A black bolete strain AC2266 and its application

Abstract

The present invention discloses a Boletus aereus strain AC2266 and its application. This strain is obtained by self-crossing of YL1701-2, and was deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 3, 2021. The deposit number is CGMCC NO.21923, and the taxonomic name is: Boletus aereus AC2266. The present invention also discloses the application of the Boletus aereus strain in the production of Boletus aereus. This strain has a relatively fast mycelial growth rate, a relatively low optimal growth temperature, a darker cap color, a thicker cap, and a yellower stipe, and is an excellent new variety that can be used for the industrial production of Boletus aereus.

Description

Technical Field

[0001] The present invention belongs to the technical field of artificial breeding of edible fungi, and specifically relates to a new strain of Boletus aereus and its application. Background Art

[0002] Boletus aereus is a large fungus belonging to the genus Phlebopus in the family Boletinellaceae of the order Boletales, commonly known as Phlebopus portentosus. Currently, Boletus aereus is a type of bolete that can be cultivated to produce mature fruiting bodies without a host tree, and has broad market prospects.

[0003] Large-scale production is inseparable from excellent strains. Currently, the main methods for obtaining excellent strains are wild domestication and crossbreeding. Among them, crossbreeding is the most effective way to obtain excellent strains, and the specific methods include: self-crossing, related crossbreeding, allogenic crossbreeding, distant crossbreeding, etc. The process of crossbreeding work includes steps such as mate selection, identification, screening, and large-scale verification. During the crossbreeding process, the genes of the parents will be recombined, resulting in many new traits that are quite different from the parents.

[0004] Boletus aereus belongs to a tetrapolar mating system. Therefore, the self-crossing method of selecting closely related gametes for hybridization requires much more work than crossbreeding with distantly related gametes. Compared with other crossbreeding methods, self-crossing has the advantage of aggregating and homozygosing excellent genes, and at the same time, many new traits that are quite different from the parents appear due to the recombination of genes. Therefore, it is also selected by many breeders. Summary of the Invention

[0005] In view of this, an embodiment of the present invention provides a strain of Boletus aereus AC2266 with excellent cultivation traits and suitable for industrial production, and provides the application of the AC2266 strain in the production of Boletus aereus.

[0006] To achieve the above object, the present invention mainly provides the following technical solutions:

[0007] On the one hand, an embodiment of the present invention provides a strain of Boletus aereus, which is obtained by self-crossing the strain of Boletus aereus YL1701-2, and is taxonomically named: Boletus aereus AC2266, which was deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 3, 2021, with the deposit number CGMCC NO.21923.

[0008] The strain of the present invention is obtained by self-crossing the monosporic hyphae of the Boletus aereus strain YL1701-2. Its fruiting body morphological characteristics include a brownish cap, a yellow stipe, cylindrical shape, relatively short, and the diameter of the cap is slightly larger than or equal to the diameter of the stipe. The pores are yellowish-brown, the flesh is dense and firm, the flesh is golden yellow, without color change after injury, and has a faint earthy smell. Identified by SSR markers, it has certain genetic differences from the strain YL1701-2 and other strains, and is determined to be a new strain of gene recombination.

[0009] On the other hand, an embodiment of the present invention provides the application of the above-mentioned Boletus aereus AC2266 strain in the production of Boletus aereus.

[0010] The advantages of the present invention are as follows:

[0011] Compared with the strain YL1701-2 used in the current industrial cultivation of Boletus aereus, the mycelium of the AC2266 strain grows faster, the optimum growth temperature is lower, the cap color is darker, the cap is thicker, and the stipe is yellower. It is an excellent new variety that can be used for the industrial production of Boletus aereus. Description of the Drawings

[0012] Figure 1 It is a dendrogram of the unweighted pair group method with arithmetic mean (UPGMA) cluster analysis described in Example 3. Detailed Embodiments

[0013] The following further elaborates on the present invention in conjunction with embodiments, but the embodiments do not limit the technical solutions of the present invention. All equivalent substitutions or changes made based on the teachings of the present invention should fall within the protection scope of the present invention.

[0014] The raw materials used in the present invention and the like are all conventional products that can be obtained by purchasing in the market.

[0015] A strain of Boletus aereus described in the present invention is obtained by self-crossing the monosporic hyphae of the Boletus aereus strain YL1701-2, and is classified and named as: Boletus aereus AC2266. It was deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 3, 2021, with the deposit number CGMCC NO.21923.

[0016] The following will further elaborate on a strain of Boletus aereus and its application in the present invention in conjunction with specific embodiments:

[0017] Example 1:

[0018] Breeding and production method of the Boletus aereus AC2266 strain:

[0019] The present invention is obtained by using the method of self-crossing breeding.

[0020] 1.1 Cultivation of fruiting bodies and collection of spores of parental strain YL1701-2:

[0021] Select the fruiting bodies of strain YL1701-2 with good growth in the mushroom house. When the caps of the fruiting bodies are fully expanded and the pores at the edges of the caps are obvious, pick them to collect spores.

[0022] 1.2 Spore coating, subculture and identification of single colonies of parental strain YL1701-2:

[0023] The spores obtained in the above steps are coated on a medium and subcultured to obtain monokaryotic hyphae. Among them, M1 medium is more suitable for this medium. The composition of this medium includes 200 g of potato (boiled in water), 20 g of glucose, 2 g of yeast extract, 1 g of MgSO4, 1 g of KH2PO4, 16 g of agar, and made up to 1000 mL.

[0024] Specifically, the collected spores are diluted with sterile water and then coated on M1 medium. After the spores germinate, they are subcultured on M1 medium. After the hyphae grow full, the monokaryotic hyphae are identified and preserved.

[0025] 1.3 Identification of mating types of monokaryotic hyphae.

[0026] 1.4 Pairing of compatible spore monokaryotic hyphae, microscopic examination, cultivation and fruiting verification:

[0027] Pair the compatible monokaryotic hyphae among the monokaryotic hyphae obtained in the above steps, and transfer the successfully paired hyphae to a medium for cultivation and then conduct fruiting verification, so as to breed strain AC2266 of Boletus aereus.

[0028] Specifically, after the compatible spore monokaryotic hyphae are paired and cultured until the hyphae come into contact with each other, microscopic examination is carried out, and strains that do not form clamp connections under special circumstances are excluded. Transfer the successfully paired hyphae to a slant medium for cultivation. After the hyphae grow full, fruiting verification can be carried out. The fruiting verification records the fruiting rate, fruiting uniformity, yield, product characteristics, etc. of the self-crossed strains. Through primary screening, re-screening and production verification, strains with high fruiting rate, low contamination rate, high average yield, complete and non-deformed fruiting body morphology, and hard and preservable flesh are repeatedly screened, and finally strain AC2266 of the present invention is obtained.

[0029] The strain AC2266 of Boletus aereus of the present invention is applied to the production of Boletus aereus. The Boletus aereus produced by strain AC2266 has a high yield, a darker cap color, a thicker, larger cap, a yellower and shorter stipe, and is suitable for industrial production of Boletus aereus. The following examples provide the application of strain AC2266 in the production of Boletus aereus.

[0030] Example 2

[0031] Basic information of strain AC2266 of the present invention and morphological characteristics of its fruiting bodies:

[0032] 2.1 Culture medium

[0033] The most suitable culture medium for strain AC2266 is Medium M1, and the formula is as follows: 200 g of potatoes (boiled for water), 20 g of glucose, 2 g of yeast extract, 1 g of MgSO4, 1 g of KH2PO4, 16 g of agar, and made up to 1000 mL.

[0034] 2.2 Mycelial growth

[0035] Strain AC2266 grows on Medium M1 for 25 days. The colony is dark yellowish-brown, the mycelium is dense, the growth is vigorous, there are no sclerotia, no guttation, and the daily growth amount at 30 °C reaches: 3.2 mm / day, which is 0.67 mm / day faster than that of the parental strain YL1701-2. The optimal growth temperature range is AC2266: 25 - 30 °C, YL1701-2: 30 - 35 °C.

[0036] Specific data are shown in Table 1.

[0037] Table 1 Mycelial growth at different temperatures for 25 days

[0038]

[0039]

[0040] 2.3 Morphological characteristics of fruiting bodies

[0041] Compared with the parental strain YL1701-2, the fruiting bodies of strain AC2266 have the following similarities and differences:

[0042] Similarities: The cap is hemispherical, there is no earthy smell in taste, and there is no color change after injury.

[0043] Differences: ① The color of the cap of AC2266 is darker and the thickness is thicker; ② The color of the stipe of AC2266 is yellower.

[0044] Specific data are shown in Table 2.

[0045] Table 2 Comparison of morphological characteristics of fruiting bodies of Boletus aereus strains AC2266 and YL1701-2

[0046]

[0047] Example 3

[0048] SSR molecular marker identification.

[0049] Collect the mycelia of strains AC2266, YL1701-2 and other 6 Boletus aereus strains. After extracting DNA, perform PCR amplification on the DNA samples with 21 pairs of primers described in the text. Add FAM or HEX fluorescent labels to the ends of the forward primers when synthesizing the primers. Genotype the PCR products using an ABI 3730 sequencer. Use the software Gene mapper 4.1 (Applied Biosystems Co., Ltd., USA) to analyze the accurate loci of the data with reference to the core base repeat numbers corresponding to the primers. Calculate the number of alleles, effective number of alleles, Shannon's information index, expected heterozygosity, and observed heterozygosity using Popgene 32. Calculate the genetic similarity coefficient using the NTSYSpc-2.10 software and perform cluster analysis using the unweighted pair-group method with arithmetic mean (UPGMA). The UPGMA cluster analysis results are as Figure 1 shown.

[0050] The information of the 21 pairs of SSR primers used is shown in the following table.

[0051] Table 3 SSR primer information for strain identification

[0052]

[0053]

[0054] The analysis results show that strain AC2266 has genetic differences from other strains including its parent strain YL1701-2 at the SSR loci used for identification, and it is a new strain.

[0055] As mentioned above, it is only a preferred embodiment of the present invention, and it does not impose any form of limitation on the present invention. Any simple modification, equivalent change, and modification made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A Boletus aereus ([ Phlebopus portentosus Phlebopus portentosus ) strain AC2266, characterized in that, It was deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 3, 2021, with the deposit number CGMCC NO. 21923.

2. Use of the Boletus aereus strain AC2266 according to claim 1 in the production of Boletus aereus.

Images