivating 3d, 5d, 7d, 9d, 11d respectively, then remove and be positioned over 25 DEG C of renewal cultivation 7d, each processing sets 3 repetitions, observes bacterium
Silk growing state, measures mycelial growth rate；
S103：After strains tested activation 5d, picking colony edge mycelia block disk is inoculated in PDA synthesis plating mediums
Centre, is placed at 40 DEG C and cultivates 5d, take out the renewal cultivation 7d at 28 DEG C；The preferable mycelia block of germination and growth is selected to be inoculated in
On PDA synthetic mediums, 42 DEG C of culture 3d are put into, take out the renewal cultivation 7d at 28 DEG C, then select the preferable bacterium of germination and growth
Silk block is inoculated on PDA synthetic mediums, after being placed in 45 DEG C of culture 5h, is taken out and is cultivated 7d at 28 DEG C；Each Temperature Treatment is extensive
After the long 7d of demutation, mycelial growth situation is observed, measures mycelial growth rate；
S104：After strains tested activation culture 8d, cultivation strain is made；After cultivation strain purseful, cultigen is accessed out
50d or so is cultivated in mushroom bag culture medium, fruiting bag is put into respectively 25 DEG C, 28 DEG C, carry out produce agaric under 31 DEG C of Cultivation conditions, observe
Budding situation.
The application effect of the present invention is explained in detail with reference to experiment.
1 materials and methods
1.1 test material
1.1.1 strains tested
Strains tested 15,916 be Guangxi main breed, and its introduces a collection derives from Lishui of Zhejiang；Remaining bacterial strain is Guangxi agriculture
Institute of microbiology of the industry academy of sciences is wild separating obtained, and passes through 2015-2016 fruiting experiment preliminary screenings.Specific bacterial strain is compiled
Number it is：YE1, YE2, HME2, TY013, TY024, TY026, TY041, TY051, TY063, TY071, TY072, TY073, YE (my god
2), YE (day 3), 916.(916 strains belong to existing, local Cotton Varieties by Small Farming Households post-sampling separation.Remaining bacterial strain is wild separation
Kind, in preservation collected by sheet.)
Strain source situation table
1.1.2 for examination culture medium
PDA synthetic mediums：Potato 200g, glucose 20g, agar 20g, epsom salt 2g, potassium dihydrogen phosphate 3g, steaming
Distilled water 1000mL, pH value are nature.
Cultigen, fruiting bag culture medium：Weed tree sawdust 78%, rice bran 20%, land plaster 1%, calcium superphosphate 1%, water content
60%, pH=7.0.
1.3 test method
1.3.1 the colony growth situation of strains tested at different temperatures compares.
Experiment sets 6 thermogrades, and each processing sets 3 repetitions.5d bacterial strain mycelia disk (diameter 6mm) will be activated
PDA synthetic mediums (culture dish diameter 9cm) center is inoculated into, is respectively placed in 25 DEG C, 28 DEG C, 31 DEG C, 34 DEG C, 37 DEG C, 40 DEG C
Incubator in cultivate 7d, observe mycelial growth situation, measure mycelial growth rate.Mycelial growth rate (mm/d)=bacterium colony half
Footpath (mm)/growth number of days (d).
1.3.2 recovery situation compares after strains tested cultivates different time at 40 DEG C
The bacterial strain mycelia disk activated is inoculated into PDA synthesis plating mediums center, is placed in 40 DEG C of incubators and divides
Pei Yang not be after 3d, 5d, 7d, 9d, 11d, then remove and be positioned over 25 DEG C of renewal cultivation 7d, each processing sets 3 repetitions, observes mycelia
Growing state, measure mycelial growth rate.
1.3.3 cloud ear bacterial strain high temperature resistant is taken exercise
After strains tested activation 5d, picking colony edge mycelia block disk is inoculated in PDA synthesis plating mediums center, puts
5d is cultivated at 40 DEG C, takes out the renewal cultivation 7d at 28 DEG C, the preferable mycelia block of germination and growth is then selected and is inoculated in PDA
On synthetic medium, 42 DEG C of culture 3d are put into, take out the renewal cultivation 7d at 28 DEG C, then select the preferable mycelia block of germination and growth
It is inoculated on PDA synthetic mediums, after being placed in 45 DEG C of culture 5h, takes out and cultivate 7d at 28 DEG C.Each Temperature Treatment recovers life
After long 7d, mycelial growth situation is observed, measures mycelial growth rate.
1.3.4 cloud ear bacterial strain different temperatures fruiting, which is buddingged, compares
Agaric bagging method routinely carries out strain, fruiting bag production.Strain bag specification 15cm × 26cm × 0.06cm,
Fruiting bag specification：15cm×55cm×0.04cm.After strains tested activation culture 8d, picking mycelia block is connected to Cultivar culture medium
On, make cultivation strain.After cultivation strain purseful, 50d or so is cultivated by 28 DEG C in cultigen access fruiting bag culture medium, by length
After the fruiting bag of full mycelia is punched with automatic punching device, be put into respectively 25 DEG C, 28 DEG C, carry out produce agaric under 31 DEG C of Cultivation conditions, see
Examine the situation of buddingging.10 fruiting bags of each processing inoculation.
2 results and analysis
2.1 strains testeds colony growth situation under different temperatures condition of culture compares
Experiment is provided with 6 thermogrades, and the mycelial growth situation description of strains tested at different temperatures is shown in Table 1, bacterium
The silk speed of growth is shown in Fig. 1.
Mycelium growth vigor of the strains tested of table 1 under different cultivation temperatures
Remarks：" ++++" the pure white densification of mycelial growth is represented, " +++ " represents that mycelial growth is pure white intensive, and " ++ " represents bacterium
Silk growth is pure white finer and close, and "+" represents that growth is shallow white sparse,
From Fig. 2, table 1, strains tested the mycelia equal normal germination and growth of energy, culture under 25-37 DEG C of condition of culture
Mycelia can sprout when temperature is 40 DEG C, but be unable to normal growth.At 25-31 DEG C major part strain growth speed with temperature raise and
Accelerate, the 31-37 DEG C of speed of growth is gradually reduced.
At 31 DEG C, mycelial growth is HME2, TY026, YE1, TY071, YE2, YE days 2 faster；But colony growth is preferable
Bacterial strain be HME2, TY026, YE1, TY071, YE2, TY041, therefore 31 DEG C of well-grown bacterial strains be HME2, TY026,
YE1、TY071、YE2。
At 34 DEG C, mycelial growth be faster HME2, TY041, TY026, TY013, YE2, YE days 2, YE days 3；Bacterium colony is given birth to
Long preferable bacterial strain is HME2, TY026, YE1, TY071, YE2, TY041, YE days 3,916, TY013, thus 34 DEG C of growths compared with
Good bacterial strain is HME2, TY026, TY041, TY013, YE2, YE days 3.
At 37 DEG C, colony growth situation difference is obvious, mycelial growth be faster HME2, TY041, YE days 2, YE days 3, its
Secondary is TY072, TY026, YE1；The preferable bacterial strain of colony growth be TY041, HME2, TY026, YE1, YE days 2, YE days 3, therefore
37 DEG C of well-grown bacterial strains be HME2, TY026, TY041, YE1, YE2, YE days 2, YE days 3.
Each bacterial strain colony growth situation and the growing way of mycelia draw and given birth under the high temperature conditions between 31-37 DEG C of integral high temperature area
Length is preferable and stable bacterial strain is：HME2, TY026, YE1, TY041, YE2, YE days 3.
Recovery situation of the 2.2 strains tested mycelia after culture different time under the conditions of 40 DEG C
The different cloud ear bacterial strains of table 2 cultivate mycelia restoration ecosystem situation after different time under the conditions of 40 DEG C
Remarks：" ++++" the pure white densification of mycelial growth is represented, " +++ " represents that mycelial growth is pure white intensive, and " ++ " represents bacterium
Silk growth is pure white finer and close, and "+" represents that growth is shallow white sparse.
From table 2, Fig. 3, bacterial strain YE1, TY071, TY041, TY026, HME2 cultivated at 40 DEG C 3d, 5d, 7d, 9d,
After 11d, mycelial growth rate and growing way are better than other bacterial strains and stably；Bacterial strain YE (day 3) though, TY013, TY063, YE2
So mycelial growth rate is fast also fast in indivedual process, but unstable or mycelium growth vigor is poor, the mycelial growth of control strain 916
Speed and mycelium growth vigor are general；Therefore, by cultivating different number of days processing under the conditions of 40 DEG C, screen mycelial growth rate it is fast and
The pure white fine and close bacterial strain 5 of mycelia, it is respectively：YE1、TY071、TY041、TY026、HME2.
2.3 cloud ear bacterial strain high temperature resistant exercise regimes
Mycelium growth vigor after the strains tested high-temperature exercise of table 3 compares
From table 3, Fig. 4, after the high-temperature exercise method processing set under 40 DEG C of condition of culture, most of bacterial strain bacterium
It is born long very fast, difference is little, and the colony growth of only totally 2 bacterial strains of bacterial strain TY072,916 is slower；Bacterial strain TY041, YE1,
TY026, TY071, HME2 mycelial growth is very fast and the pure white densification of bacterium colony, though TY063 mycelial growth rates are fast, mycelium growth vigor
It is not good enough, therefore be in 40 DEG C of good bacterial strains of exercise performance：TY041、YE1、TY026、TY071、HME2.
As shown in figure 5, after 42 DEG C of high-temperature cultivations are taken exercise, bacterial strain TY026, TY073 mycelial growth rate is most fast, next to that
TY071, TY041, YE days 3, HME2, and the good bacterial strain of mycelium growth vigor have TY026, YE1, YE2, HME2, TY071, TY041,
TY073, therefore be in 42 DEG C of good bacterial strains of exercise performance：TY026、HME2、TY071、TY041、TY073.
As shown in fig. 6, after 45 DEG C of high-temperature cultivations are taken exercise, bacterial strain TY026, TY041 colony growth rate is most fast, secondly
TY071, TY073, HME2, and mycelia also grows pure white densification, therefore be in 45 DEG C of good bacterial strains of exercise performance：TY026、
TY041、TY071、TY073、HME2。
After 40 DEG C, 42 DEG C, 45 DEG C of high-temperature exercise processing, bacterial strain TY041, TY026, TY071, HME2, TY073 are remained to
Good restoration ecosystem under 28 DEG C of suitable culture conditions, illustrate that there is preferable heat-resisting quantity.
2.4 cloud ear bacterial strain different temperatures fruitings, which are buddingged, to be compared
From table 4, at 25 DEG C, major part bacterial strain can also normally be buddingged, and with the rise of temperature, can normally be buddingged
Bacterial strain gradually decreases, and at 31 DEG C, only YE1, HME2, TY026, TY041, TY071 can normally budding, and illustrate these bacterial strain energy
High temperature resistant fruiting.
The final purpose of the Breeding of Edible Mushroom is to obtain the excellent new varieties of Comprehensive Traits.And resistance is the excellent product of edible mushroom
The indispensable important character of kind, it is directly connected to success or failure and the economic benefit of cultivation.Therefore the breeding objective of edible mushroom, except rich
Outside production property and composite merchandise character, screening, the exercise of the resistance such as high temperature resistant, low temperature properties, disease resistance, matrix adaptability are edible
The important goal of germplasm innovation in bacterium breeding.In the cultivation of edible mushroom, the heat-resisting quantity of edible mushroom is extremely important, and it is related to
The determination of producing region, the selection of cultivating facility, cultivation season.The high temperature resistant character of edible mushroom mycelium closely connects with yielding ability
Lock, therefore the screening of high temperature resistant character is also the screening of high yield struture simultaneously.In heat-resisting quantity experiment in the present invention, ratio is showed
Preferable bacterial strain YE1, HME2, TY026, TY041, TY071 yield in 2015-2016 fruiting experiments is also of a relatively high
Kind.In test most of cloud ear bacterial strain mycelia moves on to 28 DEG C of positive normal temperature in culture dish after 40 DEG C of high-temperature process
Degree culture, can restore normal growth, and this explanation mycelial heat-resisting quantity of cloud ear is stronger.And in production when running into 35 DEG C
In the case of too high temperature and environmental condition regulate and control improperly above, easily cause the rotten bag generation of heat evil, its reason is probably to try
The mycelia tested recovers normal temperature culture, can radiated in time in culture dish after by high-temperature process certain time, and is producing
Middle bacterium bag is by after high temperature damage, it is impossible to which dredging radiating in time restore normal growth condition, when mycelium exceedes endured journey
When spending, vigor declines even dead.In test, strains tested is cultivated under the high temperature conditions and hot conditions handle restoration ecosystem
Ability is better than check variety 916, and simply mycelia restoration ecosystem situation is each variant, illustrates the mycelium of strains tested in high temperature resistant
Really show advantageous in property, also there is higher promotional value in southern area.Different types of edible mushroom, it is same kind of
Different strains, the different growing of same bacterial strain, the tolerance of high temperature is not quite similar.The heat-resisting quantity of kind is taken exercise and sieve
Choosing, first have to that since being screened mycelial heat-resisting quantity, the strong of economical character is carried out again after obtaining relatively resistant to elevated temperatures material
Change and stability test.
The result of the test of the present invention shows：Culture and the processing of a series of hot conditions by different hot conditions,
The mycelial speeds of growth of cloud ear bacterial strain TY026, TY041, HME2 and mycelium growth vigor show relatively stable；YE1 is in 25-37
Under the conditions of DEG C culture and 40 DEG C under the conditions of cultivate different number of days after recovery situation show preferably；TY071 is in 37 DEG C of cultures
Performance is general, but bacterium performance is preferably and stably in 40 DEG C of different cultivated days and 40-45 DEG C of high-temperature exercise；At 28-31 DEG C
Under hot conditions, TY026, TY041, HME2, YE1, TY071 can normally budding.Can with Preliminary Identification TY026, TY041,
HME2, YE1, TY071 are high temperature resistant kind.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (4)
- A kind of 1. lab screening method of high temperature resistant cloud ear bacterial strain, it is characterised in that the indoor sieve of the high temperature resistant cloud ear bacterial strain Choosing method：Bacterial strain is set to cultivate 7d under 6 thermogrades, each processing sets 3 repetitions；5d bacterial strain mycelia disk will be activated PDA synthetic mediums center is inoculated into, is placed at 40 DEG C and takes out renewal cultivation after different incubation times；Strains tested activates 5d Afterwards, picking colony edge mycelia block disk is inoculated in PDA synthesis plating medium center cultures, high-temperature exercise；Strains tested is lived After changing culture 8d, for making cultivation strain；After cultivation strain purseful, cultigen is accessed in fruiting bag culture medium and cultivates 50d, Fruiting bag is put into produce agaric under Cultivation condition respectively.
- 2. the lab screening method of high temperature resistant cloud ear bacterial strain as claimed in claim 1, it is characterised in that the high temperature resistant cloud ear The lab screening method of bacterial strain comprises the following steps：Step 1, sets 6 thermogrades, and each processing sets 3 repetitions；It is comprehensive that activation 5d bacterial strain mycelia disk is inoculated into PDA Culture medium central is closed, is respectively placed in 25 DEG C, 28 DEG C, 31 DEG C, 34 DEG C, 37 DEG C, cultivates 7d, observation mycelia life in 40 DEG C of incubator Long situation, measure mycelial growth rate；Step 2, the bacterial strain mycelia disk activated are inoculated into PDA synthesis plating mediums center, are placed in 40 DEG C of incubators and divide Pei Yang not be after 3d, 5d, 7d, 9d, 11d, then remove and be positioned over 25 DEG C of renewal cultivation 7d, each processing sets 3 repetitions, observes mycelia Growing state, measure mycelial growth rate；Step 3, after strains tested activates 5d, picking colony edge mycelia block disk is inoculated in PDA synthesis plating mediums Centre, is placed at 40 DEG C and cultivates 5d, take out the renewal cultivation 7d at 28 DEG C；The preferable mycelia block of germination and growth is selected to be inoculated in On PDA synthetic mediums, 42 DEG C of culture 3d are put into, take out the renewal cultivation 7d at 28 DEG C, then select the preferable bacterium of germination and growth Silk block is inoculated on PDA synthetic mediums, after being placed in 45 DEG C of culture 5h；Taking-up cultivates 7d at 28 DEG C；Each Temperature Treatment is extensive After the long 7d of demutation, mycelial growth situation is observed, measures mycelial growth rate；Step 4：After strains tested activation culture 8d, cultivation strain is made；After cultivation strain purseful, cultigen is accessed into fruiting 50d is cultivated in bag culture medium, fruiting bag is put into respectively 25 DEG C, 28 DEG C, carry out produce agaric under 31 DEG C of Cultivation conditions, observation is buddingged feelings Condition.
- 3. the lab screening method of high temperature resistant cloud ear bacterial strain as claimed in claim 2, it is characterised in that bacterium in the step 1 Silk growth rate (mm/d)=colony radius (mm)/growth number of days (d).
- A kind of 4. high temperature resistant of the lab screening method screening of high temperature resistant cloud ear bacterial strain as described in claims 1 to 3 any one Cloud ear bacterial strain, it is characterised in that the high temperature resistant cloud ear bacterial strain is TY041, TY026, TY071, HME2, YE1.
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| CN112243798A (en) * | 2020-10-22 | 2021-01-22 | 山东同康农业开发有限公司 | Energy-saving and efficient tremella strain breeding and planting method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109517754A (en) * | 2018-11-20 | 2019-03-26 | 上海交通大学 | A method of high temperature bacterial strain is isolated and purified using common biochemical equipment |
| CN110402758A (en) * | 2019-08-02 | 2019-11-05 | 黑龙江省科学院微生物研究所 | A kind of method of the screening and culturing medium and the lab screening black fungus strain of resistance to heat shock of the black fungus strain of resistance to heat shock |
| CN112243798A (en) * | 2020-10-22 | 2021-01-22 | 山东同康农业开发有限公司 | Energy-saving and efficient tremella strain breeding and planting method |
| CN113322300A (en) * | 2021-07-21 | 2021-08-31 | 西充星河生物科技有限公司 | Bacterium activity detection method |
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