Method for constructing DNA fingerprint of chrysosporium keratinophilum and identifying chrysosporium keratinophilum variety by using SNP molecular marker combination in missense coding region

Abstract

The application discloses a method for constructing a DNA fingerprint of Tremella aurantialba and identifying Tremella aurantialba varieties by using a SNP molecular marker combination located in a missense coding region, wherein the SNP molecular marker combination is composed of 54 SNP molecular markers, and the method for constructing the DNA fingerprint of Tremella aurantialba is as follows: first, extracting DNA of Tremella aurantialba germplasm resources or varieties used for constructing the fingerprint, and detecting genotypes of 54 SNP sites; and then, combining and sorting specific sequences of genotypes of each Tremella aurantialba germplasm resource or variety at the 54 SNP sites according to site number order to form a DNA fingerprint of Tremella aurantialba. Further, the DNA fingerprint of Tremella aurantialba is used for identifying Tremella aurantialba varieties. The SNP molecular marker combination has high specificity and resolution, and can reveal a real level of genetic diversity of Tremella aurantialba. The established DNA fingerprint database can correctly reflect DNA fingerprint information of different Tremella aurantialba varieties, and can identify Tremella aurantialba varieties, and the identification result is accurate and efficient.

Description

Technical Field

[0001] This invention relates to the field of molecular biology technology of *Auricularia auricula-judae*, and in particular to a method for constructing a DNA fingerprint map of *Auricularia auricula-judae* using a combination of SNP molecular markers located in the missense coding region and for identifying *Auricularia auricula-judae* varieties using the constructed map. Background Technology

[0002] Golden ear fungus is an edible and medicinal fungus. Its fruiting body is rich in polysaccharides, carotenoids, amino acids, minerals, and vitamins, and it has now been domesticated and industrialized. However, as more and more people cultivate golden ear fungus, the current identification of varieties mainly relies on ITS sequences and SSR markers, which suffers from poor universality, limited coverage, and a lack of standardized management. This leads to unclear understanding of varieties, often resulting in "synonyms," and also presents some problems in defining and protecting selected varieties. There is an urgent need to develop a rapid and convenient identification method.

[0003] DNA fingerprinting, constructed based on molecular markers, offers significant advantages for the identification of germplasm resources. Commonly used molecular markers include restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), simple repeat sequence (SSR), single nucleotide polymorphism (SNP), insertion / deletion (INDEL), and multinucleotide polymorphism (MNP). However, there are few reports on using SNP marker genotyping technology to construct fingerprint profiles for *Auricularia auricula-judae* (Golden Ear) and to identify its germplasm using SNP markers. Therefore, constructing a DNA fingerprint database of SNP sites located in missense coding regions is of great significance for the efficient identification of *Auricularia auricula-judae* varieties and further breeding of new varieties. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for constructing a DNA fingerprint of *Auricularia auricula-judae* using a combination of SNP molecular markers located in the missense coding region, and a method for identifying *Auricularia auricula-judae* varieties using the fingerprint.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A method for constructing a DNA fingerprint of *Auricularia auricula-judae* using SNP molecular marker combinations located in missense coding regions, wherein the SNP molecular marker combinations consist of 54 SNP molecular markers, numbered Na-001 to Na-054, and the molecular marker information is as follows:

[0007] Na-001 1 253697 G / C 0.50 Misinterpretation NAU27000088 Na-002 1 454898 C / G 0.50 Misinterpretation NAU27000159 Na-003 1 756076 A / G 0.49 Misinterpretation NAU27000262 Na-004 1 857971 T / C 0.48 Misinterpretation NAU27000292 Na-005 1 909639 C / A 0.49 Misinterpretation NAU27000309 Na-006 1 960691 A / C 0.50 Misinterpretation NAU27000330 Na-007 1 1462841 T / C 0.50 Misinterpretation NAU27000490 Na-008 1 1512880 A / G 0.50 Misinterpretation NAU27000507 Na-009 1 2064653 T / G 0.45 Misinterpretation NAU27000684 Na-010 1 2114833 C / A 0.48 Misinterpretation NAU27000699 Na-011 1 2365559 A / T 0.49 Misinterpretation NAU27000773 Na-012 1 2516050 T / C 0.50 Misinterpretation NAU27000832 Na-013 2 52532 G / C 0.40 Misinterpretation NAU27003583 Na-014 2 102769 G / C 0.49 Misinterpretation NAU27003601 Na-015 2 554273 G / A 0.50 Misinterpretation NAU27003743 Na-016 2 755289 T / A 0.49 Misinterpretation NAU27003820 Na-017 2 1711098 C / G 0.49 Misinterpretation NAU27004102 Na-018 2 1962078 A / G 0.50 Misinterpretation NAU27004181 Na-019 3 418629 T / C 0.48 Misinterpretation NAU27002380 Na-020 3 619217 A / G 0.49 Misinterpretation NAU27002317 Na-021 3 2118847 C / T 0.47 Misinterpretation NAU27001880 Na-022 4 492 T / A 0.50 Misinterpretation NAU27003006 Na-023 4 50576 G / T 0.50 Misinterpretation NAU27003025 Na-024 4 803556 C / A 0.50 Misinterpretation NAU27003241 Na-025 4 1304596 G / C 0.49 Misinterpretation NAU27003410 Na-026 4 1455131 T / G 0.50 Misinterpretation NAU27003452 Na-027 5 51465 C / A 0.47 Misinterpretation NAU27004304 Na-028 5 154390 A / G 0.49 Misinterpretation NAU27004338 Na-029 5 1366854 G / T 0.49 Misinterpretation NAU27004727 Na-030 6 52332 G / T 0.49 Misinterpretation NAU27005336 Na-031 6 403906 T / G 0.49 Misinterpretation NAU27005438 Na-032 6 654616 G / T 0.50 Misinterpretation NAU27005510 Na-033 6 1208098 C / T 0.49 Misinterpretation NAU27005694 Na-034 6 1258241 A / G 0.49 Misinterpretation NAU27005707 Na-035 6 1309115 C / T 0.50 Misinterpretation NAU27005724 Na-036 6 1411486 T / G 0.50 Misinterpretation NAU27005751 Na-037 8 351894 G / C 0.49 Misinterpretation NAU27004974 Na-038 8 853015 G / C 0.49 Misinterpretation NAU27005129 Na-039 9 1427494 A / C 0.50 Misinterpretation NAU27002996 Na-040 10 106311 A / G 0.50 Misinterpretation NAU27001239 Na-041 10 508343 A / G 0.50 Misinterpretation NAU27001109 Na-042 10 809163 G / A 0.50 Misinterpretation NAU27001015 Na-043 10 959559 C / G 0.49 Missense variant NAU27000969 Na-044 10 1062995 G / C 0.47 Missense variant NAU27000933 Na-045 11 304323 C / G 0.50 Missense variant NAU27006302 Na-046 11 505871 G / A 0.50 Missense variant NAU27006375 Na-047 11 606208 A / C 0.50 Missense variant NAU27006402 Na-048 11 756690 A / G 0.47 Missense variant NAU27006448 Na-049 11 958639 T / G 0.47 Missense variant NAU27006523 Na-050 11 1110652 T / G 0.48 Missense variant NAU27006566 Na-051 12 152186 C / G 0.50 Missense variant NAU27005903 Na-052 12 352913 A / G 0.49 Missense variant NAU27005965 Na-053 12 455822 A / C 0.49 Missense variant NAU27006008 Na-054 12 556163 A / G 0.49 Missense variant NAU27006037

[0008] In the above information, the markers are defined as follows: the marker with the number Na-001 is located on chromosome 1, with a physical location of 253697, an allele of G / C, a polymorphism of 0.50, a missense coding region, and a coding gene of NAU27000088; the meanings of other molecular markers are explained in the same way as those with the number Na-001.

[0009] The method for constructing a DNA fingerprint of *Eurydon edulis* using the aforementioned SNP molecular marker combination is as follows:

[0010] S1. Extract DNA from the golden ear germplasm resources or varieties used to construct fingerprint maps and detect the genotypes of 54 SNP sites from Na-001 to Na-054.

[0011] S2. For each Golden Ear germplasm resource or variety, the genotypes at the 54 SNP loci are combined and sorted according to the locus numbering order to form a specific sequence. This specific sequence is the Golden Ear DNA fingerprint map. The map number, name, and DNA fingerprint are as follows:

[0012] 1 18A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 2 1A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T A / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 3 27A G / G G / G G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C G / G G / G G / G T / T G / G A / A C / CA / A C / C T / T G / G C / C G / G T / T A / A G / G G / G T / T G / G G / G C / C A / A T / T T / T G / G G / GC / C G / G A / A G / G G / G C / C C / C A / A A / A A / A T / T T / T C / C G / G C / C A / A 4 29A C / C G / G A / A T / T C / C A / A T / T G / G T / T C / C A / A C / C G / G G / G A / A T / T C / C G / G C / CA / A C / C T / T G / G C / C C / C T / T C / C G / G G / G G / G T / T G / G C / C A / A C / C G / G C / C G / GC / C G / G A / A G / G C / C C / C G / G G / G C / C G / G T / T G / G C / C A / A A / A A / A 5 45A C / C C / C A / A T / T C / C A / A T / T A / A T / T A / A A / A C / C G / G G / G A / A A / A C / C G / G C / CG / G C / C T / A T / T C / C C / C G / G A / A G / G G / G T / T G / G G / G T / T G / G T / T T / T G / G G / GA / A A / A G / G A / A C / C G / G C / C A / A C / C G / G G / G G / G C / C G / G C / C A / A 6 58A G / G G / G A / A C / C C / C A / A T / T G / G G / G C / C A / A C / C G / G G / G G / G A / A G / G G / G T / TG / G C / C T / A T / T A / A G / G G / G A / A G / G T / T G / G G / G T / T T / T G / G T / T G / G G / G G / GA / A G / G A / A A / A C / C G / G G / G A / A A / A A / A T / T T / T C / C G / G C / C G / G 7 5A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 8 9A G / C G / C G / A T / C C / A A / C C / T G / A T / G C / A A / T C / T G / C G / C G / A T / A G / C A / G C / TA / G C / C T / A G / T C / A G / C T / G A / C G / A G / T T / G G / T G / T C / T A / G T / C T / G G / C G / CC / A G / A A / G G / A C / C C / G C / G A / G A / C A / G T / G T / G C / G G / A C / A A / G 9 BY1A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 10 D2A C / C G / G A / A C / C A / A C / C C / C G / G G / G A / A T / T T / T G / G G / G A / A A / A C / C A / A T / TG / G C / C T / T G / G C / C G / G T / T C / C G / G G / G G / G G / G T / T T / T A / A C / C T / T C / C C / CA / A G / G A / A G / G C / C G / G G / G G / G A / A G / G T / T T / T C / C A / A A / A G / G 11 D3A C / C C / C A / A C / C A / A C / C T / T A / A T / T C / C A / A T / T G / G G / G G / G A / A C / C G / G C / CA / A C / C T / T G / G A / A G / G G / G C / C A / A G / G G / G T / T G / G C / C A / A C / C T / T G / G G / GA / A A / A A / A A / A C / C G / G C / C G / G C / C G / G G / G G / G C / C G / G C / C A / A 12 D51A G / GG / GA / AC / CA / AC / CC / CG / GT / TC / CT / TT / TG / GG / GG / GT / TC / CG / GT / TA / AC / CT / TG / GA / AC / CT / TA / A G / GG / GG / GG / GG / GC / CG / GT / TT / TG / GG / GA / AA / AG / GG / GC / CG / GC / CG / GA / AA / AT / TG / GG / GA / AA / AG / G 13 D60A G / GC / CG / GT / TA / AC / CT / TA / AT / TC / CA / AT / TG / GG / GA / AT / TC / CG / GT / TA / AC / CT / TT / TA / AG / GG / GA / A G / GT / TG / GG / GG / GT / TG / GT / TT / TG / GG / GC / CG / GG / GG / GC / CG / GG / GA / AC / CA / AT / TG / GC / CA / AA / AG / G 14 D9A G / GG / GG / GT / TC / CA / AC / CG / GT / TC / CA / AC / CC / CC / CG / GA / AC / CA / AT / TA / AC / CT / AG / GC / CG / GT / TA / AG / GT / TT / TT / TT / TC / CA / AC / CG / GC / CG / GC / CG / GA / AG / GC / CG / GC / GC / CG / AA / GA / CA / CA 15 F1_2-9 G / CC / CG / AT / CC / CA / CC / CC / CG / AT / TC / CA / AC / CC / CC / CG / AT / AG / CA / GC / TA / GC / CT / TT / TC / AG / GT / GA / CG / GG / TT / GG / TT / TT / TG / GT / TT / TT / TG / GG / GC / CG / GA / GG / GA / AC / AC / AG / CA / GC / GC 16 F1-21A G / G G / C G / A T / C C / A A / C C / T G / A T / G C / A A / T C / T G / C G / C G / A T / A G / C A / G C / TA / G C / C T / A G / T C / A G / C T / G A / C G / A G / T T / G G / T G / T C / C A / G T / C T / G G / C G / CC / A G / A A / G G / A G / C C / G C / G A / A C / C A / G T / G T / G C / G G / A C / A A / G 17 F2A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 18 K18-21A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C T / T T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 19 KF1_23 C / C G / G A / A C / C A / A C / C T / T A / A T / T C / C T / T T / T C / C C / C G / G T / T G / G A / A C / CG / G C / C T / T T / T A / A G / G T / T A / A G / G T / T T / T T / T G / G C / C G / G T / T T / T G / G C / CC / C A / A A / A G / G C / C G / G C / C A / A C / C A / A G / G G / G C / C A / A A / A G / G 20 LLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 21 MD G / G G / G A / A T / T C / C A / A C / C G / G T / T C / C T / T T / T G / G G / G G / G T / T C / C A / A T / TA / A C / C T / A T / T C / C C / C G / G C / C A / A G / G G / G G / G G / G C / C G / G T / T T / T G / G G / GA / A A / A A / A G / G G / G C / C G / G G / G A / A A / A G / G G / G G / G G / G C / C A / A 22 MX9A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 23 NLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 24 X18A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 25 XDA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 26 XR11A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 27 YLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 28 YRA G / GG / GA / AC / CA / AC / CT / TA / AG / GA / AT / TT / TG / GG / GG / GT / TG / GA / AC / CA / AT / TT / AG / GC / CC / CG / GC / CA / AT / TG / GG / GG / GC / CA / AC / CG / GC / CC / CA / AA / AG / GA / AG / GC / CG / GG / GA / AA / AT / TT / TC / CG / GC / CA / A

[0013] The method for identifying *Auricularia auricula-judae* varieties using the DNA fingerprinting constructed in this invention involves extracting DNA from the spores of the *Auricularia auricula-judae* germplasm resources or varieties to be tested, detecting the genotypes of 54 SNP loci (Na-001~Na-054) in the genomic DNA of the germplasm resources, determining the genotype of the SNP molecular markers based on the SNP loci detection results, comparing them with the constructed DNA fingerprinting, and calculating the genetic similarity GS. When the genetic similarity GS of the *Auricularia auricula-judae* germplasm resources or varieties to be tested is greater than or equal to 96%, they are identified as the same germplasm or variety. The genetic similarity GS = n / N × 100%, where N is the number of SNP loci, and n is the number of SNP loci with the same genotype for each *Auricularia auricula-judae* germplasm resource or variety.

[0014] Compared with the prior art, the present invention has the following effects:

[0015] This invention provides a combination of SNP molecular markers for constructing DNA fingerprinting of *Auricularia auricula-judae*, consisting of 54 missense coding region (SNP) molecular markers. This combination exhibits high specificity and resolution, enabling the accurate identification of the genetic diversity of *Auricularia auricula-judae*. The DNA fingerprinting library established using this SNP molecular marker combination accurately reflects the DNA fingerprinting information of different *Auricularia auricula-judae* varieties, facilitating variety identification. The detection results are precise and efficient, effectively distinguishing 28 *Auricularia auricula-judae* germplasm resource varieties and providing technical support for the protection of *Auricularia auricula-judae* germplasm resources. Attached Figure Description

[0016] Figure 1This invention presents a phylogenetic tree of 28 golden ear germplasm resources constructed based on 131,632 SNPs;

[0017] Figure 2 This invention presents a phylogenetic tree of 28 golden ear germplasm resources constructed based on 54 missense coded SNPs.

[0018] Figure 3 This is the genotyping result of 28 *Auricularia auricula-judae* materials in Example 2 of this invention. Each row represents one material, and each column represents one SNP genotype. Blue, red, purple, and orange represent alleles C / C, A / A, G / G, and T / T, respectively. Unknown alleles are shown in white, and alleles are represented by 0 / 0; heterozygous sites are shown in gray.

[0019] Figure 4 This shows the genetic similarity results of 28 golden ear fungus materials in Example 2 of this invention. The genetic similarity changes from blue to red.

[0020] Figure 5 This table shows the genotyping results of the golden ear fungus materials D13A and 3-13, D25A and 3-25, D40A and 3-40, and D47A and 3-47 in Example 3 of this invention. Each row represents one material, and each column represents one SNP genotype. Blue, red, purple, and orange represent alleles C / C, A / A, G / G, and T / T, respectively. Missing alleles are shown in green, with alleles indicated by - / -; unknown alleles are shown in white, with alleles indicated by 0 / 0; heterozygous sites are shown in gray.

[0021] Figure 6 The genetic similarity results of D13A and 3-13, D25A and 3-25, D40A and 3-40, and D47A and 3-47 golden ear materials in Example 3 of the present invention are shown, where the genetic similarity gradually changes from blue to red. Detailed Implementation

[0022] The specific embodiments of the present invention will be described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0023] Example 1. Missense coding region SNP sites (molecular marker combinatorial) used to construct a DNA fingerprint of *Eurydon edulis*.

[0024] This invention uses 28 Auricularia auricula germplasm resource samples (sample information is shown in Table 1) to extract genomic DNA from Auricularia auricula buds. Whole genome resequencing was completed at BGI Genomics Co., Ltd. in Shenzhen. Based on the sequencing data, 54 missense coding region (SNP) sites were obtained.

[0025] Table 1. Information on 28 Golden Ear Germplasm Resource Samples

[0026] 1 18A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 2 1A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 3 27A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 4 29A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 5 45A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 6 58A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 7 5A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 8 9A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 9 BY1A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 10 D2A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 11 D3A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 12 D51A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 13 D60A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 14 D9A Single spore Yunnan Junshijie Biotechnology Co., Ltd. 15 F1_2-9 Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 16 F1-21A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 17 F2A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 18 K18-21A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 19 KF1_23 Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 20 LLA Cultivated varieties Gutian area, Fujian 21 MD Single spore Yunnan Junshijie Biotechnology Co., Ltd. 22 MX9A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 23 NLA Cultivated varieties Ninglang area of ​​Lijiang, Yunnan 24 X18A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 25 XDA Cultivated varieties Xundian area of ​​Kunming, Yunnan 26 XR11A Cultivated varieties Yunnan Junshijie Biotechnology Co., Ltd. 27 YLA Cultivated varieties Yunlong area of ​​Dali, Yunnan 28 YRA Cultivated varieties Yongren area of ​​Chuxiong, Yunnan

[0027] The SNP site screening method is as follows:

[0028] (1) Trimmomatic-0.39 was used for quality control of sequencing data, with default parameters;

[0029] (2) The quality-controlled data were compared with the reference genome Ns_27 (NCBI project number: PRJNA1264744) using BWA 0.7.17 software to generate a sam file;

[0030] (3) Use SAMtools 1.22 to convert the SAM file to a BAM file, and then sort and remove duplicates. Analyze each material separately to ensure the accuracy of individual variation;

[0031] (4) Use GATK 4.5 to merge the bam files of each strain, and then perform mutation detection to obtain SNP data;

[0032] (5) The SNP data were further screened using VCFtools 0.1.17, with the criteria being a deletion rate of more than 20% and a minimum allele frequency of less than 0.05, resulting in 131,632 SNPs.

[0033] (6) The 131,632 SNPs obtained were filtered according to the criteria of polymorphism information content (PIC) greater than 0.4 and the distance between adjacent SNPs being at least 50kb, resulting in 343 SNPs;

[0034] (7) SnpEff 5.2 was used to annotate the 343 SNPs obtained. Sites located in the coding region and which are missense variants were screened, and finally 54 SNPs were obtained. The specific information is shown in Table 2.

[0035] Table 2. Molecular marker information for 54 coding variant SNPs

[0036] Na-001 1 253697 G / C 0.50 Misinterpretation NAU27000088 Na-002 1 454898 C / G 0.50 Misinterpretation NAU27000159 Na-003 1 756076 A / G 0.49 Misinterpretation NAU27000262 Na-004 1 857971 T / C 0.48 Misinterpretation NAU27000292 Na-005 1 909639 C / A 0.49 Misinterpretation NAU27000309 Na-006 1 960691 A / C 0.50 Misinterpretation NAU27000330 Na-007 1 1462841 T / C 0.50 Misinterpretation NAU27000490 Na-008 1 1512880 A / G 0.50 Misinterpretation NAU27000507 Na-009 1 2064653 T / G 0.45 Misinterpretation NAU27000684 Na-010 1 2114833 C / A 0.48 Misinterpretation NAU27000699 Na-011 1 2365559 A / T 0.49 Misinterpretation NAU27000773 Na-012 1 2516050 T / C 0.50 Misinterpretation NAU27000832 Na-013 2 52532 G / C 0.40 Misinterpretation NAU27003583 Na-014 2 102769 G / C 0.49 Misinterpretation NAU27003601 Na-015 2 554273 G / A 0.50 Misinterpretation NAU27003743 Na-016 2 755289 T / A 0.49 Misinterpretation NAU27003820 Na-017 2 1711098 C / G 0.49 Misinterpretation NAU27004102 Na-018 2 1962078 A / G 0.50 Misinterpretation NAU27004181 Na-019 3 418629 T / C 0.48 Misinterpretation NAU27002380 Na-020 3 619217 A / G 0.49 Misinterpretation NAU27002317 Na-021 3 2118847 C / T 0.47 Misinterpretation NAU27001880 Na-022 4 492 T / A 0.50 Misinterpretation NAU27003006 Na-023 4 50576 G / T 0.50 Misinterpretation NAU27003025 Na-024 4 803556 C / A 0.50 Misinterpretation NAU27003241 Na-025 4 1304596 G / C 0.49 Misinterpretation NAU27003410 Na-026 4 1455131 T / G 0.50 Misinterpretation NAU27003452 Na-027 5 51465 C / A 0.47 Misinterpretation NAU27004304 Na-028 5 154390 A / G 0.49 Misinterpretation NAU27004338 Na-029 5 1366854 G / T 0.49 Misinterpretation NAU27004727 Na-030 6 52332 G / T 0.49 Misinterpretation NAU27005336 Na-031 6 403906 T / G 0.49 Misinterpretation NAU27005438 Na-032 6 654616 G / T 0.50 Misinterpretation NAU27005510 Na-033 6 1208098 C / T 0.49 Misinterpretation NAU27005694 Na-034 6 1258241 A / G 0.49 Misinterpretation NAU27005707 Na-035 6 1309115 C / T 0.50 Misinterpretation NAU27005724 Na-036 6 1411486 T / G 0.50 Misinterpretation NAU27005751 Na-037 8 351894 G / C 0.49 Misinterpretation NAU27004974 Na-038 8 853015 G / C 0.49 Misinterpretation NAU27005129 Na-039 9 1427494 A / C 0.50 Misinterpretation NAU27002996 Na-040 10 106311 A / G 0.50 Misinterpretation NAU27001239 Na-041 10 508343 A / G 0.50 Misinterpretation NAU27001109 Na-042 10 809163 G / A 0.50 Misinterpretation NAU27001015 Na-043 10 959559 C / G 0.49 Misinterpretation NAU27000969 Na-044 10 1062995 G / C 0.47 Misinterpretation NAU27000933 Na-045 11 304323 C / G 0.50 Misinterpretation NAU27006302 Na-046 11 505871 G / A 0.50 Misinterpretation NAU27006375 Na-047 11 606208 A / C 0.50 Misinterpretation NAU27006402 Na-048 11 756690 A / G 0.47 Misinterpretation NAU27006448 Na-049 11 958639 T / G 0.47 Misinterpretation NAU27006523 Na-050 11 1110652 T / G 0.48 Misinterpretation NAU27006566 Na-051 12 152186 C / G 0.50 Misinterpretation NAU27005903 Na-052 12 352913 A / G 0.49 Misinterpretation NAU27005965 Na-053 12 455822 A / C 0.49 Misinterpretation NAU27006008 Na-054 12 556163 A / G 0.49 Misinterpretation NAU27006037

[0037] In the above information, the markers are defined as follows: the marker with the number Na-001 is located on chromosome 1, with a physical location of 253697, an allele of G / C, a polymorphism of 0.50, a missense coding region, and a coding gene of NAU27000088; the meanings of other molecular markers are explained in the same way as those with the number Na-001.

[0038] Example 2. Genotyping and fingerprinting of *Auricularia auricula-judae* germplasm resources.

[0039] Constructing a DNA fingerprint: DNA was extracted from spore samples of *Auricularia auricula-judae* germplasm resources used for constructing the DNA fingerprint. Genotypes of 54 SNP loci in the *Auricularia auricula-judae* germplasm resources were obtained through whole-genome resequencing at BGI Genomics Co., Ltd. in Shenzhen. Based on the genotyping results, the genotypes of each *Auricularia auricula-judae* germplasm resource at the 54 SNP loci were combined and sorted according to locus numbering to form a specific sequence. This specific sequence is the DNA fingerprint of the *Auricularia auricula-judae* germplasm resource or variety, as shown in Table 3. A phylogenetic tree was constructed using 131,632 SNPs and 54 SNPs located in the coding region and representing missense variants. Both showed similar trends, indicating that these 54 SNPs can reflect the true population diversity, as shown in Table 3. Figure 1 The phylogenetic tree of 28 *Auricularia auricula-judae* germplasm resources constructed based on 131,632 SNPs, as shown in this invention, and Figure 2 The phylogenetic tree shown is based on 54 missense SNPs constructed from 28 golden ear germplasm resources according to this invention. Genotyping results of the 28 golden ear materials are shown below. Figure 3 Each row represents one material, and each column represents a SNP genotype. Blue, red, purple, orange, and white represent alleles C / C, A / A, G / G, T / T, and 0 / 0, respectively. Heterozygous sites are indicated in gray. The genetic similarity results of the 28 golden ear materials are shown below. Figure 4 The genetic similarity gradient changes from blue to red. Among the 28 golden ear fungus materials, the similarity of cultivated strains 18A, 5A, LLA, NLA, X18A, XDA, YLA and YRA are all above 96%, which means they are likely obtained by asexual reproduction of the same strain. The same is true for cultivated strains 1A, BY1A, F2A, K18-21A, MX9A and XR11A.

[0040] Table 3. Fingerprint patterns of golden ear germplasm resources

[0041] 1 18A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 2 1A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T A / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 3 27A G / G G / G G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C G / G G / G G / G T / T G / G A / A C / CA / A C / C T / T G / G C / C G / G T / T A / A G / G G / G T / T G / G G / G C / C A / A T / T T / T G / G G / GC / C G / G A / A G / G G / G C / C C / C A / A A / A A / A T / T T / T C / C G / G C / C A / A 4 29A C / C G / G A / A T / T C / C A / A T / T G / G T / T C / C A / A C / C G / G G / G A / A T / T C / C G / G C / CA / A C / C T / T G / G C / C C / C T / T C / C G / G G / G G / G T / T G / G C / C A / A C / C G / G C / C G / GC / C G / G A / A G / G C / C C / C G / G G / G C / C G / G T / T G / G C / C A / A A / A A / A 5 45A C / C C / C A / A T / T C / C A / A T / T A / A T / T A / A A / A C / C G / G G / G A / A A / A C / C G / G C / CG / G C / C T / A T / T C / C C / C G / G A / A G / G G / G T / T G / G G / G T / T G / G T / T T / T G / G G / GA / A A / A G / G A / A C / C G / G C / C A / A C / C G / G G / G G / G C / C G / G C / C A / A 6 58A G / G G / G A / A C / C C / C A / A T / T G / G G / G C / C A / A C / C G / G G / G G / G A / A G / G G / G T / TG / G C / C T / A T / T A / A G / G G / G A / A G / G T / T G / G G / G T / T T / T G / G T / T G / G G / G G / GA / A G / G A / A A / A C / C G / G G / G A / A A / A A / A T / T T / T C / C G / G C / C G / G 7 5A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 8 9A G / C G / C G / A T / C C / A A / C C / T G / A T / G C / A A / T C / T G / C G / C G / A T / A G / C A / G C / TA / G C / C T / A G / T C / A G / C T / G A / C G / A G / T T / G G / T G / T C / T A / G T / C T / G G / C G / CC / A G / A A / G G / A C / C C / G C / G A / G A / C A / G T / G T / G C / G G / A C / A A / G 9 BY1A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 10 D2A C / C G / G A / A C / C A / A C / C C / C G / G G / G A / A T / T T / T G / G G / G A / A A / A C / C A / A T / TG / G C / C T / T G / G C / C G / G T / T C / C G / G G / G G / G G / G T / T T / T A / A C / C T / T C / C C / CA / A G / G A / A G / G C / C G / G G / G G / G A / A G / G T / T T / T C / C A / A A / A G / G 11 D3A C / C C / C A / A C / C A / A C / C T / T A / A T / T C / C A / A T / T G / G G / G G / G A / A C / C G / G C / CA / A C / C T / T G / G A / A G / G G / G C / C A / A G / G G / G T / T G / G C / C A / A C / C T / T G / G G / GA / A A / A A / A A / A C / C G / G C / C G / G C / C G / G G / G G / G C / C G / G C / C A / A 12 D51A G / G G / G A / A C / C A / A C / C C / C G / G T / T C / C T / T T / T G / G G / G G / G T / T C / C G / G T / TA / A C / C T / T G / G A / A C / C T / T A / A G / G G / G G / G G / G G / G C / C G / G T / T T / T G / G G / GA / A A / A G / G G / G C / C G / G C / C G / G A / A A / A T / T G / G G / G A / A A / A G / G 13 D60A G / G C / C G / G T / T A / A C / C T / T A / A T / T C / C A / A T / T G / G G / G A / A T / T C / C G / G T / TA / A C / C T / T T / T A / A G / G G / G A / A G / G T / T G / G G / G G / G T / T G / G T / T T / T G / G G / GC / C G / G G / G G / G C / C G / G G / G A / A C / C A / A T / T G / G C / C A / A A / A G / G 14 D9A G / G G / G G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C G / G A / A C / C A / A T / TA / A C / C T / A G / G C / C G / G T / T A / A G / G T / T T / T T / T T / T C / C A / A C / C G / G C / C G / GC / C G / G A / A G / G C / C G / G C / C G / G A / A A / A T / T T / T C / C G / G C / C A / A 15 F1_2-9 G / C C / C G / A T / C C / C A / C C / C G / A T / T C / C A / A C / C C / C C / C G / A T / A G / C A / G C / TA / G C / C T / T T / T C / A G / G T / G A / C G / G G / T T / G G / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / G G / A C / C G / G C / C A / A C / C A / G G / G T / G C / G A / A A / A G / G 16 F1-21A G / G G / C G / A T / C C / A A / C C / T G / A T / G C / A A / T C / T G / C G / C G / A T / A G / C A / G C / TA / G C / C T / A G / T C / A G / C T / G A / C G / A G / T T / G G / T G / T C / C A / G T / C T / G G / C G / CC / A G / A A / G G / A G / C C / G C / G A / A C / C A / G T / G T / G C / G G / A C / A A / G 17 F2A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 18 K18-21A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C T / T T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 19 KF1_23 C / C G / G A / A C / C A / A C / C T / T A / A T / T C / C T / T T / T C / C C / C G / G T / T G / G A / A C / CG / G C / C T / T T / T A / A G / G T / T A / A G / G T / T T / T T / T G / G C / C G / G T / T T / T G / G C / CC / C A / A A / A G / G C / C G / G C / C A / A C / C A / A G / G G / G C / C A / A A / A G / G 20 LLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 21 MD G / G G / G A / A T / T C / C A / A C / C G / G T / T C / C T / T T / T G / G G / G G / G T / T C / C A / A T / TA / A C / C T / A T / T C / C C / C G / G C / C A / A G / G G / G G / G G / G C / C G / G T / T T / T G / G G / GA / A A / A A / A G / G G / G C / C G / G G / G A / A A / A G / G G / G G / G G / G C / C A / A 22 MX9A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 23 NLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 24 X18A G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A C / C T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 25 XDA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 26 XR11A C / C C / C G / G T / T C / C A / A C / C G / G T / T C / C A / A C / C C / C C / C A / A A / A C / C G / G T / TG / G C / C 0 / 0 T / T A / A G / G T / T A / A G / G G / G T / T T / T T / T T / T G / G T / T T / T G / G G / GC / C G / G A / A G / G C / C G / G C / C A / A C / C G / G G / G G / G G / G A / A A / A G / G 27 YLA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A 28 YRA G / G G / G A / A C / C A / A C / C T / T A / A G / G A / A T / T T / T G / G G / G G / G T / T G / G A / A C / CA / A T / T T / A G / G C / C C / C G / G C / C A / A T / T G / G G / G G / G C / C A / A C / C G / G C / C C / CA / A A / A G / G A / A G / G C / C G / G G / G A / A A / A T / T T / T C / C G / G C / C A / A

[0042] Example 3. Identification and genetic similarity calculation of golden ear germplasm resources.

[0043] A method for identifying a golden ear fungus variety includes extracting DNA from the golden ear fungus germplasm to be tested, using whole-genome resequencing to detect the genotypes of 54 SNP loci in the genomic DNA of the golden ear fungus germplasm, comparing it with the golden ear fungus DNA fingerprint database in Table 4, calculating genetic similarity, and identifying the golden ear fungus germplasm. Detailed results are available in [link to results]. Figure 5 and Figure 6Taking samples 3-13, 3-25, 3-40, and 3-47 as examples, the strain information is shown in Table 5. DNA was re-extracted from these samples and named D13A, D25A, D40A, and D47A. Whole-genome resequencing was performed on these four samples, and the genotypes of 54 SNP loci were extracted. The results showed that D13A was identical to 3-13, D25A to 3-25, D40A to 3-40, and D47A to 3-47 in terms of alleles. The genetic similarity was then calculated using the formula GS = n / N × 100%, where N is the number of SNP loci, and n is the number of SNP loci with identical genotypes for each *Auricularia auricula-judae* germplasm resource or variety. For example, D13A and 3-13, the number of SNP loci mentioned by N is 54, each locus has 2 alleles, for a total of 108 alleles. n is the number of SNP loci with the same genotype when comparing D13A and 3-13. There are a total of 108 identical alleles, that is, the genetic similarity GS is 108 / 108×100%=100%. Therefore, the genetic similarity of D13A and 3-13 is 100%. D25A and 3-25, D40A and 3-40, D47A and 3-47 are calculated using the same method, and the genetic similarity GS all reach 100%. It is determined that D13A and 3-13, D25A and 3-25, D40A and 3-40, and D47A and 3-47 are all of the same type, which is consistent with the facts.

[0044] Table 4. Variations of D13A and 3-13, D25A and 3-25, D40A and 3-40, D47A and 3-47 in the golden ear fungus.

[0045] DNA fingerprinting

[0046] 1 3-13 C / C C / C G / G T / T C / C A / A T / T A / A T / T C / C A / A T / T G / G G / G A / A A / A G / G G / G T / TA / A C / C T / T G / G C / C G / G T / T C / C A / A G / G G / G T / T T / T T / T A / A C / C G / G G / G G / GA / A A / A G / G G / G C / C G / G C / C G / G A / A A / A T / T T / T G / G A / A A / A G / G 2 3-25 C / C C / C G / G T / T C / C A / A C / C G / G T / T A / A T / T T / T G / G G / G G / G T / T G / G A / A T / TA / A C / C T / T G / G A / A G / G G / G C / C A / A G / G T / T T / T G / G T / T G / G T / T T / T G / G G / GC / C A / A G / G A / A C / C G / G C / C G / G A / A G / G G / G G / G G / G G / G C / C A / A 3 3-40 G / G C / C G / G T / T C / C A / A T / T A / A T / T C / C A / A C / C C / C C / C A / A T / T C / C A / A T / TG / G C / C 0 / 0 T / T A / A G / G T / T C / C A / A T / T T / T T / T G / G C / C A / A C / C G / G G / G C / CA / A A / A A / A G / G G / G C / C G / G A / A C / C A / A T / T T / T G / G A / A A / A A / A 4 3-47 G / G C / C A / A C / C A / A C / C T / T A / A T / T C / C A / A T / T G / G G / G G / G T / T C / C A / A C / CA / A C / C T / A T / T C / C G / G T / T C / C A / A T / T G / G T / T T / T C / C A / A C / C T / T C / C C / CC / C A / A A / A A / A G / G C / C G / G A / A C / C A / A T / T T / T C / C A / A A / A G / G 5 D13A C / CC / CG / GT / TC / CA / AT / TA / AT / TC / CA / AT / TG / GG / GA / AA / AG / GG / GT / TA / AC / CT / TG / GC / CG / GT / TC / CA / AG / GG / GT / TT / TT / TA / AC / CG / GG / GG / GA / AA / AG / GG / GC / CG / GC / CG / GA / AA / AT / TT / TG / GA / AA / AG / G 6 D25A C / CC / CG / GT / TC / CA / AC / CG / GT / TA / AT / TT / TG / GG / GG / GT / TG / GA / AT / TA / AC / CT / TG / GA / AG / GG / GC / CA / AG / GT / TT / TG / GT / TG / GT / TT / TG / GG / GC / CA / AG / GA / AC / CG / GC / CG / GA / AG / GG / GG / GG / GG / GC / CA / A 7 D40A G / GC / CG / GT / TC / CA / AT / TA / AT / TC / CA / AC / CC / CC / CA / AT / TC / CA / AT / TG / GC / C 0 / 0 T / TA / AG / GT / TC / CA / AT / TT / TT / TG / GC / CA / AC / CG / GG / GC / CA / AA / AA / AG / GG / GC / CG / GA / AC / CA / AT / TT / TG / GA / AA / AA / A 8 D47A G / GC / CA / AC / CA / AC / CT / TA / AT / TC / CA / AT / TG / GG / GG / GT / TC / CA / AC / CA / AC / CT / AT / TC / CG / GT / TC / CA / AT / TG / GT / TT / TC / CA / AC / CT / TC / CC / CC / CA / AA / AA / AG / GC / CG / GA / AC / CA / AT / TT / TC / CA / AA / AG / G

[0047] Table 5. Information on strains of *Auricularia auricula-judae* D13A and 3-13, D25A and 3-25, D40A and 3-40, and D47A and 3-47.

[0048] 1 3-13 Single spore Yunnan Junshijie Biotechnology Co., Ltd. 2 3-25 Single spore Yunnan Junshijie Biotechnology Co., Ltd. 3 3-40 Single spore Yunnan Junshijie Biotechnology Co., Ltd. 4 3-47 Single spore Yunnan Junshijie Biotechnology Co., Ltd. 5 D13A 3-13 Obtained through asexual reproduction Yunnan Junshijie Biotechnology Co., Ltd. 6 D25A Obtained through asexual reproduction on March 25th. Yunnan Junshijie Biotechnology Co., Ltd. 7 D40A Obtained through asexual reproduction between 3 and 40 Yunnan Junshijie Biotechnology Co., Ltd. 8 D47A Obtained through asexual reproduction between 3 and 40 Yunnan Junshijie Biotechnology Co., Ltd.

[0049] The foregoing description of specific exemplary embodiments of the present invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various exemplary embodiments of the invention, as well as various choices and modifications. Therefore, the scope of protection of the present invention is not limited to the embodiments.

[0050] The SNP molecular marker combinations and the constructed *Gynostemma pentaphyllum* DNA map provided by this invention can be applied to any of the following aspects:

[0051] (1) Used for constructing DNA fingerprinting of golden ear fungus;

[0052] (2) Used for genotyping of golden ear fungus;

[0053] (3) Used for the identification of golden ear germplasm resources and varieties;

[0054] (4) Used for the breeding of golden ear germplasm resources;

[0055] (5) Used for molecular marker-assisted breeding of golden ear fungus.

Claims

1. A method for constructing a DNA fingerprint of *Eurydon edulis* using SNP molecular marker combinations located in missense coding regions, characterized in that... The SNP molecular marker assemblage consists of 54 SNP molecular markers, numbered Na-001 to Na-054, with Ns_27 as the reference genome and NCBI project number PRJNA1264744; the molecular marker information is as follows: In the above information, the markers are defined as follows: the marker with the number Na-001 is located on chromosome 1, with a physical location of 253697, an allele of G / C, a polymorphism of 0.50, a missense coding region, and a coding gene of NAU27000088; the meanings of other molecular markers are explained in the same way as those with the number Na-001. The method for constructing a DNA fingerprint of *Eurydon edulis* using the aforementioned SNP molecular marker combination is as follows: S1. Extract DNA from the golden ear germplasm resources or varieties used to construct fingerprint maps and detect the genotypes of 54 SNP sites from Na-001 to Na-054. S2. For each Golden Ear germplasm resource or variety, the genotypes at the 54 SNP loci are combined and sorted according to the locus numbering order to form a specific sequence. This specific sequence is the Golden Ear DNA fingerprint map. The map number, name, and DNA fingerprint are as follows:

2. A method for identifying *Auricularia auricula-judae* varieties using a DNA fingerprint constructed according to claim 1, characterized in that, DNA was extracted from the spores of the *Auricularia auricula-judae* germplasm resources or varieties to be tested. The genotypes of 54 SNP loci (Na-001~Na-054) in the genomic DNA of the germplasm resources were detected. The genotyping of the SNP molecular markers was determined based on the SNP loci detection results. The results were compared with the constructed DNA fingerprint and the genetic similarity GS was calculated. When the genetic similarity GS of the *Auricularia auricula-judae* germplasm resources or varieties was greater than or equal to 96%, they were identified as the same germplasm or variety. The genetic similarity GS = n / N × 100%, where N is the number of SNP loci and n is the number of SNP loci with the same genotype for each *Auricularia auricula-judae* germplasm resource or variety.

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