CN111727806A - Rapid germination method for phlebopus portentosus sclerotium - Google Patents
Rapid germination method for phlebopus portentosus sclerotium Download PDFInfo
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- CN111727806A CN111727806A CN202010555325.1A CN202010555325A CN111727806A CN 111727806 A CN111727806 A CN 111727806A CN 202010555325 A CN202010555325 A CN 202010555325A CN 111727806 A CN111727806 A CN 111727806A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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Abstract
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.
Description
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.
Example 1 Germination method of Phlebopus portentosus sclerotia collected in test tube
(1) Collecting sclerotium in the phlebopus portentosus test tube strain, cleaning mycelium on the surface of the sclerotium (avoiding influencing the observation of germination condition of the sclerotium) and dividing into 5 parts, wherein 4 parts are respectively put into a refrigerator with the temperature of-20 ℃, 4 ℃ and an incubator with the temperature of 10 ℃ and 15 ℃ and are stimulated for 24 hours at low temperature; and the other 1 part of the extract is used as a control group and is placed in a culture room for 24 hours at the temperature of 28-30 ℃ for the Phlebopus portentosus strains. Respectively putting 5 parts of sclerotium into test tubes filled with soil (vegetable garden soil and turf are uniformly mixed according to the ratio of 2: 1) with the thickness of 2-3 cm, spraying water, adjusting the water content of the soil to 45% -55%, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug; each set of experiments was set up in 5 replicates.
(2) And (3) placing the test tube in an incubator at 28 ℃ for dark light culture, observing the germination condition of sclerotium in due time, and germinating by taking mycelium appearing on the surface of sclerotium as sclerotium. The results are shown in Table 1.
TABLE 1 Sucus portentosus sclerotium germination status collected from test tubes
The results in table 1 show that after the phlebopus portentosus sclerotium collected from the test tube is subjected to low-temperature stimulation at 4 ℃, mycelium appears on the surface of the sclerotium after being cultured in an incubator for 16 hours, the sclerotium begins to germinate, and finally the germination rate of the sclerotium reaches 96%; covering soil for sclerotium treated at 10 ℃ for 21h to start germination, wherein the germination rate is 91%; covering soil for sclerotium treated at 15 ℃ for 28h to start germination, wherein the germination rate is 84%; the sclerotium treated at the ultralow temperature of minus 20 ℃ and the sclerotium of the control group do not germinate after being covered with soil.
Example 2 Germination method of Phlebopus portentosus sclerotia collected from cultivation fungal bags
(1) Collecting sclerotium formed in the culture bag of Phlebopus portentosus, cleaning mycelium on the surface of sclerotium, dividing into 5 parts, respectively placing 4 parts in refrigerator at-20 deg.C and 4 deg.C and incubator at 10 deg.C and 15 deg.C, and stimulating at low temperature for 24 hr; and the other 1 part of the extract is used as a control group and is placed in a culture room for 24 hours at the temperature of 28-30 ℃ for the Phlebopus portentosus strains. Respectively putting 5 parts of sclerotium into test tubes filled with soil (vegetable garden soil and turf are uniformly mixed according to the ratio of 2: 1) with the thickness of 2-3 cm, spraying water, adjusting the water content of the soil to 45% -55%, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug; each set of experiments was set up in 5 replicates.
(2) And (3) placing the test tube in an incubator at 28 ℃ for dark light culture, observing the germination condition of sclerotium in due time, and germinating by taking mycelium appearing on the surface of sclerotium as sclerotium. The results are shown in Table 2.
TABLE 2 Suillus portentosus sclerotium germination status collected from cultivation bags
The results in table 2 show that after the phlebopus portentosus sclerotium collected from the cultivation fungus bag is subjected to low-temperature stimulation at 4 ℃, mycelium appears on the surface of the sclerotium after the sclerotium is cultured in an incubator for 19 hours, the sclerotium begins to germinate, and finally the germination rate of the sclerotium reaches 100%; covering soil for 18h on sclerotium treated at 10 ℃ to start germination, wherein the germination rate is 93%; covering soil for sclerotium treated at 15 ℃ for 22h to start germination, wherein the germination rate is 86%; the sclerotium treated at the ultralow temperature of minus 20 ℃ and the sclerotium of the control group do not germinate after being covered with soil.
Example 3 Germination method of Phlebopus portentosus sclerotia collected from a culture dish
(1) Collecting sclerotium in a culture dish of Phlebopus portentosus, cleaning mycelium on the surface of sclerotium, dividing into 5 parts, respectively placing 4 parts in a refrigerator at-20 deg.C and 4 deg.C and an incubator at 10 deg.C and 15 deg.C, and stimulating at low temperature for 24 hr; and the other 1 part of the extract is used as a control group and is placed in a culture room for 24 hours at the temperature of 28-30 ℃ for the Phlebopus portentosus strains. Respectively putting 5 parts of sclerotium into test tubes filled with soil (vegetable garden soil and turf are uniformly mixed according to the ratio of 2: 1) with the thickness of 2-3 cm, spraying water, adjusting the water content of the soil to 45% -55%, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug; each set of experiments was set up in 5 replicates.
(2) And (3) placing the test tube in an incubator at 28 ℃ for dark light culture, observing the germination condition of sclerotium in due time, and germinating by taking mycelium appearing on the surface of sclerotium as sclerotium. The results are shown in Table 3.
TABLE 3 Suillus portentosus sclerotium germination status collected from Petri dishes
The results in table 3 show that after the Phlebopus portentosus sclerotium collected from the culture dish is subjected to low-temperature stimulation at 4 ℃, mycelium appears on the surface of the sclerotium after the sclerotium is cultured in an incubator for 17 hours, the sclerotium starts to germinate, and finally the germination rate of the sclerotium reaches 97%; covering soil for sclerotium treated at 10 ℃ for 17h to start germination, wherein the germination rate is 92%; covering soil for sclerotium treated at 15 ℃ for 24 hours to start germination, wherein the germination rate is 84%; the sclerotium treated at the ultralow temperature of minus 20 ℃ and the sclerotium of the control group do not germinate after being covered with soil.
Example 4 Germination method of Phlebopus portentosus sclerotia collected from the field
(1) Collecting wild Phlebopus portentosus sclerotium, cleaning mycelium on sclerotium surface, dividing into 5 parts, respectively placing 4 parts in refrigerator at-20 deg.C and 4 deg.C and incubator at 10 deg.C and 15 deg.C, and stimulating at low temperature for 24 hr; the other 1 part was used as a control group and left at room temperature for 24 hours. Respectively putting 5 parts of sclerotium into test tubes filled with soil (vegetable garden soil and turf are uniformly mixed according to the ratio of 2: 1) with the thickness of 2-3 cm, spraying water, adjusting the water content of the soil to 45% -55%, covering a small amount of fine soil on the surface of the sclerotium, and covering a test tube plug; each set of experiments was set up in 5 replicates.
(2) And (3) placing the test tube in an incubator at 28 ℃ for dark light culture, observing the germination condition of sclerotium in due time, and germinating by taking mycelium appearing on the surface of sclerotium as sclerotium. The results are shown in Table 3.
TABLE 4 Sclerotium ochrolepis sclerotium germination status from field
The results in table 4 show that after the wild Phlebopus portentosus sclerotium is subjected to low-temperature stimulation at 4 ℃, mycelium appears on the surface of the sclerotium after the sclerotium is cultured in an incubator for 20 hours, the sclerotium begins to germinate, and finally the germination rate of the sclerotium reaches 94%; covering soil for sclerotium treated at 10 ℃ for 23h to start germination, wherein the germination rate is 92%; covering soil for 30 hours on sclerotium treated at 15 ℃ to start germination, wherein the germination rate is 87%; covering soil for 72h to start germination of control sclerotium, wherein the germination rate is 81%; the sclerotium processed at the ultralow temperature of minus 20 ℃ does not germinate after being covered with soil.
The results show that sclerotium formed in the artificial cultivation process can be promoted to rapidly germinate through proper low-temperature stimulation, the sclerotium can germinate within 24 hours after low-temperature (4-10 ℃) stimulation, and the germination rate reaches over 90%; the germination time of the sclerotium collected in the field after low-temperature (4-10 ℃) stimulation is shortened from 72 hours to 24 hours, and the germination rate is improved to more than 90%.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (3)
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%.
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