CN114868599A - Method for accelerating induction of formation of pleurotus ostreatus primordium by using maltose - Google Patents
Method for accelerating induction of formation of pleurotus ostreatus primordium by using maltose Download PDFInfo
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- CN114868599A CN114868599A CN202210653315.0A CN202210653315A CN114868599A CN 114868599 A CN114868599 A CN 114868599A CN 202210653315 A CN202210653315 A CN 202210653315A CN 114868599 A CN114868599 A CN 114868599A
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- maltose
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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
- A01G18/40—Cultivation of spawn
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Abstract
The invention discloses a method for accelerating induction of formation of an oyster mushroom primordium by using maltose, and belongs to the technical field of edible fungi. The invention discloses a method for accelerating induction of oyster mushroom primordium formation by using maltose, which is characterized in that 1.0-2.5 g/L of maltose is added into a PDA culture medium. The method for accelerating induction of the formation of the primordium of the oyster mushroom by using maltose, disclosed by the invention, can accelerate induction of the primordium in an oyster mushroom plate culture medium and improve the proportion of the primordium of an oyster mushroom plate.
Description
Technical Field
The invention relates to the technical field of edible fungi, in particular to a method for accelerating induction of formation of an oyster mushroom primordium by utilizing maltose.
Background
The primordium formation of oyster mushroom (also called pleurotus ostreatus) is an important mark for the transition from vegetative growth (hyphal growth) to reproductive growth (fruiting body growth and development), and is also a key step for finally forming yield and economic benefit. The external conditions for forming the primordium mainly comprise nutritional conditions and environmental conditions, wherein the nutritional conditions mainly comprise carbon, a nitrogen source, a carbon-nitrogen ratio, inorganic salt, growth hormone and the like, and the environmental conditions mainly comprise temperature, illumination, humidity, pH value, oxygen, carbon dioxide and the like. It has been found that many chemicals in Pleurotus ostreatus, such as resveratrol, beta-adenylate, cellular haemolysin, auxin, etc., can induce primordial formation. The carbon source is an important energy source for growth and development of the oyster mushrooms and is a raw material for synthesizing carbohydrates and amino acids. In many studies, carbohydrate metabolic pathways are abnormally active during primordial formation, maltose being one of the most common disaccharides and reducing sugars, and being very soluble in water. Maltose has not been reported to promote the formation of Pleurotus ostreatus primordium.
Therefore, it is an urgent problem to be solved by those skilled in the art to provide a method for accelerating induction of primordium formation of Pleurotus ostreatus using maltose.
Disclosure of Invention
In view of the above, the present invention provides a method for accelerating the induction of the formation of primordia of Pleurotus Ostreatus by using maltose.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for accelerating and inducing formation of oyster mushroom primordium by utilizing maltose is characterized in that 1.0-2.5 g/L of maltose is added into a PDA culture medium.
Further, the method for accelerating and inducing formation of the primordium of the oyster mushroom by using maltose comprises the following steps:
(1) activated culture of strain
Taking out the oyster mushroom strain P99 preserved at low temperature of 4 ℃ or preserved by liquid nitrogen, inoculating the oyster mushroom strain P99 on a flat plate containing a PDA culture medium, and performing activated culture at 25 ℃ for 5-7 days; activating for 1 time;
(2) preparation of culture Medium
Peeling 200g of potatoes, cutting the potatoes into small blocks of 0.5-1 cm, putting the small blocks into a pot filled with 800ml of boiled deionized water, and boiling for 10-15 min; then filtering with 4 layers of gauze, adding 20g of glucose and 15g of agar powder into the filtrate, and heating to completely dissolve the glucose and the agar powder; adding deionized water to a constant volume of 1000mL, and stirring and uniformly mixing to obtain a PDA culture medium; then adding 100ml of PDA culture medium into a triangular flask with the volume of 250ml, adding 1.0-2.5 g/L of maltose into the triangular flask, and sealing a breathable film to obtain the PDA culture medium containing maltose; sterilizing at 121 deg.C for 30 min; placing the sterilized triangular flask in a super clean bench, pouring 20ml of sterilized PDA culture medium containing maltose into a sterile flat plate, and cooling for later use;
(3) hypha culture
Punching a strain block on the edge of the hypha flat plate activated in the step (1) by using a puncher with the diameter of 5mm, then inoculating the strain block into the center of the flat plate culture medium cooled in the step (2), and placing the flat plate culture medium in an incubator at 25 ℃ in the dark for culturing for 5-7 days until the hypha has the diameter of 4-5 cm;
(4) primordial induction
And (4) moving the flat plate cultured in the step (3) until the hypha diameter is 4-5 cm to an incubator at 15 ℃ to induce the primordium, illuminating for 10 hours and darkness for 14 hours, and observing the formation of the primordium.
Further, the oyster mushroom is oyster mushroom strain P99.
According to the technical scheme, compared with the prior art, the method for accelerating induction of primordium formation of oyster mushrooms by utilizing maltose can accelerate induction of primordium in oyster mushroom plate culture medium (for example, the primordium can appear in 8-day oyster mushroom culture medium by adding 1.5g/L maltose into the basal medium after M2 treatment), and can improve the proportion of primordium appearing on oyster mushroom plates (for example, the proportion of the primordium appearing in M1-M4 treatment is between 20-60% after 18 days of induction, and the proportion of the primordium appearing in the contrast is 0 without adding maltose).
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a graph showing 18 days of primordia induction by the addition of maltose (M2) at a concentration of 1.5g/L in accordance with the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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
A method for accelerating and inducing formation of oyster mushroom primordium by using maltose comprises the following steps:
(1) activated culture of strain
Taking out the oyster mushroom strain P99 preserved at low temperature of 4 ℃ or preserved by liquid nitrogen, inoculating the oyster mushroom strain P99 on a flat plate containing a PDA culture medium, and performing activated culture at 25 ℃ for 5-7 days; activating for 1 time;
(2) preparation of culture Medium
Peeling 200g of potatoes, cutting the potatoes into small blocks of 0.5-1 cm, putting the small blocks into a pot filled with 800ml of boiled deionized water, and boiling for 10-15 min; then filtering with 4 layers of gauze, adding 20g of glucose and 15g of agar powder into the filtrate, and heating to completely dissolve the glucose and the agar powder; adding deionized water to a constant volume of 1000mL, and stirring and uniformly mixing to obtain a PDA culture medium; then adding 100ml of PDA culture medium into a triangular flask with the volume of 250ml, respectively adding maltose with the concentration of 0, 1.0, 1.5, 2.0 and 2.5g/L into different triangular flasks, and sealing the breathable films to obtain PDA culture medium containing maltose with different concentrations; sterilizing at 121 deg.C for 30 min; placing the sterilized triangular flask in a super clean bench, pouring 20ml of sterilized PDA culture medium containing maltose with different concentrations into a sterile flat plate, and cooling for later use;
(3) hypha culture
Punching a strain block on the edge of the hypha flat plate activated in the step (1) by using a puncher with the diameter of 5mm, then inoculating the strain block into the center of the flat plate culture medium cooled in the step (2), and placing the flat plate culture medium in an incubator at 25 ℃ in the dark for culturing for 5-7 days until the hypha has the diameter of 4-5 cm;
(4) primordial induction
And (4) moving the flat plate cultured to the hypha diameter of 4-5 cm in the step (3) to an incubator at 15 ℃ to induce the primordium, illuminating for 10 hours and darkness for 14 hours, and observing the formation of the primordium, wherein the results are shown in Table 1.
TABLE 1 proportion of primordia appearing on the oyster Mushroom plates under different treatment conditions
| Treatment of | Inducing for 8 days | Inducing for 13 days | Inducing for 18 days | Inducing for 25 days |
| CK(0g/L) | 0 | 0 | 0 | 10％ |
| M1(1.0g/L) | 0 | 0 | 20％ | 40％ |
| M2(1.5g/L) | 40％ | 50％ | 60％ | 80％ |
| M3(2.0g/L) | 0 | 20％ | 40％ | 80％ |
| M4(2.5g/L) | 0 | 0 | 40％ | 60％ |
The results in Table 1 show that 40% of the plates were induced to develop primordia after 8 days with maltose (M2); 60% of the plates developed primordia after 18 days of induction (FIG. 1). After 18 days of induction, all plates added with maltose showed an ortho ratio of 20-60%, and after 25 days of induction, only 10% of the plates showed ortho in the Control (CK).
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 method for accelerating induction of formation of an oyster mushroom primordium by using maltose is characterized in that 1.0-2.5 g/L of maltose is added into a PDA culture medium.
2. The method for accelerating the induction of the formation of primordia of Pleurotus ostreatus using maltose as claimed in claim 1 comprising the steps of:
(1) activated culture of strain
Taking out the oyster mushroom strain P99 preserved at low temperature of 4 ℃ or preserved by liquid nitrogen, inoculating the oyster mushroom strain P99 on a flat plate containing a PDA culture medium, and performing activated culture at 25 ℃ for 5-7 days; activating for 1 time;
(2) preparation of culture Medium
Peeling 200g of potatoes, cutting the potatoes into small blocks of 0.5-1 cm, putting the small blocks into a pot filled with 800ml of boiled deionized water, and boiling for 10-15 min; then filtering with 4 layers of gauze, adding 20g of glucose and 15g of agar powder into the filtrate, and heating to completely dissolve the glucose and the agar powder; adding deionized water to a constant volume of 1000mL, and stirring and uniformly mixing to obtain a PDA culture medium; then adding 100ml of the PDA culture medium into a triangular flask with the volume of 250ml, adding 1.0-2.5 g/L of maltose into the triangular flask, and sealing a breathable film to obtain the PDA culture medium containing maltose; sterilizing at 121 deg.C for 30 min; placing the sterilized triangular flask in a super clean bench, pouring 20ml of sterilized PDA culture medium containing maltose into a sterile flat plate, and cooling for later use;
(3) hypha culture
Punching a strain block on the edge of the hypha flat plate activated in the step (1) by using a puncher with the diameter of 5mm, then inoculating the strain block into the center of the flat plate culture medium cooled in the step (2), and placing the flat plate culture medium in an incubator at 25 ℃ in the dark for culturing for 5-7 days until the hypha has the diameter of 4-5 cm;
(4) primordial induction
And (4) moving the flat plate cultured in the step (3) until the hypha diameter is 4-5 cm to an incubator at 15 ℃ to induce the primordium, illuminating for 10 hours and darkness for 14 hours, and observing the formation of the primordium.
3. The method for accelerating induction of primordium formation of Pleurotus ostreatus using maltose according to claim 1 or 2, wherein the Pleurotus ostreatus is Pleurotus ostreatus strain P99.
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Citations (7)
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| CN101659578A (en) * | 2009-09-18 | 2010-03-03 | 鲁东大学 | Artificial culture method of Kyushu cordyceps sinensis and culture medium thereof |
| WO2015196734A1 (en) * | 2014-06-25 | 2015-12-30 | 广东省昆虫研究所 | An ophiocordyceps sinensis sporocarp artificial cultivation method |
| CN106119131A (en) * | 2016-06-28 | 2016-11-16 | 河南省农业科学院植物营养与资源环境研究所 | A kind of method alleviating excess copper suppression Growth of Pleurotus Mycelium |
| JP2017046687A (en) * | 2015-09-01 | 2017-03-09 | 学校法人甲南学園 | Mushroom culture medium liquids, mushroom culture media, and production methods of mushroom culture medium liquids, and mushroom cultivation methods |
| CN107189948A (en) * | 2017-06-07 | 2017-09-22 | 河南省农业科学院植物营养与资源环境研究所 | A kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth |
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- 2022-06-09 CN CN202210653315.0A patent/CN114868599A/en active Pending
Patent Citations (7)
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