TW200819060A - Culture improvement of lepista nuda and cultivation method thereof - Google Patents

Abstract

The culture improvement of Lepista nuda and cultivation method thereof are disclosed. An Inter-strain mating of the parental strains LNE2 and LNG is performed and the resulted hybrid dikaryons are subjected to a series of fruiting tests to select a better new strain. The new strain has been deposited in Food Industry Research and Development Institute as a deposition number of BCRC 930083. The new strain has advantages of fast growth rate, big mushroom body, firm and fine texture, and flexible mouthfeel when tasted. Moreover, both the production quantity and quality can be effectively improved by the cultivation method of the present invention.

Description

200819060 IX. Description of invention:

Technical Field] A modification of a variety of lilac mushrooms. [First technology]

Long, among them, the technological level of modern science and technology with agricultural production is constantly improving. The productivity and consumption power of agriculture is increasing at an alarming rate. The food production industry is very heavy and edible fungi. The cultivation technology of various mushroom strains is now exquisite. Chemical agriculture, one of the most important technologies, and the general products of the categorical products are very good in the need of excellent strains or varieties, and the combination of culture techniques: to the scale of commercial production. Bacterial culture technique

The corpus is named for its exemption from color and has a special aroma. It is considered to be the best food in France. It is one of the top ten favorite edible fungi. This genus is a saprophytic edible fungus. It can be found in broad-leaved mixed forests and forest margins in the autumn and winter seasons. It is mainly distributed in Europe, North America and Asia, and is known as the mushroom-shaped disc cap. Wood Blewit or Blewit. The fruiting body is purple or lilac-purple at the beginning, the color is lighter in the later stage, the umbrella is thick, crisp and delicious, and the sub-aroma has a strong aroma. In Europe, the annual output can be 250-300. The formation of hernia is mainly based on mushroom alcohol (Buocten-3-ol), of which the content of pleats 5 200819060 is the most, and the stipe and umbrella are less. The fruiting body of the lilac mushroom contains vitamin A1. Other nutrients are for further analysis. In addition, it has an inhibition rate of more than 90% for mouse sarcoma and Ehrlich cancer, and has the value of developing into a health food such as anti-cancer and anti-cancer. However, the fresh mushroom body contains a certain component that can decompose red blood cells, and further cooking is required to neutralize its toxicity. The early cultivation of the lilac mushroom is based on the beech and apple leaves. The time for the mushrooming is very long, about 6 months, and the temperature required for the mushroom is low, and the cultivation is not easy. Therefore, the present invention mainly performs the improvement of the species of the lilac mushroom, and selects the product for the production of 13⁄4 and has a short time. SUMMARY OF THE INVENTION The main object of the present invention is to improve the variety of the syringa mushroom to breed a microorganism with a unique edible taste, and cooperate with the cultivation method to effectively increase the production yield of the syringa mushroom and improve the sage of the sage mushroom. In order to achieve the above purposes, the present invention utilizes hybrid double-nuclear culture technology to hybridize LNE2 and LNG strains as hybrid parents, and selects a new purple lilac mushroom new line from the hybrid progeny. The edible microorganisms of the lilac mushroom bred by the present invention have been deposited in the Food Industry Development Research Institute under the registration number BCRC 930083. The lilac mushroom has the advantages of fast growth, full and thick mushroom shape, firm and delicate tissue, and elastic mouthfeel. The method for cultivating the syringa mushroom of the present invention comprises the steps of: (1) cultivating and preserving and breeding the hybrid parental original species; (2) cultivating the hybrid parent; (3) performing single spore separation and single spore of the sclerotium Identification and culture of strains; (4) 6 200819060 Confirmation, culture and preservation of hybridization and hybridization of single-spore strains; (5) Screening of first-time cultivation of hybrid dinuclears; and (6) Excellent hybridization Re-cultivation of dinuclear bodies and investigation of mushroom characteristics. Among them, the step (1) further includes the production of a·original species; b. cultivation of the hybrid parental line; and the collection of c. The cultivation method of the lilac mushroom of the present invention comprises the steps of: (1) parental culture; (2) compost production; (3) management of the next and the next species; (4) soil and mushroom stimulation operation; ) Harvesting and packaging. Among them, the composting system is a straw compost, which can be obtained by the traditional mushroom straw composting method or by fermentation in a tunnel type fermentation chamber. [Embodiment] Some exemplary embodiments embodying the features and advantages of the present invention will be described in detail in the following description. It is to be understood that the present invention is capable of various modifications in various embodiments, and is not intended to limit the scope of the invention. In order to further improve the varieties of the lilac mushroom, the inventor of the present invention introduced four strains of lilac mushroom from France, namely LNE2, LNP, LNG and LNB, and observed the growth of the four strains in the mushroom compost, and found LNE2 and LNP. And LNG three strains can produce mushrooms, while LNB strains only have hyphae growth. Among them, LNE2 was the earliest mushroom, and the yield was also the highest, with an average of 435.4 g / 2 kg compost per bag. There was no significant difference between LNP and LNG. However, because the LNG strain was larger, LNE2 and LNG were selected. The strains were subjected to single-spore hybridization to screen out the excellent new lines. 7 200819060 About this case The details are as follows. The breeding method of Ziding spice secret line will be carried out in step-by-step (1). The breeding and breeding of the original parent species of the hybrid parent: The hybrid parent LNE2 and LNG are respectively composted corn-yang-liated base (the formula is Air-cured compost 25 grams, crushed corn 1 gram, Yanke and steamed i liters) should be periodically transplanted, cultured in test medium at a fixed temperature of about one month, and then transferred to 5 its constant temperature box for preservation. (2) Cultivation of hybrid parents: a. Production of the original species: using the original wheat kernels, wheat (speaking to the feed), washing with water, placing in cold water, boiling for 2 minutes, removing water, and Add 1% calcium carbonate, add to the four-necked flask (6 3x3 5xl5cm) after cooling, and stuff the tampon, then sterilize (12rc, 12 kg/cm2) for 1 hour in high temperature and high pressure steam boiler. After cooling, it will be cultivated in compost. The culture medium of the clove mushroom hyphae was inoculated into a four-necked flask at 24. (: After 3 days of constant temperature culture, it is a kind of wheat grain. 6. 6 · Miscellaneous parents, cultivation of π mouth · · will be purchased Yangzi compost (the formula is 5,000 kg of dried straw, 500 kg of chicken manure, lime 4〇kg, 100kg of superphosphate, 125kg of gypsum, 1(10)kg of ammonium sulfate, 50kg of soy flour and 200kg of rice bran), put 15kg into the plastic box, and evenly mix the above two strains of wheat , the temperature is set at 24 ° C, the relative micro-95 % 'carbon dioxide concentration 3000ppm, after the mycelium is full, the soil is covered, and the temperature of the mushroom is reduced to 13t, the carbon dioxide concentration is reduced to 500ppm, and the daylight is illuminated for 8 hours. c. Collection: The lilac mushroom body for the supply of spores is first removed in 8200819060 dust-free sterile filter box with a sterilized scorpion knife. The stalk is inserted downwards in the sterilized::: skin, and the ring is used. On the protruding wire, it is placed under the iron of the killing type. The above device is moved into the filter paper, and the spores are dropped to the core of the paper to be carefully placed. People kill g = = (four) two inoculation into the 5 C constant temperature box Save the spare. In the soil-breaking test tube, (3) into the (four) scorpion of the μ (four) using the scorpion dilution culture separation method separation == confirmation and culture: the hybrid parent _ the spores to no 8 = strain, 'will be collected The homologous sentence is applied to the malt yeast of the 9em culture dish = the appropriate concentration. The base formula is 2G grams of malt extract and yeast nutrient (culture, ===:: r only see alone: 兮, In this case, it is true that 5 other spores, sputum, sputum, and sputum are carefully removed from the parsley containing the above colonies, and then, 35 I knives, 糸 糸 棉 棉 棉 棉 棉 棉 棉 棉 棉 棉 棉Microscopic examination showed that there were no deductions on the hyphae, and if there were non-singular spores, the 8 filaments were normally caught on the test tube of the composted corn medium, which should be a single spore strain. Confirmation of hybridization and hybridization of dinuclear cells, and transmission of the monosporic strains after culture were carried out in a composting corn medium slant test tube 9 200819060, and the single-spore strains of the two hybrid parents were simultaneously inoculated into the same test medium at a distance of 5 to 1 cm. On the top, after culturing at 25 ° C for 20 to 25 days, Observing the changes of the colonies of the two hybrid strains, if the two colonies fuse with each other and lose their individuality and cannot be distinguished, that is, there is no boundary between the two colonies, that is, there may be affinity between the two strains, respectively. At the top of the two colonies or at the top and bottom of the contact, the hyphae are taken out. If the hyphae are taken out, the hybridization can be confirmed. At this time, the hybrid mycelium can be removed and cultured. The new medium is transplanted and preserved regularly according to the above method. (5) Screening for the first cultivation of the hybrid dinuclear: After confirming the hybrid dinuclear, the original wheat kernel is prepared, and each double-nucleus is inoculated with 3 boxes. The culture substrate formula used is the same as the formula of the cultivated parent. From the comparison of different dinuclear bodies, the screening efficiency of the mushroom production is high, the appearance of the mushroom body is good, the firmness of the mushroom body tissue is good, and the mushroom is early and neat, for further cultivation test. Comparison. • (6) Investigation of re-cultivation and mushroom characteristics of excellent hybrid dinuclear: The superior dinuclear body was obtained from the first cultivation test, and then the cultivation strain was prepared. Each double-nuclei was cultivated in 10 boxes, each frame was The unit compares the morning and evening of the mushroom, the uniformity of the mushroom, the characteristics of the mushroom, the yield and the efficiency of the mushroom production, so as to select the hybrid dinucleus with high characteristic and cultivation economic value, and then expand the cultivation scale, multiple times with the hybrid parent or current Commercial cultivars are compared to obtain the most excellent edible microorganisms of the lilac mushroom. It is worth mentioning that in the cultivation, reproduction and preservation of the original parent species of the hybrid parent, the permanent preservation and genetic characteristics of the strain are stable and reliable, and the liquid nitrogen storage method can be used in 200819060, and the program cooling controller is used. The preserved species of S. cerevisiae decreased from room temperature to -4 〇t at a rate of 1 ° C per minute: and then dropped to -100 ° C, and immediately stored in liquid nitrogen. In addition, the strains to be stored in liquid nitrogen can be cultured in the sterilized millet. After 20 days of incubation at a fixed temperature, carefully transfer the sterilized ultra-low temperature glass ampoules (1.2 ml, boron) in a dust-free sterile filter box.矽 acid glass material) or plastic bottle (2ml). φ Also when using ampoules, check for seals after sealing, otherwise ampoules may occur. When the strain is placed in an ampoule or a plastic bottle, a 10% glycerin solution can be used as a frost protection agent. When the strain is taken out and thawed, the strained ampoule which is taken out from the liquid nitrogen drum can be directly thawed in 37t: to 38t: warm water, and the strain is thawed and then transferred to the compost corn acacia medium for cultivation. In the cultivation of the hybrid parent, when the mushroom body grows to about 3 cm high, and before the spores are not produced, in order to avoid the contamination of the other spores or bacteria of the clove mushroom, and the spores of the pathogen, the mushroom can be selected. The shape of the cultivation bottle should be bagged at the place where the spirulina mushroom matures until the pleats are well developed, and the stalks are collected for collection. On the other hand, in the cultivation of strains and the collection of the spores, different varieties are preferably cultivated separately in different mushroom chambers, and do not operate in the same sterile dust-free filter box on the same day to reduce the mutual contamination of spores of different varieties. opportunity. The cultivation and management method of the lilac mushroom cultivated by the invention is the same as that of the cultivated parent strain, and the mushroom should be investigated after the mushroom is harvested, the mushroom uniformity, the small mushroom number, the stem diameter and the tissue firming situation, etc. After maturity, investigate the appearance of the mushroom body, including the surface color, thickness, length and tissue of the mushroom handle. 11 200819060 degrees, the size, color, surface of the mushroom, or any hydatid mass, and observe the presence or absence of the mushroom body. Yellow phenomenon and record the weight of each frame of the mushroom, and calculate the biological efficiency of each double core (biological efficiency is the percentage of the fresh weight of the mushroom divided by the dry weight of the cultivation substrate at the time of inoculation). Through the repeated trials and screenings of the above-mentioned breeding methods, the present invention selects an excellent strain of purple lilac mushroom, named as Tai Nong No. 1 lilac mushroom (L-heart (ARI-LN1)), and on April 25, 1995.曰 Deposited at the Food Industry Development Research Institute of Hsinchu, Taiwan, with the registration number BCRC 930083. The mushroom shape of Tai Nong No. 1 is very rich and coarse, the microorganisms are firm and tender, the taste of the food is elastic, and it has the characteristics of rich aroma, smooth and tender umbrella, good resistance to transportation and long shelf life, so it has considerable market competitiveness. . The following Tables 1 to 3 are the comparison of the mycelial growth characteristics, the comparison of the size of the fruiting body and the characteristics of the fruiting of the new strain of the syringa mushroom ARI-LN1 bred by the present invention and the parental lines LNE2 and LNG. • Table 1. Comparison of mycelial growth characteristics of new RIS-LN1 and parental lines (LNE2, LNG). Mycelial growth characteristics S. Lilac mushroom line LNE2 LNG ARI-LN1 Mycelium color transparent transparent bacteria Silk surface color lavender lilac lavender temperature adaptability (°c) 5-25 5-30 5-25 hyphae optimum growth temperature rc) 25 20 20-25 growth of mycelium at different temperatures (mm/day 0.62 0.32 0.62 12 200819060 10°c ———— 1.08 1.03 1.08 15°C 1.50 1.56 L68 20°C 1.91 1.99 2.08 25〇C 1.93 1.94 2.08 30°C 0 0.04 0 Table 2, Lilac Mushroom Comparison of average sub-entity sizes of new strains ARI_LN1 and parental lines (LNE2, LNG)

Fruit body size lilac mushroom strain (cm) LNE2 LNG ARI-LN1 umbrella length 8.09 7.91 7.85 umbrella width 7.44 7.19 7.42 umbrella height 1.81 1.99 1.66 pleat length 2.93 2.93 2.80 pleat width 0.74 0.72 0.61 umbrella thickness 1.14 L25 1.44 stipe Length 8.31 10.67 8.38 Stipe width 2.81 2.74 2.89 Average yield of lilac mushroom 衫 (2, LNG) (kg/bed) Bio-efficiency (%) From covering soil to mushrooming jinyunxu LNE2 9933.75 52.72 35.75 LNG 8754.00 46.46 39.50 ARI-LN1 11009.25 58.43 34.25 4氺16kg compost 3% spawn(3/4/5*16Kg c〇mpost), every _ mouth Η Η) bed, per bed area W, compost 64 kg, Composting|^|74Gf6&'Mother one weight 13 200819060 Table 3 bis, comparison of the characteristics of the new AI-LN1 and the parental line (LNE2, LNG) of the lilac mushroom series Lilac mushroom strain LNE2

LNG ARI-LN1 Average yield ί kg%j ^298833^ 1916.83 3177.67 Bioefficiency from covering soil to mushroom (%) Days required 63.44 35.67 ~ 40.70 46.17 67.46 35Ό0 f product 58*39cm, compost 16kg compost water content 70.56% . Same as above

.A and II, the lilac introduced by the present invention is a new parental product LNE2, lng, which has a higher yield and a shorter number of days, and is indeed superior to the parental product (four) tender:: bred The mushroom shape of the lilac mushroom is full and large, and the microbes (the growth conditions of the second edible microbe:: listening, not more than 15/., length = 24 °c. 'The development temperature of the body is 65-67%. , Γί. The fermented compost is maintained at 90-93%^, growth and growth stage of the air line, silk: no need for growth, no special ventilation, dioxin, 曲() Ventilation. When walking, the mushroom and mushroom body need to pass through the toilet. The "disgusting" is controlled at about 300 〇 PPm, and the ppm is good. The money is good. The dioxobic concentration is maintained below, which will further illustrate the lilac mill of the present invention. Planting of edible microbes 14 200819060 The production method of culture is as follows: (1) Parental culture step · · Transplanting the lilac genus u (ARI_LN1) of the present invention into compost corn acacia medium, followed by aseptic operation The lilac mushroom cultivated in the compost corn acacia medium on the stage The hyphae are inoculated into a bottled wheat cultivating substrate sterilized by high pressure steam, and then at a temperature of 24 〇c.

After the cultivation of the mycelium of the clove mushroom is full of the inner matrix of the bottle, the seed of the purple scented grinding is obtained. (2) Composting process: a·Materials and serving size: add 5,000 chicken per 5,000 kilograms of dried straw, 80 kg of ammonium sulfate, 150 kg of superphosphate, 1 kg of acid, 250 kg of gypsum, 50 kg Soy flour and 200 kg of rice ^. Production process: composting can be carried out by using a transecting straw stacking method or a modern tunnel-type straw composting indoor fermentation method. ...+ The grass composting method is pre-fermented in the outdoor, and then in the room, the yeast is used in the tunnel-type straw composting room, and the 1st and the females are taken. After adding the water to the city, the water is fully added, and then the material is added, and the stacker is scooped. Pre-fermentation is carried out in a private tunneling fermentation chamber. When the heap is removed from the compost for two days, the fermentation is good and the mixture is not mixed. (4) The ratio is about 5%. The mixture is fully mixed with the composting machine and the fermentation chamber is fixed. The post-fermentation is continued. Temperature. When the composting temperature is above the machine, carry out the low of six hours. 15 200819060 Adjust the compost temperature to 52 °C for one day, then reduce it to 48-50X: maintain three days, and finally reduce it to 45-48 °C. After two days of maintenance, if there is no ammonia in the compost, it can be cooled down to 35 °C. The next day, the compost can be removed for the undergrowth of the purple clove mushroom. 0 (3) Management of the next and the next species·· After composting and fermentation When the compost temperature drops to about 25 °C, it can be planted. Before the next planting, the compost should be loosened. The amount of the seed is 3% (w/w) of the compost, and the grain and compost are evenly mixed, and then the surface is leveled. The compost temperature was kept at 24 °C during the subsequent colonization. (4) Surgery operation of covering soil and mushroom: After composting, when the mycelium of the wheat grain is full of compost, it can be covered with l-2cm clay soil (pH 7.0), and it is cooled and greatly ventilated. During the stimulation of the mushrooming, a small amount of water is applied to the surface of the mushroom bed about two to three days to maintain the humidity of the mushroom bed. (5) Harvesting and packaging steps: When the mushroom body develops to the appropriate stage, the mushroom umbrella is flattened, and the edge is not taken up before rewinding. When harvesting, it should be noted that it should not interfere with the surrounding growth of the mushroom body and will carry mud carbon. The mushroom feet of the soil are partially removed and packaged for refrigerated sales. After each cycle of harvesting, the mushroom bed needs to be properly replenished and hydrated to facilitate the formation of the mushroom body in the next cycle. Through the combination of the above steps, it is possible to mass produce edible microorganisms with better quality, and can effectively stimulate the speed of the mushroom, which not only makes the finished product of the mushroom tidy, but also has higher biological efficiency. By using the above five steps, the yield can be increased, and with appropriate mushroom management, the mushroom body produced in each cycle of 16 200819060 can be effectively controlled to ensure the stability of production quality and yield. The invention not only can greatly increase the production yield of the lilac mushroom by 10%, but also uses less labor, reduces the production cost, and can improve the production quality of the lilac mushroom, so that the mushroom shape of the lilac mushroom is full and thick, the microorganism is firm and delicate, and the taste is tasted. Flexible. In addition, the lilac mushroom is a relatively high-grade edible mushroom in Europe, such as France, and the price is 5-10 times that of the mushroom, so it has great market potential in the future. Therefore, the present invention does provide a very competitive industry. A new variety of lilac mushrooms with a high economic value of cultivation. The lilac mushroom cultivated can be further processed and used. For example, the fresh mushroom body can be made into various kinds of dishes, and can be made into a vacuum package and can be eaten by microwave heating. In addition, it can also be killed. After cooling, add appropriate seasonings to make cans. It can be placed in the refrigerator and ready to be taken out as a cold dish. In addition, the body can be dried to make a gift box. After the blisters are opened, the brittleness of the umbrella is still maintained, and the handle is more tough. In summary, the present invention utilizes the advantage of hybridization to breed a new φ species of purple lilac mushroom, and the germinated purple lilac mushroom has a unique edible taste, and the cultivation production method can effectively increase the production yield of the gerbera mushroom and improve the quality of the lilac mushroom body. . Therefore, the present invention is extremely valuable for industrial use and is submitted in accordance with the law. This case has been modified by people who are familiar with the technology, and is not intended to be protected by the scope of the patent application. 17 200819060 [Description of the diagram]

[Main component symbol description]

Claims (1)

200819060 .j X. Patent application scope: 1. A fungus fungus microorganism, which has the identification characteristics of the strain deposited in the Food Industry Development Research Institute and registered as BCRC 930083. 2. The edible fungus microorganism of claim 1, wherein the fungus microorganism is a lilac mushroom. 3. A method for cultivating lilac, which is a hybrid parental culture technique, using LNE2 and LNG strains as hybrid parents for hybridization, and selected from the hybrids of the miscellaneous 10 to be deposited in the Institute of Food Industry Development. , the registration number is BCRC 930083 bacteria; ^ identification features. 4. The method for cultivating the lilac mushroom according to claim 3, wherein the breeding method comprises the steps of: (1) cultivating, breeding and preserving the original parent of the hybrid parent; (2) carrying out the cultivation of the hybrid parent (3) performing single spore isolation of the spores and confirmation and culture of the single spore strain; (4) performing hybridization between the single spore strain and confirmation of the hybrid dinuclear, culture mw and preservation; (5) performing hybridization of the dinuclear body; The first cultivation of the mushroom screening; and (6) the re-cultivation of the excellent hybrid dinuclear and the investigation of the characteristics of the mushroom. 5. The method for cultivating the lilac mushroom according to item 4 of the patent application, wherein the step (1) further comprises the steps of: a. production of the original species; b. cultivation of the hybrid female line; and c. collect. 19 200819060 $ Ψ 6. A method for cultivating a lilac mushroom having an identification characteristic of a strain deposited under the Food Industry Development Research Institute and having the registration number BCRC 930083, the cultivation method comprising the steps of: (1) Maternal culture; (2) composting; (3) management of the next and next species; (4) soiling and stimulation operations; • (5) harvesting and packaging. 7. The method for cultivating a lilac mushroom according to claim 6, wherein the compost is a straw compost. 8. The method for cultivating a lilac mushroom according to claim 6, wherein the compost is obtained by fermentation in a tunnel type fermentation chamber. 9. The method for cultivating a lilac mill according to claim 6, wherein the compost is obtained by pre-fermentation in the outdoor and post-fermentation in a room. 20 200819060 VII. Designated representative map: (1) The representative representative of the case is: Figure (2) A brief description of the symbol of the representative figure: 8. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention = 200819060 f綷mjm is the supplement of the invention patent specification (the format, order and bold text of this manual, please do not change it at all, please do not fill in the mark part) ※Application No.: 幻" ※Application date: Θ·" to .1 PC classification: I. Name of the invention: (Chinese / English) Lilac mushroom and its cultivation and cultivation methods / CULTURE IMPROVEMENT OF LEPISTANUDA AND CULTIVATION METHOD THEREOF II. Applicant: (1 in total) Name: (Chinese/English) (Signature) ID: Agricultural Research Institute of the Agricultural Research Institute / AGRICULTURAL RESEARCH INSTITUTE □ Designated as the representative of the person to be served: (Chinese / English) (Signature) Lin Junyi/LIN, CHUN ΥΙ Residence or business address: (Chinese/English) zhongzheng Road, Wanfeng Village, Wufeng Township, Taichung County 189 f/189Chimg-ChengRd., Wufeng Taichung, Taiwan 413, ROC· Nationality: (Chinese/English) Republic of China/TW Telephone/Fax/Mobile: 04-23302301 e-mail: III. Inventor: (Total 4) Name: (Chinese/English) ID : 1 ·Chen Jintong / CHEN,JIN TONG 2·Chen Meixing/CHEN,MEEIHSING 3·Peng Jinteng/PENG,JINT〇RNG 〆4·Lin Junyi/LI& CHIE^YIH Nationality: (Chinese/English) 1 - 4. Republic of China / TW 1