CN112063534A - Hypsizigus marmoreus strain - Google Patents

Abstract

The invention provides a hypsizigus marmoreus strain Nongshizhen No.1, which is preserved in China general microbiological culture Collection center (CGMCC) in 6-15.2020, with the preservation number as follows: CGMCC No. 20201. The 'Nonwangzhen No. 1' strain provided by the invention has the quality characteristics of excellent hypsizigus marmoreus (white beech mushroom) strains, such as more buds in a germination period, full and uniform whole harvest period, thin and short stipe, good hardness, storage resistance, white mushroom bodies and the like. In addition, the average fresh mushroom yield of 'Nonmanni No. 1' in a productivity test reaches 275.9 g/bottle, the average yield per unit is higher than that of a control variety by more than 10% (the difference is very obvious), the ratio of the thickness of a pileus and the diameter of a stipe is smaller than that of B01 (the difference is obvious), and the method is more suitable for producing white beech mushrooms.

Description

Hypsizigus marmoreus strain

Technical Field

The invention relates to a hypsizigus marmoreus strain nong Wanzhen No.1, belonging to the field of edible fungi.

Background

Hypsizigus marmoreus [ 2 ]Hypsizygus marmoreus (Peck) H.E.Bigelow]Belonging to Basidiomycotina, Hymenomycetes, Agaricales, Tricholomataceae, Hypsizygus, and is called Hypsizygus marmoreus, Pleurotus geesteranus, and Hypsizygus marmoreus because it has unique flavor of crab. Fujian province is the main cultivation region of hypsizygus marmoreus in southern China, the daily yield of Fujian Shunchang reaches 200 tons, and 56 seafood mushroom factory production enterprises account for more than 65% of the national daily yield. At present, the hypsizigus marmoreus is divided into two types according to market requirements, wherein one type is the thick hypsizigus marmoreus with longer mushroom stems, and the other type is the white beech mushroom or crab-flavor mushroom with shorter and thinner mushroom stems. At present, uniform stipe, more mushroom bodies and white mushroom bodies become the main trend of the white beech mushroom market. The main cultivar B01 of Fujian Wanchen white beech mushroom has the advantages of white mushroom body, thick stipe and the like, but the yield per unit is low. Along with the development of the market, the quality requirement of consumers on the white beech mushrooms is higher and higher, the problem of insufficient backup varieties of production enterprises is increasingly highlighted, and the selection of excellent varieties with high quality, high yield, good commodity and good market prospect is more important.

Disclosure of Invention

The invention aims to solve the technical problems of loose mushroom body, small bud number, low yield per unit and the like of the white beech mushroom (hypsizigus marmoreus) variety used in the existing production, thereby providing a hypsizigus marmoreus strain with more buds, regular mushroom bodies, white mushroom bodies, good hardness of mushroom stems, high quality and high yield.

The hypsizigus marmoreus strain 'nong Wan Zhen No. 1' of the present invention belongs to hypsizigus marmoreusHypsizygus marmoreus) The culture medium is preserved in the China general microbiological culture Collection center, the preservation address is No. 3 Xilu No.1 of Beijing, Chaoyang, the preservation date is 6 months and 15 days in 2020, and the preservation number is CGMCC No. 20201.

The 'Nonmuzhen No. 1' strain has the fastest growth of hyphae on a PDA culture medium at 25 ℃, the hyphae are thickest and fluffy, the edges of bacterial colonies are round, and the hyphae at the tip are neat and thick. The fruiting body has slightly more buds, slightly thin stipe, less buds and good stipe hardness, and is suitable for factory production as white beech mushroom.

'Nonwangzhen No. 1' was bred from hypsizigus marmoreus 'B01' and 'B02' by crossing, and the hybrid strain 'Nonwangzhen No. 1' was numbered 'Hy 5046', and was subjected to cultivation test after selection. 'B01' is the main cultivated species of white beech mushroom of Fujian Wanchen Biotechnology GmbH, and has the advantages of white mushroom body and thick stipe, but the yield per unit is low. 'B02' has the characteristics of uniform stipe thickness, high yield and the like, but the stipe is thin and yellow, and is a hypsizigus marmoreus test variety of the Fujian Tengchen Biotechnology GmbH. Cultivation tests show that the 'Hy 5046' has the characteristics of more buds, plump and uniform whole body, thin and short stipe, good hardness, storage resistance, small difference of the whiteness of the mushroom body from that of a contrast variety 'B01', and the like, and the overall appearance character is more suitable for producing the white beech mushroom. 'Hy 5046' improves the defects that the yield per unit of a parent 'B01' is low and the stipe of another parent 'B02' is thin and yellow, integrates the advantages of white mushroom body, uniform stipe and thick stipe of 'B02' and high yield of the parent 'B01', has excellent comprehensive properties, is suitable for producing white beech mushroom, and has the yield higher than 'B01', and is named as 'Nonmuzhen No. 1' (figure 1).

The judgment of the difference between 'Nonmamel No. 1' and the parent strains 'B01' and 'B02' by the antagonistic reaction of the strains shows that hypha at the junction between 3 tested hypsizigus marmoreus strains has obvious antagonistic lines, and the strain to be detected, namely the hypsizigus marmoreus 'Nonmamel No. 1', is further presumed to be a different variety from two control strains.

The hypsizigus marmoreus strain disclosed by the invention adopts a Kmer subtraction assembly technology according to genome sequencing data to obtain a differential fragment (SV), 15 SV sequences (with sequences shown as SEQ ID No. 1-15) are selected to design specific primers (A1-A13 and B1-B5) to amplify Nonweikan No.1, B01 and B02, and except that A4 is not amplified successfully, other primers obtain PCR products. The PCR amplification results for each specific fragment are shown in fig. 3 and 4: a1, A2, A3 and A5 are consensus sequences of B02 and Nonwanzhen No.1, and are different from B01. A6, A9 are unique sequences that B02 differs from B01 and Nonworhenn No. 1. A7, A8, a10 are B02 and B01 consensus sequences, differing from Hy 5046; A11-A13 are consensus sequences of Hy5046, B01 and B02; B1-B5 are unique sequences that distinguish B01 from Hy5046 and B02. By combining the detection results, the DNA fingerprints of the Nongwan Zhen No.1, B01 and B02 have obvious difference and are determined to be different varieties.

The invention has the advantages that:

the 'Nonwangzhen No. 1' strain provided by the invention has the quality characteristics of excellent hypsizigus marmoreus (white beech mushroom) strains, such as more buds in a germination period, full and uniform whole harvest period, thin and short stipe, good hardness, storage resistance, white mushroom bodies and the like. In addition, the average fresh mushroom yield of 'Nonmanni No. 1' in a productivity test reaches 275.9 g/bottle, the average yield per unit is higher than that of a control variety by more than 10% (the difference is very obvious), the ratio of the thickness of a pileus and the diameter of a stipe is smaller than that of B01 (the difference is obvious), and the method is more suitable for producing white beech mushrooms.

Drawings

FIG. 1 Standard fruiting pictures of "Nonmanni No. 1' (Hy5046) and the parents B01 and B02.

FIG. 2 ` Nonmanni No.1 ` (Hy5046) was tested antagonistically with ` B01 ` and ` B02 `.

FIG. 3 shows an electrophoretogram for identifying the SV strain of "Nonmamel No. 1" (in the figure, M represents a 1000bp Marker, a blank represents a negative control, and the other lanes are marked with the numbers of the SV strains).

FIG. 4 is a schematic diagram showing the amplification results of each specific fragment.

FIG. 5 Standard and fruiting pictures of "Nonmamel No. 1' (Hy5046) and control B01 (left is" Nonmamel No.1 "and right is B01).

Detailed Description

Example 1

The hypsizigus marmoreus 'B01' is a white beech mushroom main cultivated variety of the biological science and technology company Limited in Fujian Tengchen, has the advantages of white mushroom body, thick stipe and the like, but has low yield per unit. Hypsizigus marmoreus ` B02 ` is a test variety of Hypsizigus marmoreus from the Biotechnology GmbH of Fujian Tengchen.

The method comprises the following steps of selecting main cultured hypsizigus marmoreus 'B01' and 'B02' as hybridization parents, pairing 20 parent B01 single spore strains and 14B 02 single spore strains with higher growth speed and more developed aerial hyphae according to a full combination mode, pairing 140 combinations in total to obtain 100 hybrid strains with lock combination, screening 78 hybrid strains with better growth vigor and lock combination for primary screening fruiting experiments, screening 45 hybrid strains capable of fruiting, re-screening, enabling 29 strains to normally fruiting, and enabling the yield of the hybrid strains to be 22 compared with that of a control variety B01: wherein 11 strains show that the number of buds is excessive, the mushroom stems are thin and the mushroom stems are bent; 7 strains had tumors of different degrees; in addition, the mushroom caps are screened out to be yellow, and are concave with the mushroom caps, so that the variety is poor in taste; the remaining 2 strains grew better: hy5046, Hy 5085. Wherein Hy5046 stipe is short and has more buds, and is suitable for producing white beech mushroom; according to the comprehensive re-screening yield and character performance, Hy5046 and the hybrid strain named as 'Nonmanni No. 1' are finally selected (FIG. 1).

And carrying out difference comparison on the 'Nonmamel No. 1' strain and the parent strain by an antagonistic reaction method.

The detection basis is as follows: inoculating, culturing and observing the sample to be detected according to the agricultural industry standard NY/T1845-2010 edible fungus strain differential identification antagonistic reaction of the people's republic of China.

Detection materials: 3 strains of hypsizigus marmoreus, wherein the strain to be detected is 'Nonmamex No. 1', and the control strain 2 is 'B01' and 'B02', respectively.

The detection method comprises the following steps: and (3) according to aseptic operation, respectively punching edges of activated bacterial colonies of the strains of the proper age by using a phi 8 mm puncher to serve as inoculation blocks, wherein the distance between the two inoculation blocks is 10 mm, and hyphae face upwards. Culturing at 20 deg.C under ventilation and dark condition until inoculating lump mycelium contacts, and culturing under natural light for 14 d.

Results and analysis: the hyphal reaction at the interface between the two strains on the surface of the culture was observed under light or natural light, and the results are shown in the figure (FIG. 2).

And (4) detection conclusion: as can be seen from the antagonistic results in fig. 2, there was significant antagonism among Hy5046, B01 and B02, and the hypha morphologies of the three were not very different, and Hy5046 hyphae were sparse. There was no significant difference in hyphal morphology between B01 and B02.

Example 2: and comparing the DNA difference of the strain with that of the parent strain by using an SV detection technology.

Detection materials: 3 strains of hypsizigus marmoreus, wherein the strain to be detected is 'Nonmamex No. 1', and the control strain 2 is 'B01' and 'B02', respectively.

The detection method comprises the following steps: according to genome sequencing data, a Kmer subtraction assembly technology is adopted to obtain differential fragments (SV), and a plurality of SV sequences (with the sequences shown as SEQ ID NO. 1-17) are correspondingly selected to design specific primers to amplify a sample to be detected. (see Alkan, C., Coe, B.P. & Eichler, E.E. Genome structural variation and genetic analysis. Nature reviews. Genetics, 2011 (12): 363-. The SV test primer sequence is synthesized by Optisco Biotechnology Limited.

The activation, culture, genome DNA extraction and electrophoresis detection of the mycelium of the strain to be detected are carried out according to the agricultural industry standard of the people's republic of China.

18 pairs of SV primers used for identifying the authenticity of the strain are as follows:

primer sequences for SV of Table 1

Results and analysis: the SV detection technology establishes a DNA fingerprint of the hypsizigus marmoreus strain: according to genome sequencing data, a Kmer differential subtraction assembly technology is adopted to obtain differential fragments (SV), 17 SV sequences are selected to design specific primers (A1-A13, B1-B5) to amplify Hy5046, B01 and B02, except that A4 is not successfully amplified, other primers obtain PCR products, the amplification result is detailed in figure 3, and the sequencing result of each specific fragment is detailed in a sequence table. The PCR amplification results for each specific fragment are shown in fig. 4: a1, a2, A3, a5 are consensus sequences of B02 and Hy5046, differing from B01. A6, a9 are unique sequences that distinguish B02 from B01 and Hy 5046. A7, A8, a10 are B02 and B01 consensus sequences, differing from Hy 5046; A11-A13 are consensus sequences of Hy5046, B01 and B02; B1-B5 are unique sequences that distinguish B01 from Hy5046 and B02. By combining the detection results, the DNA fingerprints of 'Nonmanni No. 1', B01 and B02 have obvious difference and are different varieties.

And (4) detection conclusion: SV analysis results show that the hypsizigus marmoreus 'Nonmamel No. 1' to be detected is a hybrid of 'B02' and 'B01', has obvious difference with DNA fingerprints of parent varieties, and is different varieties.

Example 3: the 'Nonmannianzhen No. 1' is subjected to multiple stable yield tests, and the result shows that the 'Nonmannianzhen No. 1' has the requirement of industrial cultivation and has higher yield than that of the parent 'B01'.

Stable yield test reference strains: 'Nonwan Zhen No. 1', control strain: 'B01'.

Test site: fujian Wanchen Biotechnology GmbH.

Test arrangement: the strains are manufactured uniformly, and the test points are cultivated and managed according to a local production management mode.

And (3) test results: experiments show that hyphae of 'Nonweishen No. 1' are thicker in PDA, and the optimal growth temperature is 25 ℃. The cultivation results of "Nonworhen No. 1" are shown in FIG. 5.

Cultivation finds that: (1) the time from inoculation to harvest, 'nong Wan Zhen No. 1' is the same as the parent; (2) the average yield of ` Nonmanni No.1 ` was 275.9 g/vial, and the average yield of control ` B01 ` was 245.5 g/vial. (3) 'Nonwan Zhen No. 1' has an average pileus diameter of 16.2 mm, a thickness of 6.6 mm, an average stipe length of 62.0 mm and a diameter of 9.0 mm. (4) The disease and insect resistance of 'Nonweisheng No. 1' is equivalent to that of the main cultivated variety (B01), and no mixed bacteria pollution, disease and insect damage occur in the stability test and the productivity test of multiple batches.

Statistics of yield results of stable yield test strains

TABLE 2 Multi-batch test results

Note: ' indicates P <0.01, the difference is very significant; ' indicates 0.01< P <0.05, the difference was significant.

Statistical analysis of the test strain yields: through the statistical analysis of yield, the cultivation test result shows that the average fresh mushroom yield of 'nong Wan Zhen No. 1' is 275.9 g/bottle, and the average yield is higher than that of B01 by more than 10% (the difference is very obvious). The mushroom cap of 'nong Wan Zhen No. 1' is moderate in size and mushroom stem thickness, is white beech mushroom quality required by the market, is uniform in whole, hard in mushroom stem, good in dryness and durable in storage.

Example 4: the results of the 'Nonworhen No. 1' productivity test show that the method is suitable for industrial production.

The test strains were: 'Nonwan Zhen No. 1' (Hy5046), control strain: 'B01'

Test site: fujian Wanchen Biotechnology GmbH.

Test arrangement: the strains are uniformly prepared, the total number of the bottles of the crossbreed 'Nonmanni No. 1' is 1000, the total number of the bottles of the control parent 'B01' is 1000, and the test point is cultivated and managed according to a local production management mode.

And (3) counting the spawn running speed of the strains: the productivity test has little difference between the culture spawn running state and the previous batch of cultivation tests. By day 2 of culture after inoculation, Hy5046 and B01 both germinated normally, with Hy5046 germinating slightly more strongly. After 11 days of culture, both Hy5046 and B01 showed a bottom, and the bottom was better. After 21 days of culture, Hy5046 and B01 are filled at the bottom, a small amount of hypha begins to appear on the bottle body, the hypha on the Hy5046 bottle body is strong, and the hypha on the tip of the B01 bottle body is slightly thin. Culturing for 35 days, wherein the growth rates of Hy5046 and B01 are not greatly different, and the hypha of the bottle bodies of the two bottles are filled in the bottle, and the bottle is moved to a storehouse for after-ripening culture. After 55 days of after-ripening culture, the culture materials Hy5046 and B01 are changed into dark yellow, and part of bottle shoulder hypha of both varieties slightly yellow, at the moment, the culture is ripe and mycelium stimulation is carried out. The culture was carried out for a total of 90 days.

Productivity test strain yield results statistics: the average yield of ` Nonmanni No.1 ` (Hy5046) was 264.3 g/vial, and the yield of control ` B01 ` was 239.9 g/vial. Hy5046 produced 10.2% more than B01 (the difference was very significant).

TABLE 3 recovery record for productive testing

Note: ' indicates P <0.01, the difference is very significant; ' indicates 0.01< P <0.05, the difference was significant.

The productivity test results show that: ' Nonmamel No.1 ' has a lid diameter and thickness and a stalk diameter and length slightly smaller than the parent relative to the control ' B01 ' (see FIG. 5 for representative photographs of Nonmamel No.1 ' and parent ' B01 ').

TABLE 4 Productivity test pileus stipe measurement results

Note: unit, pileus diameter (mm), pileus thickness (mm), stipe diameter (mm), stipe length (cm).

Statistical analysis of the test strain yields: through the analysis of yield statistics, the result of a productive test shows that the average yield of the 'nong Wan Zhen No. 1' is improved by 10.2 percent compared with that of the parent (the difference is extremely obvious). Hy5046 measured cap and stipe data less than B01. Overall appearance: hy5046 has slightly more buds, thinner stipe, less small buds and good stipe hardness, and is suitable for industrial production of white beech mushroom.

The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.

SEQUENCE LISTING

<110> Fujian agriculture and forestry university

FUJIAN VANCHEN BIOTECHNOLOGY Co.,Ltd.

<120> a Hypsizigus marmoreus strain Nongshizhen No.1

<130> 55

<160> 53

<170> PatentIn version 3.3

<210> 1

<211> 876

<212> DNA

<213> A1

<400> 1

tgtgaaagtg agatgacggg tgtggtagat gagcgaagga atgcaactgc agtcaaggcg 60

agtgctcttg ctctagagca gaactgacga atattgtcca gtattgtcac ggtctccagc 120

tgctcacacg acccgatctg ggtgccttgg tctgtgggaa tggctacggg gttgttcaga 180

tgtgtgctcg cctatattgc cgagcatcgc caccctttcc accgttggtc gttccattgt 240

agtacttact tgactatgaa cagaattgaa tcattattat ccttctcctt ggaacttcag 300

ttattgagct caatggccaa cttgatgcat tctgaatatt tataagtaac tccctgtcca 360

atatgtgcta ggaactagta cttactgact ttgcagacac ttcaccagtt tgccccactc 420

tcccctccaa atggcctccg tcgtcggtat ggatgaaata cctaaacgtc ttttaacgca 480

agcagaggca atgcaggaac ttgtactggc ggcagaatca gttaaggact atgattgata 540

ctttatatcc cacaactacc ctcgcggtga ccaagaatag catcctacta ttttgtcgga 600

gtcagtgact aacattgcat ctaccactga ttgcatagct tggtgaacac tgctgaacaa 660

gtaaagaaac ttcaatctcc actgagcgca cctctcacgg gcctcctctc ccacgtactg 720

gcctttactt actattagta tacaggcgag caagccgcga ggtgactatc atttggctcg 780

gtactaagga ttggatccga agtagtgagc atgtttgtgg ctgtacgggt gtccatgtaa 840

atttgccagt agtaagcagt accccttcgg agtcaa 876

<210> 2

<211> 724

<212> DNA

<213> A2

<400> 2

tgtgggcatt ttagcaacga atcttagtac aagaacaagc aatcatttat cgtcgagggc 60

atcgacactt cgttaaacag cttgacggcg aaacatcgat ttttggagac ttggggaaca 120

ccagttcatt tccgtgagtt cctcttaatt ttcttgcatc ctggccattt tatcaaccat 180

ctgtcttcgt acatgacgga taataggtgc tttcaggctc ctcaaagtgc gattgcctgc 240

gacgaagtca gttggtaaat ttacgtgcct tcgcaaaacg gtctcgatca tagtgcgttt 300

tctgttgttg ttttttcatc gtagccatct gttaacgtat cagactcaca gatgatgcac 360

acttcgccta agaacgccgc caataccccg gtctattgca cgacggcgcg ccaacttcct 420

ccaccatgtc atccactcca cctctccctc gcagacttca tccccgcctt ttcaccaaaa 480

cctactaggt ccactctcag aaggaatgaa ggcactgtcc accaaacctc acccgacaat 540

ttcacgcgtt cagccgactc tacctactgg gaactcgaaa acgtcaacct ccgttccgtc 600

acatatggcg gtggcttcgt ttccaggaaa agtccaaagc ggtggtcaag ctgtttccct 660

gcgaacgcca gtaccatgag ccgcggtaac aacgttcgga gtcgcaggaa atacgagcac 720

aggg 724

<210> 3

<211> 966

<212> DNA

<213> A3

<400> 3

ttcaactcgc atgggtacat acgccagcca cccgagcgtt attaaatttc aaatttgccg 60

aagcaaaacg gtaaatccat accacggaat tgcgaatcca tgaacaattt cgagatcttt 120

cggatgcaga ataagatcat ccgagccgcc acactatgat agccgacata catctgaagt 180

gtaccccttg attacctagt aattatcccc gctgatgtca tggttggatt accagagaaa 240

cgaagtccga gttcgaagcc tacttgcctc gtcacctagt cagagacagc catgtccttc 300

attacaggat agggttcagt gtggaaggag agttccttcc aatctttcgc tgctttgcca 360

gccgaggtgc ccagatttcg atcgacacga ggactttcaa cagctgacgc caaatctcga 420

tctcggaagc aaatgatcca tccagaacaa ggcgtcggag cggcaagtac ctgaccaatg 480

tatggaatgc ctctctcaac tcacgagtgt tcaaaggttt cgaacttgac acacacatac 540

ttcaagatat tcaagagact aatcaaggat ggctgcctaa acttttgttt gcccaacttc 600

aaagttcatc gatagcggtg cccgtactag tactgtttcg ccatcaacat gtgttcggcg 660

cttaatgtca tcggcgcgtg tctgcagctg tgtctctgct ccttcaaagg aattattatt 720

ttttaatccg ttcataattg aatctatgcg ggagatttct atatcatggc taacacttcc 780

ctttcagtca ctgtctggcg atcatggcct gattttttgg atgatttttc ttacaaaaaa 840

ctttgcgagt ctggatcgac cagacatgag gggattaatc ctggattaca actatgcctc 900

gtgtacagcc tgactacata taaattcact ctagttttgt gccgcagacg ccctacaacc 960

tctgat 966

<210> 4

<211> 844

<212> DNA

<213> A5

<400> 4

gtgttgccca agtgatggtt gtattaaagc cgaagtaaaa gtgatataca gaatatgggt 60

acatattatg atgtatttag tagccgaata tttggcgggt ataacaaaac tagcccagaa 120

tgaaggagct ctatgcttcg cgccaaccat gcgtggctca agaagctttt ggaagcgtga 180

caggcacgta ggcagagaaa gcgagggttg taggagaatt gtcgcttggg aatgacccat 240

cggagcaaac tgtccttaat gtcgagacag actcggagtg gaactgagag acttcagttt 300

tgagatgtat ccggagagca gtatcgtgct tgtcatcgtt gtcatgtttg agtctagcag 360

aagcgaattc tttcggtgag gtggttctgg aatatgagga cattgtcagc caggtagcga 420

gacgtgatgt agacctggca ctgaccgagg caaggacgac atgggcagga ttgtctcatt 480

ctttcgcctg aataggggcg tgatccattg gaaacacgct ttatattcct ttacggcgtc 540

atcgccaaag gagaagaatg caaagaagaa gatcgagctg atgggcacag tccaccaggt 600

gaaatgcgtc cagcggagaa tgtcaggcgg gatgaggcga gaaggaaaga cgccaacgcg 660

agagaagttg ctgtggacat cagtccagcc tgtccacggg cgtaggccgt tacgacaggt 720

aaaccagacg tgaattgaca tcacgacgag cccccagaac atttgaacaa tggccattgc 780

catcaaacgg aagtaacgcg atgtggttag cgcagaattt gagtcacgaa gatgacgagc 840

gaaa 844

<210> 5

<211> 907

<212> DNA

<213> A6

<400> 5

cttcggcaac ttcagacgag atactgcgat ctgcacggtg gattagggag aaggaggcca 60

ctcgcgggct ttgttggaga ataggttgtt ttgtgcccat ggctatgtgc acaagtcttt 120

cttacaggat acgcaacacc acataaggta cgtcgtcctt gaaccctctg ccttttcgag 180

tatatgctta catccttttt gccccttctt ctctgccgtt tcatctcgtt gttccgcttt 240

cccgtttgta ccttgtacct ccttcgcttc aacgccgtcc tcgacaccca ggactctctg 300

aaaacttatc cgtggaaaac atgcacttca ctcgggcctt cgtatgtcac gagtcgctat 360

ccgcagtttg aatactgacc aaaatgagtg tagattgtct tgcagaatga taatgagcgc 420

tcggtcgact tgctactgtt ggcccgtatt caccagcagc atagaatgcc gccgttcggg 480

aggcctctcg acatcgatcg gccgttcctc gccaactcac gctgaacttc ccagtttttg 540

cttcaccgaa tacaatgact acaatatttt tctgtatgtg tgctacatgg tcgtatcaaa 600

gtcctgcgtc ttccacactg ggtgagtacc ttgacagcat tgtatgatta tcgtcgtgac 660

gtggctgaca attagatctt tacaggctct caatgaacag aaaatatatc tttttaaccc 720

tttccacaac tgatcggtta gctgacagtc atcgccggcg ccgagcgtta tcgatcccgg 780

tgcgtcggtt gcctcctctc gcgtggttac ggatgctcat cgtggacaga gcgactgggc 840

acatcatgct ccttcgaaga tatcaacata accgtagagg ccgcgaagaa tgtctgtaac 900

gccgctc 907

<210> 6

<211> 938

<212> DNA

<213> A7

<400> 6

gtcaccttca caagtcgcag ctgtcatgga ggaagaggac gagcagcagg acgacgtcag 60

gcgtggcccg ctcgatgcgg tcgtcagcct ccaggtagcg gagatgctcg ggttgaaccc 120

gaccatgatc gagatggtgt tggccgcggc acgcacattg tcagaatccg aggcagagca 180

gctcatgctc ctcacggacg accagatcgc tcttgggttc cgccacattg ccacccatcc 240

ggaaggtttc gccgacgacg acgaaccgct accgtctgaa caagagatct cccctcgtcc 300

tccaaagcgt aagagtggag agcaagcatg caaggcaagg gctgagacac gtgcccggca 360

ccacgatgct caacggagat cgagcacgga gccatcccag accgggggaa gccagtccga 420

cacgccgtgc ccaccctcgt ccaagaaaag actccggcgt cgggggaaca ggtcgtcgtc 480

ggaggatgaa tcgagccagt agggtcgcca gtaaccgtag ttgacctgtc gtaattcaca 540

agtttttttt atctttttgc tcgaatcatg gcctgaagat gcgcagggtc ccgcaaggta 600

cacttcaaca ccggtctgag ggccatcata agctctgcaa ccatcgccgg gttttatacg 660

ctcagaaaga ggcttctggg ccccgctggg ggctgcgtcg ggctcgttca aggtctccgc 720

ataacctgtc caggtatgca ttcgaaatcc accacccgag gtgctcaggc tgtgggcgtc 780

caccatcctc gccacgctgc atcgacccgg ggaactgttt gcatcgcccc cagtgtctgg 840

ctcatgcgtt gctcttctaa aagcaagtcc tgtctgctca attgtttttc gactaccaag 900

aaaacattta ctctacagtc aaaacgctct ctccaagt 938

<210> 7

<211> 749

<212> DNA

<213> A8

<400> 7

ggcgtccctc tcctacaata atgccacctc ctgttccttc aatcgaaata ctggtcaccg 60

tgactgttac tatatcaaaa catagagtag ctgtctaggt gctgtaagag tggcaccagt 120

gcctaatcaa ataatgagtt ttgtcctctg aagggtgcgt agctgtttac gccaaagcca 180

ggaagtaggt caaagagtat attattatct aaacatatct gttgtgagct gagttgctac 240

gctgattctg tatactgcgt aataagtgat gaaacatgaa ttgaaacaaa agaaggctgc 300

tgaccatcga gttcattacc aagtaattat aggtgtcaag tgagatatag gctcattttt 360

gtgagattag ctgagatggc caattgggtc aaactcgctg catttctctc acttgcatca 420

agagatgcag gattaggagc ttttcctgtc gcaactggtc catcgggtcc ccaaaaatat 480

acgactttgt tccaaatggc aaagagcctt ttctcctcgg catctgaaat acatgtctta 540

gagtggaata attgtgatag gtaaagaaac acgaacctgc aggacaaccc ttccagtgca 600

tttttgcagc ctgggcagcc caggtatacc aggagagctt atacccatac gatttgggtt 660

tccatccaaa ggaaatgacg acgtctttcg ctgtcaatac tctgccgttg ggtagcgtaa 720

tctgttgtac cttgcccttg attgaaagt 749

<210> 8

<211> 867

<212> DNA

<213> A9

<400> 8

ggcttctgac aagaccacct gtctcaatat cccaaacttc tgcaatataa aaacgcccag 60

caatgacaag aaacttatca tcaggtgaaa actctacacg ataagggtgt gttttagttg 120

gacatgtaag aagacactct ccagtgattg tactgaaaac ttccaccttg tcatatccat 180

caaggaggca taacatcttt ccaagatttg agaacctggt attgcgtcct cctcttgcat 240

agattgtaat tggaatagga ttccagtatt caagaacacc atatatgaca ttgacttcat 300

ttttaatgtc atcactatgc aagttgcgca agagactttt cattggagtc ataggtagag 360

ctgagtggta gatatgggga acagaagact gcacacactc tcggaactct cgcacaaaat 420

gaaggccatc attaacccac tcaataagac tgtccaagtc cttagaagac aaagagccat 480

gagtttgggc agtttcaagc caggtacata tccggtgaag gaagtgaatg gcagatcgat 540

tctctacaag gcttagcact tcaaaccaga acagctgttt ttgctttaca aaaatatgaa 600

gaagaggcca gacatttggc aaggttgcaa atgctgaagt ataggtaaga tgatcaaccc 660

agaaacaaca tgcatattca agtactcctc caatgcactt ctttcggtct tgaagtggta 720

agacagaatt catagcatag cgaggaagtt cacatatatt tctgcgcaaa tttgcaacca 780

tgatctcaag gcagtgcact gcaagcatgg gatggtaaac agcaggatca ataaaaaacc 840

tcggatcaac acagcgagat tcacaag 867

<210> 9

<211> 782

<212> DNA

<213> A10

<400> 9

tcatcctcgg ctacctttca gattgtagtt ccagtgaatt ttttagattt tgactgccgc 60

aaagaggtat gtaaattatc cttgatgttc atatttgata tctctctgat gaagatctac 120

agtcaccgtc attgttccct gggtcatcaa atgctccaat ctcactggct actgttccca 180

gtggtccagg cacattgaac tccatacaca ttgaagagga ggcggacatt agtggagacc 240

cagatgaccc atccgacgag tcagattctg tagagttaga tatgcttgag gctcactcac 300

aagttgagag ttcgaaaggt ctgtaggtgt ttttctacac ccacatattc agtcctaatg 360

caaaccaccg atcacaggca aaaagcggca gttggatgat tcagatccaa ccttgtccgg 420

ccctctcgca acccttcaga acttgatcaa ctcaccacct gatgtgggtg atgagtatac 480

acgtgtcaag aaggatatct ttcatgcatt tcatatgata cctacgccga tcaatcatgg 540

catgagacct gcttttcttc gagctctcca tgatcatatg ctgcggtggg atcctgcttc 600

tcgtgacatt gttgacagaa cttgccgcga ggttttcaag ctcacgttcg atcaaatgct 660

tgcccgaaat ccacagttca ttgctgaatg aaccccacgg catgtcccgc ctccaagtgt 720

tcttgttcct gccatgcgcc atgtcttcga gacgttcgga aacaagactg atgcgaagac 780

ga 782

<210> 10

<211> 459

<212> DNA

<213> A11

<400> 10

ttcccttggt ccttttcgca ggcggcggtg cctcttcctc ctcctcgtcc tcctccgact 60

catcgctgtc gtgctcgctc acctcgtgat gcgccttccc cttctccagc tcctcttcct 120

cctcagagtc cgtttcggat gcctcggacg agttggagct gggggacgtc gttgcgacgg 180

ggggtgcgag cacgggggca acagcagcag cttttggggc acgcgtgcgg ggtgccgctt 240

tctttgcctg gttgtaggaa gtacaagtca gcaatcaaca ttcttgacat ttctcacgtg 300

tatggtaaat ccacatacag caatggcagc ggcggtggca gcgttggcag cggcgacttc 360

ctcgggggat cgtgagtcgt aggctttctt ttcttgcgag tactgctcct ttgcactggc 420

aacctgactg ttgtacacct agacgcgacg gagtcagta 459

<210> 11

<211> 579

<212> DNA

<213> A12

<400> 11

ctgccagtca tgtggatggt tcatatgaaa gtgaagtttt acaaataccg cttcttctcc 60

tgacacttgt tcctcatctc tcaatcatcc gcggatggag acaaaggtcg agaccacggt 120

ggtcgtacct ctcgatggcg agaaatccga aggttctacc attgaacaga gccaacagga 180

gaatcatgag gaagaatttc aagaaggagg cctcgtggga tgggctacag tagctggggc 240

gtaagaaatg tttctcatgt tcatcggaaa tgttctgatg atctgcattc ttgtctaggt 300

tcatcgttca attctgcggc ttcgggtatg ttgtaatacg ttcgccggga aagcgagctg 360

aattcactca cttctgtgca gatacacgac gtcttttggg gtgtaccaag gtaaaccgcg 420

gccgaaatgt tcacatgacg cctgcatgac actcgtctaa ataaactgtt gaaatatcac 480

ccctagattt ctacacccgg atctacatca ccaatgagtc gccttctgcg atatcgtgag 540

taatgccgta cataatacat tcgtcgcgat tcttccgat 579

<210> 12

<211> 612

<212> DNA

<213> A13

<400> 12

aggtcaactt gtgcaccgat tacagcaatt gtcggctttt cgttcctgct cacttggata 60

tttatactct atcaaccctc cccaggtcct ggcatagtac aaaagcttgg gtggcaatcc 120

tgggatgtgg tcactatgcc tagcgagaat cagagtcctg cgccggggaa cgtatcttcc 180

gatctcggca cagatatcga cgttccggaa ggagttgact ggtggaatgt cagtgtccca 240

gataccaagg ttgattcctc caatttccca ttagacgtat gggcaccact gctaccccac 300

actgccggtt gtgcgttgag tttgactttt gtaatgaggc aacttattaa cagagcatga 360

gtagtgtccg aaatcaccgt cacgcggtgt ttacttgacc ccagctatgg caaggacttg 420

tgcgctcccg attccacaag tgagcaggac gcaataaagg gaaagtgggt acgggtggac 480

cgaaacctga acctcgaggc gaactatata tctggatggc tggtgcgtga tcatcgaaga 540

ggcgttccac ttgaacctga agagctatat tacagaatat atattatcgc agaacgcgca 600

ggcaggatat aa 612

<210> 13

<211> 932

<212> DNA

<213> B1

<400> 13

ggatcgctca tgctactcaa aatctgggat gatactgaaa aattgaaatt gattgtgcta 60

gcagtacaag tgaggagtac tatcgaggtt caagggaaaa aaatgatcgt ggaggttgtt 120

acgcgtaatg gctacaagct tgtaaggagc gacgtattga gatacaccaa agccgaagtg 180

aataaaaata tatgaatgct gcgaggctcg atgctcggaa aagcggaagg gtgaatatgg 240

cgctgggcca ataagttaat cggagccacc aaaagaggct aatgtgttgg gtatctttga 300

gctaagattt agcgaggagg gagctcaaga gaggacgagc aaagcacctt aggattatgc 360

aatggtgcaa aaaccaccgt atgcacctga acggctatcg gcgtcgacga gcaatgattc 420

atcgttctcg agagttatgg cggcggcatc ttcgccgagg gtgaaggaga acgatgtgaa 480

tgaatccatt tgttgagtgc ggatctgagg ttgcttagcc gatggtggac tcgaattaac 540

ggactgcacg ctgtcgacct tgcccggatt tatagtccca gcgggttgtt gtccgccgat 600

gttggccagt agttcggcca cgaggcgatg gtgactttcc ggtctcattg tgaaaaaaac 660

actgaacaat gtctccatga ttagttggag ttgttgacta ttattgaaca cattgttgtc 720

ctcacatgcc acatatgccc gtagtcacct acggtccgaa attccattcc tcaccatgaa 780

taactgccag atctgtcagt gtgagcaact gcgagtctgc gactgaaaga actgcatcct 840

ccgcactacc gtgactcacc ggtcattttc tggcccccca tccctaagcc gaatatatga 900

ataccagttt ccacagatca actccccaga gt 932

<210> 14

<211> 773

<212> DNA

<213> B2

<400> 14

agtcacctct gcttggattt tgaagtacat cccatggctg acgaccgctc tcgaaagtgc 60

cccaaactgg gtgatcgatc ttttgcctcg cgaaaaccag ttttttcgtg acctgcgtat 120

ttactcatca gcccaaatca gtaaatactt ggacgacccc tccgctcttg atctcgctga 180

ccaaaccact atttttcatc acctcctcaa tccgaaggcg gccactgtcc catcgcgcac 240

ttccctcatg ggagaagtga tggcgttgct cgcagcaggt catgatacag tgggcggcgc 300

gtgcaccgtc ggggcttacc atgtcctcaa gaaccactct gttcaggcaa ggctccgagc 360

cgagttgcat gaagcctggc ccgatcctaa cgcgccggcg acgtaccaga ctttggagaa 420

attaccatac ctggtgtgta gcctaatttt tgttcgtctt gactctacct cacatagtgc 480

agactgccgt catcaaggag tcattgcgtc tttcccatgg tgggttactg tcacatttga 540

cttctatcgc taacggataa tcaggcgtag tatctaatct acctcgagtc gctggtgaag 600

cattaacaat tgccggttat aatgtacctc cacgcgcaag taccctcttt ttccctaatt 660

aattcagctc ctaagcgacc ccatagactg tcgtttctat gtctgccatc tatctacacc 720

tgaattgcga catctttcct gatccccgca cattcaaccc cgatagatgg ctt 773

<210> 15

<211> 897

<212> DNA

<213> B3

<400> 15

agttatcctg tttcctcgca cgatcgcagt catctatttt gtcttctacc ttatcaaaca 60

cacccgttcc tgcccaaagt cgatccgtag ccgctgagca cgatacgagc aaaggaaagg 120

acgagaggag aggataagga taagctggac ccggagctgt gaatggtagg cgccgaggac 180

gagattgaga ctgcgaggtt ctgtcacgag aagaatgggt tcgaggtggg ttttgttcta 240

cgtttcgatt ttttttcttc cactggactc ttcttttttg accagacaat cttttggcga 300

catgccttcc actgttcgga ccccgtgctc tccactacca tcctactttt caattctccg 360

cacaaaacgg atgcggaccg ggaagtcctg tgcagcctac ggtccgtaat ccatctttct 420

ctggaaatcg acggggtggc gcgcctctgg atgacctaga cccgcgaaca tgatatgtac 480

ggttcggaca tcgaggcgcg ttcataattt ttgggtctag gtttgccacg gagatcatgg 540

cgaaatacaa tgttttcttc tgatttagta gccaaatatc gcaaatcaga tcagatcgat 600

ggcatccgaa ggttgctccg cactaccatc gtcctctttt ccttccagcc ctacggacgt 660

tgctcagagg ccagtgagtc agatatgtac gaagtattga ccagtgtatt caaagtatac 720

tgcagctgct ttcgatttca agctcttatc atcaacaatg gatccgtccc ccgcgatgcc 780

acaggaactc attgacaaga tcgtagacga tcttcacgac gatccgattg cgctgagagc 840

ctgtgcactg tcatcacact ctctcctttt acgtagccag ggccacatat tttccat 897

<210> 16

<211> 785

<212> DNA

<213> B4

<400> 16

ccagtaatgt tcgtgtgcct ctgatgagat cgctggacca actacaagaa agttacattt 60

ttcccgacgt tcctctggag agccaacgat ctggatagcc tcaattacac ggccaacgcg 120

aggccttggt atgattggaa actggccccc taaatccgat gtccagataa tccaagagcc 180

tgcatggcaa atgtccccgt taaataccac ggtgtcttgt ggtgtataga ccagagcatc 240

tggttgtagg accgaggaca agccaggaga ggtggctgct tgtgttgttg accagcgctt 300

ttctgggccg cgccgggagg aatgacctgg tcaatcctgt cagcggttcg actcgcaaat 360

tctgaatcca catgacactt acctggctca agcttcacgt cttcatttcg ttccagtcca 420

agaatgcgtg gaccaaatcc ttggagattc aggatgttca aaggctcttg agctgcatgc 480

tgccagtgaa gatcatcggg gttctgaaca cgctgcatcc aggtagaaat gctctgcgaa 540

gcccgctgtt gctttctttt gcgacgagat acactggggg gttcaggcgt gaggcgattc 600

atcagaaaca gccctccact aagcagatgg cgaacacggt tccccttggc cattcggcgt 660

gcaatatcgt atgagggtgc atgtcgattg ctatgaatac tggcagatct gatgatcgcg 720

ttgaatgatt cgaacccttc tgttgcgaag agcatcgcag gcccgaaccg tcgaatatgc 780

tctgg 785

<210> 17

<211> 867

<212> DNA

<213> B5

<400> 17

gaaaacgcga acgacaatga cgggaacgtc agcgaacgtc tgcccaggtc gtaactgaga 60

tacgatctcg cgccactcag gattgcacgt catggtgatg aaaaatgttg gcggacctcc 120

catggcagca atggccagag aatcagcaat ttgttcactt gcccatcgac gtgatccaag 180

aaacgaagac ggcaaataga cattttctga actgtcaatg tcgtctgacc ccatgagtgc 240

cgcatcttca acacaaaccc gctgctggtt atagcgaatg aagttcaagc gggactccaa 300

gtcacggctg aacatgtcca caatgtattc atttgcgagg cggccgaaga tctgaaagcg 360

tggttcacga agcaggcgag cacgataatg ccacatttgt gtcgtgggtg aatctgcatt 420

tccatcaacg gcgacgtttg catggaaatt gtcatgtgag ccaataaggc cccagcctag 480

agttccttga ggaaagagaa gaggatatgc aagtggttcc cacagacggc tgacagtggc 540

gatcgtatga ttctcgccgt cgtctttctt gatgaccatg gtacgtgatc tgatttgtga 600

ctgcgtagta ttgtcatacg acattattgc tgcaatctca gcagaccctc tatcttcgag 660

gatgagtgaa gcatgtggag tggtgtcatg gcgtaggaac atgagagcgg cagcgaatgg 720

attggtgcgg aggagtgtgt cgcgtaacgt atggatccat tgtggaggca aagtggctgc 780

ccattcctca aagggtgttt gggtgtccat aaatccatcg tacagcagcc atcgaactgc 840

agaattcttg tgattgggtc gaagacg 867

<210> 18

<211> 20

<212> DNA

<213> Artificial sequence

<400> 18

tgtgaaagtg agatgacggg 20

<210> 19

<211> 20

<212> DNA

<213> Artificial sequence

<400> 19

ttgactccga aggggtactg 20

<210> 20

<211> 20

<212> DNA

<213> Artificial sequence

<400> 20

tgtgggcatt ttagcaacga 20

<210> 21

<211> 20

<212> DNA

<213> Artificial sequence

<400> 21

ccctgtgctc gtatttcctg 20

<210> 22

<211> 20

<212> DNA

<213> Artificial sequence

<400> 22

ttcaactcgc atgggtacat 20

<210> 23

<211> 19

<212> DNA

<213> Artificial sequence

<400> 23

atcagaggtt gtagggcgt 19

<210> 24

<211> 21

<212> DNA

<213> Artificial sequence

<400> 24

aactcagggt cggctatttt c 21

<210> 25

<211> 21

<212> DNA

<213> Artificial sequence

<400> 25

acaacatcac tcaccacaga a 21

<210> 26

<211> 20

<212> DNA

<213> Artificial sequence

<400> 26

gtgttgccca agtgatggtt 20

<210> 27

<211> 20

<212> DNA

<213> Artificial sequence

<400> 27

tttcgctcgt catcttcgtg 20

<210> 28

<211> 20

<212> DNA

<213> Artificial sequence

<400> 28

cttcggcaac ttcagacgag 20

<210> 29

<211> 19

<212> DNA

<213> Artificial sequence

<400> 29

gagcggcgtt acagacatt 19

<210> 30

<211> 19

<212> DNA

<213> Artificial sequence

<400> 30

gtcaccttca caagtcgca 19

<210> 31

<211> 20

<212> DNA

<213> Artificial sequence

<400> 31

acttggagag agcgttttga 20

<210> 32

<211> 21

<212> DNA

<213> Artificial sequence

<400> 32

ggcgtccctc tcctacaata a 21

<210> 33

<211> 21

<212> DNA

<213> Artificial sequence

<400> 33

actttcaatc aagggcaagg t 21

<210> 34

<211> 19

<212> DNA

<213> Artificial sequence

<400> 34

ggcttctgac aagaccacc 19

<210> 35

<211> 21

<212> DNA

<213> Artificial sequence

<400> 35

cttgtgaatc tcgctgtgtt g 21

<210> 36

<211> 20

<212> DNA

<213> Artificial sequence

<400> 36

tcatcctcgg ctacctttca 20

<210> 37

<211> 20

<212> DNA

<213> Artificial sequence

<400> 37

tcgtcttcgc atcagtcttg 20

<210> 38

<211> 20

<212> DNA

<213> Artificial sequence

<400> 38

ttcccttggt ccttttcgca 20

<210> 39

<211> 20

<212> DNA

<213> Artificial sequence

<400> 39

tactgactcc gtcgcgtcta 20

<210> 40

<211> 20

<212> DNA

<213> Artificial sequence

<400> 40

ctgccagtca tgtggatggt 20

<210> 41

<211> 20

<212> DNA

<213> Artificial sequence

<400> 41

atcggaagaa tcgcgacgaa 20

<210> 42

<211> 20

<212> DNA

<213> Artificial sequence

<400> 42

aggtcaactt gtgcaccgat 20

<210> 43

<211> 20

<212> DNA

<213> Artificial sequence

<400> 43

ttatatcctg cctgcgcgtt 20

<210> 44

<211> 20

<212> DNA

<213> Artificial sequence

<400> 44

ggatcgctca tgctactcaa 20

<210> 45

<211> 20

<212> DNA

<213> Artificial sequence

<400> 45

actctgggga gttgatctgt 20

<210> 46

<211> 20

<212> DNA

<213> Artificial sequence

<400> 46

agtcacctct gcttggattt 20

<210> 47

<211> 20

<212> DNA

<213> Artificial sequence

<400> 47

aagccatcta tcggggttga 20

<210> 48

<211> 21

<212> DNA

<213> Artificial sequence

<400> 48

agttatcctg tttcctcgca c 21

<210> 49

<211> 21

<212> DNA

<213> Artificial sequence

<400> 49

atggaaaata tgtggccctg g 21

<210> 50

<211> 20

<212> DNA

<213> Artificial sequence

<400> 50

ccagtaatgt tcgtgtgcct 20

<210> 51

<211> 20

<212> DNA

<213> Artificial sequence

<400> 51

ccagagcata ttcgacggtt 20

<210> 52

<211> 20

<212> DNA

<213> Artificial sequence

<400> 52

gaaaacgcga acgacaatga 20

<210> 53

<211> 20

<212> DNA

<213> Artificial sequence

<400> 53

cgtcttcgac ccaatcacaa 20

Claims (1)

1. A hypsizigus marmoreus strain, characterized in that: the Hypsizigus marmoreus is Hypsizigus marmoreus (A) and (B)Hypsizygus marmoreus) "nong Wan Zhen No. 1" was deposited in China general microbiological culture Collection center (CGMCC) at 6/15 in 2020 with the following deposition numbers: CGMCC No. 20201.

Images