RU2378821C2 - Method for intense growth of oyster mushrooms and production of substrate planting mycelium for their extensive cultivation - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: proposed invention is related to agriculture and biotechnology, namely to mushroom growing. Objective of the present invention is development of method for growth of oyster mushrooms, which provides for guaranteed increase of crops and simplification and cheapening of method for production of substrate planting mycelium for their extensive cultivation. Stated objective is achieved by combination of two technologies - intensive growth of oyster mushrooms and production of substrate planting mycelium for their cultivation in extensive mushroom growing. As for intensive growth of oyster mushrooms, stated objective is achieved by the fact that growth of mushrooms is carried out in liter glass jars (it is possible to use jars with capacity of 0.7-1.5 l), filled with substrate, which are closed with two layers of crimped foil, without tying, and after sterilisation in autoclave at the pressure of at least 1.6-1.8 atm. for 3 hours and after cooling, substrate surface is inoculated with mother mycelium, leaving at the same time distance from 1 cm (0.5-1 cm) between inoculated surface of substrate and foil coating. Silting of substrate surface is carried out for 32-38 days at the temperature of 20-22°C. After silting jar covers are not removed, and mushroom outgrowths that come out of cover outside are carefully picked by downward hand motion, holding cover. There are two main fruiting "waves", with interval of 8-16 days. In the first two main "waves" fruit yield is very high. After the second main fruiting, cover is removed. As for method for production of substrate planting mycelium for provision of mass extensive mushroom growing - as substrate planting mycelium, they use chaff-sawdust nutrient substrate spent in two main fruiting "waves" in process of intensive oyster mushroom growth, with ratio of 2:1, from which upper 2^nd centimetre layer is removed (which is older physiologically with remains of air mycelium and outgrowths bases, possible surface infection, laying of flies and mushroom mosquitoes). Substrate is accumulated and stored in perforated plastic bags in refrigerating chambers at low positive temperatures (3-5°C) until spring selling to mushroom growers and farmers. In process of summer mushroom fruiting with intense cultivation based on heat resistant lemon cap oyster mushroom, four fruiting "waves" are produced, and spent substrate is used to prepare fungus compost (organic fertiliser for vegetable growing). Proposed method is tested on the basis of biotechnologies and plant protection department GNU-DV awarded with order of the Red Banner of Labour in Research Institute of Agriculture of Russian Agricultural Academy, and substrate planting mycelium is tested by amateur mushroom growers and farmers under conditions of Khabarovsk neighborhood for growing of mushrooms by means of extensive method.

EFFECT: preservation of internal aseptic property of substrate, reduced labour expenses and improved profitability of specialised mushroom farms operation.

3 cl, 3 tbl, 2 dwg

Description

The present invention relates to agriculture and biotechnology. The method allows you to combine two separate technological processes - the intensive production of marketable mushrooms in liter glass jars, with high fruit yield in the first two (main) “waves”, and the production of inexpensive substrate (floor-sawdust) planting mycelium for mass supply of extensive mushroom growing.

Oyster mushroom (Pleurotus ostreatus) is one of the most valuable objects of world and domestic mushroom growing. Oyster mushroom breeding is most beneficial compared to other cultivated mushrooms. Other types of wood oyster mushrooms (Florida, lemon-hat, pulmonary) are also promising for wide cultivation / A. Morozov Industrial production of oyster mushrooms. - M .: ATS, 2004 .-- 112 s .; Annenkov B.G., Azarova V.A. Introduction of new original species of oyster mushrooms into the Amur mushroom growing // Modern scientific support of the Far Eastern agrarian industry (Mat. 5 x Kazminsky readings, 11/29/2006). - Khabarovsk: DVNIISKH RAAS, 2007. - P.24-30 /.

There are two main methods for the artificial cultivation of winter-hardy wood oyster mushrooms: extensive (in summer on chocks and felled trunks, in three-year revolutions) and intensive (usually in two-month revolutions from late summer to early next summer under controlled indoor conditions, on straw or sawdust substrates ) / Annenkov B.G., Azarova V.A. Scientific fundamentals of mushroom growing and intensive cultivation of oyster mushrooms in the Amur region // Scientific fundamentals of increasing the efficiency of agricultural production production in the Far East of Russia (Mat. 4 Kazminsky readings, 11.29.2005). - Khabarovsk: DVNIISKH RAAS, 2006. S.130-140 /.

With all the variety of methods and techniques for intensive cultivation of oyster mushrooms, they are all divided into two groups of technologies:

- Euro-technology (semi-sterile), when fruiting occurs on large mycelium-infested and freed from formwork blocks or on perforated plastic bags with pasteurized or fermented agricultural substrates;

- Asian (absolutely sterile) with the fruiting of mushrooms from the openings (after the sterile period of surface inoculation and overgrowing of the substrate) of the holes of small vessels (ceramic, glass, polypropylene) or heat-resistant plastic bags with compacted and enriched with nutritive additives (bran, dust, flour, etc.) d.) a sterile sawdust substrate. / Annenkov B.G., Azarova V.A. Comparative evaluation of Hungarian and Chinese technologies for the intensive cultivation of oyster mushrooms // Energy-saving technologies for the cultivation of crops in the Far East (Mat. Scientific session, on the occasion of the 70th anniversary of the DalNIISH, July 14-15, 2005, Khabarovsk). - Vladivostok: Dalnauka, 2006. - P.126-139; Tishenkov A.D. Mushroom growing in China // School of mushroom growing, 2006, No. 1 (S.29-35), No. 2 (S.29-36), No. 3 (S.25-31) /.

In Euro technology of intensive mushroom growing, inoculation of nutritious agricultural substrates uses only sterile seed mycelium grown in cans on boiled / Annenkov B.G. Production of edible mushroom cereal mycelium: informalists of the Khabarovsk Central Scientific Research Institute, 1999, No. 34. - 3 p. / Or in heat-resistant bags on soaked malt / Kuznetsov O.Yu., pat. RF No. 2192119, 2002 / and autoclaved grains (oats, rye, wheat, millet, etc.), which, with a high starting growth coefficient for grains infected with fungal hyphae, allows grains (with a relatively high inoculum dose of more than 5%) quickly , i.e. before competing molds, colonize the substrate in semi-sterile conditions and achieve high yield of mushrooms.

Known methods for the cultivation of edible xylotroph mushrooms: in separable vessels with a removable throat / Application of Japan No. 49-17099.1974, /; in designs consisting of a flexible plastic bag and a solid can neck / Japanese application No. 54-17666, 1979 /; in special vessels / Japanese application No. 57-39604, 1982 /; in plastic bags / Japanese application No. 62-11030, 1987 / and in wide-necked bottle vessels / Japanese application No. 63-16992, 1987 /. In China, wood oyster mushrooms (ordinary, lemon-bonnet, etc.) are cultivated in heat-resistant plastic bags, the open end of which is passed through a soft leather ring, which acts as a neck filled with cotton wool. This method of intensive culture of oyster mushroom was introduced by Chinese specialists in Turkmenistan / Orazov Kh.N. Oyster mushroom in Turkmenistan // School of mushroom growing, 2006, No. 4. - S. 38-39 /.

Closest to the claimed invention regarding intensive cultivation of mushrooms is Japan's application No. 49-6939, 1974. The method is based on filling cans of any configuration with a nutrient substrate suitable for growing mushrooms. The neck of the vessel is tightened with a heat-resistant breathable film, which is still tightened with a hoop or rubber band for sealing purposes. Then carry out the process of sterilization and seeding of mycelium. Growing is carried out for a predetermined time (until the mushroom mycelium reaches the bottom of the can). After cooling, the coating is removed, moistened and the growing process continues. Between fruiting banks remain open.

However, along with exceptional laboriousness and additional manual labor costs, a major drawback of the “Asian” sterile technologies is that after the calculated opening of the fruiting holes on the cultivation vessels, irrational losses of substrate moisture from evaporation begin, the availability of the bearing surface of the substrate for infections and pests sharply increases, death of the primordium of mushrooms from drying out (especially with instability of air humidity in the conditions of semi-artisan production, in the early summer on me, or at home).

Regarding substrate planting mycelium for extensive cultivation of oyster mushrooms - in Russia and the CIS countries expensive cereal (fully or with added components) planting mycelium is also traditionally used for inoculation in extensive mushroom growing.

However, this is not rational, since such a high-quality mycelium is, first of all, necessary to ensure intensive mushroom production. Asian mushroom growers consider grain a valuable food (and feed) raw material, and it is not necessary to use it for the manufacture of seed mycelium, which is used in fully sterile mushroom growing technologies. The sawdust enriched sterile planting mycelium is also quite adapted for successful wood grafting during extensive cultivation.

The objective of the present invention is to develop a method for growing oyster mushrooms, which ensures guaranteed harvesting and simplifies, reduces the cost of production of substrate mycelium for extensive cultivation of oyster mushrooms.

The problem is solved by combining two technologies - the intensive cultivation of oyster mushrooms and the production of substrate planting mycelium for extensive cultivation of oyster mushrooms.

Regarding the method of intensive cultivation of oyster mushrooms, the problem is solved as follows: glass liter jars filled with a substrate are closed with two pressed layers of foil without binding, and when the surface of the substrate is inoculated with uterine mycelium, a distance of 0.5-1 cm is left between the inoculated surface of the substrate and the can coating, moreover, after overgrowing, the coating from the cans is not removed, and the sprouts of mushrooms emerging from under the coating to the outside are gently pulled out with a hand movement downward, holding the coating, and they remove the coating after torogo fruiting. Cans can be 0.7-1.5 liter capacity. However, the optimal capacity is 1 liter.

Regarding the production of substrate planting mycelium for extensive cultivation of oyster mushrooms, the problem is solved in that as a substrate planting mycelium, a floor-sawdust nutrient substrate in a ratio of 2: 1, worked out in the two main “waves” of fruiting, is intensively grown, from which the upper 2- centimeter layer.

As shown by a detailed analysis of the patent and scientific and technical literature, the use of a constant coating of cans that are not tied, leaving 0.5-1 cm between the coating and the inoculated surface of the substrate, and the use of the “worked” in the first two waves of fruiting of the overgrown nutrient substrate as an inoculum for subsequent extensive mushroom cultivation meet the criteria of "novelty" and "inventive step".

A patented method for intensive cultivation of oyster mushrooms and the combined production of substrate planting mycelium for their extensive cultivation is carried out in the following sequence. Prepare a nutrient substrate, the main component (about 70%) of which in the claimed method, unlike Asian sterile technologies for cultivating mushrooms, is any cheap nutritious cellulose-containing agricultural waste: sex (empty ears) of oats, released in a large mass during the processing (sorting) of seed grains, finely chopped corn cobs, straw (ground) dust or husks of sunflower. Hardwood sawdust or fermented, long-lying, brown (i.e. with weakened fungistasis) conifers make up a smaller part - 30% - of the substrate. This composition of the nutrient substrate allows its rapid overgrowth and good yield of oyster mushrooms, and sawdust contributes to a good compaction of the substrate in banks, volumetric moisture retention and increase the effectiveness of the fungus inoculations on chocks (i.e., adapt the resulting substrate inoculum to extensive mushroom cultivation).

In the containers, the components of the nutrient substrate are mixed - oat flooring or other finely ground agricultural waste and sawdust in a ratio of 2: 1, poured with hot water and insisted for about 30 minutes. After that, the free water is drained and the raw mixture is laid out in liter cans (0.7-1.5 liters can be), the contents are compacted with a wooden “crush” so that no more than 1.5 cm remains between the densified surface of the substrate and the edges of the cans. Then the cans are covered with two layers of ordinary food-grade aluminum foil (13.5 × 13.5 cm squares), which they squeeze slightly around the neck with their hands, and put the cans in a heated vertical autoclave, covering each layer of the covered cans with cardboard and tin perforated gaskets. After filling the autoclaves, they are closed by heating the cans with the current steam, and autoclaved at an excess pressure of 1.6-1.8 atm. within 3 hours. Then they turn it off without releasing steam, leaving the cans in a closed autoclave until the next morning - for slow cooling. The next day, the autoclaves are opened, banks and pads are carefully removed and fed into a sterile box for inoculation. Inoculation is carried out with a uterine sterile grain inoculum mycelium, picking up a loosened (long metal sterile spatula) with a sterile tablespoon, lifting the foil (do this near burning alcohols) and pouring 0.5-1 cm layer from the spoon onto the surface of the grain substrate, lower the foil and squeeze its circumference around the neck of the jar. Moreover, between the inoculated surface of the substrate and the coating (foil) of the can should remain a distance of 0.5-1 cm (i.e. up to 1 cm). Inoculated cans are transferred to a clean (processed) cultivation room, equipped with a glued window and light, special air conditioning (cooling, heating, air purification) and shelving racks, which cans are put on and sprouted (Table 3) at room temperature (20-22 ° С ) more than a month (32-38 days) - depending on the type and strain of oyster mushroom, but not requiring cold shock. During the growing period, the cultivation room is treated twice - in the middle of the month and by the end of the month - with an inexpensive effective household pyrethroid insecticide. After the period of overgrowing begins spontaneous fruiting of oyster mushrooms from under the foil (figure 1, 2). Two cleanings are carried out with an interval of 8-16 days, carefully, holding the foil cover, pulling out sprouts of mushrooms with a hand movement downward. Immediately after the second cleaning, the cans are sent to another clean room for repetition of the contents in perforated plastic bags (with laser perforation), which they keep until spring in the refrigerator (or cold room) at low positive temperatures (3-5 ° C). When re-cooking, after removing the foil from the cans, remove the upper 2-centimeter layer of the substrate with a solid wooden spatula and discard it (as worked, physiologically older, with the remains of the aerial mycelium and the bases of the splices, as well as with a possible surface infection, laying flies or mushroom mosquitoes ) The accumulation of batches of substrate mycelium for spring use is carried out from late autumn to spring bookmarks. In summer mushroom rotations with an intensive culture based on heat-resistant lemon-oyster mushroom, four “waves” of fruiting and even five are obtained (Table 2), and the worked-up substrate is used in the preparation of fungocompost - an organic fertilizer for vegetable growing.

Testing of the proposed method was carried out in 2006-2007. on the basis of the Department of Biotechnology and Plant Protection GNU-DV of the Order of the TKZ NIISH Agricultural Academy. During intensive cultivation of oyster mushrooms, inoculation was performed with uterine sterile seed inoculum mycelium (own production), prepared on the basis of different types and strains of oyster mushrooms from the collection of GNU-DV of the Order of TKZ NIISH Agricultural Academy / Annenkov B.G. Khabarovsk genetic collection of cultivated tree edible and medicinal mushrooms as the basis for the development of mushroom growing in the Amur region // Forest biologically active resources (Mat. 3rd international conference. September 25-27, 2007) - Khabarovsk: DalNIIILKh, 2007. - P.228 -234./. In 2006, virulence tests of substrate seed mycelium were carried out both when using pasteurized straw substrate as an inoculum during eurotechnology, and for spring vaccinations of freshly sawed aspen and other chocks. It was found that the substrate inoculum differed, with respect to the sterile seed inoculum mycelium, in a reduced growth rate, which is clearly not enough for the rapid successful colonization of pasteurized straw substrate in intensive eurotechnology. In the spring, 20 chocks were inoculated (7 end openings with a diameter of 2 cm and a depth of 5-6 cm, with a sealing of the introduced substrate with a round stick with a diameter of 1.8 cm and setting the chocks on the soil and shading them). In the second half of September 2006, fruiting was obtained on all grafted chocks (Fig. 3), i.e. the claimed substrate mycelium was quite suitable for use in extensive mushroom growing. In the spring of 2007, lots of substrate mycelium, with a recommendation for use, were offered for testing to twenty amateurs-mushroom growers from Khabarovsk and the suburban Khabarovsk region. In September, positive reviews were received, and in some cases, documentary photos (figure 4).

Compared to others, this method has the following advantages:

firstly, they use small, liter, jars, which allows them to be placed in the autoclave more rationally, and this, in turn, allows faster fruiting, shorten the period of mushroom rotation and achieve effective fruiting in the main first "waves" (Fig.2 and table 1);

secondly, they use a coating of foil, slightly squeezed around the neck of the can, but not covered with a rope or elastic band, which makes it easier and faster to open the cans and conduct inoculation, it is more guaranteed to maintain sterility in the first stages;

thirdly, after complete overgrowing of the substrate, the foil coating is not removed, the formation of primordia occurs under the foil under favorable conditions of high humidity and the required concentration of CO ₂ around the neck. Two layers of foil, unlike one, are better held on the bank and are not thrown off the neck by the primordia of mushrooms that go outside from under the lower edge of the foil and mature.

Due to the combination of two technological processes - intensive cultivation of oyster mushrooms and the production of substrate planting mycelium for their extensive cultivation - the proposed method allows you to save internal asepsis of the substrate, reduce labor costs and increase the profitability of specialized mushroom farms, massively provide an important (popular) direction of domestic mushroom farming with an inexpensive inoculum, with acceptable quality for the extensive method.

It is planned to use the new method in the GNU-DV of the Order of the TKZ NIISH Agricultural Academy, where a mini-plant (i.e. elitkhoz) has already been created, intended for the annual production of up to 10 tons of high-quality sterile grain mycelium for the needs of local intensive mushroom growing. It is possible to organize a new unit - a specialized mushroom farm (special farm) for the cultivation of edible oyster mushrooms and seed for the development of mass extensive mushroom cultivation in the Amur Region.

Table 1 The productivity of the first wave of oyster mushrooms on different substrates in different volume glass jars under the foil (early summer 2005) Type of substrate and volume of glass jars The initial mass of raw autoclaved substrate, g The period from inoculation to harvesting the first wave, days Harvest of mushrooms with cans, g Fruit-giving, in% The average number of mushrooms on the bank, pcs. The average mass of one mushroom, g I. Oat grain (oyster mushroom lemon-hat - strain IIH-1) 1 liter 675 36-38 108 ± 9 16,0 25.7 4.2 2 liters 1350 40-42 190 ± 15 14.1 32,0 5.9 3 liter 2025 44-46 232 ± 12 11.5 34.3 6.8 II. A mixture (1: 1) of grinding corn kernels and soya dust (IIH-1) 1 liter 600 32-34 115 19.2 27 4.3 2 liters 1300 35-36 202 15,5 35 5.8 3 liter 1900 37-39 225 11.8 38 5.9 III. A mixture (1: 1) of grinding corn kernels and soya dust (P. ostreatus - strain NK-35) 1 liter 600 34-36 125 20.8 twenty 6.3 2 liters 1300 42-46 235 18.1 27 8.7 3 liter 1900 52-54 250 13,2 29th 8.6

Table 3 Comparative intensity of autoclaved oat grains and floor-sawdust substrate in mycelium glass jars under foil, surface inoculated with uterine culture of various species and oyster mushroom strains Type and strain of oyster mushroom Depth (cm) of substrate overgrowth with mycelium on day 25 after surface inoculation Oat (spring 2006) Floor sawdust substrate spring 2006 fall 2006 1. Oyster mushroom (R. ostreatus) but. strain NK-35 14.9 11.8 11.9 b. strain P-77 14.3 11,2 - at. Pingu 14.5 11.5 11.8 city "Florida" 15.0 11.9 - 2. Oyster mushroom lemon hat 14.7 10.9 12.1 (P. citrinopileatus) Note. The height of the substrate in a liter jar = 15 cm

Claims (3)

1. A method of intensively growing oyster mushrooms, including preparing and disinfecting the substrate, filling glass jars with it, adding mycelium to the substrate, moistening and compacting the substrate and forming mushrooms, characterized in that the glass jars are covered with two pressed layers of foil and sterilized in an autoclave at a pressure of at least 1.6-1.8 atm for 3 hours, leaving a distance of 0.5-1 cm between the inoculated surface of the substrate and the coating of the cans, and after overgrowing, the coating from the cans is not removed, and the sprouts of fungi emerging from under covering of outward movement of the hand gently pulled down, holding the coating, and the coating is removed after the second fruiting. 2. The method according to claim 1, characterized in that the cultivation of the substrate is carried out for 32-38 days at a temperature of 20-22 ° C. 3. A method for the production of substrate planting mycelium for extensive cultivation of oyster mushrooms, characterized in that the substrate planting mycelium used in two main “waves” of fruiting during intensive cultivation of oyster mushrooms is a sawdust nutrient substrate in a 2: 1 ratio, from which the upper 2-centimeter is removed layer.

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