New tremella auramtialba strain JSJ-J2HD2-HD3 bred by space breeding technology and application of new tremella auramtialba strain JSJ-J2HD2-HD3

Abstract

The invention relates to a new strain JSJ-J2HD2-HD3 of Naematia auramtiala, which is bred by a space breeding technology, and an application of the new strain JSJ-J2HD2-HD3, and belongs to the technical field of edible mushroom breeding. The new strain of the tremella aurantialba is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.41788. The new strain of the tremella aurantialba and a stereum hirsutum strain JSJ-G2YX001C which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.41790 are cultured and bred under a dark culture condition to obtain a yellow tremella aurantialba sporocarp strain. The novel tremella aurantialba strain can omit color changing conditions, save production cost and improve production benefits, can be used as an excellent strain resource for genetic breeding of tremella aurantialba species, researches color-related gene information, and provides an important theoretical basis for gene positioning breeding.

Description

Technical Field

[0001] The present invention belongs to the technical field of edible mushroom breeding, and particularly relates to a new strain of Tremella aurantialba JSJ-J2HD2-HD3 selected by space breeding technology and its application. Background Art

[0002] Tremella aurantialba Naematelia aurantialba belongs to Basidiomycota, Tremellomycetes, Tremellales, Naemateliaceae, and Naematelia Naematelia . The fruiting body of Tremella aurantialba is a heterogeneous complex jointly formed by Tremella aurantialba and Stereum hirsutum Stereum hirsutum . It has a smooth and tender taste, is rich in nutrients, and contains amino acids, dietary fiber, β-carotene, vitamin B2, etc. The biological characteristics of Tremella aurantialba itself are relatively special, consisting of two species, so the entire breeding system is relatively complex and difficult. Currently, most of the newly selected strains of Tremella aurantialba reported are systematically selected, that is, new varieties are selected through the domestication and cultivation of wild resources. Systematic selection of new varieties is one of the common methods for variety breeding. Although this method is simple to operate and low in cost, its genetic basis range is small, the breeding cycle is long, and the efficiency is low. And currently, for the Tremella aurantialba strains cultivated artificially in the market, the primordium is white under dark culture conditions, and the primordium expands and grows to form a fruiting body that also remains white. After giving light color-changing conditions during the growth process, the fruiting body will turn yellow and then mature. Recently, a report on using space breeding technology to select new varieties of Tremella aurantialba was published (CN119464086A), which disclosed that the total flavonoid content of 1 strain of Tremella aurantialba fruiting body selected by the Chinese Academy of Sciences Kunming Institute of Botany through spaceflight was higher than that of Tremella aurantialba without spaceflight.

[0003] Due to the complex biological characteristics of Tremella aurantialba, the development speed of the industry is relatively late compared to other major edible mushrooms, and the development of strain resources is correspondingly less. To ensure the sustainable and healthy development of the Tremella aurantialba industry, it is necessary to have more excellent trait strains for subsequent gene-directed breeding and other work. Therefore, it is extremely important to develop new varieties of Tremella aurantialba through different breeding methods and screen excellent trait strains.

[0004] References related to Tremella aurantialba can be found in: [1] Yunnan Junsijie Biotechnology Co., Ltd. A strain of Tremella aurantialba JSJ-J2X1001W and its application and molecular marker identification method: CN118272238A. 2024-07-02. [2] Yunnan Junsijie Biotechnology Co., Ltd. A strain of Tremella aurantialba JSJ-J2F1001C and its application and molecular marker identification method: CN118207101A. 2024-06-18. [3] Yunnan Junshijie Biotechnology Co., Ltd. A Tremella aurantialba strain suitable for industrial cultivation: CN112592835A. April 2, 2021. [4] Kunming Edible Fungi Research Institute, All China Federation of Supply and Marketing Cooperatives. Two new Tremella aurantialba strains and their SSR molecular marker identification methods: CN115261236A. November 1, 2022. [5] Kunming Edible Fungi Research Institute, Yunnan Supply and Marketing Cooperatives Scientific Research Institute. A new Tremella aurantialba strain, Junjin'er No. 1 strain, and its cultivation method: CN111990164A[P]. November 27, 2020. [6] Kunming Institute of Botany, Chinese Academy of Sciences. A space-borne Tremella cinnabarina strain, Hang 8, and its application: CN119464086A. February 18, 2025. [7] Cao Yao, Shen Zhenhui, Yang Linlei, etc. Determination of the polarity of Tremella aurantialba using single spore hybridization technology [J]. Acta Edulis Fungi. 2024, 31(01): 38-44. Summary of the Invention The object of the present invention is to provide a new Tremella aurantialba strain JSJ-J2HD2-HD3 selected by space breeding technology and its application.

[0005] The technical solution adopted by the present invention is as follows: A new strain JSJ-J2HD2-HD3 of Tremella aurantialba selected by space breeding technology, the strain is preserved in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 41788. Naematelia aurantialba

[0006] Naematelia aurantialba The application of the new strain JSJ-J2HD2-HD3 of Tremella aurantialba selected by space breeding technology of the present invention is to use the new strain JSJ-J2HD2-HD3 to cultivate and produce Tremella aurantialba seeds with a yellow fruiting body from primordium to maturity under dark culture conditions with the strain JSJ-G2YX001C of Stereum hirsutum; the strain JSJ-G2YX001C of Stereum hirsutum is preserved in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 41790. stereum hirsutum

[0007] ​​​The strain provided by the present invention is yellow from the primordium stage to the mature stage, and does not require additional color-changing conditions to turn yellow. The color-changing conditions can be omitted, saving production costs and improving production efficiency. Moreover, it can be used as an excellent strain resource for the genetic breeding of Tremella aurantialba, studying gene information related to color, and providing an important theoretical basis for gene mapping breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 It is a photograph of the biological characteristics of the Tremella aurantialba strain JSJ-J2HD2-HD3; Figure 2 It is a photograph of the biological characteristics of the Stereum hirsutum strain JSJ-G2YX001C; Figure 3 It is the fruiting body of the cultured fungal seeds of the two compatible monokaryotic spore strains D2 and D3 of Tremella aurantialba carried and Stereum hirsutum JSJ-G2YX001C; Figure 4 It is the fruiting body of the cultured fungal seeds of the two compatible monokaryotic spore strains D2 and D3 of Tremella aurantialba without carrying and Stereum hirsutum JSJ-G2YX001C.

[0009] Figure 5 It is the fruiting body of the cultured fungal seeds of the Tremella aurantialba strain JSJ-J2HD2-HD3 and Stereum hirsutum JSJ-G2YX001C. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0010] The content of the present invention will be further elaborated below in conjunction with specific embodiments.

[0011] A new strain JSJ-J2HD2-HD3 of Tremella aurantialba obtained by space breeding technology, the Tremella aurantialba strain JSJ-J2HD2-HD3 is deposited in the China General Microbiological Culture Collection Center (CGMCC), the deposit number is: CGMCC No. 41788, the deposit date is February 19, 2025, and the taxonomic name is Tremella aurantialba Naematelia aurantialba , which belongs to the family Naemateliaceae and the genus Naematelia Naematelia aurantialba . It is an asexual spore, the colony is smooth and milky white. See Naematelia . Figure 1 .

[0012] The said strains were obtained by smearing 44 Tremella aurantialba spore materials (see Table 1) from Yunnan Junsijie Biotechnology Co., Ltd. on weighing paper. After air-drying the weighing paper, it was placed into a 0.2 mL PCR tube and carried on Shenzhou-15 for space mutagenesis on November 29, 2022. Among them, there were 10 monokaryotic strains and 34 heterokaryotic strains. The control materials of the above 44 Tremella aurantialba strains were stored in a 4°C refrigerator. All the carried materials were returned on July 5, 2023. The carried and control materials were simultaneously activated under the culture conditions of PDA medium, 20°C, and darkness. Among them, a total of 64 control and carried materials were successfully activated (see Table 1). All the successfully activated control and carried materials were cultured and compared with the Stereum hirsutum strain JSJ-G2YX001C stereum hirsutum ), and the fruiting body traits of each control and carried material during cultivation were observed for any changes.

[0013] Table 1 List of Materials of Tremella aurantialba Carried on Shenzhou-15 by Yunnan Junsijie Biotechnology Co., Ltd. and Activation Status

[0014]

[0015]

[0016]

[0017] In the table, "√" indicates successful activation; "×" indicates failed activation.

[0018] The Stereum hirsutum strain JSJ-G2YX001C was isolated from the fruiting body of Stereum hirsutum collected in Yuxi City, Yunnan Province, and is preserved in the China General Microbiological Culture Collection Center (CGMCC). The preservation number is: CGMCC No. 41790, and the preservation date is February 19, 2025. The taxonomic name is Stereum hirsutum stereum hirsutum , which belongs to the family Stereaceae (Stereum hirsutum) and the genus Stereum Stereum ). It is in the mycelial state, with a circular and white colony. See Figure 2 .

[0019] The steps of the culture comparison experiment of the control and carried materials with Stereum hirsutum JSJ-G2YX001C are as follows: Prepare the liquid spawn of *Stereum hirsutum*. Medium: 200 g of potatoes (boiled juice), 20 g of glucose, 3 g of potassium dihydrogen phosphate, 1.5 g of magnesium sulfate, 1000 mL of water. After autoclaving and cooling, inoculate 20 mycelial agar blocks of strain JSJ-G2YX001C with a size of 5 mm×5 mm×5 mm, and place them on a magnetic stirrer at 21 °C with a rotation speed of 180 for 5 days; All successfully activated materials contain Tremella aurantialba monokaryotic spores and asexual spore materials. For the monokaryotic spore materials, 2 compatible strains need to be cultured with *Stereum hirsutum* JSJ-G2YX001C simultaneously. Pick 0.5 g of each of the 2 compatible monokaryotic spores that have been successfully activated and cultured for 10 d on PDA medium and mix them with 4 mL of the liquid spawn of *Stereum hirsutum*, then inoculate them into the culture substrate (formula: 30% cottonseed hulls, 56% wood chips, 12% wheat bran, 1% potassium dihydrogen phosphate, 1% calcium sulfate, water content 55% - 65%, pH 6 - 6.5; 130 g of culture substrate is filled in a 240 mL original seed bottle), and culture them at 18 °C, relative humidity 50%, CO₂ mass concentration ≤ 800 mg·L -1 , observe the occurrence of primordia under dark conditions during the culture period, and transfer the primordia to a new original seed bottle for continuous culture; for the asexual spore materials, directly pick 0.5 g of the spores after the culture is completed on PDA medium, mix them with 4 mL of the liquid spawn of *Stereum hirsutum*, and then inoculate them into the culture substrate and culture according to the above conditions; all carried and control materials are cultured and cultivated for comparative experiments according to the above method; A total of 64 strains of all control and carried materials were cultured and cultivated with *Stereum hirsutum*, and 26 fruiting bodies were obtained. Among them, from the materials carried by spaceflight, Tremella aurantialba fruiting bodies obtained by culturing two compatible Tremella aurantialba monokaryotic spore strains D2 and D3 with *Stereum hirsutum* JSJ-G2YX001C were yellow from primordium growth to mature fruiting bodies under dark culture conditions (see Figure 3 ); while the Tremella aurantialba fruiting bodies obtained by culturing the control materials D2 and D3 without spaceflight with *Stereum hirsutum* JSJ-G2YX001C were white from primordium growth to mature fruiting bodies under dark conditions (see Figure 4 ), that is, Tremella aurantialba fruiting bodies with a yellow color throughout the growth cycle were obtained by culturing two compatible monokaryotic spore strains D2 and D3 carried by spaceflight with *Stereum hirsutum* JSJ-G2YX001C.

[0020] Extract and isolate the asexual spores from the yellow Tremella aurantialba fruiting bodies obtained by culturing two compatible monokaryotic spore strains D2 and D3 with *Stereum hirsutum* JSJ-G2YX001C, and then culture and verify the asexual spores with *Stereum hirsutum* JSJ-G2YX001C. The relevant extraction, separation, and culture steps are as follows: 1) Harvest the obtained yellow Tremella aurantialba fruit bodies, clean them, wipe the mushroom bodies with 75% alcohol, then rinse with sterile water and place them in a laminar flow hood to dry. 2) Take 10 pieces of fruit body tissue blocks and place them in 250 mL of liquid medium (the liquid medium is: 200 g of potato juice filtered after boiling, 20 g of glucose, 1000 mL of water); culture under the conditions of a rotation speed of 150 r / min and a temperature of 23 °C for 4 - 5 days; after the culture is completed, transfer 0.5 mL to a PDA medium, culture for 4 - 5 days, then separate the asexual spores in the medium to a new PDA medium and continue to culture for 4 - 5 days, and transfer the asexual spores to a new PDA medium again. After ITS identification, it is identified as Tremella aurantialba, and the asexual spores of Tremella aurantialba are obtained, named JSJ-J2HD2-HD3. 3) Pick 0.5 g of the asexual spores of the JSJ-J2HD2-HD3 strain and mix them with 4 mL of the liquid strain of Stereum hirsutum JSJ-G2YX001C, then inoculate them into the culture substrate (formula: 30% cottonseed hulls, 56% wood chips, 12% wheat bran, 1% potassium dihydrogen phosphate, 1% calcium sulfate, water content 55% - 65%, pH 6 - 6.5; 150 g of culture substrate is filled in a 240 mL original seed bottle), and culture at 18 °C, relative humidity 50%, CO2 mass concentration ≤ 800 mg·L -1 , and observe the occurrence of primordia under dark conditions. During this period, transfer the primordia to a new original seed bottle for continuous culture.

[0021] 4) After 20 days of transfer culture through the occurrence of primordia, the asexual spores of the JSJ-J2HD2-HD3 strain and the Stereum hirsutum JSJ-G2YX001C strain also obtained yellow Tremella aurantialba fruit bodies under dark culture conditions, as shown in Figure 5 .

[0022] The growth time of the yellow Tremella aurantialba fruit bodies obtained by culturing the JSJ-J2HD2-HD3 strain is shortened by 5 days compared with the control Tremella aurantialba fruit bodies. It is used to prepare the mushroom seed fruit bodies, and the mushroom seed fruit bodies can be used in about 40 days, while the control mushroom seed fruit bodies need about 45 days to be used for cultivation. And the crude polysaccharide nutrient components of the two are detected, indicating that the crude polysaccharide content of the yellow Tremella aurantialba fruit bodies obtained by being carried is significantly increased compared with the control Tremella aurantialba fruit bodies (Table 2).

[0023] Table 2 Record table of relevant indicators of mushroom seed fruit bodies cultured with and without carried materials .

Claims

1. Tremella aurantialba selected by space breeding technology ( Naematelia aurantialba ), the new strain JSJ-J2HD2-HD3, characterized in that The strain is deposited in the China General Microbiological Culture Collection Center, with the deposit number CGMCC No. 41788.

2. Use of the new strain JSJ-J2HD2-HD3 of Tremella aurantialba selected by space breeding technology as claimed in claim 1, characterized in that, Naematelia aurantialba Use the new strain JSJ-J2HD2-HD3 to culture and produce Tremella aurantialba spawn with the strain Stereum hirsutum ( stereum hirsutum ), and the Tremella aurantialba spawn is a spawn fruiting body that is yellow from the primordium to the mature stage under dark culture conditions; the Stereum hirsutum strain JSJ-G2YX001C is preserved in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 41790.

Images