CN111321238A - Method for identifying mating types of twenty-two species in yellow morchella flora - Google Patents
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- 230000013011 mating Effects 0.000 title claims abstract description 80
- 241000221638 Morchella Species 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000894007 species Species 0.000 title claims abstract description 22
- 238000012408 PCR amplification Methods 0.000 claims abstract description 16
- 239000002773 nucleotide Substances 0.000 claims description 8
- 125000003729 nucleotide group Chemical group 0.000 claims description 8
- 101100512904 Caenorhabditis elegans mes-6 gene Proteins 0.000 claims description 6
- 241000098761 Eutropis clivicola Species 0.000 claims description 6
- 241001223337 Medicago granadensis Species 0.000 claims description 6
- 241001161346 Morchella dunensis Species 0.000 claims description 6
- 241001219984 Morchella palazonii Species 0.000 claims description 6
- 238000000137 annealing Methods 0.000 claims description 5
- 238000004925 denaturation Methods 0.000 claims description 5
- 230000036425 denaturation Effects 0.000 claims description 5
- 238000012257 pre-denaturation Methods 0.000 claims description 5
- BLRBOMBBUUGKFU-SREVYHEPSA-N (z)-4-[[4-(4-chlorophenyl)-5-(2-methoxy-2-oxoethyl)-1,3-thiazol-2-yl]amino]-4-oxobut-2-enoic acid Chemical compound S1C(NC(=O)\C=C/C(O)=O)=NC(C=2C=CC(Cl)=CC=2)=C1CC(=O)OC BLRBOMBBUUGKFU-SREVYHEPSA-N 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 230000001568 sexual effect Effects 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 17
- 238000001514 detection method Methods 0.000 abstract description 9
- 108020004414 DNA Proteins 0.000 description 12
- 230000014639 sexual reproduction Effects 0.000 description 11
- 241000233866 Fungi Species 0.000 description 6
- 240000002769 Morchella esculenta Species 0.000 description 6
- 230000003321 amplification Effects 0.000 description 5
- 238000003199 nucleic acid amplification method Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 239000011543 agarose gel Substances 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 241000235349 Ascomycota Species 0.000 description 1
- 101100306017 Caenorhabditis elegans rpb-2 gene Proteins 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 102100021429 DNA-directed RNA polymerase II subunit RPB1 Human genes 0.000 description 1
- 101001106401 Homo sapiens DNA-directed RNA polymerase II subunit RPB1 Proteins 0.000 description 1
- 240000005984 Mammea americana Species 0.000 description 1
- 235000002779 Morchella esculenta Nutrition 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
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Abstract
The invention provides a method for identifying mating types of twenty-two species in a yellow morchella group, which mainly comprises the following steps: extracting total DNA of strains of a strain to be detected as a PCR amplification template; the strains with expected bands are obtained by carrying out PCR amplification on EMAT1-1L/EMAT1-1R through primers, MAT1-1 mating types are indicated, strains with expected bands are obtained by carrying out PCR amplification on EMAT1-2L/EMAT1-2R through primers, MAT1-2 mating types are indicated, if only one mating type strain is detected, only a male parent or a female parent is indicated, the fruiting capacity is poor or not good, and if 2 strains with expected amplified bands can be detected, the strains with two mating types of the male parent and the female parent have the fruiting capacity. By using the primers and the detection method for detecting the mating type genes of the morchella strains, the reliability of high yield and stable yield in the cultivation process of the morchella can be greatly improved.
Description
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to a method for identifying twenty-two morchella species, which comprises the following steps: (M.steppcola, M.yangii, M.yishuica, M.clivicola, M.dunensis, M.palazonii, M.amelicana, M.esculenta, M.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27) mating type genes. The technology of the invention is suitable for the research on the aspects of selective breeding, cultivation, mating type gene, species identification, sexual reproduction evolution and phylogeny of morchella strains.
Background
O' Donnell, Taskin and Du Huai et al use polygenic lineage-consistent phylogenetic species recognition (GCPSR) to classify Morchella fungi into yellow Morchella branch (Esculenta Clade), yellow Morchella branch (Elata Clade) and Red Morchella variegates branch (Rufobrunnea Clade) based on the nucleotide sequences of 5 genes in total, LSU, ITS, EF1-a, RPB1 and RPB 2. The twenty-two species related by the invention belong to a yellow morchella strain.
Mating-type loci (MAT) are key factors for regulating the sexual reproduction mode of ascomycetes, and divide sexual reproduction into heterozygote (hetetosalbum), homozygote (homozygote) and sublevel homozygote (subcordary homozygote). The heterozygote fungi are self-sterile, the haploid nucleus of the heterozygote fungi only carries one mating type gene (MAT1-1 or MAT1-2), and the fusion of haploid of complementary mating type is necessary to complete sexual reproduction; the haploid nucleus of the homologous mating fungus self-fertile (self-fertile) carries two compatible mating type genes (MAT1-1 and MAT1-2), and a single strain can complete sexual reproduction; secondary sibling fungi also appear to be selfing fertile, but unlike sibling fungi, the individual sexual germ cells contain haploid nuclei of two complementary mating types at the same time, appearing to "independently" complete sexual reproduction.
A large number of previous experiments prove that twenty-two morchella species related to the invention are all heterozygotic fungi, the sexual reproduction process (fruiting) of the morchella species can be completed only by combining two mating types, and meanwhile, the ascospores of the morchella species are all multinuclear single homokaryons for the first time. Therefore, the detection of the mating type of the strains is a prerequisite for obtaining an effective cultivated strain, and the detection of the mating type of the strains is a necessary and primary step for ensuring the reliability of the strains.
Disclosure of Invention
The invention aims to solve the problems of artificial cultivation and strain identification of yellow morchella, and provides a method for identifying the mating types of twenty-two species in a yellow morchella group and a related universal specific primer pair. The provided primer pair can be universally used for the mating type identification of the twenty-two morchella species, the method is simple and convenient, and the result is stable, reliable and easy to observe.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: the mating type identification method for twenty-two species in the yellow morchella colony comprises a specific primer pair EMAT1-1L/EMAT1-1R for detecting mating type MAT1-1, a specific primer pair EMAT1-2L/EMAT1-2R for detecting mating type MAT1-2, a corresponding PCR amplification system and a corresponding PCR amplification program; comprises 1) extracting total DNA of strains to be detected as PCR amplification template; 2) PCR amplification is carried out on EMAT1-1L/EMAT1-1R by using a primer pair to obtain a strain with an expected band, showing that the strain contains MAT1-1 mating type, PCR amplification is carried out on EMAT1-2L/EMAT1-2R by using a primer pair to obtain a strain with an expected band, showing that the strain contains MAT1-2 mating type, if only one mating type strain is detected, showing that the strain only contains a male parent or a female parent,
the specific primer pair of the EMAT1-1L/EMAT1-1R is as follows:
the nucleotide sequence of EMAT1-1L is 5'-TAGGTAGGTCCCAAGAACACC-3'; (SEQ ID No.1)
The nucleotide sequence of EMAT1-1R is 5'-GATACCATGGCGAACATTCTG-3'; (SEQ ID No.2)
The MAT1-2L/MAT1-2R specific primer pair comprises:
the nucleotide sequence of EMAT1-2L is 5'-CTTGCCACTACGCGGTCTAT-3', (SEQ ID No.3)
The nucleotide sequence of EMAT1-2R is 5'-CACGGCTCTGGTATCCATTC-3'; (SEQ ID No.4)
Furthermore, PCR amplification systems for identifying mating types MAT1-1 and MAT1-2 are the same, and both comprise 2 × PCRnix12.5 muL, 1 muL of forward and reverse primers with the concentration of 5 muM, 10ng of DNA template and ddH2Make up to 25. mu.l of O.
Further, preferred are PCR amplification programs of the specific primer pair EMAT1-1L/EMAT1-1R for identifying mating type MAT1-1 and the specific primer pair EMAT1-2L/EMAT1-2R for identifying mating type MAT1-2, wherein the amplification programs of the four pairs of specific primers are the same and are all as follows: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 1min, annealing at 50 ℃ for 30sec, extension at 72 ℃ for 1min, and 35 cycles; extension at 72 ℃ for 10 min.
When the specific primer pair EMAT1-1L/EMAT1-1R is used for detecting the mating type of MAT1-1 of the strain, a unique band with the size of about 0.7kb can be obtained.
When the specific primer pair EMAT1-2L/EMAT1-2R is used for detecting the mating type of the strain MAT1-2, a unique band with the size of about 0.8kb can be obtained.
The invention provides a PCR specific primer pair for identifying the mating types of twenty two species in a yellow morchella group: the specific primer pair for detecting mating type MAT1-1 is EMAT1-1L/EMAT1-1R, and the specific primer pair for detecting mating type MAT1-2 is EMAT1-2L/EMAT 1-2R.
The invention also provides application of the PCR primer pair, the reagent or the kit in identification of mating types of twenty-two species in the yellow morchella group, cultivation of morchella, strain breeding, species identification, sexual reproduction evolution and phylogenetic research.
In the invention, an EMAT1-1L/EMAT1-1R primer pair is used for PCR amplification to obtain a unique band of about 0.7kb, which indicates that the strain contains MAT1-1 mating type; an EMAT1-2L/EMAT1-2R primer pair is used for PCR amplification to obtain a unique band of about 0.8kb, which indicates that the strain contains MAT1-2 mating type; the detection of the strain indicates that the strain is a strain having two mating types and can be used as an effective cultivation strain if two expected bands are detected, and indicates that the strain is a strain having only one mating type and cannot be used for cultivation if only one expected band is detected.
The invention designs four pairs of specific primers based on the characteristics of two mating gene sequences of M.steppcola, M.yangii, M.yishuica, M.clivicola, M.dunensis, M.palazonii, M.antiarcica, M.esculenta, M.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27 obtained by previous research work, and can be universally used for the detection of the mating types of the twenty-two morchella strains MAT1-1 and MAT 1-2. The amplified band is a single band, the mating type of the strain can be determined by the presence or absence of the band, and the result is stable and reliable.
The mating gene sequences and detection methods of twenty-two morchella related in the invention are not reported at home and abroad at present. The four pairs of specific primer pairs can be universally used for detecting the mating types of twenty-two morchella, and have important significance for researching sexual reproduction and system evolution of morchella.
The primer pair EMAT1-1L/EMAT1-1R can be universally used for MAT1-1 mating type detection of the twenty-two morchella.
The primer pair EMAT1-2L/EMAT1-2R can be universally used for MAT1-2 mating type detection of the twenty-two morchella.
Compared with the prior art, the invention has the innovation points that: m. steppcola, M.yangii, M.yishuica, M.clivicola, M.dunensis, M.palazonii, M.americana, M.esculenta, M.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27 gene sequences are not reported at present at home and abroad, four pairs of specific primers of the invention can be commonly used for amplification of conserved segments of the twenty two toadstool mating genes, and the mating type of strains can be identified by the existence of amplified bands, and the method is simple and the result is stable and reliable. The method can be further applied to scientific researches such as morel cultivation, strain breeding, species identification, sexual reproduction evolution, phylogeny and the like.
Drawings
FIG. 1 is an electrophoretogram for determining the mating type of Mes-21 monospora strain. MAT1-1, EMAT1-1L/EMAT1-1R primer pair amplifies mating gene MAT 1-1; MAT 1-2: the EMAT1-2L/EMAT1-2R primer pair amplifies the mating gene MAT 1-2. 1-11 represent Mes-21 ascospore monospore strains L110-1-L110-11.
FIG. 2 is a 22 yellow Morchella species Maxillus (MP) tree constructed based on MAT1-1 mating type genes. These twenty-two species are: m.steppcola, M.yangii, M.yishuica, M.clivicola, M.dunensis, M.palazonii, M.ericana, M.esculenta, M.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27, including wild and cultivated strains from different regions.
FIG. 3 is a 22 yellow Morchella species Maxillus (MP) tree constructed based on MAT1-2 mating type genes. These twenty-two species are: m.steppcola, M.yangii, M.yishuica, M.clivicola, M.dunensis, M.palazonii, M.ericana, M.esculenta, M.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27, including wild and cultivated strains from different regions.
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available by purchase.
Example 1Mes-21L110 Monosporozoite Strain mating type identification Mes-21 Monosporozoite Strain L110-1-L110-11 was inoculated on PDA medium, cultured at 24 ℃ for 7 days, an appropriate amount of mycelium was picked, DNA was extracted by CTAB method, 4. mu.L was taken and DNA quality and concentration were determined by electrophoresis on 1% (W/V) agarose gel.
Respectively taking DNA of L110-1-L110-11 as templates, and carrying out PCR amplification by using a primer pair EMAT1-1L/EMAT1-1R, wherein a 25 mu L amplification system comprises 12.5 mu L of 2 × PCR mix (TIANGENBIOCI), 1 mu L of each 5 mu M EMAT1-1L, EMAT1-1R primer, 1 mu L of DNA template and ddH2O9.5. mu.L. The PCR amplification procedure was: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 1min, annealing at 50 ℃ for 30sec, extension at 72 ℃ for 1min, and 35 cycles; extension at 72 ℃ for 10 min.
Respectively taking DNA of L110-1-L110-11 as a template, carrying out PCR amplification by using a primer pair EMAT1-2L/EMAT1-2R, wherein a 25 mu L amplification system comprises 12.5 mu L of 2 × PCR mix (TIANGENBIO Inc), 1 mu L of each 5 mu M EMAT1-2L, EMAT1-2R primer, 1 mu L of DNA template and ddH2O9.5. mu.L. The PCR amplification procedure was: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 1min, annealing at 50 ℃ for 30sec, extension at 72 ℃ for 1min, and 35 cycles; extension at 72 ℃ for 10 min.
All amplification products were electrophoretically detected on a 1% (W/V) agarose gel, and the results are shown in FIG. 1, and strains L110-3, L110-5, L110-7 and L110-10 had a unique band of about 0.7kb when the primer pair MAT1-1L/MAT1-1R was amplified, whereas no band was detected when the primer pair MAT1-2L/MAT1-2R was amplified, and they were identified as mating type MAT1-1 strain. While strains L110-1, L110-2, L110-4, L110-6, L110-8, L110-9 and L110-11 had no band when the primer pair MAT1-1L/MAT1-1R was amplified, and had a unique band of about 0.8kb when the primer pair MAT1-2L/MAT1-2R was amplified, and were identified as mating type MAT1-2 strains.
By using the primers and the detection method for detecting the mating type genes of the morchella strains, the reliability of high yield and stable yield in the cultivation process of the morchella can be greatly improved. The molecular marker of the mating type gene is simple, convenient and easy to operate, and can be applied to morchella mating type gene identification, cultivation, strain breeding, species identification, sexual reproduction evolution and phylogenetic research.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Sequence listing
SEQ IDNo.1
SEQ IDNo.2
SEQ IDNo.3
SEQ IDNo.4
Sequence listing
<110> university of Chongqing teacher
<120> a method for identifying the mating types of twenty-two species in yellow morchella flora
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gataccatgg cgaacattct g 21
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cttgccacta cgcggtctat 20
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<213> Artificial sequence (Esculenta Clade)
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cacggctctg gtatccattc 20
Claims (10)
1. A mating type method for identifying twenty-two species in a yellow morchella group, wherein the twenty-two species are m.steppcola, m.yangii, m.yishuica, m.clivicola, m.dunensis, m.palazonii, m.amelicana, m.esculena, m.galilaea, Mes-6, Mes-9, Mes-10, Mes-15, Mes-19, Mes-20, Mes-21, Mes-22, Mes-23, Mes-24, Mes-25, Mes-26 and Mes-27, characterized in that a specific primer pair EMAT1-1L/EMAT1-1R for detecting mating type 1-1, a specific primer pair EMAT 1-57L/EMAT 1-1R for detecting mating type MAT1-2, and a PCR amplification system corresponding thereto;
wherein the EMAT1-1L/EMAT1-1R specific primer pair,
the nucleotide sequence of EMAT1-1L is 5'-TAGGTAGGTCCCAAGAACACC-3',
the nucleotide sequence of EMAT1-1R is 5'-GATACCATGGCGAACATTCTG-3';
the EMAT1-2L/EMAT1-2R specific primer pair,
the nucleotide sequence of EMAT1-2L is 5'-CTTGCCACTACGCGGTCTAT-3',
the nucleotide sequence of EMAT1-2R is 5'-CACGGCTCTGGTATCCATTC-3'.
2. The method for identifying the mating types of twenty-two of morchella yellow flora according to claim 1, wherein the PCR amplification systems for identifying the mating types MAT1-1 and MAT1-2 are the same, and each PCR amplification system comprises 2 × PCR mix12.5 μ L, 1 μ L of 5 μ M forward and reverse primers, 20ng DNA template and ddH2Make up to 25. mu.L of O.
3. The method for identifying the mating types of twenty-two in the yellow morchella group as claimed in claim 2, wherein the PCR amplification program for identifying the MAT1-1 mating type of the strain by using the specific primer pair MAT1-1L/MAT1-1R is as follows: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 1min, annealing at 50 ℃ for 30sec, extension at 72 ℃ for 1min, 35 cycles; extension at 72 ℃ for 10 min.
4. The method for identifying the mating types of twenty-two of yellow morchella populations according to claim 3, wherein a unique band of about 0.7kb in size can be obtained when the specific primer pair MAT1-1L/MAT1-1R detects the MAT1-1 mating type of the strain.
5. The method for identifying the mating types of twenty-two in the yellow morchella group as claimed in claim 2, wherein the PCR amplification program for identifying the MAT1-2 mating type of the strain by using the specific primer pair MAT1-2L/MAT1-2R is as follows: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 1min, annealing at 50 ℃ for 30sec, extension at 72 ℃ for 1min, 35 cycles; extension at 72 ℃ for 10 min.
6. The method for identifying the mating types of twenty-two of yellow morchella populations according to claim 5, wherein a unique band of about 0.8kb can be obtained by detecting the mating type of the strain MAT1-2R with a specific primer pair MAT1-2L/MAT 1-2R.
7. The method for identifying the mating types of twenty-two members of the yellow morchella group according to claim 2, wherein: the PCR primer pair for identifying the mating types of twenty-two species in the yellow morchella flora comprises a specific primer pair for detecting the mating type MAT1-1, namely EMAT1-1L/EMAT1-1R, and a specific primer pair for detecting the mating type MAT1-2, namely EMAT1-2L/EMAT 1-2R.
8. A reagent for identifying the mating types of twenty-two of the yellow morchella group, which comprises the PCR primer pair of claim 7.
9. A kit for identifying the mating types of twenty-two of yellow morchella populations, comprising the PCR primer pair of claim 7.
10. Use of a primer set comprising the sequence of claim 4 or claim 5, the PCR primer set of claim 7 or the reagent of claim 8 or the kit of claim 9 for the identification of the mating type, species selection, cultivation, species identification, sexual evolution and phylogenetic studies of twenty-two species of the yellow morchella group.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107151698A (en) * | 2017-04-01 | 2017-09-12 | 中国科学院昆明植物研究所 | Identify the mating type method of 11 kinds in black hickory chick monoid |
| CN108828103A (en) * | 2018-08-21 | 2018-11-16 | 辽宁省农业科学院 | Hickory chick HPCE fingerprint and its standard finger-print |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107151698A (en) * | 2017-04-01 | 2017-09-12 | 中国科学院昆明植物研究所 | Identify the mating type method of 11 kinds in black hickory chick monoid |
| CN108828103A (en) * | 2018-08-21 | 2018-11-16 | 辽宁省农业科学院 | Hickory chick HPCE fingerprint and its standard finger-print |
Non-Patent Citations (2)
| Title |
|---|
| XI-HUI DU等: "Correction to: Heterothallism and potential hybridization events inferred for twenty-two yellow morel species", IMA FUNGUS, vol. 13, no. 1, 19 May 2022 (2022-05-19), pages 2 * |
| 熊川等: "四川秋季发生的两种羊肚菌生境调查与鉴定", 菌物学报, vol. 35, no. 1, 22 January 2016 (2016-01-22), pages 2 * |
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