CN109837335A - A method of joint ATAC-seq and RNA-seq screens edible and medical fungi functional gene - Google Patents

Abstract

A kind of method that the present invention discloses joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene, using ATAC-seq method test out edible and medical fungi environmental factor induction before and after the cultural hypha stage mycelia and by environmental factor induction former base differentiation after former base opening Chromatin domains and analyze open area encoding gene, edible and medical fungi is tested out in the mycelia in mycelia stage and the mRNA sequence of the former base after former base is broken up using RNA-seq method, and the shared differential gene between the gene order obtained by ATAC-seq method and RNA-seq method is found using Venn figure, the critical functionality gene order for influencing the differentiation of edible and medical fungi former base is found in shared differential gene eventually by biological analysis.

Description

A method of joint ATAC-seq and RNA-seq screens edible and medical fungi functional gene

Technical field

The invention belongs to biology techniques fields, more particularly, to a kind of joint ATAC-seq and RNA-seq screening food medicine With the method for bacterium functional gene.

Background technique

Edible and medical fungi is the higher fungus that a major class has large-scale fructification, is commonly called as mushroom or gill fungus, and there are about 1000 in China at present Multiple eating bacterium, largely belongs to basidiomycetes, and fraction belongs to sac fungus.Edible and medical fungi polysaccharide rich in, protein, three Terpene, polypeptide, dietary fiber, taurine, mannitol, unsaturated fatty acid, adenosine, lactones, minerals and vitamins Deng, while having the characteristics that less salt, low fat, low sugar and high protein, one of ten big healthy food are recommended as by nutritionist, are had There are huge market prospects and potentiality to be exploited.Wherein, Sparassis crispa is a kind of rare edible and medical fungi, is mainly distributed on the sub- heat in East Asia The ground such as band and Temperate Region in China, including China Jilin, Heilungkiang, Yunnan contain protein, essential amino acid in Sparassis crispa fructification It is surveyed with several functions compositions, main active beta glucan rich content, japanese food analytical tests such as vitamins Its fixed content is 43.6%, is the 2 times or more of ganoderma lucidum, Agricus blazei, be mushroom class most, and medicinal, healthcare function with higher, There is the title of " mushroom of illusion " in Japan.Modern study confirms that Sparassis crispa has the physiology such as antibacterial, antitumor, immunological regulation, hematopoiesis Biochemical activity.As a kind of rare edible and medicinal fungi, Sparassis crispa is received due to its good nutritive value and health-care efficacy The extensive concern of scholar and consumer.

In recent years, with the rising of China's edible and medical fungi proportion of output value and in the raising of international market share, about Edible and medical fungi cultivation and the research of yield-increasing technology are concerned.Currently, each research institution is mostly by the research collection to edible and medical fungi In in terms of artificial cultivation either Crop Physiology, biochemical activity research, capture emphatically with good nutritive value and guarantor It is good for effect and wild resource is few, acquire domestication's work of difficult edible and medical fungi.But the cultivation skill about edible and medical fungi Art is still not mature enough, cannot achieve extensive cultivation.As genetic engineering constantly develops, Biotechnology in Genetic Breeding is introduced into food The important channel of an edible and medical fungi artificial cultivation and volume increase will be become in the culture of medicinal fungus.It is applied to edible mushroom heredity at present The mode of breeding mainly includes that artificial selection breeding, mutation breeding, crossbreeding, protoplast fusion breeding and genetic engineering are educated Kind.Wherein, three kinds of artificially breeding, mutation breeding and crossbreeding methods are more common at Development of Molecular Biology initial stage, although The excellent bacterial strain of character at energy breeding, but screen more blindly, and labor intensive and time；Protoplast fusion breeding The character of fusant enhancing is only limitted to the character that two parents have had in the process, it is difficult to obtain new character.And gene work Journey breeding is genetic manipulation at the genetic level, can will replicate table in any one functional channel genes recipient cell It reaches, to select new varieties.Genetic engineering breeding is that the edible and medical fungi strain of breeding more merit brings dawn.

Mostly important in the overall process of gene breeding is exactly from being filtered out in edible and medical fungi to edible and medical fungi growth To the functioning gene of key effect.Functioning gene screening technique mainly passes through proteomics, functional genomics etc. Biological omics technology is screened, and the elementary tactics of research is: (1) by express spectra variance analysis, bioinformatic analysis Obtain candidate gene with approach such as gene clonings: (2) carry out Evaluation of Functional to candidate gene, make a definite diagnosis its function to be screened Gene.These researchs are currently limited to the fields such as biology, medicine and disease treatment, do not have also in edible and medical fungi and are generally applicable in Property.Genescreen about edible and medical fungi is only limitted to sieve using single technologies such as transcript profile sequencing, proteomic techniques Choosing, but the method that single technology screening goes out difference expression gene in edible and medical fungi former base atomization in this way would generally A large amount of differential gene is found, cause the workload of subsequent gene Function Identification and verifying very big, and monotechnics are found out The specificity of specific gene is lower, is easy to appear more false positive, can not efficiently filter out edible and medical fungi former base differentiation rank The key gene of section.Genescreen and Function Identification are carried out by the biology techniques of multiple combinations, and then finds and can influence The critical functionality gene of edible and medical fungi former base differential period improves edible and medical fungi artificial cultivation efficiency, stabilization and new varieties Cultivation is the trend of the following edible and medical fungi cultivation technique development.

Summary of the invention

To solve the above-mentioned problems, the present invention provides a kind of joint ATAC-seq and RNA-seq screening edible and medical fungi function The method of gene can be opened wherein having combined two kinds of gene order surveying methods of ATAC-seq and RNA-seq using ATAC-seq The information of Chromatin domains is put, then cooperates RNA-seq to obtain corresponding transcript information, finds the upstream regulation of related gene Sequence, the regulated and control network under the specific space-time of global analysis, and being mutually authenticated, in combination with former base differentiation critical environments because Son efficiently filters out the key gene of former base differentiation.

The purpose of the present invention is what is be achieved through the following technical solutions:

A method of joint ATAC-seq and RNA-seq screens edible and medical fungi functional gene, utilizes ATAC-seq method Test out the mycelia in cultural hypha stage of the edible and medical fungi before and after environmental factor induction and by environmental factor induction former base point The opening Chromatin domains of former base and open area encoding gene is analyzed after change, tests out edible and medical fungi using RNA-seq method In the mycelia in mycelia stage and the mRNA sequence of the former base after former base is broken up, and is found using Venn figure and pass through ATAC-seq The shared differential gene between gene order that method and RNA-seq method obtain, eventually by biological analysis shared poor The critical functionality gene for influencing the differentiation of edible and medical fungi former base is found in allogene.

It is a kind of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene method screening Sparassis crispa function base Because of upper application.

Further, the mycelia in the cultural hypha stage of the Sparassis crispa takes the mycelia cultivated under dark condition and warp It crosses light induction but does not go out the mycelia of former base also.

Further, joint ATAC-seq and RNA-seq is in the method for screening edible and medical fungi functional gene in screening silk ball Application on bacterium functional gene, the specific steps are as follows:

S1, the mycelia for cultivating Sparassis crispa under dark condition and by light induction but also do not go out former base mycelia, The mycelia cultivated under dark condition and the former base after light induction pass through ATAC experiment respectively and obtain purifying DNA progress Acquisition mRNA progress Illumina HiSeq sequencing is tested in Illumina HiSeq sequencing by RNA-seq；

S2, mycelia and pass through light that Sparassis crispa is cultivated under dark condition are obtained respectively after the sequencing of ATAC-seq method The mycelia contrast groups and the mycelia cultivated under dark condition and the original after light induction of former base are not gone out according to induction but also The lower gene dosage for reconciling and raising is expressed in base contrast groups, while is obtained respectively after the sequencing of RNA-seq method in dark item The mycelia cultivated under part and the bacterium for passing through light induction but also not going out the mycelia contrast groups of former base and being cultivated under dark condition The lower gene dosage raised that reconciles of expression in silk and the former base contrast groups after light induction；

S3, the bacterium for passing through light induction using Venn map analysis but also not going out the mycelia of former base and being cultivated under dark condition The gene dosage and common table of downward are co-expressed in silk contrast groups after the sequencing of ATAC-seq and RNA-seq integrated processes Up to the gene dosage of up-regulation；Also with former base of the Venn map analysis after light induction and the bacterium cultivated under dark condition The gene dosage for co-expressing downward in silk contrast groups after the sequencing of ATAC-seq and RNA-seq integrated processes, co-expresses The gene dosage of up-regulation；

S4, comprehensive analysis, which are passed through in two contrast groups obtained in step S3, co-expresses the lower gene dosage for reconciling and raising, And the shared differential gene of two contrast groups is found using Venn map analysis；

S5, the key that bioinformatic analysis filters out the differentiation of light induction Sparassis crispa former base is carried out to shared differential gene Functioning gene.

Further, ATAC experiment specific steps are as follows:

A1, sample preparation: it takes mycelia under the dark condition in silk ball bacteria cultivation, go out former base by light induction but also Mycelia, the former base after light induction, with liquid nitrogen flash freezer, cryopreservation；Sample is put into and is pre-cooled containing homogenate In the homogenizer of buffer, the tissue of freezing is allowed to thaw and by tissue grinder, pre-cooling centrifugation takes supernatant, utilizes Iodixanol Gradient method separating nucleus, counts nucleus, and every 50000 cell nucleus packing saves；

A2, swivel base reaction and purifying: 50000 nucleus are transferred to containing 1ml ATAC-RSB and 0.1%Tween- In 20 pipe, Omni-ATAC ATAC-seq reaction mixture is added in centrifuging and taking supernatant, and 30min purifying is incubated at 37 DEG C DNA, with 10 μ l elution buffer eluted dnas；

A3, PCR amplification: take 10 μ l dense by DNA, water, the 2.5 μ l of 10 μ l nuclease frees of step (2) swivel base after purification Degree for 25 μM PCR primer 1,2.5 μ l concentration be 25 μM the μ l NEB Next of PCR primer 2 and 25 High-Fidelity 2 × PCR Master Mix；

A4, sequencing and gene data are analyzed and then obtain the gene dosage of the different lower reconciliation up-regulations of sample expression.

Further, the RNA experiment is successively by sample preparation, mRNA extraction, library construction and sequencing and data It analyzes and obtains the lower gene order raised that reconciles of expression in turn.

Further, Sparassis crispa mycelia and former base sample need to be ground in liquid nitrogen in the RNA experiment, utilize Trizol RNA extracts reagent and extracts mRNA.

Further, the step of RNA experiment Chinese library building is as follows:

B1, with the enrichment with magnetic bead mRNA with Oligo, NEB Fragmentation is added into obtained mRNA Buffer makes its fragmentation become short-movie section；

B2, using the mRNA after fragmentation as template, synthesize the first chain of cDNA with hexabasic base random primer, and buffering be added Liquid, dNTPs, RNaseH and DNA Polymerase I synthesize the second chain of cDNA, simultaneously by the purifying of QIAQuick PCR kit EB buffer is added to elute；

B3, be purified by flash after double-strand cDNA carry out again end reparation, plus base A, plus sequence measuring joints processing, through agarose Gel electrophoresis recycling purpose size segment simultaneously carries out PCR amplification, completes library construction work.

Further, bioinformatic analysis includes carrying out GO analysis, KEGG to shared differential gene in the step S5 Analysis, GSEA analysis and signal network analysis.

Beneficial effects of the present invention:

1, the method in the present invention using joint ATAC-seq and RNA-seq carries out edible and medical fungi in former base differential period Critical functionality gene is screened, middle compared to the prior art to have higher effect using monotechnics screening function gene Rate, preferably specificity and accuracy, it is big to overcome the differential gene quantity occurred in monotechnics screening process, subsequent gene Verifying heavy workload, low efficiency are easy to appear the problems such as false positive, and two kinds of sequencing approaches of ATAC-seq and RNA-seq are for eating The environmental factor that medicinal fungus genescreen can will affect edible and medical fungi growth is taken into account, and then obtains the something lost under specific space-time Communication breath；ATAC-seq obtains the information of the open Chromatin domains of the edible and medical fungi under specific space-time, all in the specific space-time The lower gene transcribed and cis-regulating element can detecte, and obtain in combination with RNA-seq in the specific space-time Under corresponding transcript information, so that it may accurately find under the specific space-time upstream regulatory sequence of related gene and under Swim regulating and controlling sequence；The joint of two kinds of sequencing approaches of ATAC-seq and RNA-seq can help (two kinds from different perspectives of genescreen The differential gene that sequencing approach obtains) and different level (different growing environment factors) account for, global analysis is specific Regulated and control network under space-time, while ATAC-seq sequencing approach and RNA-seq sequencing approach can be under same specific space-times It is mutually authenticated, improves the accuracy of key function genescreen structure.

2, the method for combining ATAC-seq and RNA-seq is used for the sieve of the critical functionality gene of Sparassis crispa in the present invention Choosing, while the specific space-time-illumination that will affect environmental factor i.e. Sparassis crispa growth that Sparassis crispa grows is taken into account, and selects Mycelia of the Sparassis crispa under dark condition, the mycelia for by light induction but also not going out former base, the former base after light induction For test sample, it is contrasted group, by finding two contrast groups differential genes, and finds common difference base from differential gene Cause filters out the important controlling gene of Sparassis crispa mycelia growth, that is, the key gene of fruit-body formation.

Detailed description of the invention

Fig. 1 is the overall technology route map of Sparassis crispa functional gene screening in the present invention；

(a) is the Sparassis crispa mycelia of dark condition culture and light after the sequencing of RNA-seq and ATAC-seq method in Fig. 2 According to the gene dosage Venn analysis chart for the mycelia expression downward for inducing but not going out also former base；(b) pass through RNA-seq and ATAC-seq The mycelia of dark condition culture and light induction but the gene dosage that also the mycelia expression of former base is not raised out after method sequencing Venn analysis chart；(c) by RNA-seq and ATAC-seq method sequencing after dark condition culture mycelia and light induction after The gene dosage Venn analysis chart that former base expression is lowered；(d) dark condition is trained after the sequencing of RNA-seq and ATAC-seq method The gene dosage Venn analysis chart of former base expression up-regulation after feeding mycelia and light induction；

(a) is former base contrast groups of the Sparassis crispa after the mycelia of dark condition culture and light induction and dark item in Fig. 3 The mycelia of part culture and light induction but do not go out in the mycelia contrast groups of former base to co-express gene dosage Venn points of downward also Analysis figure；It (b) is former base contrast groups of the Sparassis crispa after the mycelia of dark condition culture and light induction and dark condition culture Mycelia and light induction but the gene dosage Venn analysis chart for not going out in the mycelia contrast groups of former base to co-express up-regulation also.

Specific embodiment

In order to better understand the present invention, below by embodiment to the present invention into further explanation, embodiment is served only for It explains the present invention, any restriction can't be constituted to the present invention.

A method of joint ATAC-seq and RNA-seq screens edible and medical fungi functional gene, utilizes ATAC-seq method Test out the mycelia in cultural hypha stage of the edible and medical fungi before and after environmental factor induction and by environmental factor induction former base point The opening Chromatin domains of former base and open area encoding gene is analyzed after change, tests out edible and medical fungi using RNA-seq method In the mycelia in mycelia stage and the mRNA sequence of the former base after former base is broken up, and is found using Venn figure and pass through ATAC-seq The shared differential gene between gene order that method and RNA-seq method obtain, eventually by biological analysis shared poor The critical functionality gene for influencing the differentiation of edible and medical fungi former base is found in allogene.

It is a kind of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene method screening Sparassis crispa function base Because of upper application.

Further, the mycelia in the cultural hypha stage of the Sparassis crispa takes the mycelia cultivated under dark condition and warp It crosses light induction but does not go out the mycelia of former base also.

Further, joint ATAC-seq and RNA-seq is in the method for screening edible and medical fungi functional gene in screening silk ball Application on bacterium functional gene, the specific steps are as follows:

S1, the mycelia that Sparassis crispa is cultivated under dark condition and the mycelia, black for not going out former base by light induction but also The mycelia of dark condition culture and the former base after light induction pass through ATAC experiment respectively and obtain purifying DNA progress Illumina Acquisition mRNA progress Illumina HiSeq sequencing is tested in HiSeq sequencing by RNA；

S2, by ATAC-seq technology sequencing after obtain respectively Sparassis crispa in the mycelia of dark condition culture, by illumination Induction but the gene dosage lowered that do not go out also to reconcile under expression in the mycelia of former base and the former base after light induction, pass through simultaneously It crosses after the sequencing of RNA-seq technology and obtains the mycelia cultivated under dark condition respectively, do not go out former base by light induction but also The lower gene dosage for reconciling up-regulation of expression in mycelia and former base after light induction；

Referring to fig. 2, what is obtained after the sequencing of ATAC-seq technology lures in the mycelia of dark condition culture and by illumination Lead but also do not go out former base mycelia contrast groups in express downward gene dosage have altogether be 198, by RNA-seq technology be sequenced The mycelia cultivated under dark condition that obtains afterwards and under being expressed in the mycelia contrast groups for not going out former base by light induction but also It is 830 that the gene dosage of tune, which has altogether,；After the sequencing of ATAC-seq technology the mycelia cultivated under dark condition that obtains and It is 182 that the gene dosage for not going out in the mycelia contrast groups of former base to express up-regulation by light induction but also, which has altogether, by RNA- The mycelia cultivated under dark condition and pass through light induction but do not go out the mycelia expression of former base also that seq technology obtains after being sequenced It is 795 that the gene dosage of downward, which has altogether,；

Referring to fig. 2, the mycelia cultivated under dark condition that obtains and by illumination after the sequencing of ATAC-seq technology It is 1227 that the gene dosage that downward is expressed in former base contrast groups after induction, which has altogether, is obtained after the sequencing of RNA-seq technology The mycelia cultivated under dark condition and the gene dosage that downward is expressed in former base contrast groups after light induction have altogether It is 1435；By ATAC-seq technology sequencing after obtain in the mycelia of dark condition culture and the original after light induction Expressed in base contrast groups up-regulation gene dosage have altogether be 1241, by RNA-seq technology sequencing after obtain in dark item It is 1469 that the mycelia cultivated under part and the gene dosage that up-regulation is expressed in the former base contrast groups after light induction, which have altogether,；

S3, using the above-mentioned differential gene data of Venn map analysis, obtain by light induction but do not go out also the mycelia of former base Under being co-expressed after the sequencing of ATAC-seq and RNA-seq combination technology in the mycelia contrast groups cultivated under dark condition The differential gene quantity of tune has 45, and the differential gene quantity for co-expressing up-regulation has 32；It is obtained also with Venn map analysis It the former base after light induction and is combined in the mycelia contrast groups of dark condition culture by ATAC-seq and RNA-seq out The differential gene quantity for co-expressing downward after technology sequencing has 174, and the differential gene quantity for co-expressing up-regulation has 350 (as shown in Figure 2)；

S4, comprehensive analysis, which are passed through in two contrast groups obtained in step S3, co-expresses the lower differential gene number for reconciling and raising Amount finds the shared differential gene of two contrast groups using Venn map analysis；

Referring to Fig. 3, the former base after light induction and the mycelia contrast groups cultivated under dark condition are co-expressed The gene of downward and the mycelia and the mycelia contrast groups cultivated under dark condition that do not go out former base by light induction but also are common The shared differential gene quantity 13 for obtaining and co-expressing downward in wherein two contrast groups is compared in the gene that expression is lowered It is a, shared differential gene quantity 17 of up-regulation are co-expressed, therefore shared differential gene quantity is 30, specifically shares difference Gene is shown in sequence table；

S5, GO analysis, KEGG analysis, GSEA analysis and signal net are carried out to the shared differential gene obtained by step S4 The bioinformatic analysis such as network analysis filter out the critical functionality gene of light induction Sparassis crispa former base differentiation.

Wherein, specific step is as follows for the ATAC experiment:

A1, sample preparation: it takes mycelia under the dark condition in silk ball bacteria cultivation, go out former base by light induction but also Mycelia, the former base after light induction, with liquid nitrogen flash freezer, cryopreservation；Sample is put into and is pre-cooled containing homogenate In the homogenizer of buffer, the tissue of freezing is allowed to thaw and by tissue grinder, pre-cooling centrifugation takes supernatant, utilizes Iodixanol Gradient method separating nucleus, counts nucleus, and every 50000 cell nucleus packing saves；

A2, swivel base reaction and purifying: 50000 nucleus are transferred to containing 1ml ATAC-RSB and 0.1%Tween- In 20 pipe, Omni-ATAC ATAC-seq reaction mixture is added in centrifuging and taking supernatant, and 30min purifying is incubated at 37 DEG C DNA, with 10 μ l elution buffer eluted dnas；

A3, PCR amplification: take 10 μ l dense by DNA, water, the 2.5 μ l of 10 μ l nuclease frees of step (2) swivel base after purification Degree for 25 μM PCR primer 1,2.5 μ l concentration be 25 μM the μ l NEB Next of PCR primer 2 and 25 High-Fidelity2 × PCR Master Mix；

A4, sequencing and gene data are analyzed and then obtain the gene dosage of the different lower reconciliation up-regulations of sample expression.

Wherein, the RNA experiment is successively extracted by sample preparation, mRNA, library construction and sequencing are analyzed with data And then obtain the different lower gene dosages raised that reconcile of sample expression.

The preparation of sample in RNA experiment: the mycelia cultivated under the dark condition in cultivation, by light induction but also not is taken The mycelia of former base, the former base after light induction out, with liquid nitrogen flash freezer, cryopreservation.

MRNA is extracted: after sample is ground in liquid nitrogen, being extracted according to the operating instruction that Trizol RNA extracts reagent total RNA.Utilize NanoDrop2000 ultramicrospectrophotometer, Ago-Gel, Agilent 2100RNA Nano 6000Assay Kit carries out pattern detection.

Steps are as follows for library construction:

B1, with the enrichment with magnetic bead mRNA with Oligo, NEB Fragmentation is added into obtained mRNA Buffer makes its fragmentation become short-movie section；

B2, using the mRNA after fragmentation as template, synthesize the first chain of cDNA with hexabasic base random primer, and buffering be added Liquid, dNTPs, RNaseH and DNA Polymerase I synthesize the second chain of cDNA, simultaneously by the purifying of QIAQuick PCR kit EB buffer is added to elute；

B3, be purified by flash after double-strand cDNA carry out again end reparation, plus base A, plus sequence measuring joints processing, through agarose Gel electrophoresis recycling purpose size segment simultaneously carries out PCR amplification, completes library construction work.

Wherein, after the completion of library construction, tentatively quantitative, dilution library to 1ng/ μ l is first carried out using Qubit2.0, then It is detected using Insert Fragment length of the Agilent 2100 to library, after Insert Fragment meets expection, uses Q-PCR method Accurate quantitative analysis (library effective concentration > 2nM) is carried out to the effective concentration in library, to guarantee Library Quality.

The above description is only an embodiment of the present invention, is not intended to limit the scope of the invention, all to utilize this hair Equivalent structure or equivalent flow shift made by bright description is applied directly or indirectly in other relevant technology necks Domain is included within the scope of the present invention.

Sequence table

<110>Edible Fungus Research Institute, Fujian Academy of Agricultural Sciences (Fujian Mushroom Fungal Research Station)

<120>a kind of method of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene

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cccctcccaa aggcccagct cttcaccctc tgtactgtaa gactggtcga tcccatcgca 180

ttcacacaga ttttccctta tgtcaacgag atgatggagc ggctacgcct caccaacgat 240

ccgtcgaaga ttggtttcta cagtgggatg gtggaaagca gttttgccat ctctcagctg 300

tgttgcatat accactgggc acgcctttcc gatagaatag gtcgtcgccc ggtggttctc 360

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gaagaaacac tgccgagtaa acggtggaag aatgactatg tacaaatgcg gcaaatgggt 780

tcagaacata gaggcggtga tccagaaaag ccgccagagc ccaagagtct caaattctta 840

ctatcaatac ccgtcatccg tgccctgtgc ctcagtggat tcgctttgac cttcctcaat 900

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ttgacaccat atttcagctg gaaaattaca caccatcgac accacattgc ccatgcatct 180

atggaacggg atcagctatt tgttccgagg accaggagcg accttggtat tccttcgaaa 240

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agcgagtccg agttataccg caccgcgaga ggcttggccc atgccaacaa gcctttcagg 360

agctacattg gcatgggcta tcacaatgcg gtagtgccgc cggttattct ccgcaacatc 420

ttggagagtc ctgcttggta tacgccgtat acgccttacc agcctgagat tgcccaagga 480

cgtcttgagt cacttgtcaa ctaccagaca atggtcatgt ctctgacggc aatggaaatt 540

gcgaacgcgt cgcttctgga cgaagctacc gctgccgcag aagcgatggt catggcatat 600

gcttcgtccg gtcacaagaa gcatactttc ttcgcggaca aaggtgtatc gcctcagact 660

ctggccgttc tccagacgcg cgcgaatggg tttggtctcc ggcttgttgt aggaaacgta 720

ttttcgatta ccaaggacga aactctcatt tcggaccttt gcggcgtact ggtgcaatat 780

ccggatgtta atggcagcat caaagattac ggtgacctgg tccgagccat ccatggctcg 840

ggcgcattag ttgtggctgc gacggacctt ctggcttgac agtgctcaag ccgcctggga 900

atggggtgct gacattgtcc tcggtaactc tgcccgcttt ggtgtcccag ctggctatgg 960

tggtccccac ggagcgttct tcgcgtgcac ggataaactg aaacgtaaga tgcctggaag 1020

gcttattggt aggagccgag atgtgtacgg gaagcccgcg taccgattgg cattgcaaac 1080

tcgggaacag cacatccgac gggagaaggc gacgagcaat atctgtacca gtcaggctct 1140

tctcgccaac atggccgcga tgtatgctgt gtaccatggt cccacaggtt tgcaacgcat 1200

cgcgagcaag gtccacgcgc ttacacaagt gctgaagtct tcggtcgagg cctacggtta 1260

caaggcgata aacacagatt tcttcgatac actgacattc gacgtgactg gtgccgcgaa 1320

ggacgtgaat gctgtgcatt cagcagcttt ggcctcgagc atcaatttcc gcaaggtcga 1380

tgacaagctc gtcggagtga ccttggacga gagcgtcagg cctgaagaca tcgtcgacat 1440

cgccaacgtc ttcgcggcag cagcatcggc caagccaata tcgctttctg ccatcgcgcc 1500

cgtgcaatcc tccgccgttc cccaacccct tcagcggaca tcggagtacc ttccccacca 1560

ggtcttcaac tcgcaccact cggaaacaga gatgcttcgc tacatctacc atcttcagtc 1620

gaaggacctc ggcctcgagc acgctatgat ccccctcggc agttgcacca tgaagcttaa 1680

cagcacgagc agcatgatcc ctctgacatg gcccgagttc ggcaacgtgc atccgttcgt 1740

ccctgctgat caagtcaagg gttacgaaca gttgatcaag gaactcgagt ctgacctctg 1800

taaggtcact ggcttccatg cttgctctct acaacccaac tctggcgctg ctggtgaata 1860

tgctggtctg tcggtcatca gggcatacca ccagtcccgc ggcgagagcc atcgcgatat 1920

ctgtctcatt cctgttagcg cacacggaac aaaccctgct tcggctgtta tggctggcct 1980

caaggtcgtg cccgtaaaga ctctgacgga cggcaaccta gacttgggag acctgaacgc 2040

taaggctgag aagtacaagg acaatctcgc tgcgttcatg attacatacc cgtcgacgtt 2100

cggagtattc gagcacggtg tgcaggacgc ttgcaagatc atccacgaca acggcggaca 2160

ggtctacctc gatggtgcaa acctcaatgc tcagattggc ttgacgaatc ccgcgacatg 2220

tggtggcgat atttgtcata tgaatttgca caagaccttc gctatccccc atggtggcgg 2280

tggtccaggt gtcggaccaa tctgtgccgc cgagcatctc gcacctttct tgcccggcca 2340

cccagttatt gcaactggcg gagataatgc aatcaacgcg gttgcagctg caccgtacgg 2400

aagcgcgtcc attcttctca tctcctgggc atacatcaag atgctcggtg gtgacggtct 2460

cgcacaagcc tcaaagatcg cgctgctgaa cgccaactac atggcgcatc gcctgtccgc 2520

acattacaac cttcgcttca agaacggcaa cggccgcgtt gcgcacgagc tcctgcttga 2580

cctcgctgag ttcgacaagg ctgcgggcat caaggtgaac gacttcgcga agcgtcttca 2640

ggactacggc ttccacccgc ctacatgttc ttggcccatc tcgacgtgca tgctcattga 2700

gcccacggag tcagaaacgt tgcaagaaat tgacaggttc tgtgatgcca tgatccaaat 2760

ccgccaagaa gcagaagaca tcatcacgaa caagcagccc aaagacaaca atgtgttgaa 2820

gaatgcccca catcctgtct ctattatcgc cctctctgaa gaggagtgga accgcccgta 2880

ttctcgccac caagccgcgt atcctgtgcc agggctactg gagaggaaat tctggcccac 2940

ggtgtcgagg attgacgatg catatggtga tctgcatctc atctgcgatt gcccttctgt 3000

agaggagctg gctgctcctt ccccgtga 3028

<210> 4

<211> 939

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 4

atgggcgcat tcgggttgaa tgctagcatt ctcgactccg ctaacctcgc gtggaagatg 60

ggactctgcg cacgtgggtt ggccgatacc gagaagctga tgcccactta cgaacacgaa 120

cgccgtcggc atgctgtgcg cattatcgaa acttccggca cttacttgcg ctttgtctgc 180

gcttcaaacg cacccattgt tcgcctggat ggcgtgggta cggaggctcg cgaggacgat 240

gacgatagac cggccctccc gaaggagata accgaagaag cggatccaga caggcgattc 300

ttgaaggagt tcttcgctaa gctcggggcg ttcttgctgg gcgtggactt cgagtacgga 360

cacaacctgc tcaacccgcc tcagcagatg aggaacgacg tctctctgtc ccgcgttttg 420

ctgaagcctg cgacggaagt cgcgtggggt gtgcgcgctc ccagcccgcg cgtcacgctc 480

agccagcaga agacgggcta tctgtatgac gtctgtggcg gggcggacaa gttcacgctc 540

ctcgtcttcg cgtccaactt ccagggcccc atcctgcgcg gtcttaccgc gttggatgcc 600

catctcgcct catctcagtc gttctacaat cggtttggcc ggtcgaacat gtttaagatc 660

gtcctgatca ccaacctcct cccactcgat tacgaggacc atttcaccgg cgatgccacg 720

ggcactctgg agtaccttcg caagattgcg acgctcgtct gggacgacca gctgccaggc 780

agggatgcgc acacggtgta cggagtcgat cacgcaaagg gcgctgtcgc cgtcgtgcgg 840

cctgacttgt ggaccgggat atccgttctg cccggggaag cagagatgct ggacgaatat 900

ttcgaaaggt tcttgacggt ccctgggaag aaatcgtag 939

<210> 5

<211> 1596

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 5

atggttgagg tcaagaacaa agtctgtctg atctggtgga atcgtcgctc accccgtcaa 60

gatagtgtgc acgacggctg gggtatcgcc cccgagaaga acatgaaggg cgatgccatc 120

tccgccgggg acacgcccaa catggacgcc ctcgccaagg agcacaacta ccgcacgttg 180

cacgcccacg ggatctccgt cgggctcaac gacgggctca tgggcaactc ggaagtcgga 240

cacctgaaca tcggcgcggg ccgtattgta tggcaggaca tcgtcaggat cgatgtgtcc 300

atcaagaagc ggcagttcca caagaacgag accattgtcg cgagctgtaa acacgcgaaa 360

gagggcaatg gccgtttgca cctcctcgga ttggtttctg acgggggtgt tcactcgcac 420

attcgccacc tatatgcgct cctcgagacc gcgaaagagc agggcgttcc gcacgcatac 480

atccactttt tcggtgatgg tcgcgacacc gcaccgcgct ccgcggcggg ctacgccaag 540

gaactgctcg agttcatcga gaaggagaag tacggcgaga tcgcgaccgt cgtcgggcgg 600

tactacgcga tggaccgcga caagcgctgg gagcgaatca agatcgccgt agacgggctt 660

gtgaagggtg tgggcgagaa gggtgaggat gccgtcaagg ccatcgagga gcgctatgcg 720

aaggacgaga cggacgagtt cttgaagcct attatcgtca atggcgacga gggccgcatt 780

aaagatggcg ataccctgtt cttcttcaac taccggtctg accgtatgcg cgagatcgcc 840

acggttctcg gcctccccga caagcccatg gaggtagacg tgcccaagga tctccacata 900

tctacaatgt cgaggtacaa cgccgacttc cccttcccag tcgccttccc gccgcaggcc 960

atgacgaacg tgctcgccga gtggctctcc aagcagggca tcaagcaggc gcacgtagcc 1020

gagacggaga agtatgcgca cgtcacgttt ttcttcaacg gcggcgtcga gaagcagttc 1080

acggcggagg agcggcacat gatcccgtcg cccaaggtcg cgacgtatga caagcaacca 1140

aagatgagcg tgcagggcgt cgcggacaag gtcgcggagc tcgtcaagca gggcaagcat 1200

gagttcgtca tgtgcaactt cgcgccgccg gacatggttg ggcacacagg cgtttacgac 1260

gccgcggtcg aggcaatcgg cttcacagac aaggcagtcg cgacggtata taaggcctgc 1320

gaggcggcgg ggtacgttct gctcgtgacc gcggaccacg ggaacgcgga gcagatgatc 1380

aacccggaga cgggcgcgcc gcacaccgcg cacacgacga acgccgtgcc cttcatcatg 1440

accggcccca aggagctgaa gttcgcagag gataagaagg atggtgagga agaagagggt 1500

gcgctgtgcg acgtcgcgcc gaccgtgctt gctgttatgg gactacccca gccagaagaa 1560

atgaccggcc gttccctact tgcgaaggcg tcttag 1596

<210> 6

<211> 1521

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 6

atgtcagtca ctcaccaaga tacgacgtct gttgaagaga aagctgctgg ggctttccag 60

accgcgacgt tccttgtcga gccgttacat ggggagagga gactcgtgag gaggttggac 120

gcccggatca tgccgatcgt ctgcctgctt tacctgttcg cctatctgga cagaacaaac 180

ctagggaatg cacgcttgca gggactcccc ggggacgctc taaatggaga ccctacgggt 240

gttctgtttg aatgggtcaa ctccgtgttt ttcttctcct acattatttt ccaaattccg 300

gcaacaatta cctcgaagct tttccctcca cgcatatggc taggttgctc tgcgcttgga 360

tgggggcttt gttcaactct gtcggcctgc ggtttcaact tcgccggttt aacagttgca 420

aggatcggac tcggtatgtt cgaagcagga tttggtccag gaattccgct ttacatgtca 480

ttctattaca ccaagcaaga gatcgggatg cgattagcat acctgtttgg cttcggtgct 540

gtcgcaggag cctgtggagg gctcatcgcg tttggcgtcc agaatgcaca cacggcactg 600

gcgaattgga ggctgttatt tatcattgag ggtttgccga cgatcttcct tggtgttctg 660

tgcttgttcc tcctgcccga ccgtcccgag gagacaacat tcttgaacga acgagaacgc 720

gagttagcta tggagcgagt taatagagga atcaaggcag acatagggag gacaattaag 780

aaaaaacaca ttgcggcggc ttttctggat tggaaggtgt gcatctcatc gccgcttcgg 840

gtctgtcaat cgactcattg tccttcacct cagatttacg ctgcaggtgt gatctacttt 900

ggtgcaaact gtgcactggc atcaatatct gctttcctcc caaccatcat cacgaccttc 960

ggatactcga atgcgctcgc acagttgctc acggtccctc catacgctgt tgctgccatt 1020

attctttgca gtacttcata ttgctcagat aggttgcaaa gtcgtggctt ttccgtcgtc 1080

gctgcgagta tgctaggcgt tctgggctat gttctgttac tcgctgttcc ttataacgac 1140

catgtccggt attttgcaac attctgcatc acaagtggaa catacacgat tattggcgtt 1200

gtcattgctt ggttcacaca taatcttggg tcggagacta aaaaggctac gggcatcccg 1260

atgtacattg ccattggaca gtgcggaagc gtactgggct cccatttatt cccaactaca 1320

gacggtcctc gattcatcac tggatttgca gtttcatgtg ccctggagtt cgtttccgcg 1380

gtaacagcta ttattctcac catgtattat cgcatggaga acaagcgaag agacggtaag 1440

tttgggaagc ccattaggga tgcaatggta gatacgtcag aactggctga tatggccccc 1500

gattttcgtt acgtcccgtg a 1521

<210> 7

<211> 889

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 7

atgtcgaacc gtgaagaatt cctaacctcg accgacggca cgaagctctg ggcagaatcg 60

gccggcgacc cttccaaacc ggcgctggtc ttcatccacg gcctggcatg cactgccttg 120

ggattcgaga ggcagttcac cgatccgaaa ctgctcgaga acttgcatct ggtgcggtac 180

gagatgcggg gacacgggcg gagcgatatg ccagaattcc cggaggcgta tggttccatc 240

caccatgcag aggattttcg caccgtgtgt gaggcgtttg ggctgcacag accattcgtg 300

ctgggatgga gccttggtgg ctgtatcccc gtggacatcg tcgccgcgta cggcgcggac 360

ttcatctccg gcgtagtcta cgtcggcgga cccgtcctat ccttcaggct gaacgaagaa 420

tgctggcatc cgctcttcca cgccatgatc cccctgattt gctctgagga ctcagaagtt 480

gtagccaaga gcgcgtccat gttcttcgac tcgtgcgtcg cggatcccat cccgtaccct 540

acgaagctgc tgtggatggg cggctttggc tcgcagccgc cgaagattag gcactattcg 600

ctccgcgcgg gcaggacgac acgcggtggc ggccgaggcg gggaagctgc ccgtgctgat 660

cctgcagggg acggaagata cgcactgttt gtatgagagg atgctcgagc aggcgagggc 720

ggtctatgac gatgtggaag tgcagttgat ggagggggtc gggcacaccc cgcattacga 780

acggcatgaa gacaccaatc ggcatattct tgcttttgtc gagaagaaag cgtcccaagc 840

gcaggaacga gaggctcgga ggagcagatg tagggtctca ccaggataa 889

<210> 8

<211> 2442

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 8

atgaccgtca ctcgcgggtt cacggaggaa gcggacaaac ttctagacta cctcgaacac 60

ggaatttttg atgctttgca gaagcaatac ctccgcagtt tcatattcgc aatctacctc 120

gataatcaag accccaacaa tatcgtggaa gcatatacct tcaacttcaa ggtatgctcc 180

tgcgctagct gctatcttcg gaacactgag atgatccgcc ctcagtacta tactcttcct 240

gggactgaca ctgtagtgcc tgtgatgtct cttgacgcag accttgagaa actatccctc 300

accaaaacta agaaagttgc cgatccagtg gcagacgcaa tgaagaatgg aagggtgcct 360

accttgggag aagtgaagag aagcctcaag aactccagcg ctgatattgg catcacccca 420

gagcgccgat ttgccacgtt caagctcttc tatcaagatc acactccgga cgattacgag 480

ccgaagcact ttcgttcggg tgatgtgaag aaagataagt ggttcttcac cacacatgac 540

aaaggagaag tcccagagag gtgcagtgtg ggttctctcg agaccggcta tcacggcgtc 600

aacgtacgcg tgacttcggt ctctgcgtac ttgccctcag ccgaggataa caatgcgccg 660

ttcttgggca cgaccagcgg acacgcccat gcagcgccga tcctttcgcc agcggaagaa 720

gccgccttgc ggaaccagca aacgcagata cagcgccaag atgctataga caggcgggtc 780

gtttgggatg cagacgatgg tcttggcgat gcggacgctg acggtgagga tgacccagat 840

gttacgtcag gtgaaaatgc gagcttctac cttggaagtt ggcgagtgca tgattctggt 900

gtcgagttcg tcatgcccat tggtataagg gacgacgaag ggaatatcaa gcctttctct 960

aaggaagaaa gacaggagcg tgcagttagc cgagccgaag aagcgcaata tactggaaag 1020

caggacagcg tcccaagtca cgtagctcaa ctggcgcaag ctgaacgcca ccccgcggaa 1080

gcgataatgc gaacgcaaca actggaccat acccaggtga tcgagacacc cctcagcgcg 1140

ctctcccgcc gtccgtcccc gtctccttct cgctcgccat cccctactcc taagcagagc 1200

aagcgaggga cttccgggcg tataccgtca gcagggcctt ctcttcctcc ttcagaccta 1260

gatgcacagt cgttctcttc gctgtccagc ctcgcggacg gtatccagtc catcgatacg 1320

cagatggtga aagatctcat tgcgaatgct gcggctacgg aaattgaaga taccgagatg 1380

ttagacatgg aaactcaggt tgttcctcaa gcctcagctg agaatgattc tattcagtcg 1440

ttctccaaca tcgccggcac ccagcgatca cctcatattc gacctccaag gagtacagca 1500

gataaagaac gcagcgacgc cgaagacgat tcttcgctgg attgtgaatg cggcgtaacg 1560

aacgaagata acgactgctg catgtgcgag ggtggatgca agaggtggtt ccatatatgg 1620

tgtatgggct tccactcggc tcaggacgaa cgcataccag cagtctttgt ctgctttgac 1680

tgccgtgtga gggcggacaa gaactgggac ctcatcatgg tgcatgatct gtacccgagg 1740

atgctcgcgc ggttcaggga cctggccata ttcaggagag cgatcaagat cttcgagatg 1800

tataacccgg agaacttgtc ttcgttcacg aagttgatgg attgcgattc tgtcgtcgca 1860

gggcagctgt tcaagcgtct cgagacagaa ggcttcatcg cgaaggagac catagaggtg 1920

gacgacctcg gtctgatgga gacgagcacc cgtaccgcca agggaaagca aaaggggaag 1980

gctggggcga agacgagaca gtcgcagcgc cggaagacca tgcagaagcc aaagtacgtg 2040

tttgtgcggg ctaccacgaa gggtcaggcg tacgacgact atttcaatcc ggatcctgag 2100

gtcgagaaga gactgcttgg gttatctaac ctgaaaccag cgcgcagatc gcacaagaag 2160

agagtagtgg aggacatgga tgtcgaaatg gacgtaccgc tctcccagcc acggctggtg 2220

gacgttgata tcaacatggc ggtggattcg aacacagatg gcttgcctac aattctcgca 2280

gagtctcaga cccaagcaga aactcagcag atcgatgcgc atggcgaggg cgaaggtggc 2340

caaatcaggc gcaagacgac gaggacgttc agtcaagagg tgccccggac gagcaaaaaa 2400

atcaagatgt ccgtcggacc agcggtggac cttggggact ga 2442

<210> 9

<211> 1288

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 9

atgtcccttc cgcttcgcaa aattggaacc acgccggtga ccgccatcgg ttacggagct 60

atgggaatat ctgccttcta cggaaaggcg ctgcctgacg aagaacgctt aaagattctt 120

gatgccgtgt acgagaacgg ttgcaccatg tgggataccg ccaatatata cgatgacagc 180

gaggtgctca tcggaaaatg gttcaaacgc actggcaagc ggaacgagat tttcctcgcg 240

accaagttcg gaattttcaa ctccggcggc agagctatca acggtgatcc ggagtatgtc 300

caccaggcat tcgaaaatcg ctgaagcggc ttggtgttga caaggtcgag cttactatct 360

gcatcgtcca gactccactg tcccgatcga gcgcaccgtt ggtgccatgg cggagctcgt 420

aaaagccggc aaagtgaagt acctcggtct ctccgagtgt tccgccgaaa cgctgcgccg 480

cgcacacgcg gtgcacccga tttccgcact tcaagtcgag tactcgcctt tcaccctaga 540

cattgaggac gagaagatcg ggctgctaaa gacggcgcgt gagctcggcg tcaccatcat 600

cgcgtactcg ccactcggtc gtggtctcat cacgggaaaa taccggtctc ccgacgactt 660

cgantatgtc cagcgcaccg ttggtgccat ggccgagctc gtaaaagccg gcaaagtgaa 720

gtacctcggt ctctccgagt gctccgccga aacgctgcgc cgcgcacacg cggtgcaccc 780

gatctccgca cttcaagtgg agtactcgcc gttcacgctg gacatcgagg acgagaagat 840

tggtctgcta aagacggcgc gtgagctagg cgtcaccatc atcgcgtact cgccactcgg 900

ccgtggcctt atcacgggca aataccggtc ccccgacgac ttcgatgagg acgacttccg 960

gcttacggtg ccgcgatact cgaaggagaa cttccccaac atcctcaaac tcgcggatgg 1020

cctcaagagg atcggggatc gccacggcgc gacggctggc caagtctcgc tcgcgtggct 1080

cctcgcgcag ggccccgacg tcatccccat ccctggaact accaagattc cctcactgaa 1140

ggagaatttg ggcgcggcga agcttacgct gacgccggag gagatccagg aggtgcggga 1200

gatcgcaaag gcagcagatg catcgaacgg ccctcgttac cctggcggta tggagtccct 1260

cttgtttgct gacaccccgc cgttgtaa 1288

<210> 10

<211> 1350

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 10

atggctgacc gtgttgcctc gtcgaaatgg tctgcgttca tgagggactc gtatgtcaag 60

cgcagaatcc ctgttggcgg tacaattgat gtcaagaagc tcgaagaaac tgcacgacag 120

aagctgaagg atcgacaaga tgcatttctc tatgccttcg gtagtgcagg aacgagttct 180

acgtgtgacg ccaatttagt agagtttact aagtggagga tcataccccg aatgctgcgc 240

gacgtcacaa tgcgcaattt agaggcaagt acatcatccg catgcctatc tcacacgttg 300

ttgaatcagg attccgctcc acagaccact atctttggag tcaaatatcc atcccctctg 360

ttcctatctc caatcggtgt ccaaaccctt tatcatgctg atggagaact tgcttctgct 420

gcagccgcgg cgcaagtcgg tgttcccttc atcatgagca ccgcttcgac ccgcagtatc 480

gaggcggtcg ccaaagccag cgggtccggt ccccgctggt accagctgta ctggcccgtg 540

tcgcccgatc tgacactatc gttgatatct cgtgcaaagg cgaacggctt cagtgcgctc 600

gtgattacgc tcgataccat gcttctgggt tggcgcccac atgaccttga caccgcctat 660

ttacccttct tccacggggt cggcacgcag attgggttca cggacccggt gttcatggcg 720

aagcatggcc tccagccgtt cccggaagac gatgcaccat cgtttcccta tgacccagag 780

gagatggaaa ggcgtatcaa ggctggggac gagacgttga agcagcgcgc gtacttgggc 840

gtgaagtaca tgggagagac tgcttcgggc gttttcagaa gctgggacga cctgaagttc 900

atccgtgata actgggatgg accgctaatt ttgaaaggca tcctatccgt tgaaagattg 960

caatggacaa tggcattgat ggaatcgtcg tgtctaatca cggcaagcgg ccgtcaggtc 1020

gacggcgccg ttcccgcact gtatgcgctc gagcaaatca tgaagagccc aagggtgaag 1080

gaagcccagg caagcggcaa gttgacaatt ctcttcgact ccggcatccg caccggcagt 1140

gacgtcttca aagcgattgc gctgggcgca caaggtattt tatatgcaag gccgtacatc 1200

tacgggctca tagttggcgg ccaagcgggc gtcgagaacg ttctcagaag tgttctcgcc 1260

gacactgaga tcactctcgg tcttagcggt tatgagagtt tgcagcaaat acatgggaaa 1320

gctgacaagg ttatggtcaa gctggggtaa 1350

<210> 11

<211> 2298

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 11

atggtcgcct ccagcgtttt gggtttcccc cgtatcggtg caaaccgtga ggtgaaaaaa 60

gccgtggaaa gctactgggc tggcaaaatc tctgccgaag ccttaaccaa ggttgctgcc 120

gacgtcaaga agactacctg gaccagcctc aaggccagag gcgtagatct catccccagc 180

ggtgatttct ccctctatga tcacgtcctc gatcactccg ccgctttcaa cgccattccc 240

aagcgttacc tcggccatgg cctctccgcg ctcgatgtat atttcgccat gggccgcggt 300

cgtcaggcgg acggcgtcga cgtgcctgcg tctgagatga agaagtggtt cgactccaat 360

taccacttcg tcgtccccga gtttgcagag gacaccgatt tcaagctcaa cttcaacaag 420

gccctcgagg agtacaacga ggcgaaggcc atcggcatcg tcacccgccc cgtcgtcctt 480

gggcccctct ccttccttgc cctcggtaag gctgccaagg aggcgaaggc cggtttcgag 540

cccatcacac tcctccccag gctcctcccc gtctacaagc agcttcttgc tgatctcaag 600

gcggccggcg tggagtccgt ccagatcgat gaacccgtgt tggtcctcga ctctgctgct 660

ggtctggaaa agcggttctc tgctgcctac gtggaacttg agcccgtctc tcccaagatc 720

gtgctgacga cgtactacgg gcgcctcgac tccaacctca cgttcgtcgc caagctcccc 780

gtcgccggtc tccacatcga cctcgatcgc gctcctggtc aacttgacca ggtgatcgca 840

gctgttaagc ccacgagcat cgtcctgtcc ctcggtgttg tctccggccg caacatctgg 900

aagaccgacc tccaggcagc aatcaagctc ggccagaaga cgatcgatgc cctcggcgct 960

gaccgcgtca tcatcgcaac ctcctcttcg ctcctgcata cgcccgttac gcttgatgct 1020

gagaataagc tcacgccgga tcagaaggac tggttctcgt tcgcgctcga gaaggccagc 1080

gaggtcgccg tcgtcgccgc tgttctctcc ggctcccagg accccaaggt agccgaggcc 1140

ctcgcggcga acaccgcgtc catcgctaag cgtcgcgagt tcgagcgtac atcggacgat 1200

gcggtacgga agcgcgtcgc tgctatcacc cctgatatgc tggagcgcaa gagtcccttt 1260

gctgtccgca aggaggtgca gcagaagtat ctcaacctgc ctaagttccc cactactacc 1320

atcggttcct tcccgcaaac gaaggagatc cgtaccgctc gtgcgaagct caacaagaag 1380

gaaatcaccg tggagcagta caacgagttc atcaagaagg agatcgagag cgtcgtacgc 1440

ttccaggagt cgatcggcct tgacctcctt gtccacggtg agccggagcg gaacgacatg 1500

gtgcagtact tcggcgagca gctcgagggc ttcgtcttca cccagaatgg ttgggtgcag 1560

agctacggct ctcgctacgt ccgtccccca atcgtcgtct ccgatgttcg ccgttctggc 1620

ccgatgaccg tcgcgtggag cagctacgcg cagagcctca cgaccaaacc catgaagggt 1680

atgttgactg gccccgtcac tatcctcaac tggtccttcc cccgtgcgga tgtctcccgt 1740

gagctccagt gcaagcagct tgcgcttgcc ctccgcgacg aagtcggcga cctcgagaag 1800

gccggcatca aggctgtcca ggtggacgag cccgctcttc gtgagggcct gcctctccgt 1860

aaggccgact gggagcctta cctaaactgg gcgctgccgt catttaagat cgcgacggcg 1920

ggcgtgtccg acgcgctgca aacgcactcg cacttctgct actccgactt cgacgacatc 1980

ttcccgtcga tccaggcgct cgatgcggac gtcatctcca tcgaggcctc caagagcgac 2040

atgaagctgc tcaacacgtt caagcactac gggtactcga accagattgg gcccggtgtc 2100

tacgacatcc actcaccgcg tgtccccggc gagcaggaga tcaaggaccg catagcgtct 2160

atgctccaga tccttcccga gtcgctcctt ttcgtcaacc ctgactgtgg tctgaagact 2220

cgcggctgga aggagacaga ggcatcgctc aagaatgtcg tcactgccgc ccgctgggcg 2280

cgccagacat acgcttaa 2298

<210> 12

<211> 1441

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 12

atggatatga tcacctattt tatgtgggca gtcgccgtca tagcttgtgc ggcgctgctg 60

agcaggaaca agcgaaaccc actctacctc cttgtcctcc tgcagatcca ttaatcggcc 120

atctgcgtca attcacgttc gctcctcaac gtgagctctt ccagaaatgg gctacggctt 180

acggcgacgt cttccatctc aatatcttag ggcagattat cgtagttgtg aactccctcc 240

aagtcgcaac tgacctcctc gagaaacgga gtgcgaacta cagtgacagg cctgcttgta 300

ctgccctgga gatgatgggc tggaaggcga acgttgcctt tatgcggtac ggcgcgcggt 360

ggaggaagca cagaaagctt ttccaagagt actttggcca gagtcaaagc ctgacatacc 420

gcgcccatca gactgaggaa gcgcgtatgc tcctgaaaga tttgctactt cgcccgcgga 480

gttcaaggca tgtacgcaga agtatgctct tctgagcatc ataggcattg catacggaca 540

tcaggtaaag ttggatgacg acgtctatgt caaaatcgca caaggcgcaa tccacggact 600

cgaagaagcc gctgcaggaa ccacgctgct cgacctctta cccttccccg aacaggaagc 660

ggggaccgcg cagaattcgt tctttgtgtc tcatttggag cgcctcggca cggacggcgt 720

ggacgaagaa gacctcgagg acatccaggg tgccgctgcg acgatactta catcctcgct 780

catccagagt ttcctcatgt tcatgctgct gttccccgac gtgcagcgca aggcgcagga 840

agagatcgac caagtcgtcg gctccgggcg gctcccagac ttcgatgacc gcgatacgct 900

gccgttcgtg gaatgcgtgc tccaggagac catacgatgg tacccaatcg ctccgttcgg 960

tagagaaccc ccgatagatc ggcactcgga tgttgctgac cttagacccc tagctattcc 1020

tcaccgaagt ctcgacgacg acatatacaa cggaatgctt attccgaaag gctctgtaat 1080

catacctaac gtgatggcaa tgagccgcga tgagacaatg tacaaagatg cagacaagtt 1140

cctccctgag cgcttcctac cggctcctgc cggtcgcggc gagccgcatc tcggcgtcat 1200

ctttgggttc ggtcggcgga tctgccccgg gcagtatttt gcagacaaca gcgcatggat 1260

cgtcatcgcg agtatcctcg cggctttcga catctgcaag cccgttggag cggacgggat 1320

agcagttgag ccccacatgg agtacaccac tgacgggatt gcaagccgtc cgatgccttt 1380

cgagtgtgtt ctgcgaccgc gctcagagac ggcgaaggcg ctggttgagc agcacacgta 1440

a 1441

<210> 13

<211> 519

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 13

atggctctca aggtcgacaa tattccccgc ctcagtggaa cagaccttgt tcatgctgat 60

gatcagttgc acggtcctga agtcgctcct tccatctcag taacttccac attccgtgct 120

gagcagccgc tcaccagcac aacggtggga agtgatgacc acgatttcga ccccctcaac 180

ccgatcaacc acgtgtactc tcgctacacc caaaacgtca gcagcagggc cgagaaagtc 240

ctcagcaaaa tcaacggagg ttatgctatt acctttgctt cgggactggc tgcgtcatat 300

gctgccttag tacacctgaa gcccaagcgt gtcgccatca caggtggtta tcacggatgc 360

catctcacca tacaagtgta caagcagagc cgaggcgagg ggttgcccat tatcggcata 420

gacgacagtt tccagcctgg tgatctatgc tggctggaga cgcctctgaa cccgacaggt 480

gaagccagag atattcagta ttatgctgac aaggcatga 519

<210> 14

<211> 2121

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 14

atgctctcac tcgcccgcgt gcggtcggtc gcagagctgc tcgcgatgct ccttgcaagc 60

gttcggacgc gtccgctcaa gctcctcggc cccgccgctg cggtgagtgc actcgtgctc 120

gtcctgcggc atctctgtgc gcgtcgggca catacgcagg cgtatactcg agcgctggcg 180

ggagacgcgg cgcgtgtggc gcgccgtatc aagagcggag acgaggagta cgatccagag 240

gagtatgacg tcattgtcgt tggcggaggc accgcgggct gtgtggtcgc ggcacggctg 300

tcggaggacc cgtcgatacg tgtcctgctt atcgaggccg gcgagagctc ccggcatatg 360

gtgttctcgc agataccatc tgcctttgct tctctatttc atggcaagca cgaatacaat 420

ttctacacca tcccgcaacc aaacgctggt gggaaggcaa agtactggcc gcgaggcacg 480

ttgcgttatt ctcgttatcg gttggacgct aacaattgtt cgatggtagc caagcttttg 540

ggtggatgtt cctcattaaa tgcgatgatt ttccaccact gcgcgccgtc cgactatgac 600

gaatgggtaa ggtaccagaa aggcgcggag ggtgcagagg gctggtcgta cagcgaattc 660

ctcccgtact tcatcaagtt cgagaatttc caccccagtg aaaaatttcc cctcgttgac 720

ccgacgctgc gtggatccgg cggtcctgtc gccagtgagt ctcctccctt gtatatgcat 780

atggagctga tcgaacctgc agctgggtat tttggaaatg ctgcggtgca tacgatgaat 840

ttcatcgagg cctgcgacag ggtcggcata ccgcgcaatc cggacctgaa cacgaagaac 900

gcgttgggcg tgagcaagac catgacaacc atcgactcca acggccgccg tgtgacgaca 960

gagaccgcat acctgactcg caaggttctc gctcggccaa acctcaaggt tgtaaccaag 1020

gctcgggtgt tgcgcattgt cttcgacgtc agcagtgaga caccgcgtgc ggtgagcgtg 1080

cagttcctcg acgagcaggg caagcgcttc gaggcgaagg cacgtaagga ggtcgtcctc 1140

agcgcgggtg cgatccatac gccccagctt ctcatgctgt ccggcgttgg gcccgccgca 1200

caccttgcgg agcatggcat ccccgtcgtc gcggacctgc ctggggttgg tgcgcacctg 1260

atggaccacc ctacgatcga cctgtacttc agggacaaga cgcgggcact ggaggctgga 1320

atgtcgcaca cgcccaaggg gttcatgaag acgctggggt tcttccgtgc gatgctgcag 1380

taccagctca cgcgccgagg accccttacg acgaaccttg ccgaggctgt tgcattcata 1440

cgttcgtccg accgctcact gttcctcaag gacgagttta tcccagatgc ggagctcgag 1500

gacacatcgt ccgcgcccga cgcgccggac atcgaactct tcagctcgcc gatggcgtat 1560

acagagcacg gatatggcca gcctttcgac gggtacacct ttgggctgca tgccacgctc 1620

ttgcggccaa cgagcacggg caccatccgc ctcagctccg ctgatccgct cgacccaccc 1680

attatcgatc ccaactatct ctccacgcgg cacgatgtcg cgctcctcgc gcgtgccgca 1740

cgcctgctcg cgcgcatcgc gtatacggag ccactcgcgg gcatgcttga ccccaccggc 1800

gaggcggagc cggcgttcaa tcataaattg cttgcgctgg acgacgcggc gctcgaggaa 1860

ctcgtgcgcg cgcgcgtgga gacgctgtac cacccggcgt gcaccgcacg catggcgccc 1920

gccgcggacg gcggtgtcgt cgacccgttt ttgcgcgtgc atggcattcc gaacttgagg 1980

gtcgcggatg cgtcggtgtt tccgacgatt gttgcgggcc acacggcact tgagattggg 2040

acacagacac ggagcgacgg gtggggctcg gtacgtgagt cgctcaagta caccatctgt 2100

gaccaccggc caccgtcgtg a 2121

<210> 15

<211> 366

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 15

atggtcttcc cagtccgagg ctttgcgaat ggcagaagaa gtgagcaagc gcgagcggca 60

cgtcactgtg ctcttcgaca acgcagggat cacgggcgca cgcgctccgc ggcctgcggc 120

gcccaccgca gcggagttcc gcaaagcctt cctcgacggg gtccccgagg aagccttccc 180

caacatgaac gccgtggggc tcttcgtcac atcgatgaac acgctgggcc aggtcgacga 240

gctcgggcgg gtaacacctc ccgcccggcc gtgagttccc gttcgaggtt cccgccgtgc 300

agcccggccc gcagtcgctc ctcgggggct cgaacaagga cgtgggcacg ctcgcgctgt 360

gcttag 366

<210> 16

<211> 366

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 16

atggtcttcc cagtccgagg ctttgcgaat ggcagaagaa gtgagcaagc gcgagcggca 60

cgtcactgtg ctcttcgaca acgcagggat cacgggcgca cgcgctccgc ggcctgcggc 120

gcccaccgca gcggagttcc gcaaagcctt cctcgacggg gtccccgagg aagccttccc 180

caacatgaac gccgtggggc tcttcgtcac atcgatgaac acgctgggcc aggtcgacga 240

gctcgggcgg gtaacacctc ccgcccggcc gtgagttccc gttcgaggtt cccgccgtgc 300

agcccggccc gcagtcgctc ctcgggggct cgaacaagga cgtgggcacg ctcgcgctgt 360

gcttag 366

<210> 17

<211> 321

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 17

atggaacgtg gttgcgcgtt ggagttgtac gtgatttcgc tgggtgctta tcgtgtattt 60

gttctaccag cgcgccacag aactttgtcg gaatgccgta ctatcgtacc agacgtcgag 120

gtgcgcaaac acgaactctg cggtgaagtt agggttccaa attggctgtt aatggcgact 180

ccaggaccgg tgaaattcca gggttggctc gaagcacaag ttgacgatcc gcgggacaca 240

cttgggcttc tctcgctcgg gatcgcgcca ctgcgcgacg tggagctccg aggacgccac 300

cacgcatcgg cgggtatctg a 321

<210> 18

<211> 820

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 18

atgcccgact accccgagtc gtcccaccgc accgtcctcg ccgatccccg cccgtccggg 60

gggccctccg cccgcccgcc gcagtggtcc cgggcgcagc agctgtacac catcgagtcc 120

gagcccgcgc ccgccgtcga ccgggcatct tcctccccag cccccgctcc ccgtccccgg 180

ccgagaacat caagccggag ccctcggacc ccgactccga cttcatcttc gagctcgagc 240

cgcctccccg cccccgaccg atcccgacca tctctcctcg tgcaccgagg tccccctccg 300

cgccaccaac gcatcccccg cgatgcgcaa gatgatgagc gtcttccgcc tgcacccctt 360

cgccgtccac aacggccgcg gcgctgccgt cgctgccgtg cccctcgtcg acgtcgggcc 420

cctcaccgag gagcccatca tggtcgagtt ccaggtctat ctcccatatc ccctcgtccc 480

ccagtctccc gagccagacg acgaccgccc ccagtacacc cgccagcgca tgcacgcccg 540

cgaacagccc cccgcttggc tccgctcccc atccccagcc tccagcacat atccagaccc 600

atccgacgcc cacgaggacc agcgcaactg gcgctttgcc gacgcggacc tcccatatcc 660

ctcgagcccc gtcgaatctc cccccttcgc gcccctcatg acccccgccc agagcctcca 720

ctggagcatg cgctacggcg tcgacggcac catccccttc ccaacacctg ccgaagcgca 780

gggtgagttg cattaccctc ctacgcgtca acgctgctga 820

<210> 19

<211> 2145

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 19

atgtccagct ccagatcttg cgctcccagc cgtccacctt accggccgaa gccgacctct 60

ccttgccacc cactccacac agcggcacct ccgactggac acccttggcg ggcgaagaga 120

acgccgagcc cgaaaatccg cgtcgttcag atgatgacaa cgaactccac gcggagaaag 180

cggaggggtt catcctcggg cctatgccca tcgaggaatt cctcgacttc ctccctgcga 240

agcacacggc acgcaggccg tcgacgcagg gagcgttcaa cgacgtcgac gagcctgatg 300

gagcttccca gctgttggcg gccatcaggg actacgaccg ctgcccaggg ttcgagttta 360

aacacaacgc cctggacttg acgtcggtca agcccgctct gatctgcgtc ccgaaggact 420

cgatcgggtc ggacaacgag gcagcggcga agtacgcagg ggagctgttc attgacgtgc 480

aaagcggcga ccagggcgat ttcttccgcg acccgaagcc gttcgccgac cgcgcgcgcc 540

acaagttcgt cctgtcgcag cccgacgcgg agaagacgca ccgggcgaag gcgcggcaag 600

ccctggggcg gaacgtccgc tgggccaccg acgcgctgat gcaccaacac cggctgcact 660

acttctcggt gtcgatccac gggctgcgcg cgcgcctgct gcgctgggac atgagcgggc 720

tcgtcgcgtc ggagtcgttc agcctccggg agaagcccga ggcgctgtgc gagttcctgt 780

ggaggttcgc gcacacgtcc accgtcgagc gcggctacga cccgacggtg gagcgcgcgt 840

cgagcggcga ggagtgtctg ttccggcagc tggtttcatg ccacgtccgg gaacagctgg 900

acctgaagct ggtggacgtt ccggaggggg aggtcaagga tgaggtgcgg gcggagtacg 960

aggcgcagtg tgcgcttcat gagaagaccg tggcgctgca ttatgaggcg gggcgcgtcg 1020

cggcgctgac catctgtgcg gaggacggga ggctcaagcg gtgtctcgtg tcgaggccga 1080

tcgctacgaa cttctcgatg aggcggggtg cgatgcgcgg gtactgggcg gtgatggaag 1140

gcagggttgt tttcttgaag gacacctggc ggcagtacgt ccatgagccc gaaggccgca 1200

tactgaggga gcttgaagcg cacaaagtca agaacatccc gcatgtggag cacgatggag 1260

atgttccgaa ggtgatgtac tgtgacggca tgctcacgcg gttctcggcc atggcgaact 1320

tcgagaggac gactcggagc aaggcaggcc aacagccgtc attccaatgc actgagacgg 1380

acttacaccg ggaggccgaa tggctatgtt atggcggcgc tcggattgtg gtgcacccgc 1440

gggtccacca ccgtctgatc ctcaaggaag tcggcttccc acttcagaca tgcgaggcac 1500

gaaggagctg ctcaacgcga cgcacgacgc gttccaagat ttatctatta acctgcgctg 1560

atcgctgcgc acgacaaggc acggaggatc caccgagaca tcagcgcggg caacatcatg 1620

ctttatcggc agccatatca ggaccatcgg ataggttatc tgatcgactg ggagctctcg 1680

tcgaaggtac aggaggacgg gcaggcgacg gacaagtata tcgtgggcac gcgcatgttc 1740

ctgtcgcagc gtctcgaggc gcgaagccag cggaggcaca tgttacagga cgacatggag 1800

gcactcctgt atgtcgtcgc gcactgtggc ctccactggc ttcagcatcc tgcgcactcc 1860

aaggacacga ggcaagtcgt tcaggctatg ttccacaacc gcaaggaggt ccacgcgggg 1920

gatgtacctg gggtgacaca cacgacattc cagacgatgg agccgggcaa aggcaagctg 1980

gacatcaagg agaatcggac ctacatcgac ctcccgaact tcaacgcgga tctcaaggat 2040

tggctggtcg atggacctga attagccgcg ggatgcaaat gcggcgacgc agtgggacac 2100

gcccgagccg tggaacgatt actggaagga attcttggac cgtaa 2145

<210> 20

<211> 1116

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 20

atggctgctt taattttcgg tctatctcca gaaagctggc agagggcaga ccagacattc 60

aagacacttc tctccaatga agaacgttgg agagatagac agcagttcct tgagtctcgc 120

ggatacatgc tgcgtccaag atatcgtccg ggttggattc cttcatggac aggtacgaag 180

aaaaaccctg tgttctgtga ggatgcgaag acacttccag cacgcccgca cctcatagat 240

gccactcgac tctcggatgg caaactggtg tacatcaaga gagttcagac aggggacaac 300

gaatcacagc tcgctactat gttatcgtcc gaaactctgc gcaaggatcc gaggaacaat 360

tgtgttcccg tcattgatct gttccaggat tcagacgatc ctaccatttc ctacatggtt 420

atgccgtttc tccgtcttgt caatgatccc ccgttcacaa ttatcgaaga cttggtagat 480

ttcattgaac agatgctgca gggtctcgtt tttcttcacg aaaatggagt ggctcatcgc 540

gattgttcgt acaagaatgt tatgatggat gcaagtgcta tgtaccccca tggctttcat 600

ccggtgaagg aggattactt gcccgatgga gaaaccggag tagaacctct cccacgttct 660

accgtctcca tccggtatta ctacgtcgat ttcggcatct cagtgctcat ccctcctgac 720

gtccatccta aactcaccgt tggagaactc ggtcgagatc gagagccgcc ggagttgtcg 780

cctgatgtcc catacgaccc gttcaaactc gatgtctttc tcattggcaa tttattccga 840

cgtcttctct acgatgaata ttccaatgtg gaatttttgt cttctctttt cggcccgatg 900

attcgagatg atcctgcatc taggcccgat gcccaagacg cactgcaaca gtggcacaca 960

attcgaaagg gactgtcgtt cttcaaccgc aggtgggagc tgcggtctcg tgaggaaacg 1020

cggagcacat tcgtcttgga tgccatgtgc ttcgtcaaag tagcgatgta cgctgtcagg 1080

cagctgttcg gatgggctgt agatgttcaa gggtga 1116

<210> 21

<211> 795

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 21

atgaaccatc tcaatttgaa gttgccgctg ctgctgtctg cagcatatat agttcaactg 60

tgtgtgctgg tcccgcaacc gccgccgaag gaagatgcac aggcgaggtt tatctccgtg 120

gaaccaagac actccaggga gacgtttcgc tggctgccga agttcatcat ggtgagtccg 180

acctctgctc gacatctggg cgctcaccgt ggttgtacct caaagcggac agtctgggcg 240

gcctgtttgt gcgagtccgc gatcctcatt gcataccaat ttccggccca ccatctctcg 300

cgccgagtac tgcaagcgct catccgcggg cccgctccag acaccttcgc tggcgtcggg 360

atccgcgtta cagtcccgtt catcatcggc tggtgtttcc tcattgcagg aaccttcatc 420

cggcagtcgt gctaccgcac actcgggcac tttttcaccg ccgagctcgc ccttcacgag 480

gaccacaagc tcgtgacctc cgggccgtac acgctcgtgc gccatccgag ctacacgggc 540

ggcatcatgg gtgcgttcgg ggtcgccctc atgcagatgt gccctggctc gtggctgagt 600

gagtgcatgg gcatttggga tacccgcagc gggacggtgt tctgctactc gtggtttgcc 660

atcctaggca tcgtgtcggt gcagatgatc cgcaggacgg gtatcgagga ccgtgtcctc 720

aaggacgagt ttgggaagca gtgggaagat tgggctcagg gagccaagtg gaggttggtg 780

cccggcattt tctga 795

<210> 22

<211> 864

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 22

atgcccccga gctacagtcc ctgtcccagg ccctgcctca aacgcccctc ccaagatcgc 60

tgcgcctcac ccttgtctcc ccgcgacgaa tccacccagc tgctcgccat cgaccccggc 120

atcctccacc ccctcgtcca cttcgcaccc tccacagagc tcgccagcac gcactccgcg 180

tactctgcag ccgtctacga ccgctctccc atcgtcgtcc ttcccaatcg ctgcgccctc 240

cccgagcgcg gctgccccgg ccgcacctac aacgtcggtg acggcacagc ccccagcacc 300

tcgcgccagc gtcagcgcag ccccgccgcg cgccatcgcc atccccgcgc caaggacctc 360

gacctccccg aggacgagga cgacgatgat ttgacaccgc gcacctcccc gctcgcccac 420

gcctacgcgc tcccgccact cgtgcccgac ctctcctccg agtcggacga gtccgacgcc 480

gcccccggcg agcacgaccg tgcgctcatg ttcgcgcagt acgcgatgac ccagcgctcg 540

ttcgccggcg cgggcgagct cgcgcagggc atgtccgcga tgtacatcaa caccgccaag 600

ccctccgcgc tctccttcct cccgcacccg ccctcgccgc gtcgcgcgca ctcgcccacg 660

ccgccgtcgc gcgaggacga gcacgaccgc cgccgccatc ctccgtccgc gtccgcgtcg 720

tcaatgtcga gacgcaagtc gaagagcggc gagaatcgca gtccggagag tcccgcgcgg 780

gcgcggtata aggcgttctc ggaaaagagc gcgctcggcg gcgggttcgg cgtcggggac 840

ctcggctgct tgggcggctt ctga 864

<210> 23

<211> 330

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 23

atgttctccc gcatcgccac cttcaccgcg gtcctcgcgc tccccttcct tgctgttgcc 60

agccccaccg gccagaccga ccagtgcaac gtcggatcgc tccagtgttg caatagcgtt 120

atctcgcccg actcaacaga gggcttgggg ctcttgggtc tcatcggtgt cgccgtcgga 180

tccgtcacag gcagcttggg tgttacttgc tcgcccatca ccgtcatcgg agttttgggc 240

ggcagcagct gcgaggccaa ccctgtctgc tgcacagata acagctacgg caacttggtt 300

tccctcggct gcatcccagt ttctctttga 330

<210> 24

<211> 1224

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 24

atgccgttac gccctactgt caccgcggtt acggctggtg tcccaccctt tcatggctac 60

gtggatacca cggaggacgc cctctgtcta atagagatct atcgagagct cgcagatcga 120

gcggacactc gtacccacca cgaagctgca acgcactctg aagcatcgca ggaggaccac 180

tctacggaag gaggcgtcat gcttaagcga tatggcctaa taaaaaagac aattactgtc 240

aaaattgacg ggtcagacca ccacctcata tcgtactaca cccaagagga tgttcgttca 300

ggcaggcttc aacgtccgtc cagtcggaca gaccttcggt ctgttgaaat cccagcggaa 360

ctccttaagt cctctagttt tcggtatccg ctgaagtatg aatcgcaacg tggcagtaag 420

ctgaaccata taaatgatgg agataggatt gataatcagc gctcagctgt cccatcttcg 480

ccttcaagga gggcgacggg cgaacccgca ccacgtttct ccccgattag tcctgctcaa 540

ttctcacaca tacctgccca atcctccaac gcgttccatc aatctccaag caggaaatat 600

gagccgtcct atccgtcctt gtcactagcg cttcctgcgg acgttcggca gtctgcatcc 660

ccatctccca cagacacggc gtactccccc ctcagcgtcg atagttcgcc gctagagcag 720

agacgatctt cgccttggcc atccgcaagc ggcgtaactc catttcccga ccctgccgca 780

gtctacgaag caaggcaaaa tgccgtgcag acggaaccta gctcgatacc tgggcgtacc 840

tggcccaatc aacagcctgc tcgattcgac cgtacattcg atcaaggcgc ttcaaatgct 900

ggagccggcg cgcaccgcgt tggagtgtct gcatggccgc cagggaccgc tacgttctct 960

gggcaaacat acacctctgc gtcgccgatg tcgcctctaa cctccagcta ccctgggtct 1020

tccacacgag gatgcgacca gacgcgagcc agggtgtcgt ctcctgcgtg ggcatggatg 1080

caggactccg cttcttctca gtactgtcaa ttgcctcagg ccgctgcctc gacgttctcg 1140

acggagcgcc aggcatctgc aacagatttg ccatcacctc gaagtccaca agtacggccc 1200

ctgggatacc catacaggca ctga 1224

<210> 25

<211> 1826

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 25

atgtctcgca tcgcggacaa gggaaaggcc cgtgcgcgag acgacgacga ggacaatccc 60

ggccagcctt cacgccgctc accgcccgca cccgagccgg gccgttcgcc gtccctccgc 120

cgccatcctc cgtcccccga cgccatcctt cctctcctgc actgccccca atgctctccc 180

tcgcgtctcc tccacgcgcc tctcactctc cgctgcggtc acactctttg ctcgcaacac 240

attctgtctc cccccacctc gtccccgtcc ccgtccaacc ccctctccgc actctatcaa 300

ggctcagccc cctcatgtcc actcccaacc tgcaacgcgc cctcctccga gacacccacc 360

accaccacca ccaccaccag cagtcccggc gtacacttct ctccggcccc cgtgtcctct 420

tcgcgtccgt ccacacccgt ttcgcgcgtc gatgtctccg taaacaagat catctctctc 480

gtcgtccgcg cccacaacaa cacagacccc gacgcgtccg ccaccgtccg cacttacccc 540

gcagacatcg acgaccaaac cgatgacagc gatgccgcgg acgacctcga cgagcatccc 600

ttgccttcca ccgacgctgt ttccgatccc gatcgacaca gcggtctagg ccccgccgcc 660

cctgcctcta gccatccccg cccgcagtcc ccaccgcgtt cgcgcaagcg tcgccgcggt 720

ccccgcccgc gccaccgccc tcacgaccac ccgcccagcc cttcttcgga tcctgccgcg 780

cgattcgaaa aggagctgct caccgagctc acctgcgaga tctgctttgc gctgctctgg 840

cagcccgtta cgaccccgtg ccagcacacc ttctgctccc cgtgtctcgc ccgcgctctc 900

gatcacaacc tcgcctgccc cctctgccgc cagatcctcc ccgggtacga ctactttcag 960

gagcacccct gcaacaagct cgtcctctcc atcctgctcc gcgcgttccc cgaggctacg 1020

ccgagcgcgg cgccgccctc gaggccgagg cccgcgactc ccgcctcgac accccgctct 1080

tcgtctgcca gctcagcttc ccgggcatgc ccaccatcct ccactttttc gagcctcggt 1140

accgcctgat gctccgccgg tgcctcgccg cgccgtaccc cgccttcggg atggtgcccc 1200

cgccgcgcgc ctccgccggc gtggtggagt tcgggacgat gctcgagatc cgcaacgtgc 1260

agatgctgcc cgacgggcgg agcgtcgtcg agacctgggg cgtgtggcgc tttcgggtca 1320

tggagcgtgg gaccctcgac gggtatggcg tcgcgcgcgt ggagcgtgtc gaggactggg 1380

aggaggacga cgaggaggcc gttctggacg tgcggcctgt tgcgagtggg gaggaggggt 1440

ctgggacgcc cggtccgtcc tcgtcgtcct cggtcgcgtc gatgtccact tctccgcctc 1500

tttcggagcc cccgggctct gcgttgacga gcgttggccc gtcgacgatc ccgcgcagtg 1560

cgccgacgaa cgtggagctg atggcgatct gccacgagtt tcttgagtac ctccgcgagg 1620

gcacgccgtg ggtcgtgcag cacctcaaca cgtcgtacgt ccccatgccg gcggatcccg 1680

ctgcgttcag tttttggatt gggatggtgg ttctctggtg gctgcgtcgt ctcgtgatgc 1740

ggctctactt ttcgcgcgct gctggcccgt acgcctgcgt cgtcgcgctg cttgcgttcg 1800

ctgtgtactc tatacaaggc ggctga 1826

<210> 26

<211> 2008

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 26

atgtcctcct cgcgccagaa ggccccgccg gtccaacgtc ggccgcgcac cgtcgacccg 60

cccttccgag ccgtgctctc actcccccgc cgcctgctgc gcccgcccag cccccgcggc 120

aagctgccct cgtggggcat cgccccgcgc ttcaccgtca ccctggacga catcctcgac 180

cgccgccacc tcccgcccct cggcctcaaa gactttgagg agtggctcct cttcgtcgag 240

cactcccccg agtccctgta cttcatccta tggctccgcg agtacaccgc ccgcttcgac 300

gcctggatcc acgcctccaa gggcgcgttc acccccaccg accctgtcca ccccagctcg 360

taccgcactc cgcccacacg tccccctccc agccccgccc tgctttcctt ctatatgcgc 420

gcaaagcaca cattcctcct ccacgacgcc ccctacgaac tcctcgtccc caccgacgcc 480

ctcgccccct tccgctcctc gcaagcctat ccgtatgatt gcaacgccga tttcgagcag 540

tgtccacacg cacaccccgt cttcaggtcc aagctgcagc caagagaaac tgatctaccc 600

ccctttgcag gcttgccacc ggaccccctc gtcttcgccg agctcgccgc cctcgtgcgc 660

acggccctcg ccgagagtct cgcccgcttc gtccgcgcga ccttcgccaa cgtcgacgca 720

ccccgcgccg tctgcgggtg ctacggcggg acggtgatct tactcgcggg cagcatcccg 780

ccgatcgtca cgagtatcgt gctgggcagc agccgatggt ggcggctgtt tgcgctcccc 840

gggatgtggc tcggcctgac ggtgctcatc gcggccatgt acggcgtctg catgatgatc 900

tacctcttcg gcgacctccg ccaattgcgc tcgttcgagc tccttcggcc gccctccgac 960

cctgcgctga ccgttgaccg ctcgaagaag gccgggtggc tgaccccgcg cgtgacaccc 1020

gcgatctcgc cgcccacgct gctcacatcc acagccgtag agagtccggg gctgctgcct 1080

gtcccgtgtg ccccccctgc acctgcagac gcgaaggcgc cccagaagcc gcagctgagc 1140

atcgtctgcg gcccccccgc gcactggccc gggccagacc gcgcgctcac accattctcg 1200

tacgactcgc gcagcatgta ctcgcagtcc acgggcggcc tcagcgagca ctacggctcg 1260

gactacgagg acgaggacga ggacgacgcg tgcagcgacg cgcgcatcca cgtctccgac 1320

gcgttcttcg acgagcaccc cgccgcgccg gacgcgccgt tccccgcgcc ccccctcacc 1380

gcgtcgttca tccggcccta tgtctaccgc gccgaggacg cgtacgcgga gggcgacgtc 1440

gagcgtggag aggccgggac cccgccgcgc gccagcccgt cgacgtgttc gactttgacg 1500

cgctcccgcc cgcgcgccgc aggctgtccg tgccgcacgc ggccgccacg ctccaggcgc 1560

ctgcgccggt gcacatcatc gcgccggacg agaagccgcg ccgcgacggg ccgcgggcgc 1620

tcctcggcgt gctgcaggcg cgatgcgcgc cgcggaatat cgtgcgcacg cgcttcgcgg 1680

gggacgtggg cgtcggtatt gggctcggtg ccgtgagggc gcgcgggcgc gttgcgatcc 1740

cggcgtttgc gctcctcccg agcccgacgc gtcgagcgcg tcgttctccg cggcggcgag 1800

cgcgaaggcc ccctccgcgc gggagaagga acgggcgagg gggtggatcc ctgctgcgcc 1860

gtttgcggtg ccggtgacgc cggtgctgag cccggtggtg tcgcgtgcgc agtgggagat 1920

tgtggtgcgg agcggggtgc tggctgcggt tgtgagcggg cttttgggtg tggttgtttt 1980

ggcgctgcct gtgcggcgca cggtgtag 2008

<210> 27

<211> 264

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 27

atgaaactga ttgttctgcg gctgaagagc agacgactca cggaggcact tcttcaccgc 60

gatttccaat tatctgtaac aatcccggac gacaggcttt gtccccctgt accgaacagg 120

ctcaactaca ttttatggat gcaagacatc gtgcatgcga cctccatcgc ggaacctgaa 180

atatcgaaga gaattatcaa gggagttgat atgtatgttc ttccgcattc tcttgcactc 240

ctggatatcg atcgctccga atag 264

<210> 28

<211> 2617

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 28

atgccctttg agcggtatct ccaggcgccg tcctcgtcct cccagcaccg tcgctcctat 60

gacaattatc ccgtcgccca tcccgacccg tctttcggca tcgatgcctc gaccttctct 120

ccaatgtcca gttccaggtc ccccgcatga tgaaccctgc tcccaccctc gctcacggcg 180

gtggtgcaga ggttcttccc ccaggtctcc ttccctcctc cggccccatc tcctcctggt 240

tccccgccgc cggttatcgt tccccggacg acttttcccc ctcctgctct gcatcccccg 300

tctatcgtag ccacccatcg ctgactgctc cagcctattc tcttccccca ggtccacact 360

cctaccgcca ccccgacttt gccgactcac ccgtgaactc tcctcccgat tctattccct 420

cctcctccgc cgccccatac gccttccctc tccagcaacc tcctcccgct gacatcctca 480

tctcccccga ttcccacatc cacctcgatg cccagctctt actccgtgct cacctctcca 540

ttccatcctc ttctctcccc ctctccatcc cctcggccat gggccttccc gtttattccg 600

cttccggctt cgatttcctc tccatcctca gccgtgtagc caaccgtcct caccccaagg 660

tccttctcgg tcccgtagac ctcacctgct cctttgtcgt cgtcgacgtc cgtcgctacg 720

actcccccat cgtgtacgcc tcccccactt ttttcaaact caccggctac gacgaacacg 780

aggtcttggg caggaactgc cgctttctcc agtctcccga tggccgcctc cagcgcggcg 840

agcagcgtct tcatactgcc cccgaggccg tcggcctcct caaacgctgt ctcgtcgccg 900

acaaagagtg ccaaaccagc atcaccaact acaagaaggg cggggccgcc ttcatcaacc 960

tcgtcaccgt cattcccatc ccaggcggcg tcaataatac tcccgcagag gcagacgacg 1020

tcgcctacca cgtcggcttc caaatcgacc tcacagagca gcccaacgcc attctcgaga 1080

agctccgcga cggtagctac gtcgttgact acagccgcaa accgcatgcc ctcccgctcc 1140

tcggaaaccc ttctgggtcc cgcaactggc gatccaactc gatatccatg agcggcgtgt 1200

ccacggaact gcgcaccctc ttggcagacc ccgcctttct tcagtcgatc tcggtcacca 1260

cttccaccac cgcctcctgc ccaccttccg gcgagaagct cgacgtatac gacggcaaca 1320

agccgctgca tatgttcctc ctcgagtgtg cccccgactt tatccacgtc gtttccctca 1380

agggcgcctt cctctacgtc gctccagccg tccgcagcgt cctcggctat gccccagacg 1440

acctcgtcgg tcgcgccgtc tccgatttct gccaccctgc cgacgtcgtc cccctcatgc 1500

gcgagctcaa agagtcctcc ataacccccg gtccaggaag cagccaccta atcgccgcat 1560

cgtcagacgc aggcccgcgc aacgtcgacc tccttttccg catgcgcgcc cagtcgggcc 1620

actttgtctg ggtcgagtgc cgaggtcgcc tccacgtcga gcccggcaag ggtcgcaaag 1680

ccatcatcct ttccggtcgc ctccgcagca tgccccgcct tgactgggat cctgtcgacc 1740

gcgccggtgg cctcgccgcg tccgcgcgca gtcccggcga cgactcgcgc aaggaacgcg 1800

agcgcgaatt ttgggcgctc ctcagcacca acggcacctg cctctttgtt ggcgctgccg 1860

tgcacgacgt gctcggctgg ggcgcgggcg aagtcatcgg cagggctctc ggcgatttta 1920

tcggcggtgc tactacggcg gatgcgcgcc gggcgttgga ggacgagctc gcgcgcgcgc 1980

tctctccgct ctcccccgtc ccgaggtgcg ccaacgcggc gtgtcgtgcg agatgatgcg 2040

caaggatggc gtgcaggtcg tcgtgcacat cactcttttc cgctcgcagg acaacagcga 2100

tgtgctcgcg cccgacgacg cgtggcaggg cgcggtggcc gttccacacg ccccgccgtg 2160

ccccgtggtc tgccaggtga agctgttcga cgcgccgtca gatggaccgc cggcggcggc 2220

ggcggggggt atcttgcacc cactgacgga cagcgtgttc gacgagctgc acatcgcgcg 2280

cgggagcagc tggcagtacg agctgcagca gctcaagttt gcgaaccagc gcataaaaga 2340

ggaagtcgcc gcgctggagg ccagcctctg ctctaaaaca cgccaacgtc ccttttcgca 2400

gagtacctcg cgggcaaacc ctgcccacga gcagcaacag cagcagtcgc agcagcaaca 2460

acagcgagag caatacgcgc atcatccgga gctactgccc gataatacca acaattggag 2520

cacgccatat gtgcagtacc cagtaaacct cgccccgttg cagccgtcgc aaacgttgaa 2580

gcggtcatgg cacgccatag acggggcacc cacctaa 2617

<210> 29

<211> 960

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 29

atgcccgaat ttagcacttc tttgggctgc tcaaatgccc cttacatcta caatgcgacc 60

gataccacct tcctcgtgcc agttggcttg aacaatgccg accacgagat cgaaatcctt 120

ggcaaggcgg tcggcacact cacattcgtg cagggcgact ctgatgcgac ggacgtcaag 180

tacgacatca ccctccgtgc tgaccaggac gggctcttcg actccattct gctgcagcac 240

tccacggcat acgaaatcga gcacggcatc tcgaacagtc acacgcgtct tgcgacaact 300

tacgaatcgt cagcgtgcat gcgctatgat atcaaagtct ctgtgccgcc gaccctcaag 360

aacttggctg taagcacgcg cacggtcacc cacatcaagt tcgacaccga ctccatgttt 420

gacttcgaca agctctcagt gagcatgcgc gcaatgatgg aaaccggcca tcttctcctc 480

ccgcatacgg gcgttcacgc gggcatgctc ttattcgata tttcccgtgg atggctcgtt 540

ggcgacgtcg cgatcgtgga caagaccacg ctgtcgacat caaagggcga cgcagtgatg 600

aacgtgcacg tgcaccccgt cccgtcaacc gcagaccttc ccgcgaccgc tgaactgcag 660

acgacaacgg gcacgggacg caccgacgtc ttcttcgtta gcagcgctgg tcacgttcac 720

cgccccattt ctagcactca cagctcgatg cgcggtggcg aaatgtacct gacgtacaag 780

gaggcggagt tcaagggcga ggtcgacctc tctgcgaagt cgtattctgc tagtggcctc 840

cagggctcaa tgatgcgcca cggtggcatg ggcggcgagc tcccttgggt tggggacaag 900

aacggcggcg ataagatcgt tgcgcagtct cccaatggct ggattgggtt gtacttctga 960

<210> 30

<211> 594

<212> DNA

<213>wide leaf Sparassis crispa (Sparassis latifolia)

<400> 30

atgaagcagc ttgcactgct tcccgaggga aagggtctct tcaatttgtg taccaccgga 60

ctcaccttcg tccctctaaa cacgatttgt gccgacacga cgacgataca gaagacggta 120

tccgagacga tcctcgcagg tatcgcgcaa aatgtgtacc cgccaggtct gcgaaagcag 180

tacacaattc aactcaaaca ccttgagcag caggtaccta gtctcgaatt aattcttggt 240

cccggcataa tgataccgcc tgccaacccc gaccccacca agaaacacat caccctcggc 300

ttcggcctca acagtccctt ctctcggggc accatccacg tcgggtccag cgacccgctg 360

gtaccacccg tcatcgaccc gcacgtattc gaagaatcat acggcgagta cctcacgact 420

atggttgaac ttgtcaaatt tctccgccgc ctggcgaaga ccgagccact gaagagcctc 480

ctcgttggta agcatgtttg atacttgatt ttgcatttct actccagctc ttgaaactcg 540

tgcgtcggac acagagacgg aggtcagccc cggtccacag gtcgaatcag atga 594

Claims (9)

1. a kind of method of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene, it is characterised in that: utilize ATAC-seq method test out edible and medical fungi environmental factor induction before and after the cultural hypha stage mycelia and by environment because Son induction former base differentiation after former base opening Chromatin domains and analyze open area encoding gene, utilize RNA-seq method survey Edible and medical fungi is tried out in the mycelia in mycelia stage and the mRNA sequence of the former base after former base is broken up, and is found using Venn figure The shared differential gene between gene order obtained by ATAC-seq method and RNA-seq method, eventually by biology Analyze the critical functionality gene found in shared differential gene and influence the differentiation of edible and medical fungi former base.

2. a kind of method of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene as described in claim 1 exists Screen the application on Sparassis crispa functional gene.

3. a kind of method of joint ATAC-seq and RNA-seq screening edible and medical fungi functional gene as claimed in claim 2 exists Screen the application on Sparassis crispa functional gene, it is characterised in that: the mycelia in the cultural hypha stage of the Sparassis crispa is taken black The mycelia cultivated under dark condition and the mycelia for not going out former base by light induction but also.

4. a kind of joint ATAC-seq and RNA-seq as claimed in claim 3 is in the method for screening edible and medical fungi functional gene Application on screening Sparassis crispa functional gene, which is characterized in that specific step is as follows:

S1, the mycelia for cultivating Sparassis crispa under dark condition and by light induction but also do not go out former base mycelia, in dark Under the conditions of the mycelia cultivated and the former base after light induction pass through ATAC experiment respectively and obtain purifying DNA and carry out Illumina Acquisition mRNA progress Illumina HiSeq sequencing is tested in HiSeq sequencing by RNA-seq；

S2, mycelia and lure by illumination that Sparassis crispa is cultivated under dark condition are obtained respectively after the sequencing of ATAC-seq method Lead but do not go out also the mycelia contrast groups and the mycelia cultivated under dark condition and the former base pair after light induction of former base It is obtained under dark condition respectively than expressing the lower gene dosage for reconciling and raising in group, while after the sequencing of RNA-seq method The mycelia of culture and do not go out by light induction but also the mycelia contrast groups of former base and the mycelia cultivated under dark condition and The lower gene dosage for reconciling and raising is expressed in former base contrast groups after light induction；

S3, the mycelia pair for passing through light induction using Venn map analysis but also not going out the mycelia of former base and being cultivated under dark condition Than in group by ATAC-seq and RNA-seq integrated processes sequencing after co-express downward gene dosage and co-expression on The gene dosage of tune；Also with former base of the Venn map analysis after light induction and the mycelia pair cultivated under dark condition Than the gene dosage for co-expressing downward in group after the sequencing of ATAC-seq and RNA-seq integrated processes, up-regulation is co-expressed Gene dosage；

S4, comprehensive analysis, which are passed through in two contrast groups obtained in step S3, co-expresses the lower gene dosage for reconciling and raising, and sharp The shared differential gene of two contrast groups is found with Venn map analysis；

S5, the key function that bioinformatic analysis filters out the differentiation of light induction Sparassis crispa former base is carried out to shared differential gene Property gene.

5. a kind of joint ATAC-seq and RNA-seq as claimed in claim 4 is being sieved in the method for screening edible and medical fungi functional gene Select the application on Sparassis crispa functional gene, which is characterized in that ATAC experiment specific steps are as follows:

A1, sample preparation: it takes mycelia under the dark condition in silk ball bacteria cultivation, go out the bacterium of former base by light induction but also Silk, the former base after light induction, with liquid nitrogen flash freezer, cryopreservation；Sample is put into pre-cool and is buffered containing homogenate In the homogenizer of liquid, the tissue of freezing is allowed to thaw and by tissue grinder, pre-cooling centrifugation takes supernatant, utilizes Iodixanol gradient Method separating nucleus, counts nucleus, and every 50000 cell nucleus packing saves；

A2, swivel base reaction and purifying: 50000 nucleus are transferred to containing 1ml ATAC-RSB and 0.1%Tween-20 Omni-ATAC ATAC-seq reaction mixture is added in Guan Zhong, centrifuging and taking supernatant, and 30min is incubated at 37 DEG C and purifies DNA, With 10 μ l elution buffer eluted dnas；

A3, PCR amplification: the 10 μ l are taken to be by step (2) swivel base DNA after purification, the water of 10 μ l nuclease frees, 2.5 μ l concentration 25 μM of PCR primer 1,2 × PCR of the μ l NEB Next High-Fidelity of PCR primer 2 and 25 that 2.5 μ l concentration are 25 μM Master Mix；

A4, sequencing and gene data are analyzed and then obtain the gene dosage of the different lower reconciliation up-regulations of sample expression.

6. a kind of joint ATAC-seq and RNA-seq as claimed in claim 4 is being sieved in the method for screening edible and medical fungi functional gene Select the application on Sparassis crispa functional gene, it is characterised in that: the RNA experiment is successively by sample preparation, mRNA extraction, library Building and sequencing are analyzed with data obtains the lower gene order raised that reconciles of expression in turn.

7. a kind of joint ATAC-seq and RNA-seq as claimed in claim 6 is in the method for screening edible and medical fungi functional gene Application on screening Sparassis crispa functional gene, it is characterised in that: Sparassis crispa mycelia and former base sample need to be in RNA experiment It is ground in liquid nitrogen, extracts reagent using Trizol RNA and extract mRNA.

8. a kind of joint ATAC-seq and RNA-seq as claimed in claim 4 is in the method for screening edible and medical fungi functional gene Application on screening Sparassis crispa functional gene, it is characterised in that: the step of RNA experiment Chinese library constructs is as follows:

B1, with the enrichment with magnetic bead mRNA with Oligo, NEB Fragmentation Buffer, which is added, into obtained mRNA makes Its fragmentation becomes short-movie section；

B2, using the mRNA after fragmentation as template, with hexabasic base random primer synthesize the first chain of cDNA, and be added buffer, DNTPs, RNaseH and DNA Polymerase I synthesize the second chain of cDNA, purify by QIAQuick PCR kit and add EB Buffer elution；

B3, be purified by flash after double-strand cDNA carry out again end reparation, plus base A, plus sequence measuring joints processing, through Ago-Gel Electrophoresis recycling purpose size segment simultaneously carries out PCR amplification, completes library construction work.

9. a kind of joint ATAC-seq and RNA-seq as claimed in claim 4 is in the method for screening edible and medical fungi functional gene Application on screening Sparassis crispa functional gene, it is characterised in that: bioinformatic analysis includes to shared in the step S5 Differential gene carries out GO analysis, KEGG analysis, GSEA analysis and signal network analysis.