CN114806897A - Method for accelerating induction of formation of oyster mushroom primordium by utilizing fructose - Google Patents

Abstract

The invention discloses a method for accelerating induction of formation of primordium of oyster mushroom by utilizing fructose, and belongs to the technical field of edible fungi. The invention discloses a method for accelerating induction of oyster mushroom primordium formation by utilizing fructose, which is characterized in that 1.5-3.5 g/L of fructose is added into a PDA culture medium. The method for accelerating the formation of the primordium of the induced oyster mushroom by utilizing the fructose can accelerate the induction of the primordium in an oyster mushroom plate culture medium and improve the proportion of the primordium of an oyster mushroom plate.

Description

Method for accelerating induction of formation of oyster mushroom primordium by utilizing fructose

Technical Field

The invention relates to the technical field of edible fungi, in particular to a method for accelerating the formation of an induced oyster mushroom primordium by utilizing fructose.

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 primordium formation, and fructose is one of the most common monosaccharides and reducing sugars, and is very soluble in water. Fructose has not been reported to promote pleurotus ostreatus primordium formation.

Therefore, it is an urgent problem to be solved by those skilled in the art to provide a method for accelerating the induction of primordium formation of Pleurotus ostreatus using fructose.

Disclosure of Invention

In view of the above, the invention provides a method for accelerating the induction of the formation of an primordium of oyster mushroom by using fructose.

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

a method for accelerating induction of oyster mushroom primordium formation by utilizing fructose is characterized in that 1.5-3.5 g/L of fructose is added into a PDA culture medium.

Further, the method for accelerating induction of formation of the primordium of the oyster mushroom by utilizing the fructose 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 the PDA culture medium into a triangular flask with the volume of 250ml, adding 1.5-3.5 g/L of fructose into the triangular flask, and sealing a breathable film to obtain the PDA culture medium containing fructose; 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 fructose 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 fructose can accelerate induction of primordium in an oyster mushroom plate culture medium (for example, 2.5G/L of fructose is added into a basal culture medium after G4 treatment, 8 balance mushrooms can be induced to generate primordium), and can improve the proportion of primordium generation of the oyster mushroom plate (for example, 18 days of induction, the proportion of the plate with primordium generation in G2-G6 treatment is between 20 and 100 percent, and the proportion of the plate with primordium generation is 0 percent when a contrast is not added with fructose).

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 embodiments or the prior art descriptions 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 the 18-day primordium induction by the addition of 2.5G/L fructose (G4) according to 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 utilizing fructose 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 0, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5 and 4.0g/L of fructose into different triangular flasks, and sealing the air-permeable membrane to obtain PDA culture medium containing fructose 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 fructose 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 (3) moving the 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, 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％
G1(1.0g/L)	0	0	0	10％
					G2(1.5g/L)	0	0	40％	60％
G3(2.0g/L)	0	0	40％	60％
					G4(2.5g/L)	40％	60％	100％	100％
G5(3.0g/L)	0	20％	40％	60％
					G6(3.5g/L)	0	10％	20％	30％
G7(4.0g/L)	0	0	0	10％

The results in Table 1 show that 40% of the plates were primordial induced by fructose addition (G4) for 8 days. After 18 days of induction, 100 percent of the treated plate has the primordium (figure 1), the proportion of the primordium of G2-G4 is between 40 percent and 100 percent, and the proportion of the primordium of G5-G6 is reduced to 20 percent to 40 percent. After 25 days of induction, only 10% of the plates in the Control (CK) showed primordia.

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 oyster mushroom primordium formation by utilizing fructose is characterized in that 1.5-3.5 g/L of fructose is added into a PDA culture medium.

2. The method for accelerating the induction of the formation of the primordium of the oyster mushroom by using the fructose as claimed in claim 1, which 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 the PDA culture medium into a triangular flask with the volume of 250ml, adding 1.5-3.5 g/L of fructose into the triangular flask, and sealing a breathable film to obtain the PDA culture medium containing fructose; 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 fructose 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 oyster mushroom according to claim 1 or 2, wherein the oyster mushroom is Pleurotus ostreatus strain P99.

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