阅读说明：本技术 一种暗褐网柄牛肝菌菌核快速萌发方法 (Rapid germination method for phlebopus portentosus sclerotium ) 是由 杨天伟 张春霞 何明霞 刘静 高锋 许欣景 方艺伟 王文兵 戴利铭 于 2020-06-17 设计创作，主要内容包括：本发明公开了一种暗褐网柄牛肝菌菌核快速萌发方法,属于生物技术领域。本发明公开的一种暗褐网柄牛肝菌菌核快速萌发方法,先将人工栽培过程中形成的菌核及野外采集的暗褐网柄牛肝菌菌核清理干净,在4～10℃条件下低温处理24h,然后放入盛有土壤的试管或培养皿中28℃暗光培养。人工栽培过程中形成的菌核经低温刺激后能在24h内萌发,萌发率达到90％以上；野外采集的菌核通过低温刺激后萌发所需的时间缩短,萌发率也提高到90％以上。本发明一方面解决了暗褐网柄牛肝菌人工栽培过程中形成的菌核无法萌发的问题；另一方面提高了野外采集菌核的萌发速率和萌发率；该方法操作简便、经济实用。(The invention discloses a rapid germination method for phlebopus portentosus sclerotia, and belongs to the technical field of biology. The invention discloses a rapid germination method of phlebopus portentosus sclerotia, which comprises the steps of cleaning sclerotia formed in the artificial cultivation process and phlebopus portentosus sclerotia collected in the field, treating at a low temperature of 4-10 ℃ for 24 hours, and then placing into a test tube or a culture dish filled with soil for dark light culture at a temperature of 28 ℃. Sclerotium formed in the artificial cultivation process can germinate within 24 hours after low-temperature stimulation, and the germination rate reaches more than 90%; the time required for the sclerotium collected in the field to germinate after low-temperature stimulation is shortened, and the germination rate is also improved to more than 90 percent. On one hand, the invention solves the problem that sclerotium formed in the artificial cultivation process of phlebopus portentosus can not germinate; on the other hand, the germination rate and the germination rate of the wild sclerotium collected are improved; the method is simple and convenient to operate, and is economical and practical.)

1. A rapid germination method for phlebopus portentosus sclerotium is characterized by comprising the following specific steps:

(1) collecting the phlebopus portentosus test tube strains, sclerotium formed in a culture dish and a culture bag and field phlebopus portentosus sclerotium, and cleaning mycelium on the surface of the sclerotium;

(2) stimulating the cleaned sclerotia in the step (1) at a low temperature of 4-10 ℃ for 24 hours;

(3) respectively putting the sclerotium stimulated at the low temperature in the step (2) into a test tube or a culture dish filled with soil, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug or a culture dish cover; and (3) placing the test tube or the culture dish in an incubator at 28 ℃ for culturing in dark light, and observing the germination condition of sclerotium.

2. The rapid germination method of phlebopus portentosus sclerotium according to claim 1, characterized in that 2-3 cm thick soil is filled in the test tube; and soil with the thickness of 1cm is filled in the culture dish.

3. The rapid germination method of phlebopus portentosus sclerotium as claimed in claim 1, characterized in that the soil is formed by mixing vegetable garden soil and turf in a ratio of 2:1, and the water content of the soil is 45% -55%.

Technical Field

The invention relates to the technical field of biology, in particular to a rapid germination method of phlebopus portentosus sclerotia.

Background

Phlebopus portentosus (Phlebopus portentosus) is commonly known as Phlebopus nigricans, and belongs to Boletinellaceae, Phlebopus; china is mainly distributed in Yunnan, Guangxi, Hainan and Sichuan, and abroad is distributed in Thailand, New Zealand and other places. The phlebopus portentosus is the only phlebopus portentosus edible fungus which can realize the artificial cultivation of mushroom houses and can be produced all the year round at present. The Phlebopus portentosus has delicious taste and rich nutrition, is one of important edible fungi which are eaten and traded by people, and has important edible value, research significance and wide development and utilization prospects.

Through field investigation, the low-temperature and drought season, the phlebopus portentosus mycelium in the soil can form a large amount of sclerotia through cell differentiation so as to resist the influence of adverse environment. In the artificial cultivation of mushroom houses, a large number of sclerotia also appear in the whole process from the separation and screening of strains to the culture of mushroom bags, especially when the cultivation of the mushroom bags are carried out, sclerotia with dense and numb and different sizes are generated on the inner walls of the mushroom bags, and the phenomena show that the phlebopus portentosus is a stone-producing edible mushroom.

The sclerotium is a special structure formed by the differentiation and repeated branching of mycelium cells, has the functions of storing nutrients and resisting adverse environments such as low temperature, drought and the like, is considered as a static body or a dormant body of hyphae, and can germinate into hyphae or fruiting bodies under proper conditions; has important position and function in the growth and development process of edible fungi.

At present, no report is found about relevant researches on the sclerotium germination of the phlebopus portentosus. However, researchers of the method study sclerotium formed in the artificial cultivation process of the phlebopus portentosus, and find that the sclerotium formed in the test tube, the culture dish and the cultivation fungi bag in the artificial cultivation process can not germinate even if the sclerotium is placed under the conditions of proper temperature, humidity and nutrition without human intervention; the phlebopus portentosus sclerotium collected in the field can germinate into hypha and form fruit bodies under the conditions of proper temperature and humidity, but the germination speed is slow and the germination rate is low.

Therefore, the problem to be solved by the technical personnel in the field is to provide a rapid germination method of phlebopus portentosus sclerotia.

Disclosure of Invention

In view of the above, the invention provides a rapid germination method for phlebopus portentosus sclerotia, which solves the problem that sclerotia formed in the artificial cultivation process of phlebopus portentosus cannot germinate on one hand; on the other hand, the germination rate and germination rate of the wild sclerotium collected are improved.

In order to achieve the purpose, the invention adopts the following technical scheme:

a rapid germination method of phlebopus portentosus sclerotium is used for carrying out low-temperature treatment on phlebopus portentosus sclerotium.

Further, a rapid germination method of phlebopus portentosus sclerotium is characterized in that the phlebopus portentosus sclerotium is treated for 24 hours at the temperature of 4-10 ℃.

Further, a rapid germination method for phlebopus portentosus sclerotium comprises the following specific steps:

(1) collecting the phlebopus portentosus test tube strains, sclerotium formed in a culture dish and a culture bag and field phlebopus portentosus sclerotium, and cleaning mycelium on the surface of the sclerotium;

(2) stimulating the cleaned sclerotia in the step (1) at a low temperature of 4-10 ℃ for 24 hours;

(3) respectively putting the sclerotium stimulated at the low temperature in the step (2) into a test tube or a culture dish filled with soil, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug or a culture dish cover; and (3) placing the test tube or the culture dish in an incubator at 28 ℃ for culturing in dark light, and observing the germination condition of sclerotium.

Further, soil with the thickness of 2-3 cm is filled in the test tube; and soil with the thickness of 1cm is filled in the culture dish.

Further, the soil is formed by mixing vegetable garden soil and turf according to the ratio of 2:1, and the water content of the soil is 45% -55%.

Because the artificial cultivation process of the phlebopus portentosus is always in the proper temperature range: sclerotium formed in the process is not subjected to low-temperature treatment and does not germinate at the temperature of 28-30 ℃; after proper low-temperature stimulation, the germination can be promoted, and the germination rate is improved. Wild Phlebopus portentosus sclerotium is formed in a low-temperature drought season, and is collected back to a laboratory, part of sclerotium can germinate, but the required germination time is long, the germination rate is low, and the germination speed and the germination rate can be improved after low-temperature stimulation.

According to the technical scheme, compared with the prior art, the invention discloses the rapid germination method of the phlebopus portentosus sclerotium, and the phlebopus portentosus sclerotium is treated for 24 hours at the temperature of 4-10 ℃. On one hand, the invention solves the problem that sclerotium formed in the artificial cultivation process of phlebopus portentosus can not germinate; on the other hand, the germination rate and the germination rate of the wild sclerotium collected are improved; the method is simple and convenient to operate, and is economical and practical.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.