Method and specific primer pairs for authenticating Morchella importuna mating types

A kind of technology of Morchella latifolia and specific primer pair, applied in the field of biology, can solve problems such as loss of mating type, and achieve the effect of obvious social benefit

Active Publication Date: 2017-01-04
云南省农业科学院生物技术与种质资源研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Both the spore isolation and tissue isolation of Morchella may only obtain a strain wit

Method used

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  • Method and specific primer pairs for authenticating Morchella importuna mating types
  • Method and specific primer pairs for authenticating Morchella importuna mating types
  • Method and specific primer pairs for authenticating Morchella importuna mating types

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Identification of the mating type of the monoascospore population of Morchella spp. YPL6

[0033] The monoascospore strain YPL6-1—YPL6-24 of Morchella rhizome YPL6 was inoculated on PDA medium, cultured at 23°C for 10 days, picked appropriate amount of mycelia, extracted DNA by CTAB method, and took 3 μL in 1% (W / V) Agarose gel electrophoresis to detect DNA quality and concentration.

[0034] Using the DNA of YPL6-1—YPL6-24 as a template, PCR amplification was performed on P8-4F / P8-4R with primers. The 25 µL amplification system included 2×PCRmix (TSINGKE Bio Inc) 12.5 µL, 10 µM / L 1 µL each of P8-4F and P8-4R primers, 1 µL of DNA template, ddH 2 O 9.5 µL. The PCR amplification program was as follows: pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 sec, annealing at 60°C for 30 sec, extension at 72°C for 90 sec, 30 cycles; extension at 72°C for 10 min.

[0035] The DNA of YPL6-1—YPL6-24 was used as a template respectively, and the P6-1F / P6-1...

Embodiment 2

[0037] Embodiment 2 The identification of the effectively cultivated strains of Morchella thalamus

[0038] The strains to be tested are Morchella sp. sp1, SP2, YPL6, and YPL7. The strain cultivation, DNA extraction, PCR amplification system and amplification procedure are the same as in Example 1.

[0039] The amplified product was detected by electrophoresis on a 1.2% (W / V) agarose gel, and the results are shown in image 3 , SP2, YPL6, YPL7 strains have bands in PCR amplification with P8-4F / P8-4R, P6-1F / P6-1R primers, indicating that they have two mating types and are effective cultivation strains. The SP1 strain has no band when amplified by the P8-4F / P8-4R primer pair, but has a band amplified by the P6-1F / P6-1R primer pair. It is a MAT1-2 mating type strain and cannot be used as a cultivated strain. Strains YPL6 and YPL7 had fruited after cultivation.

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Abstract

The invention relates to a method and specific primer pairs for authenticating Morchella importuna mating types, and belongs to the technical field of biology. The primer pairs comprise the specific primer pair P8-4F/P8-4R for detecting the mating type MAT1-1 and the specific primer pair P6-1F/P6-1R for detecting the mating type MAT1-2. The method includes the steps that strain DNA is extracted, and PCR amplification is carried out with the two primer pairs; if expected bands can be obtained in all times of electrophoresis detection, the strain contains two kinds of mating type genes and is an effective cultivation strain; if only one amplified band can be detected, the strain contains only one mating type and can not be used for cultivation. The detection result is high in specificity, detection is stable, reliable and fast, and the method and the specific primer pairs are suitable for research on the aspects of Morchella strain breeding, cultivation, mating type genes and phylogeny.

Description

technical field [0001] The invention belongs to the technical field of biology, and in particular relates to a method for identifying the morel ( Morchella importuna ) method of mating type and specific primer pairs. The technology of the invention is applicable to the research on the selection, cultivation, mating type gene and phylogeny of hickory chick species. Background technique [0002] The sexual reproduction of Ascomycota is currently thought to be controlled by a single mating site, MAT1, which includes the MAT1-1 mating type characterized by the MAT1-1-1 gene and the MAT1-2 characterized by the MAT1-2-1 gene mating type. Ascomycetes with homothallic mating are self-fertile, while ascomycetes with heterothallic mating are self-fertile. They must be mated by individuals with mating type MAT1-1 and mating type MAT1-2 to complete sexual reproduction. [0003] Morel is a delicious edible fungus with high economic value. The current price of wild dried products is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895
Inventor 柴红梅赵永昌陈卫民张小雷苏开美刘萍陈立佼
Owner 云南省农业科学院生物技术与种质资源研究所
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