Method for identifying or assisting in identifying mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-818 thereof

A technology of protoplasts and mushrooms, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc., can solve the problems of narrow adaptation temperature range, fragile fruit body, difficult transportation and storage, etc. Achieve high accuracy and reliability

Active Publication Date: 2013-07-03
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
View PDF1 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in actual production, there are problems such as low yield, poor insect resistance,

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying or assisting in identifying mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-818 thereof
  • Method for identifying or assisting in identifying mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-818 thereof
  • Method for identifying or assisting in identifying mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-818 thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0029] The preparation method of the solid regeneration plate (RM): add sorbitol and agar to the liquid MCM medium so that the concentrations are 1M and 20g / L, respectively.

[0030] 1.2 Protoplast preparation

[0031] 1) Take the mycelia of Azalea variegata in liquid MCM medium, culture at 160rpm, 25°C for 4 days (3-5 days are acceptable), filter with 3 layers of sterile lens paper to collect the mycelia, and then wash with 0.6M mannitol Wash with aqueous solution 2-3 times.

[0032] 2) Suspend the mycelia (about 1 g) of step 1) in 1.5% lysozyme solution (dissolve 1.5 g lysozyme with 0.6M mannitol aqueous solution and set the volume to 100 mL; lysozyme was purchased from Guangdong Bi German Biotechnology Co., Ltd.

[0033] Division, product catalog number: Bd_8110001023), in a water-bath shaker (32 ° C, 60 rpm) for 2 hours, filter with 3 layers of sterile lens tissue and collect the filtrate.

[0034] 3) Centrifuge the filtrate of step 2) at 3000 rpm for 10 min and collect...

Embodiment 1

[0047] Example 1. Using PCR primer pair IS-818 to identify the mating type of the protoplast monokaryon of Pleurotus chinensis

[0048] 1. PCR reagents for identifying or assisting in identifying the mating type of monokaryon protoplasts

[0049] The reagents for identifying or assisting in identifying the mating type of protoplast monokaryon in this embodiment consist of PCR primer pair IS-818, 10 × Taq buffer, dNTP mix, Taq DNA polymerase and ddH 2 O composition.

[0050] Among them, the PCR primer pair IS-818 is composed of two single-stranded DNAs, IS-818-pre-F and IS-818-pre-R, and its sequence is as follows:

[0051] IS-818-pre-F:5'-CAGAGAAAGAATTGGCAAAGGT-3' (SEQ ID No.1)

[0052] IS-818-pre-R: 5'-CAATCAACAGGAGGTCTACGA-3' (SEQ ID No. 2).

[0053] 10×Taq buffer, dNTP mix and Taq DNA polymerase were purchased from Beijing Shengxu Baichuan Company (CNS).

[0054] 2. Identify or assist in the identification of monokaryotic mating types of protoplasts

[0055] Inoculate ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for identifying or assisting in identifying the mating type of Lepista sordid protoplast monokaryons and special primer pairs IS-818 thereof. The method for identifying or assisting in identifying the mating types of the Lepista sordid protoplast monokaryons comprises the following steps: respectively taking the genome DNA of two Lepista sordid protoplast monokaryons to be identified as templates; performing polymerase chain reaction (PCR) amplification by using PCR primer pairs shown in SEQ ID No. 1 and 2; detecting the size of the obtained PCR products; if the PCR products of the two Lepista sordid protoplast monokaryons to be identified comprise 800 to 1,000 bp of DNA segments or do not comprise 800 to 1,000 bp of DNA segments, the mating types of two Lepista sordid protoplast monokaryons are identical; and if the PCR product of one of the two Lepista sordid protoplast monokaryons to be identified comprises 800 to 1,000 bp of DNA segments and the PCR product of the other one of the two Lepista sordid protoplast monokaryons to be identified does not comprise 800 to 1,000 bp of DNA segments, the mating types of two Lepista sordid protoplast monokaryons are different.

Description

technical field [0001] The invention relates to a method for identifying or assisting in identifying the monokaryon mating type of protoplasts of variegated mushrooms and the special primer pair IS-818. Background technique [0002] The nutrient incompatibility system is a genetic system for identifying aliens. In most fungi, this system can cause genetically distinct individuals to produce characteristic somatic incompatibility in the junction zone (Qi Yuancheng et al., Chinese cultivated pellagra Comparison of somatic cell incompatibility test and RAPD analysis results of Pleurotus species. Acta Mycophyta Sinica, 2010, 29 (3)) reaction, the intensity of somatic cell incompatibility reaction varies from individual to individual. In fungi, nutritional incompatibility is also known as somatic incompatibility or heterokaryotic incompatibility. fusion to maintain individual genetic stability. The mating type is the type of combination determined according to whether the indiv...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12N15/11
Inventor 许峰刘宇李登进王守现赵爽耿小丽王兰青
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap