Fingerprint of an artificially domesticated Boletus nigricans strain hz18006 and its ssr markers

A technology of HZ18006, black boletus, applied to fungi, methods based on microorganisms, methods using microorganisms, etc., can solve problems such as difficult to obtain strains and large individual differences

Active Publication Date: 2021-12-07
JINGHONG HONGZHEN AGRI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the different genetic backgrounds among individuals of wild fruiting bodies, there are great differences between individuals. Our long-term strain screening practice has proved that only 5% of the strains directly isolated from wild fruiting bodies can reach the fruiting rate of 50%--75%, while the strains with a fruiting rate of more than 95% are only 0.5%-1%
Therefore, with a simple screening method, it is actually difficult to obtain strains that meet the production requirements

Method used

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  • Fingerprint of an artificially domesticated Boletus nigricans strain hz18006 and its ssr markers
  • Fingerprint of an artificially domesticated Boletus nigricans strain hz18006 and its ssr markers
  • Fingerprint of an artificially domesticated Boletus nigricans strain hz18006 and its ssr markers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Obtain black boletus (Phlebopus portentosus) HZ18006 bacterial strain of the present invention after carrying out artificial domestication to wild black boletus fruiting body isolate bacterial strain:

[0039] 1. Specific methods of artificial domestication:

[0040] 1.1 Collection of wild fruiting bodies and strain acquisition: wild black cattle were collected in Jinghong, Mengla, Menghai of Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, and Simao and Jinggu in Pu'er area (host trees include phoenix trees, pomelo trees and jackfruit trees) There were 100 fruiting bodies of Phlebopus portentosus, and the tissue pieces of the fruiting bodies were separated with PDA medium, and 100 wild strains were obtained after the mycelium was overgrown with the medium.

[0041] 1.2 Verification of fruiting ability: The 100 wild strains obtained are cultivated and fruited in a factory cultivation environment, and the strains with a fruiting rate of 50%-75% are selected for do...

Embodiment 2

[0051] The application of the strain HZ18006 in the industrial cultivation of edible fungi: The strain HZ18006 can easily complete the life cycle from the strain to the formation of basidiocarp on the artificial substrate, that is, the artificial cultivation of the fruiting body can be carried out through the strain under artificial conditions, and the Reach the purpose of harvesting black boletus fruiting bodies. Compared with other cultivated strains, it has excellent biological growth characteristics such as convenient fruiting body cultivation method, extensive management, fast fruiting, and high yield. The specific industrial cultivation application conditions are as follows:

[0052] 1. Make the most suitable medium M1 solid medium for HZ18006 strain: 20.0g potato, 20.0g glucose, 2.0g yeast extract, MgSO 4 1.0g, KH 2 PO 4 1.0g medium, 16.0g agar, 1000mL distilled water. M1 liquid medium does not add agar, and the other components are the same as M1 solid medium. On...

Embodiment 3

[0064] Embodiment 3: Construction of the SSR marker fingerprint of Boletus niger HZ18006 bacterial strain:

[0065] Through the development of microsatellite fingerprints, the genetic relationship analysis and species definition of HZ18006 strain and other black boletus strains can be accurately and quickly carried out:

[0066] 1. Extraction and detection of genomic DNA of Boletus nigricans HZ18006 strain: Omega mag-bind DNA kit was used to extract the total DNA of HZ18006 strain, and the operation method was carried out according to the instructions provided by the kit. The extracted DNA sample was mixed with 10XLoadingBuffer at a ratio of 10:1 and loaded on a 1% agarose gel for electrophoresis, and the quality of the extracted total DNA was detected using a gel imager. The concentration of total DNA was measured using a micro-volume ultraviolet spectrophotometer Nanodrop, and the concentration of sample DNA was adjusted to 50-100 ng / μL for PCR amplification reaction.

[00...

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Abstract

The invention discloses a strain HZ18006 of artificially domesticated Boletus nigricans and its SSR marker fingerprint. The strain has the ability to significantly increase the yield of fruiting bodies, and has been preserved in the General Microorganism Center CGMCC of the China Microbiological Culture Collection Management Committee on March 18, 2019, with the preservation number CGMCC No.17080. The laboratory and production practice have proved that the strain of the present invention has strong vitality, stable genetic characteristics, fast fruiting speed in the later stage and neat fruiting, and fast growth rate, which meets the needs of large-scale industrial cultivation, and can be harvested at one time during the best harvest period. It greatly simplifies the harvesting procedure and labor force; it also has excellent biological growth characteristics such as convenient fruiting body cultivation method, extensive management, fast fruiting, and high yield. The SSR marker fingerprint of HZ18006 strain consists of 10 pairs of SSR markers, which can be used for rapid identification of HZ18006 strain in combination with accurate quantitative capillary electrophoresis fluorescence detection method.

Description

technical field [0001] The invention belongs to the technical field of edible fungus artificial breeding and identification of strains, in particular to a black boletus strain obtained through artificial domestication, which has excellent biological characteristics and is suitable for industrialized cultivation conditions. Simultaneously, the invention also relates to a specific SSR marker fingerprint that can accurately identify the boletus niger strain. Background technique [0002] Black boletus (Phlebopus portentosus) is a common name for Boletus genus Phlebopus dark brown reticulate stalk in the order Boletales Boletales family Boletinellaceae. In Yunnan Xishuangbanna Dai Autonomous Prefecture, Honghe Hani and Yi Autonomous Prefecture, Dehong Dai and Jingpo Autonomous Prefecture, Lincang, Pu'er and other regions, as well as Thailand, it is deeply loved by consumers and is a rare edible fungus. The nutritional mode of black boletus is complex, and it has a lifestyle of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N1/36C12Q1/6895C12Q1/04C12R1/645
CPCC12N1/14C12N1/36C12Q1/6895C12Q2600/156C12N1/145C12R2001/645
Inventor 纪开萍纪光燕罗顺珍纪光玉高丽霞范春梦王秋兰陈建华
Owner JINGHONG HONGZHEN AGRI TECH CO LTD
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