Artificially domesticated phlcbopus portentosus strain HZ18006 and SSR marker fingerprint thereof

A technology of HZ18006, black boletus, applied in fungi, microorganism-based methods, and methods of using microorganisms, etc., can solve the problems of difficult to obtain strains, large individual differences, etc., to simplify harvesting procedures and labor, The effect of good image and stable genetic characteristics

Active Publication Date: 2019-11-08
JINGHONG HONGZHEN AGRI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 fr

Method used

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  • Artificially domesticated phlcbopus portentosus strain HZ18006 and SSR marker fingerprint thereof
  • Artificially domesticated phlcbopus portentosus strain HZ18006 and SSR marker fingerprint thereof
  • Artificially domesticated phlcbopus portentosus strain HZ18006 and SSR marker fingerprint thereof

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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 10X LoadingBuffer 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.

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Abstract

The invention discloses an artificially domesticated phlcbopus portentosus strain HZ18006 and a SSR marker fingerprint thereof. The strain has the ability to significantly increase the yield of fruiting bodies, is deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 18, 2019 with the preservation number of CGMCC No.17080. The laboratory and production practiceconfirm that the strain has high strain vigor and stable genetic characteristics, the fruiting speed is high and the fruiting is tidy in the late stage, the growth speed is high, the needs of large-scale factory cultivation are met, harvesting can be completed at one time in a best harvesting period, and the harvesting process is greatly simplified and labor is saved; and excellent biological growth characteristics such as simple fruiting body cultivation method, extensive management, high fruiting and high yield are further achieved. The SSR marker fingerprint of the HZ18006 strain is composed of 10 pairs of SSR markers, and is combined with an accurate quantitative capillary electrophoresis fluorescence detection method for rapid identification of the HZ18006 strain.

Description

technical field [0001] The invention belongs to the technical field of edible fungus artificial breeding and identification of strains, and in particular relates to a new strain of boletus nigras which is obtained through artificial domestication, has excellent biological characteristics and is suitable for industrialized cultivation conditions. At the same time, the invention also relates to a specific SSR marker fingerprint that can accurately identify the new strain of Boletus nigricans. 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...

Claims

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Application Information

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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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