CN108739045A - No. 7 new strains of pleurotus cornucopiae and its selection and breeding and propagation method - Google Patents
No. 7 new strains of pleurotus cornucopiae and its selection and breeding and propagation method Download PDFInfo
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- CN108739045A CN108739045A CN201810253183.6A CN201810253183A CN108739045A CN 108739045 A CN108739045 A CN 108739045A CN 201810253183 A CN201810253183 A CN 201810253183A CN 108739045 A CN108739045 A CN 108739045A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The invention belongs to microbial agriculture technical fields, and in particular to a kind of No. 7 new strains of pleurotus cornucopiae and its selection and breeding and propagation method.It is few for existing pleurotus cornucopiae cultivar, merit is gradually degenerated, per unit area yield constantly declines, the problems such as production needs cannot be met, the present invention provides a kind of No. 7 pleurotus cornucopiae new strains of pleurotus cornucopiae, the bacterial strain is parent with pleurotus cornucopiae 258 and Jinfeng 2-1, is obtained through multiple compatibility test screening using only cross breeding method.No. 7 commodity of pleurotus cornucopiae bacterial strain pleurotus cornucopiae of the present invention are good, inheritance stability, and yield is high, are the new strains with large-scale promotion application value.
Description
Technical field
The invention belongs to microbial agriculture technical fields, and in particular to a kind of No. 7 new strains of pleurotus cornucopiae and its selection and breeding and breeding
Method.
Background technology
Pleurotus cornucopiae is also known as small mushroom, is trade name (Luo Xinchang, the old sections of scholar's fine jade the 15th pleurotus cornucopiae of a kind of " tiny " oyster mushroom
The Beijing mushroom sparetime university of [M] China allusion quotation：Publishing house of Tsinghua University, 2010.9), scientific and technical literature, public document and edible mushroom in recent years
Pleurotus cornucopiae this title is widely used in production and operation.Pleurotus cornucopiae actually refers to that the fructification bacteria cover diameter of harvesting is flat less than 3cm
Mushroom class product includes mainly yellowish-white (P.cornucopiae), the oyster cap fungus of picking up the ears in Pleurotus (Pleurotus)
(P.ostreatus) and Pleurolus sapidus (P.sapidus) (《Edible fungi of china will》) etc..Document report, pleurotus cornucopiae (such as yellowish-white side
Ear) there is strengthening by means of tonics effect to human body, the diseases such as kidney deficiency, impotence, dysentery can be treated；Contained polysaccharide material is to small white mouse sarcoma S-
180 inhibiting rate is 60%~80%, and the inhibiting rate to Emhorn tumor is 60%~70%, and body's immunity can be improved.Pleurotus cornucopiae
Product is favored by domestic and international consumer.
Pleurotus cornucopiae is the important culturing edible fungus in China.Existing pleurotus cornucopiae cultivar is few and by being used for a long time, excellent
Character is gradually degenerated, and per unit area yield constantly declines, and therefore, there is an urgent need to inheritance stability, high yield, the new bacterium of good pleurotus cornucopiae in production
Strain.The selection and breeding of high yield and high quality pleurotus cornucopiae new strains can further promote the cultivation benefit of pleurotus cornucopiae, and the plantation of vast mushroom grower is excited to accumulate
Polarity meets domestic and international market consumer demand.
Invention content
The technical problem to be solved in the present invention is：Existing pleurotus cornucopiae cultivar is few, and merit is gradually degenerated, and per unit area yield is not
It is disconnected to decline, the problems such as production needs cannot be met.
The present invention solve above-mentioned technical problem technical solution be：A kind of No. 7 pleurotus cornucopiae new strains of pleurotus cornucopiae and its selection and breeding are provided
And propagation method.
The present invention provides a kind of No. 7 pleurotus cornucopiae new strains of pleurotus cornucopiae, the preserving number of the bacterial strain is CGMCC No.14553.Preservation
Time is on 09 15th, 2017, and preservation place is China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Wherein, the nucleotide sequence of the ITS1 of above-mentioned pleurotus cornucopiae bacterial strain is as shown in SEQ ID No.1, the nucleotide sequence of ITS2
As shown in SEQ ID No.2.
SEQ ID No.1：The ITS1 sequences of pleurotus cornucopiae new strains
GGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTAATGAATTCACTATGGAGTTGTTG
CTGGCCTCTAGGGGCATGTGCACGCTTCACTAGTCTTTCAACCACCTGTGAACTTTTGATAGATCTGTGAAGTCGTC
TCTCAAGTCGTCAGACTTGGTTGCTGGGATTTAAACGTCTCGGTGTGACTACGCAGTCTATTTACTTACACACCCCA
AATGTATGTCTACGAATGTTATTTAATGGGCCTTGTGCCTATAAACCATAATACAACTTTCAACAACGGATCTCTTG
GCTCTCGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTT
TGAACGCACCTTGCGCCCCTTGGTATTCCGAGGGGCATGCCTGTTTGAGTGTCATTAAATTCTCAAACTCACTTTGG
TTTCTTTCCAATTGTGATGTTTGGATTGTTGGGGGCTGCTGGCCTTGACAGGTCGGCTCCTCTTAAATGCATTAGCA
GGACTTCTCATTGCCTCTGCGCATGATGTGATAATTATCACTCATCAATAGCACGCATGAATAGAGTCCAGCTCTCT
AATCGTCCGCAAGGACAATTTGACAATTTGACCTCAAATCAGGTAGGACTACCCGCTGAACTTAAGCATATCAATAA
GCGGAGGA。
SEQ ID No.2：The ITS2 sequences of pleurotus cornucopiae new strains
GGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTAATGAATTCACTATGGAGTTGTTG
CTGGCCTCTAGGGGCATGTGCACGCTTCACTAGTCTTTCAACCACCTGTGAACTTTTGATAGATCTGTGAAGTCGTC
TCTCAAGTCGTCAGACTTGGTTGCTGGGATTTAAACGTCTCGGTGTGACTACGCAGTCTATTTACTTACACACCCCA
AATGTATGTCTACGAATGTCATTTAATGGGCCTTGTGCCTTTAAACCATAATACAACTTTCAACAACGGATCTCTTG
GCTCTCGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTT
TGAACGCACCTTGCGCCCCTTGGTATTCCGAGGGGCATGCCTGTTTGAGTGTCATTAAATTCTCAAACTCACTTTGG
TTTCTTTCCAATTGTGATGTTTGGATTGTTGGGGGCTGCTGGCCTTGACAGGTCGGCTCCTCTTAAATGCATTAGCA
GGACTTCTCATTGCCTCTGCGCATGATGTGATAATTATCACTCATCAATAGCACGCATGAATAGAGTCCAGCTCTCT
AATCGTCCGCAAGGACAATTTGACAATTTGACCTCAAATCAGGTAGGACTACCCGCTGAACTTAAGCATATCAATAA
GCGGA。
Specifically, the pleurotus cornucopiae new strains have ISSR finger-prints as shown in Figure 3.
The present invention also provides the selections of above-mentioned pleurotus cornucopiae new strains, include the following steps：
Parent's pleurotus cornucopiae 258 and Jinfeng 2-1 are chosen, the uninucleate hyphae that monospore is sprouted is obtained respectively, in PDA culture medium
" single cross is matched ", hybrid strain is obtained, hybrid strain obtains pleurotus cornucopiae new strains through primary dcreening operation, secondary screening.
PDA culture medium：200 grams of potato, 20 grams of glucose, 20 grams of agar, 1000 milliliters of water, natural pH.
" (mon-mon mating) is matched in single, single cross "：To obtaining the uninucleate hyphae of " pleurotus cornucopiae 258 " and " Jinfeng 2-1 " respectively,
Taken on PDA Tube propagation bases random pair " list × single crosses " method (bis- youths of clothing river Kennedy, 1998；Yang Xin is beautiful, and 1988), lead to
Microscope inspection is crossed, obtaining mycelium has the hybrid strain of clamp connection.
The present invention also provides a kind of propagation methods of above-mentioned pleurotus cornucopiae new strains, include the following steps：
The strain of No. 7 pleurotus cornucopiae new strains of pleurotus cornucopiae, which expands numerous production, need to follow strictly sterile working principle, and parent species production is trained with PDA
Base culture is supported, original seed and cultigen production are used using cotton seed hulls, sawdust as major ingredient, and wheat bran is the culture medium of auxiliary material composition；Culture
And cultivation condition：15~32 DEG C of mycelial growth temperature, 8~15 DEG C of fruiting temperature, growth substrate water content are 60~69%, growth
Matrix acid-base value is pH value 5.0~6.5.
Preferably, 25~28 DEG C of mycelial growth temperature.
Preferably, fruiting temperature is 9~15 DEG C.
Beneficial effects of the present invention are：
The present invention provides a kind of pleurotus cornucopiae new strains, stabilization characteristics of genetics, yield is high；Fructification is uniform in size, and stem is suitable
In, fruiting is neat, and commodity is good.The pleurotus cornucopiae wide adaptability of the present invention, suit large area to popularize plantation.
New strains provided by the invention are Pleurotaceae Pleurotus pleurotus cornucopiae kind (Pleurotus sp.), are named as pleurotus cornucopiae 7.
The bacterial strain is preserved in：China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are Chaoyang District, Beijing City north
No. 3 Institute of Microorganism, Academia Sinica of institute of occasion West Road 1, postcode：100101.Preserving number：CGMCC No.14553, biology
Credit class is named as：Pleurotus sp..
Description of the drawings
Fig. 1 show No. 7 spore shapes (A) of pleurotus cornucopiae, hypha form (B) and sporophore shape (C)；
Fig. 2 show No. 7 parent's monospore of pleurotus cornucopiae and sprouts the monokaryon bacterium colony (A) to be formed, uninucleate hyphae (B) and hybridization formation
Nucleated mycelium (C)；
Fig. 3 show pleurotus cornucopiae 7 and compares and the ISSR finger-prints of parent strain.
Specific implementation mode
The pleurotus cornucopiae bacterial strain of the present invention is for inventor from parent's pleurotus cornucopiae 258 and Jinfeng 2-1 by " (mon-mon is matched in single, single cross
Mating) " hybridization obtains, and is Pleurotaceae Pleurotus pleurotus cornucopiae kind (Pleurotus by new lines made of systematic breeding
Sp.), it is named as pleurotus cornucopiae 7.The bacterial strain is preserved in：China Committee for Culture Collection of Microorganisms's common micro-organisms center is protected
Tibetan number：CGMCC No.14553, biological classification are named as：Pleurotus sp..
The morphological feature of the bacterial strain is as follows：
1, spore feature
Observe under the microscope, in each load of pleurotus cornucopiae 7 raw 4 basidiospore, basidiospore is colourless, oval, puts down
Sliding, size is 4.5~7.5 × 3.5~5.0 μm.Spore print white (see Fig. 1 left figures).
2, mycelia body characteristics, characteristic
It observes under the microscope, the mycelium of No. 7 bacterial strains of pleurotus cornucopiae is made of the micromycelial of many branches filiform, white,
4.0~6.0 μm of hyphal diameter has clamp connection (see Fig. 1 middle graphs).
Mycelium well-grown on the culture medium based on cotton seed hulls and rice straw powder；Mycelium at 15~32 DEG C of temperature
It can grow, 25~28 DEG C of optimum growth temperature；Growth substrate appropriate aqueous amount is 65%；The optimum pH of growth substrate is 5.5；
The mycelial growth stage needs oxygen supply, the above scattering light in 10 lux of intensity of illumination.
3, sporophore shape feature
The fructification of No. 7 bacterial strains of pleurotus cornucopiae is grown thickly, cap ellipse or subcircular, lime color, 1~3cm of diameter (commodity mushroom),
1.0~2.0cm of thickness；Bacterial context white, it is fine and close；Lamella prolongs life, Length discrepancy；Column, 0.5~1.0cm of diameter are given birth in stem white, side
(Fig. 1 is right).
4, biological characteristics
Nutritional ingredient source：No. 7 kinds of pleurotus cornucopiae can be preserved in PDA culture medium mother culture media, suitable in 4 DEG C of refrigerator low temperature
It is stored refrigerated, it transfers 1 time within every 3 months.The strain that pleurotus cornucopiae 7, which expands numerous production, need to follow strictly sterile working principle, parent species production
Suitable for PDA culture medium culture；Suitable for being major ingredient to cotton seed hulls, sawdust, wheat bran forms for auxiliary material for original seed and cultigen production
Culture medium.
Culture and cultivation condition：15~32 DEG C of mycelial growth temperature, it is 25~28 DEG C most suitable；8~15 DEG C of fruiting temperature, it is most suitable
9~15 DEG C.
The selection and breeding of 1 bacterial strain of embodiment
(1) source of parent：
Few in order to solve existing pleurotus cornucopiae cultivar, merit is degenerated serious, the problem of per unit area yield constantly declines, inventor
It is parent with pleurotus cornucopiae 258 and Jinfeng 2-1, has carried out the monospore crossbreeding Test And Research Work of " 258 × Jinfeng 2-1 of pleurotus cornucopiae ".
(2) single, single cross is matched：
Collect the spore of parent respectively using spore print partition method, monospore is sprouted in PDA culture medium forms monokaryon bacterium
Silk, and observed and confirmed by sediments microscope inspection, no clamp connection person is confirmed to be uninucleate hyphae (Fig. 2).Monokaryon bacterium between parent
Silk " (mon-mon mating) is matched in single, single cross " on PDA Tube propagation bases, obtaining 81 plants of hybrid strain, (mycelia has lock shape
Joint) (Fig. 2 is right).
(3) primary dcreening operation：
Lab scale is carried out to 81 plants of monospore hybrid strains and cultivates Primary Screening Test.Culture 81 plants of cross combination bacterial strains of observation, tentatively
It is high typical pleurotus cornucopiae feature, yield are filtered out, fruiting regularity is good, and branch is more, the every more uniform preponderant strains of mushroom size
4 plants of strain, and its authenticity (Fig. 3) hybridized is confirmed from molecular biology level with ISSR methods.
(4) secondary screening：
Further secondary screening goes out 1 plant of strain excellent " P2 " from the 4 plants of bacterial strains just sifted out；P2 bacterial strains are by regional testing, life
Production experiment and field technology identification think that the bacterial strain character is excellent, stabilization characteristics of genetics, are one and are different from existing cultivar
New strains, be named as " pleurotus cornucopiae 7 ".
(5) bacterial strain authenticity identification：
Foundation《NY/T1730-2009 edible fungus species authenticity identification ISSR methods》, to just sift out No. 44, No. 6,115
Number and No. 7 i.e. 4 plants of hybrid strains, plus pleurotus cornucopiae 258 and Jinfeng 2-1 i.e. 2 plant parent strain amount to 6 plants of bacterial strains, progress bacterial strain (bacterium
Kind) authenticity identification.
1 authenticity identification primer sequence of table
The results are shown in Figure 3 for finger-print, and there is othernesses by DNA of the pleurotus cornucopiae 6 between hybrid strain and parent, this is just
It further demonstrates No. 7 bacterial strains of pleurotus cornucopiae and is different from parent strain in molecular biology level, it is thus identified that hybridization authenticity.
The pleurotus cornucopiae 7 of identification is cultivated, spore shape (Fig. 1 is left), hypha form (in Fig. 1) and fructification shape
State (Fig. 1 is right).It is observed that form is as follows：Spore：In each load of pleurotus cornucopiae 7 raw 4 basidiospore, basidiospore is colourless, ovum
Circle, smoothly, size are 4.5~7.5 × 3.5~5.0 μm, spore print white.Mycelia：Mycelium by many branches filiform fibre
Bacterium silk composition, white, 4.0~6.0 μm of hyphal diameter have clamp connection.Fructification is grown thickly, cap ellipse or subcircular,
Lime color, 1~3cm of diameter (commodity mushroom), 1.0~2.0cm of thickness；Bacterial context white, it is fine and close；Lamella prolongs life, Length discrepancy；Stem is white
Column, 0.5~1.0cm of diameter are given birth in color, side.
2 cultural area contrast test of embodiment
For the cultivation promotional value of clear new strains " pleurotus cornucopiae 7 ", according to the Crop breed audit committee of Sichuan Province《Food
With bacterium variety certification standard》, continuous two season (i.e. 2 years) two regions carried out No. 7 plastic bag cultivations of pleurotus cornucopiae experiment unjust open region
It tests (hereinafter referred to as area's examination).
For trying reference kind：In being produced with current Sichuan Province's pleurotus cornucopiae it is long-term since the cultivar " West Germany 33 " that uses always
As No. 7 controls of pleurotus cornucopiae.Cultigen with batch making West Germany 33 and pleurotus cornucopiae 7 is spare.
Plant formulation：Grass meal 60%, cotton seed hulls 35%, corn flour 2%, wheat bran 1.5%, lime 3~4%.Water content
60%, pH are natural.For the drying of examination culture material, do not go mouldy.
Cartridge bag：Using polyethylene plastic bag plastic barrel bag as the container containing of experimental cultivar culture material matrix.Cylinder
Bag specification (length × folding diameter × thickness) is 42cm × 20cm × 0.025cm.
(1) First Year regional testing
Regional testing is carried out with plastic bag cultivation, 100 bags of each variety culture carries out cultivating mushroom management indoors on bedstead.Knot
Fruit shows (table 1)：The major traits that first year of " pleurotus cornucopiae 7 " kind shows on 2 testing sites are more consistent.It is average
Bacteria developing period (number of days for being inoculated into fruiting) in 34d, grow thickly by fructification, cap subcircular, grey, shallower, the life of stem side, column,
Organize consolidation (table 2).The annual yield performance results on 2 testing sites of " pleurotus cornucopiae 7 " kind first are more consistent, average per unit area yield
For 1.10~1.12kg/ bags of per unit area yield, higher than control West Germany 33, difference reaches the pole level of signifiance, average to increase production up to 13% or more (table
3)。
Try performance results in the annual area of No. 7 kinds of 2 pleurotus cornucopiae of table first
The annual area's examination of 3 pleurotus cornucopiae of table No. 7 first is compared with compareing yield result
(2) second year regional testing
Regional testing is carried out with plastic bag cultivation, 100 bags of each variety culture is cultivated on bedstead, carried out according to a conventional method indoors
Management.The result shows that (table 4)：" pleurotus cornucopiae 7 " kind is more consistent in 2 testing site performances in the second year.Play bacteria developing period about
It it is 34 days, fructification is grown thickly, cap ellipse or subcircular, lime color, 1~3cm of diameter (commodity mushroom), 1.0~2.0cm of thickness；
Bacterial context white, it is fine and close；Lamella prolongs life, Length discrepancy；Column, 0.5~1.0cm of diameter, average product 1.11 are given birth in stem white, side
~1.12kg/ bags (table 4), compared with the control, volume variance reach the pole level of signifiance (P < 0.01), and average amount of increase in production reaches
To 15% or more (table 5).
The annual regional testing performance results of 4 pleurotus cornucopiae of table No. 7 second
The annual area's examination of 5 pleurotus cornucopiae of table No. 7 second is compared with compareing yield result
The comprehensive agronomies character such as maturity periods of No. 7 kind fructifications of pleurotus cornucopiae, morphological feature and yield is identical in areal
It is showed under the conditions of planting environment more consistent；The general effect that (two region points) is shown in different regions is more consistent.This
Show the stabilization characteristics of genetics of " pleurotus cornucopiae 7 " and adaptable wide feature.More larger than control amount of increase in production, yielding ability is good.
With the value being widely applied in production.
Embodiment No. 7 pilot productions of 3 pleurotus cornucopiae
For further confirm " pleurotus cornucopiae 7 " in big production the economical characters such as yielding ability, adaptability practical manifestation effect,
The objectively Commercial cultivation value of evaluation No. 7 kinds of pleurotus cornucopiae, provides foundation and lays the foundation to promote and apply.In Sichuan Province's pleurotus cornucopiae
Main producing region Jintang County Qingjian River town has carried out underground pilot production.With reference to " bio-statistics and field experiment " correlation principle and method
Main points, pilot production is carried out using clinker cultural method, is inoculated with 1000 bags of pleurotus cornucopiae bacterium bag or more for trying each production, therefrom at random
Sampling is divided into 3 cells, 300 bags/cell, carries out cultivation production, is control with " West Germany 33 ".No. 7 kinds of pleurotus cornucopiae are in testing site
On the result that shows be：The mycelia purseful phase is 35 days, and the fructification maturity period is 4 days, and fructification is grown thickly, cap ellipse or close
Circle, grey, color are shallow；The life of stem side, column, tissue consolidation.Yield is 1.12kg/ bags, biological transformation ratio is 110% or more
(table 6).
No. 7 2013~2014 annual pilot production performance results of 6 pleurotus cornucopiae of table
The pilot production yield performance of " pleurotus cornucopiae 7 " bacterial strain the results are shown in Table 7." pleurotus cornucopiae 7 " bacterial strain average product is
1.12kg/ bags, than control strain volume increase 16.67%, volume variance reaches the pole level of signifiance (P < 0.01).
No. 7 variety production experiments of 7 pleurotus cornucopiae of table are compared with compareing 33 yield result of West Germany
The pilot production performance results of " pleurotus cornucopiae 7 " kind averagely increase production 16.67% than check variety, and volume increase difference is aobvious
It writes.In the comprehensive agronomies characters such as mycelia growing power, fructification maturity period, sporophore shape and yield and regional testing result phase
Than the performance of total trend is more consistent, and obvious effect of increasing production, yielding ability is further confirmed in pilot production, tool
There is the value being widely applied in production.
Embodiment No. 7 attributional analysis of 4 pleurotus cornucopiae
No. 7 stems of pleurotus cornucopiae are relatively thick, consolidation, in good taste, mushroom large percentage in grade is organized to have preferable commodity.Pleurotus cornucopiae 7
Number through Institute of Analysis of academy of agricultural sciences of Sichuan Province testing result (being shown in Table 8), fructification is 21.9%, crude fat containing crude protein
1.35%, total amino acid content 15.9%；Polyoses content 8.63g/100g.
Table No. 7 nutrient composition contents of 8 pleurotus cornucopiae
Sequence table
<110>Inst. of Soil Fertilizer, Sichuan Academy of Agriculture Science
<120>No. 7 new strains of pleurotus cornucopiae and its selection and breeding and propagation method
<130> A180183K（Sequence）
<160> 13
<170> SIPOSequenceListing 1.0
<210> 1
<211> 701
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
ggaagtaaaa gtcgtaacaa ggtttccgta ggtgaacctg cggaaggatc attaatgaat 60
tcactatgga gttgttgctg gcctctaggg gcatgtgcac gcttcactag tctttcaacc 120
acctgtgaac ttttgataga tctgtgaagt cgtctctcaa gtcgtcagac ttggttgctg 180
ggatttaaac gtctcggtgt gactacgcag tctatttact tacacacccc aaatgtatgt 240
ctacgaatgt tatttaatgg gccttgtgcc tataaaccat aatacaactt tcaacaacgg 300
atctcttggc tctcgcatcg atgaagaacg cagcgaaatg cgataagtaa tgtgaattgc 360
agaattcagt gaatcatcga atctttgaac gcaccttgcg ccccttggta ttccgagggg 420
catgcctgtt tgagtgtcat taaattctca aactcacttt ggtttctttc caattgtgat 480
gtttggattg ttgggggctg ctggccttga caggtcggct cctcttaaat gcattagcag 540
gacttctcat tgcctctgcg catgatgtga taattatcac tcatcaatag cacgcatgaa 600
tagagtccag ctctctaatc gtccgcaagg acaatttgac aatttgacct caaatcaggt 660
aggactaccc gctgaactta agcatatcaa taagcggagg a 701
<210> 2
<211> 698
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
ggaagtaaaa gtcgtaacaa ggtttccgta ggtgaacctg cggaaggatc attaatgaat 60
tcactatgga gttgttgctg gcctctaggg gcatgtgcac gcttcactag tctttcaacc 120
acctgtgaac ttttgataga tctgtgaagt cgtctctcaa gtcgtcagac ttggttgctg 180
ggatttaaac gtctcggtgt gactacgcag tctatttact tacacacccc aaatgtatgt 240
ctacgaatgt catttaatgg gccttgtgcc tttaaaccat aatacaactt tcaacaacgg 300
atctcttggc tctcgcatcg atgaagaacg cagcgaaatg cgataagtaa tgtgaattgc 360
agaattcagt gaatcatcga atctttgaac gcaccttgcg ccccttggta ttccgagggg 420
catgcctgtt tgagtgtcat taaattctca aactcacttt ggtttctttc caattgtgat 480
gtttggattg ttgggggctg ctggccttga caggtcggct cctcttaaat gcattagcag 540
gacttctcat tgcctctgcg catgatgtga taattatcac tcatcaatag cacgcatgaa 600
tagagtccag ctctctaatc gtccgcaagg acaatttgac aatttgacct caaatcaggt 660
aggactaccc gctgaactta agcatatcaa taagcgga 698
<210> 3
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
cgtagtcgta caacaacaac aaca 24
<210> 4
<211> 15
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
acaacaacaa cagcc 15
<210> 5
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
cacacacaca cacacag 17
<210> 6
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
gagagagaga gagagat 17
<210> 7
<211> 19
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
gagagagaga gagagactc 19
<210> 8
<211> 19
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
gagagagaga gagagactg 19
<210> 9
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
cacacacaca cacacaa 17
<210> 10
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
tgtgtgtgtg tgtgtgg 17
<210> 11
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
atgatgatga tgatgat 17
<210> 12
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
ccgccgccgc cgccgccg 18
<210> 13
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
ggcggcggcg gcggcggc 18
Claims (6)
1. pleurotus cornucopiae bacterial strain Pleurotus sp. pleurotus cornucopiaes 7, it is characterised in that：Preserving number is CGMCC No.14553.
2. pleurotus cornucopiae bacterial strain according to claim 1, it is characterised in that：The nucleotide sequence of the ITS1 of the pleurotus cornucopiae bacterial strain is such as
Shown in SEQ ID No.1, the nucleotide sequence of ITS2 is as shown in SEQ ID No.2.
3. the selection of pleurotus cornucopiae bacterial strain described in claims 1 or 2, it is characterised in that：Parent's pleurotus cornucopiae 258 and Jinfeng 2-1 are chosen,
The uninucleate hyphae that monospore is sprouted is obtained respectively, in PDA culture medium " single cross is matched ", obtains hybrid strain, hybrid strain warp
Primary dcreening operation, secondary screening obtain pleurotus cornucopiae new strains.
4. the propagation method of pleurotus cornucopiae bacterial strain described in claims 1 or 2, it is characterised in that：It is stringent that the strain that pleurotus cornucopiae 7 expands numerous production
Sterile working principle, parent species production PDA culture medium culture are followed, original seed and cultigen production use and is with cotton seed hulls, sawdust
Major ingredient, wheat bran are the culture medium of auxiliary material composition；Culture and cultivation condition：15~32 DEG C of mycelial growth temperature, fruiting temperature 8~15
DEG C, growth substrate water content is 60~69%, and growth substrate acid-base value is pH value 5.0~6.5.
5. the propagation method of pleurotus cornucopiae bacterial strain according to claim 4, it is characterised in that：Mycelia grows optimum temperature 25~28
℃。
6. the propagation method of pleurotus cornucopiae bacterial strain according to claim 4, it is characterised in that：Fruiting optimum temperature is 9~15 DEG C.
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