CN-121817020-B - Method for breeding fruiting body by culturing under pure culture condition of Boletus sinensises

CN121817020BCN 121817020 BCN121817020 BCN 121817020BCN-121817020-B

Abstract

The invention discloses a method for breeding fruiting bodies under pure culture conditions of Boletus sinensises, belonging to the technical field of edible fungus breeding and cultivation. The method comprises the steps of activating a Chinese saprophytics strain JSJ-Bx1, inoculating the activated Chinese saprophytics strain JSJ-Bx1 into a culture medium for dark culture, performing low-temperature induction culture on a plate which is close to overgrown with mycelia, converting the mycelia from white to golden yellow, generating mycelia to be twisted, transferring the mycelia plate into a fruiting room for culture, continuing to perform induction culture and development into mature sporophores after obvious primordium differentiation is seen, collecting sporophore basidiospores, diluting, and then coating the sporophore basidiospore into a B1 culture medium for germination culture, so as to obtain monospora and polyspora mycelia serving as primordium for breeding of Chinese saprophytics. The invention can directly and quickly culture in an agar culture medium to obtain mature fruiting bodies of the Boletus sinense, and the harvested basidiospores can normally germinate and grow, so that the whole life history of the Boletus sinense can be completed under the pure culture condition.

Inventors

LI FAN
LI RONGCHUN
CHEN YONG
Cao yao
LI RONGPING
LUO XIANGYING
GUO JING

Assignees

云南菌视界生物科技有限公司

Dates

Publication Date: 20260512
Application Date: 20260313

Claims (4)

1. The method for breeding by culturing fruiting bodies under pure culture conditions of Boletus sinense (Buchwaldoboletus xylophilus) is characterized by comprising the following steps: S1, activating a Chinese saprophytic bolete strain JSJ-Bx1, inoculating the activated Chinese saprophytic bolete strain JSJ-Bx1 into a B1 culture medium, and performing dark culture for 5-7 days at a culture temperature of 30-35 ℃, wherein the B1 culture medium comprises, by mass, glucose 20 g, yeast extract 2 g, ferrous sulfate 1.5 g, 1/2MS culture medium 0.3 g, agar 20 g and water 1L; S2, performing low-temperature induction culture on the activated flat plate close to full mycelia, wherein the culture temperature is 20-30 ℃, the dark culture is performed for 5-7 days, the mycelia are changed from white to golden yellow, and mycelia kink; S3, transferring the mycelium plates cultured in the step S2 into a fruiting room for primordial induction culture, wherein the culture temperature T is 26 ℃ or less and T <30 ℃, the relative air humidity is 10-90%, the lighting conditions are 12 h/12 h, and the culture is performed alternately in a light-dark manner for 7-10 days, and after obvious primordial differentiation is seen, the culture is continued to be induced and developed into mature fruiting bodies; s4, collecting basidiospores of the obtained mature sporophore under aseptic conditions, diluting the basidiospores by 10 2 ~10 5 , coating the basidiospores in a B1 culture medium, performing germination culture at 28-30 ℃, regarding hyphae growing by germination of single colonies as monospore-forming hyphae, regarding hyphae growing by germination of a plurality of colonies as multi-spore hyphae, and regarding the monospore-forming hyphae and the multi-spore hyphae as stock seeds for breeding of the Chinese saprophytic bolete.
2. The method for breeding seed bodies in pure culture of Boletus sinense (Buchwaldoboletus xylophilus) according to claim 1, wherein in step S2, the culture temperature for low-temperature induction culture of the mycelium-grown flat plate is 26-28 ℃.
3. The method for breeding seed bodies in pure culture of Boletus sinense (Buchwaldoboletus xylophilus) according to claim 1, wherein in the step S3, the primordial induction culture temperature is 28 ℃ and the relative air humidity is 70%.
4. The method for breeding seed bodies in pure culture of Boletus sinense (Buchwaldoboletus xylophilus) according to claim 1, wherein in step S4, the basidiospores are diluted 10: 10 3 and then spread on B1 medium for germination culture.

Description

Method for breeding fruiting body by culturing under pure culture condition of Boletus sinensises Technical Field The invention belongs to the technical field of edible fungus breeding and cultivation in related industries of biological agriculture, and particularly relates to a method for rapidly obtaining a Chinese saprophytic bolete fruiting body and breeding directly on an agar medium. Background Both & B. Ortiz, petch) Both & B, belonging to Boletaceae Boletaceae, boletidae Chalciporoideae, and Boletidae Buchwaldoboletus, is a typical tropical and subtropical rare Boletus, is a non-exogenous mycorrhizal fungus, has a saprophytic capacity, is edible, has a high economic value, and is extremely deficient in wild resources. In order to solve the problem of scarcity of boletus sinensis cultivation strains, related domestic units and personnel research and study on the artificial cultivation and propagation of boletus sinensis, a boletus sinensis artificial cultivation method is typically disclosed in Chinese patent CN113348963A, a boletus sinensis mother culture medium is disclosed in Chinese patent CN120591113A, and a preparation method and application thereof, and the biological characteristics and cultivation of the boletus sinensis are disclosed in ' fungus journal of academy of sciences ' (https)// doi. Org/10.13346/j. Myctosystem.2502 '. However, the methods basically have complicated processes of stock culture, fungus package inoculation, hypha fungus growing culture, earthing fruiting and the like, have long culture time and high cost, and become an obstacle in the breeding process of the bolete of Chinese saprophyticus. Disclosure of Invention The invention aims to solve the defects of the prior breeding technology, and provides a method capable of carrying out the culture of fruiting bodies of the Boletus sinensises under pure culture conditions and carrying out efficient breeding so as to greatly shorten the breeding and breeding period of the Boletus sinensises and realize efficient fruiting body regeneration of the Boletus sinensises. The technical scheme adopted by the invention is as follows: the method for breeding by culturing fruiting bodies under pure culture conditions of Boletus sinense (Buchwaldoboletus xylophilus) comprises the following steps: S1, activating a Chinese saprophytic bolete strain JSJ-Bx1, inoculating the activated Chinese saprophytic bolete strain JSJ-Bx1 into a B1 culture medium, and performing dark culture for 5-7 days at a culture temperature of 30-35 ℃, wherein the B1 culture medium comprises, by mass, glucose 20 g, yeast extract 2 g, ferrous sulfate 1.5 g, 1/2MS culture medium 0.3 g, agar 20 g and water 1L; S2, performing low-temperature induction culture on the activated flat plate close to full mycelia, wherein the culture temperature is 20-30 ℃, the dark culture is performed for 5-7 days, the mycelia are changed from white to golden yellow, and mycelia kink; S3, transferring the mycelium plates cultured in the step S2 into a fruiting room for primordial induction culture, wherein the culture temperature T is 26 ℃ or less and T <30 ℃, the relative air humidity is 10-90%, the lighting conditions are 12 h/12 h, and the culture is performed alternately in a light-dark manner for 7-10 days, and after obvious primordial differentiation is seen, the culture is continued to be induced and developed into mature fruiting bodies; s4, collecting basidiospores of the obtained mature sporophore under aseptic conditions, diluting the basidiospores by 10 2~105, coating the basidiospores in a B1 culture medium, performing germination culture at 28-30 ℃, regarding hyphae growing by germination of single colonies as monospore-forming hyphae, regarding hyphae growing by germination of a plurality of colonies as multi-spore hyphae, and regarding the monospore-forming hyphae and the multi-spore hyphae as stock seeds for breeding of the Chinese saprophytic bolete. Preferably, in the step S2, the temperature of the low-temperature induced culture is 26-28 ℃ for the flat plate near the overgrown mycelium. Preferably, in the step S3, the primordial induction culture temperature is 28 ℃ and the air relative humidity is 70%. Preferably, in the step S4, the basidiospores are diluted to 10 3 and then spread on the B1 medium for germination culture. By utilizing the method for breeding the fruiting body under the pure culture condition, the mature fruiting body of the Boletus sinense can be directly and rapidly cultured in an agar culture medium, and the harvested basidiospore can normally germinate and grow, so that the whole life history of the Boletus sinense under the pure culture condition is completed. The method can rapidly and efficiently obtain the basidiomycete of the Boletus sinensises with physiological activity, quickens the breeding speed of the species of the Boletus sinensises, greatly shortens the breeding period of the species of the Boletus sinensises, provides a new way f