Combined tremella aurantialba strain ZWY-G1 / PNF11

Abstract

The invention relates to a combined tremella aurantialba strain ZWY-G1 / PNF11, and belongs to the field of edible mushroom breeding. The strain is a sporocarp obtained by effectively combining a tremella aurantialba strain (PNF11) in a high-altitude cold area and a stereum hirsutum strain (ZWY-G1) in a low-altitude tropical rainforest area. Wherein asexual spores of the tremella aurantialba strain PNF11 in the combined strain are milky white and yeast-shaped, and the yeast-shaped edge is in a radial hypha form; hyphae of the stereum hirsutum ZWY-G1 are thick and dense, aerial hyphae are many, and the hyphae are orange after liquid fermentation; in the industrial cultivation process, the growth period of the combined tremella aurantialba strain ZWY-G1 / PNF11 is 36-40 days, the sporocarp color is orange, the longitudinal section is light flesh color, the fruiting rate is 100%, the yield is 98%, the biological conversion rate is 85%, and the combined tremella aurantialba strain has high consistency, stability and excellent agronomic character indexes and is suitable for industrial and facility annual cultivation and liquid fermentation process cultivation.

Description

Technical Field

[0001] The invention relates to a combined golden fungus strain ZWY-G1 / PNF11, and belongs to the field of edible fungus breeding. Background Art

[0002] Golden ear is a heterogeneous fruiting body, a complex composed of Golden ear fungus and Hairy Leather Fungus. The Golden ear mentioned in daily life and the edible parts are not actually a single species. Therefore, a species or strain of Golden ear fungus that can be used for cultivation is actually composed of Golden ear fungus and Hairy Leather Fungus. Golden ear fungus cannot complete its life history independently in nature, and it does not have the ability to decompose lignin and cellulose, while Hairy Leather Fungus is a completely independent individual. Before establishing a parasitic relationship with Golden ear fungus, it can reproduce sexually and asexually like all large saprophytic fungi.

[0003] A golden ear combined strain refers to a new golden ear strain obtained by combining Golden Ear Mushroom A with Hairy Leather Mushroom B1 / B2 / B3 / B4... or combining Hairy Leather Mushroom B with Golden Ear Mushroom A1 / A2 / A3 / A4...; Golden Ear Mushroom and Hairy Leather Mushroom can be a combination of artificially cultivated strains and artificially cultivated strains, or a combination of wild strains and wild strains, or a combination of artificial strains and wild strains. This method is similar to "hybrid breeding", but is essentially different from the definition of "hybrid breeding".

[0004] Li Rongchun and Yang Linlei authorized the publication for the first time of "A method for breeding new varieties of golden ear fungus by combining golden ear asexual spores with hairy leathery fungus" (CN 112021067 B). A large number of golden ear fungus strains and hairy leathery fungus strains were obtained by separating and purifying golden ear strains from different origins and with different genetic backgrounds, which were combined and screened to form new varieties. Although many combined golden ear strains have been bred by this method, the existing combined golden ear strains still have problems such as insufficient mushroom yield, yield and bioconversion rate. At the same time, the growth period of the existing combined golden ear strains is too long. The above problems seriously restrict the factory production of golden ear fungus. Therefore, it is urgent to develop a new combined golden ear strain to meet the factory production of golden ear fungus.

[0005] The combined golden ear strain ZWY-G1 / PN-F11 applied for this time is actually basically an excellent strain obtained by applying the authorized method of (CN112021067 B). The difference is that the hairy leather fungus strain is purely wild and is isolated from the fruiting body of basswood that does not parasitize golden ear. The strain is harvested in the tropical area with an altitude of 585 meters, which does not meet the growth environment of wild golden ear, and there is no related report on wild golden ear. Based on this, the breeding method of new golden ear varieties can be greatly extended on the basis of this (CN 112021067 B) method. As a species with strong saprophytic ability, hairy leather fungus is widely distributed in nature in my country, far greater than the wild resource distribution of golden ear. The breeding of the golden ear strain ZWY-G1 / PN-F11 in this application will open up a broader space for golden ear breeding in the future. Summary of the invention

[0006] In order to solve the problems existing in the existing combined golden ear fungus, the purpose of the present invention is to provide a combined golden ear fungus strain ZWY-G1 / PNF11, which is obtained by combined cultivation of Golden ear fungus PNF11 and Lepidium pubescens ZWY-G1.

[0007] The Auricularia auricula PNF11 and the Auricularia pubescens ZWY-G1 were deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on April 30, 2024, at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit number of Auricularia auricula PNF11 is: CGMCC No. 41293, and the taxonomic name is: Naematelia aurantialba ; The deposit number of ZWY-G1 is CGMCC No. 41294, and the taxonomic name is: Stereum hirsutum .

[0008] Preferably, the specific culture method comprises the following steps: (1) The first combined culture: The liquid fermentation strain of ZWY-G1 of Psoralea corylifolia was inoculated into the central part of the induction medium for cultivation; at the same time, PNF11 of Psoralea corylifolia was picked up and shaken in sterile water to obtain a bacterial suspension, which was inoculated into the central part of the induction medium where the hyphae of Psoralea corylifolia had grown for co-cultivation. Small fruiting bodies of Psoralea corylifolia (small white hairy bumps) wrapped by Psoralea corylifolia could be clearly seen on the surface of the culture medium. At this stage, the fruiting bodies were severely wrapped by the fungus and could not be used for production.

[0009] (2) Second combined culture: The liquid fermentation strain of P. fusca ZWY-G1 was inoculated into the central part of the induction medium for culture; then the golden ear fruiting bodies formed in the first combination were picked up with a scalpel and inoculated into the central part of the P. fusca that had grown there for co-culture to obtain the combined golden ear strain ZWY-G1 / PNF11.

[0010] Preferably, the induction culture medium in step (1) contains the following percentages of substances: 80% hardwood sawdust, 3% white sugar, 15% wheat bran, 2% potato flour, and the water content of the culture medium is 60%.

[0011] Preferably, in step (1), the culture temperature of the fungus ZWY-G1 is 20-22° C., and the culture time is 4-6 days.

[0012] Preferably, in step (1), the mass ratio of Auricularia auriculariae PNF11 to sterile water is 1:100.

[0013] Preferably, in step (1), the temperature for co-culturing Auricularia auriculariae PNF11 and Leptospira pubescens ZWY-G1 is 20-22° C., the time is 35-40 days, and the culture condition is dark culture.

[0014] Preferably, in step (2), the culture temperature of the fungus ZWY-G1 is 20-22° C., and the culture time is 4-6 days.

[0015] Preferably, the diameter of the small fruiting bodies of the golden ear picked in step (2) is 0.5-1 cm.

[0016] Preferably, in step (2), the co-culture temperature of the Auricularia auricularia fruiting bodies and the fungus ZWY-G1 is 20-22° C., and the culture time is 40-50 days.

[0017] The combined Tremella fuciformis strain ZWY-G1 / PNF11 can be used for factory cultivation to obtain Tremella fuciformis fruiting bodies.

[0018] Beneficial effects of the present invention (1) The combined golden ear fungus strain ZWY-G1 / PNF11 described in the present invention is formed by combining the high-altitude golden ear fungus PNF11 with the low-altitude hairy leather fungus ZWY-G1. The combined strain is orange in color during cultivation, which is different from the current golden yellow strain.

[0019] (2) The combined golden ear strain ZWY-G1 / PNF11 of the present invention has a mushroom yield rate of 100%, a finished product rate of more than 98%, a biological conversion rate of more than 85%, and has high consistency and stability. It is suitable for factory-based, facility-based year-round cultivation and liquid fermentation process cultivation.

[0020] (3) The application of the combined golden ear strain ZWY-G1 / PNF11 can not only solve the current problem of strain degeneration in golden ear factory production, but also provide a broader space for the selection and breeding of excellent strains in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The mycelial morphology of the hairy leather fungus ZWY-G1 and the conventional hairy leather fungus SN-G in Example 1 is compared.

[0022] Figure 2 The morphology of the golden urticaria fungus PNF11 and the conventional golden urticaria fungus PN-J in Example 1 is compared.

[0023] Figure 3 The liquid fermentation state of the fungus ZWY-G1 and the conventional fungus SN-G in Example 2 is compared.

[0024] Figure 4 This is a color comparison chart of the fruiting body of the combined golden fuciformis strain ZWY-G1 / PNF11 prepared by the present invention in Example 3 and the fruiting body of the existing combined golden fuciformis strain SN-G / PN-J.

[0025] Figure 5 This is a color comparison diagram of the longitudinal section of the fruiting body of the combined golden fuciformis strain ZWY-G1 / PNF11 prepared by the present invention in Example 3 and the fruiting body of the existing combined golden fuciformis strain SN-G / PN-J. DETAILED DESCRIPTION

[0026] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following embodiments are only simple examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims. Example 1

[0027] Purification of Auricularia auriculariae PNF11 and Leptospira pubescens ZWY-G1 Golden ear fungus PNF11 was obtained by the following method: Golden ear fungus was collected from Mingqi Village, Zhuwagen Town, Zayu County, Tibet, and the alpine oak Quercus semicarpifolia The tree stump is at an altitude of 3227m, which belongs to a high-altitude cold climate area. After surface disinfection of wild golden ear fruiting bodies, the middle part is picked in a clean bench and inoculated into a liquid culture medium (glucose 20g / L, potato flour 6g / L, yeast powder 2g / L, soybean powder 6g / L, water 1L), the temperature is 21±1°C, and the shaking bottle is cultured for 4 days. 0.5 ml is drawn with a pipette and inoculated into a solid culture medium (consistent with the liquid formula, with 16g / L agar powder added). After culturing for 5 days, the yeast-like part is picked, purified and cultured twice, and the purified golden ear fungus is subjected to 18s DNA sequencing. The 18s DNA sequence of the golden ear fungus PNF11 of the present invention is shown in SEQ ID NO: 1.

[0028] The hairy leather fungus ZWY-G1 was obtained by the following method: The hairy leather fungus was collected from the fallen Menghai Lithops tree in the Xishuangbanna Tropical Botanical Garden of the Chinese Academy of Sciences. Lithocarpus fohaiensisThe invention relates to a method for preparing a wild tricholoma in a tropical rain forest, wherein the surface of the wild tricholoma is disinfected and then the tricholoma is inoculated into a solid culture medium (glucose 20 g / L, yeast powder 4 g / L, peptone 2 g / L, potassium dihydrogen phosphate 3 g / L, agar powder 16 g / L, water 1 L) through fruiting body separation, cultured at 21° C., purified 1-2 times, and the purified tricholoma is subjected to 18s DNA sequencing. The 18s DNA sequence of the tricholoma ZWY-G1 of the invention is shown in SEQ ID NO: 2.

[0029] The morphology of the hairy leather fungus ZWY-G1 obtained by the present invention and the existing hairy leather fungus SN-G is as follows: Figure 1 As shown, from Figure 1 It can be seen that the hyphae of ZWY-G1 are thicker and denser, with more aerial hyphae ( Figure 1 A) Compared with the existing creeping hyphae of the fungus SN-G ( Figure 1 B) There is a big difference.

[0030] The morphology of the golden ear fungus PNF11 obtained by the present invention and the existing golden ear fungus PN-J is as follows Figure 2 As shown, from Figure 2 It can be seen that the yeast-like spores of PNF11 are milky white, and radial hyphae are faintly visible at the edge ( Figure 2 A), while the golden ear fungus PN-J is khaki, and the overall appearance is uniform ( Figure 2 B). Example 2

[0031] A method for preparing a combined golden fungus, the specific method is as follows: (1) Preparation of liquid culture of Pseudomonas aeruginosa: The mother culture of Pseudomonas aeruginosa ZWY-G1 was inoculated into a liquid fermentation medium (glucose 20 g / L, yeast powder 4 g / L, peptone 2 g / L, potassium dihydrogen phosphate 3 g / L, water 1 L), and cultured at 21±1°C for 6 days to obtain orange liquid culture of Pseudomonas aeruginosa ZWY-G1. The obtained liquid culture of Pseudomonas aeruginosa ZWY-G1 was as follows: Figure 3 As shown, from Figure 3 It can be seen that the hairy leather fungus ZWY-G1 is orange ( Figure 3 A), while the existing SN-G is light yellow ( Figure 3 B).

[0032] (2) First combination culture: 4 ml of the liquid fungus strain ZWY-G1 obtained in step (1) was inoculated into the central part of an induction medium (80% broad-leaved sawdust, 3% sugar, 15% wheat bran, 2% potato flour, and water was added to make the water content of the induction medium reach 60%), and cultured for 5 days; Pyrrolizidine oxysporum PNF11 was selected and shaken in sterile water to obtain a bacterial suspension, the mass ratio of Pyrrolizidine oxysporum PNF11 to sterile water being 1:100, and then 0.5 ml of the bacterial suspension was pipetted with a pipette and inoculated into the central part of the induction medium on which mycelium of Pyrrolizidine oxysporum ZWY-G1 had grown, and cultured in the dark at 21°C for 37 days. Many small fruiting bodies of Pyrrolizidine oxysporum (small white hairy protrusions) wrapped by Pyrrolizidine oxysporum could be clearly seen on the surface of the induction medium, indicating that the first combination was completed. The fruiting bodies of Pyrrolizidine oxysporum in the first combination were small and scattered, and were wrapped by Pyrrolizidine oxysporum, and could not be used for production.

[0033] (3) Second combination culture: Use a scalpel to pick up the combined golden ear fruiting bodies with a diameter of 0.5-1 cm (the combined golden ear fruiting bodies obtained in the first combination culture), and inoculate them again in the central part of the induction culture medium where the hyphae of the hairy leather fungus ZWY-G1 have grown. Culture at 21°C for 45 days. At this time, evenly shaped, light-flesh-colored fruiting bodies are formed in the culture bottle, indicating that the selection and breeding of the combined golden ear strain ZWY-G1 / PNF11 is completed. Example 3

[0034] The specific steps of the combined golden ear mushroom cultivation experiment are as follows: The combined golden ear fungus ZWY-G1 / PNF11 cultured in Example 2 was inoculated into a cultivation bag (the cultivation bag formula was: 85% cottonseed hulls, 13% wheat bran, 1% gypsum, 1% potassium dihydrogen phosphate, and water was added to make the moisture content of the cultivation bag reach 60%, pH 6-7) for a mushroom fruiting experiment. At the same time, the existing combined golden ear fungus SN-G / PN-J was used for a mushroom fruiting experiment control, and relevant data were recorded.

[0035] from Figure 4 It can be seen that the fruiting body of the combined golden ear strain ZWY-G1 / PNF11 prepared by the present invention is quite different from the existing combined golden ear strain SN-G / PN-J. The fruiting body of the combined golden ear strain ZWY-G1 / PNF11 prepared by the present invention is orange ( Figure 4 A), while the fruiting body of the existing combined Auricularia auriculariae SN-G / PN-J is golden yellow ( Figure 4 B) Cut the fruiting bodies of the two, and the color of the longitudinal section is as follows Figure 5 As shown, from Figure 5It can be seen that the longitudinal section color of the fruiting body of the combined golden ear strain ZWY-G1 / PNF11 prepared by the present invention is quite different from the longitudinal section color of the existing combined golden ear strain SN-G / PN-J. The longitudinal section of the fruiting body of the golden ear strain ZWY-G1 / PNF11 prepared by the present invention is light flesh color ( Figure 5 B), while the longitudinal section of the fruiting body of the existing combined Auricularia auriculariae SN-G / PN-J is white ( Figure 5 A).

[0036] The agronomic traits of the golden ear strain ZWY-G1 / PNF11 of the present invention are compared with those of the existing combined golden ear fungus SN-G / PN-J, and the results are shown in Table 1. As can be seen from Table 1, the mushroom yield of the combined golden ear fungus prepared by the present invention reaches 100%, and the finished product rate reaches 98%, which are higher than the SN-G / PN-J strain cultivated on a large scale. The biological conversion rate of the golden ear strain ZWY-G1 / PNF11 prepared by the present invention is 35 grams higher per bag, which is 4 percentage points higher, and the growth period is shortened by 8 days. In factory cultivation, the actual benefit per bag can be increased by 1.15 yuan, which can basically offset the cost of production raw materials, and the benefit is considerable.

[0037] Table 1 Comparison of agronomic traits of the combined golden ear fungus prepared by the present invention and SN-G / PN-J

Claims

1. A combined Tremella fuciformis strain ZWY-G1 / PNF11, characterized in that: The ZWY-G1 is a strain of the fungus, with a deposit number of CGMCC No. 41294 and a classification name of: Hairy stereo ; The PNF11 is a strain of Auricularia auriculariae, with a deposit number of CGMCC No. 41293 and a classification name of: Naematelia aurantialba .

Images