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Monocaryon mating type primer set for identifying Shenxiang 215 strains and substantive derivative varieties of shiitake mushrooms, identifying method and application

A substantively derived and mating-type technology, applied in biochemical equipment and methods, recombinant DNA technology, and resistance to vector-borne diseases, etc., can solve problems such as different time and location of picking colonies, heavy workload, and experimental impact , to achieve the effect of shortening the identification time, short detection time and high accuracy

Active Publication Date: 2019-07-19
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Crossbreeding often involves a large number of monokaryons, and there are many combinations of confrontation culture between monokaryons, resulting in a large workload and time-consuming for inoculation and microscopic examination.
In addition, due to the formation time of lock-like junctions in the process of microscopic examination, the time and location of picking colonies are different, which is easy to cause human error detection, which will affect the follow-up experiments.

Method used

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  • Monocaryon mating type primer set for identifying Shenxiang 215 strains and substantive derivative varieties of shiitake mushrooms, identifying method and application
  • Monocaryon mating type primer set for identifying Shenxiang 215 strains and substantive derivative varieties of shiitake mushrooms, identifying method and application
  • Monocaryon mating type primer set for identifying Shenxiang 215 strains and substantive derivative varieties of shiitake mushrooms, identifying method and application

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Embodiment 1

[0040] This embodiment provides a method for identifying the monokaryon mating type of Lentinus edodes Shenxiang 215, which specifically includes the following steps in sequence:

[0041] (1) Mycelia culture: 176 spore monokaryon strains of Lentinus edodes Shenxiang 215 were transferred to a plate equipped with potato dextrose agar medium PDA, and cultured on a plate with a diameter of 90 cm at 25° C. in the dark.

[0042](2) Rapid preparation of genomic DNA: After mycelia were cultured for 5 days, 100 μLTE buffer was added to each sample tube of a 96-well plate, and a small amount of mycelium was picked from each monokaryon strain with the tip of an inoculation needle and suspended in In the sample tube, record the serial number, process the 96-well plate at 98°C for 5 minutes with a PCR machine, and immediately place it on ice to cool it for later use.

[0043] In the embodiment of the present invention, the method of rapidly preparing DNA from mycelium can realize rapid ide...

Embodiment 2

[0056] This embodiment provides a method for identifying the monokaryotic mating type of Lentinus edodes strain L1641 (the substantive derivative of Shenxiang 215), which specifically includes the following steps in sequence:

[0057] (1) Mycelia culture: 159 spore monokaryon strains of Lentinus edodes strain L1641 were transferred to a flat plate equipped with potato dextrose agar medium PDA, and placed on a flat plate with a diameter of 90 cm at 25° C. in the dark.

[0058] (2) Rapid preparation of genomic DNA: After mycelia were cultured for 5 days, 100 μLTE buffer was added to each sample tube of a 96-well plate, and a small amount of mycelium was picked from each monokaryon strain with the tip of an inoculation needle and suspended in In the sample tube, record the serial number, process the 96-well plate at 98°C for 5 minutes with a PCR machine, and immediately place it on ice to cool it for later use.

[0059] (3) Detecting the type of mating factor of the strain: perfo...

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Abstract

The invention discloses a monocaryon mating type primer set for identifying Shenxiang 215 strains and substantive derivative varieties of shiitake mushrooms, an identifying method and an application,and relates to the technical field of edible mushroom molecular marked auxiliary breeding. The monocaryon mating type primer set specifically comprises 4 pairs of primers. The identifying method specifically comprises the steps of culturing mycelia, performing quick preparation of genomic DNA, performing mating type PCR detection, and determining the result of electrophoresis detection. Compared with a conventional monocaryon mating type identification method, the identifying method disclosed by the invention has the advantages that detection time is shoter and accuracy is higher; and throughthe adoption of the identifying method disclosed by the invention, the identifying time can be greatly shortened, the false detection rate can be reduced, and dicaryon elimination can be realized.

Description

technical field [0001] The invention relates to the technical field of molecular marker-assisted breeding of edible fungi, in particular to a primer set for identifying the monokaryon mating type of Lentinus edodes Shenxiang 215 strain and its substantive derivatives, an identification method and an application. Background technique [0002] Obtaining monokaryon and identifying its mating type is one of the basic tasks in the study of mushroom genetics and breeding. The mating type determines the compatibility between the two monokaryotic parents of the hybrid, and the non-compatible monokaryotic cannot hybridize to form dikaryotic offspring. [0003] At present, the mating type identification of shiitake mushrooms is usually carried out by observing lock joints by microscopic examination after two-to-two confrontation culture of monokaryons. Crossbreeding work often involves a large number of monokaryons, and the combination of confrontation culture between monokaryons res...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11
CPCC12Q1/6895C12Q2600/156Y02A50/30
Inventor 张美彦章炉军尚晓冬宋春艳于海龙徐珍王瑞娟刘建雨杨慧谭琦
Owner SHANGHAI ACAD OF AGRI SCI
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