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Method for planting and propagating Ganoderma lucidum
Ganoderma lucidum is the dried fruiting body of Ganoderma lucidum, a fungus of Polyporaceae. Location: Shandong, Jilin, Hebei, Shanxi, Shaanxi, Anhui, Jiangsu, Hubei, Zhejiang, Fujian and other provinces.

I. Biological characteristics

(A) growth and development habits

Ganoderma lucidum belongs to saprophytic fungi, and its life history is as follows: basidiospore → bud tube → mononuclear mycelium → binuclear mycelium → fruiting body → basidiospore. In the life of Ganoderma lucidum, basidiospores germinate into bud tubes under suitable conditions, and after the processes of mass matching, nuclear matching and meiosis affinity, mononuclear hyphae (primary hyphae) are formed. Two different levels of mononuclear hyphae lock to form binuclear hyphae (secondary hyphae), which grow to a certain stage, and then form fruiting body primordium through specialization, aggregation and close connection. The primordium of fruiting body develops and grows upward, forming a columnar stalk. When the physiology is mature, that is, the fruiting body layer covered by fungi sends out basidiospores and begins a new development cycle.

(2) Ecological environment conditions

Ganoderma lucidum needs suitable nutrition, temperature, humidity, light, pH and other conditions during its growth and development.

1. Nutrition

Ganoderma lucidum belongs to saprophytic fungi and also belongs to facultative parasite. It can rot on rotten wood or parasitize on living trees. Most broad-leaved trees, sawdust, leaves, rice straw powder, bagasse, crop stalks, cottonseed hulls and wheat bran can be selected as artificial cultivation materials. The nutritional basis is carbohydrates and nitrogen compounds, and the ratio of carbon to nitrogen is 22∶ 1. The main carbon sources are glucose, sucrose, starch, cellulose, hemicellulose, lignin and so on. Nitrogen sources include protein, amino acids, urea and ammonium salts. It also needs a small amount of potassium, magnesium, calcium, phosphorus, vitamins, water and so on.

2. Temperature

Its growth and development need high temperature and high humidity. The adaptive temperature range is 12 ~ 32℃, and the optimum growth temperature is 25 ~ 28℃, which is higher than 35℃. Mycelia is easy to age and autolysis, and the fruiting body dies. When the temperature is lower than 12℃, not only the mycelium growth is limited, but also the fruiting body can not grow and develop normally. In addition, the temperature is not suitable, which will produce a deformed cap.

3. Humidity

In the mycelium growth stage, it is required that the water content of the culture medium is 55% ~ 65% and the relative humidity of the air is 65% ~ 70%; In the growth stage of fruiting body, the water content of culture medium is 60% ~ 65%, and the relative humidity of air is 85% ~ 95%.

4. Air

Ganoderma lucidum is an aerobic fungus. In order to ensure its normal growth and development, ventilation should be strengthened in the process of breeding to reduce harmful gases.

lamplight

Mycelium needs dark or weak light in the growth stage. In the growth stage of fruiting body, a proper amount of scattered or reflected light is needed to avoid direct light.

6.ph value

Ganoderma lucidum can grow at pH 3.0 ~ 7.5. The optimum pH value is 5.0 ~ 6.0.

Second, the cultivation techniques

(1) Cultivation of Ganoderma lucidum strains

The cultivation process of Ganoderma lucidum strain includes the separation of pure strain, the cultivation of mother strain (primary strain), the production of original strain (secondary strain) and the production of cultivated strain (tertiary strain). The production steps of culture mainly include preparation of culture medium (material), sterilization, disinfection, separation, inoculation, culture and preservation. All used instruments and tools should be disinfected to ensure aseptic operation.

1. Isolation of Pure Strain and Culture of Mother Strain

The original strain was obtained by tissue separation or spore separation, and then the mother strain was obtained by inoculation in culture medium. The mother seed was cultured in potato agar (PDA) medium.

Formula: peeled and chopped potatoes 200g, glucose 20g, agar 20g, potassium dihydrogen phosphate 3g, magnesium sulfate 1.5g, vitamin B 12 tablets and water 1000mL. 120 test tube culture medium can be prepared. Wash the fresh and mature Ganoderma lucidum with clear water, and then rinse it with 75% alcohol or cold boiled water. Under aseptic conditions, 1 piece of soybean-sized tissue was cut from the inside of the cap or stem. The tissue block was cut into small pieces with an inoculation knife, inoculated in the center of the inclined culture medium, and cultured at 24 ~ 25℃ for 7 ~ 10 days. When white, uniform and vigorous hyphae grow on the inclined plane, the mother seed is obtained. Spores can also be inoculated on inclined culture medium to cultivate mother seeds, or they can be carried out under aseptic conditions. The method is to take high-quality ganoderma lucidum fruiting bodies that have begun to release spores and disinfect them for later use. Collecting spores, inoculating the spores on the culture medium, and culturing to obtain a thin layer of lichen-like vegetative hyphae, namely the mother seed.

2. Production of original seeds and cultivated seeds

Inoculate the mother seed into the culture medium, expand the original seed, and then use the original seed to expand the cultivated seed to meet the amount of bacteria needed for culture. In small-scale production, the mother seed or the original seed can be directly inoculated and cultured.

The formula of the culture material for producing the original seed or cultivated seed is the same as that of the bag (bottle) cultivation method of fruiting body, and there are many formulas. The main raw material is sawdust or cottonseed hull, and appropriate auxiliary materials are added to make mixed culture material. Formula 1: wheat 99%, gypsum 1%, water 65%; Formula 2: sawdust 78%, bran 20%, gypsum 1%, soybean powder1%; Formula 3: bagasse 75%, bran 20%, 1% sucrose, 1% gypsum, 1% soybean powder; Formula 4: 80% cottonseed hull, 16% bran, 1% sucrose and 3% quicklime.

According to the formula, add 140 ~ 160 kg of water to every 100kg of dry material, mix well, put it in a strain bottle until the height of the bottle is 2/3, make a hole near the bottom of the bottle, clean the dirt and seal it with kraft paper. High pressure or normal pressure high temperature sterilization. Inoculate the strain after cooling, about 1 test tube mother seed inoculates 5 bottles of original seed, and 1 bottle of original seed inoculates 50-60 bottles of cultivated seed. After inoculation, put it in a culture room for culture, and it is necessary to control the conditions. After 25 ~ 30 days, the bottle is full of hyphae and can be inoculated and cultured.

(B) Ganoderma cultivation techniques

The cultivation methods of Ganoderma lucidum mainly include short cutting method (clinker short cutting method, raw material short cutting method, stump cultivation method), bag cultivation method and bottle cultivation method. The cultivation method of short-cut timber mainly adopts clinker short-cut timber method. Bottle cultivation is the earliest cultivation method, which is mainly used for the production of ganoderma lucidum spore powder and the cultivation of original or cultivated species. At present, bag cultivation is mainly used, which can be cultivated indoors, in greenhouses, greenhouses and in the open field.

1. Bag planting method

Process flow: material preparation → bagging and sterilization → inoculation → mycelium culture → Ganoderma lucidum management → harvesting and processing. Under suitable conditions, it can be cultivated all year round. It is mainly planted in spring, that is, seed production in March-April and inoculation cultivation in April-May. In autumn, seeds are produced in July and inoculated in August.

(1) Material preparation and batching: same as "Production of original seeds and cultivated seeds".

(2) Bagging and sterilization: Generally, PVC or polypropylene plastic bags are selected, the specifications are about 0.04mm thick, 36cm long and 18cm wide. Prepare the culture material and put it in a bag, about 8cm away from the mouth of the bag. The loading capacity is about 500g, so it needs to be packed. After the air in the bag mouth is exhausted, tie it tightly with a rope, put it into a sterilization pot, and sterilize it at 1.5kg/cm2 for 2 hours, or at 100℃ for 4 hours at normal pressure, then cease fire for 5 hours, and then cool it to about 25℃ and take it out of the pot.

(3) Inoculation: Inoculation is carried out under aseptic conditions. During inoculation, the strain is in close contact with the culture material, and the bag mouth is tied tightly in time. Each bottle of strain can receive 20 ~ 30 bags. Put the inoculated fungus bag into a culture room or a culture rack in a greenhouse for mycelium culture (fungus growth). The temperature should be controlled at 22 ~ 30℃, preferably at 24 ~ 28℃. Store away from light and pay attention to ventilation and cooling. 1 week or so 1 time, and discard the contaminated bacterial bags. 10 days or so, hyphae can grow full of bags.

(4) Management of fruiting: When the mycelium grows to about 30 days, white papules or protrusions of finger size are formed on the surface, that is, the primordia of fruiting bodies-Lei Zhi or mushroom buds. At this time, it is necessary to untie the plastic bag, so that Ganoderma lucidum can grow outward, and the buds of Ganoderma lucidum extend outward to form a stalk. The stipe grows on the stipe for about 15 days, and matures after 30 ~ 50 days, and spores can be harvested from the stipe. At this stage, measures such as ventilation and spraying water into the air should be taken. The temperature should be controlled at 24 ~ 28℃ and the relative humidity of the air should be 90% ~ 95%, so as to keep the air fresh and the light scattered.

Fruit body culture can be cultivated outdoors. Dig a bacterial bed with a width of 80 ~ 100 cm and a depth of 40cm, depending on the plot conditions and culture capacity. Take out the bags of cultured hyphae, put them vertically on the bacterial bed with a spacing of about 6cm, cover them with 1cm thick fine soil rich in humus, and pour enough water. Build a plastic shed on the bed to keep out direct light. Keep the temperature at 22 ~ 28℃, fresh air and relative humidity at 85% ~ 95%. After 10 days, the fruiting body primordium appeared on the bed surface, which matured after 25 days and could be harvested.

2. Segmented culture method

Process flow: material selection and preparation → bagging and sterilization → inoculation → mycelium culture → soil culture → Ganoderma lucidum management → harvesting and processing.

(1) Selection and treatment of wood-saving materials: select broad-leaved trees such as chestnut, oak, catalpa bungeana, willow, poplar, Robinia pseudoacacia and maple with a diameter of 8-10, and saw wood-saving materials with a length of 15-20 cm, and dry them in the sun for about 3 days, with a moisture content of 35.

(2) Bagging and sterilization: put the wood blocks into plastic bags, and the wood should not be too dry. Fasten the bag mouth tightly and sterilize at high pressure and high temperature for 2 hours, or at normal pressure 100℃ for 6-8 hours.

(3) Inoculation: carried out under aseptic conditions. Perforation inoculation or staged inoculation can be used. Inoculation by punching holes: punch holes in the wood, the diameter is 1 ~ 1.2 cm, the depth is 1cm, the row spacing is about 5cm, and each row has 2-3 holes, which are arranged in a zigzag manner. Inoculate under aseptic conditions immediately after punching, select and cultivate for 20 days, take out the fresh strain just formed from the primordium of fruiting body, plug it into the hole, slightly compact it, cover it with cork or bark, and gently hammer it flat with a small mallet. Cross-section inoculation is to evenly mix two pieces of wood in a bag with cold boiled water, then evenly coat the strains between the two pieces of wood and the surface of the previous piece of wood, plug a group of sterile cotton in the mouth of the bag and tie it tightly. The air temperature should be 20 ~ 26℃ and the air relative humidity should be 70%. 1m3 timber needs 60 ~ 100 bottles of strains.

(4) hypha culture: the inoculated Tricholoma bag is placed in a ventilated and dry place to culture hypha, the temperature is controlled at 22-25 DEG C, and ventilation, cooling and mildew prevention are carried out. After 30 ~ 60 days, white hyphae will cover the cavity, and the periphery of the cavity will turn white or light yellow, and then gradually turn light brown. Inoculation is successful when the cork or bark cover is covered with mycelium.

(5) Covering soil for planting: acid soil with loose soil and pH value of 5-6 and places convenient for irrigation and drainage are selected as the planting sites. Dig the soil to a depth of 25cm, remove weeds and stones, and make a frame with a width of 1.5 ~ 1.8 cm after exposure. The length of the boundary is determined according to the actual situation. Generally, it runs in the north-south direction, with drainage ditches around it and ant-killing drugs scattered. The site needs to be used for 2 ~ 3 years, and the boundary should be covered with plastic shed, which requires heat preservation, moisture preservation, ventilation and shade. When the average temperature of the day is stable at 20℃, the good hyphae are buried in the soil for culture. Digging the prepared border, spreading a layer of loose soil at the bottom of the ditch, standing the inoculated end of wood with good hyphae in the ditch about 6cm, covering the soil 1 ~ 2 cm, covering the chaff with a thickness of about 1cm to prevent the soil from splashing on the fruiting body when spraying water. Spray water after burying 1 time. If the weather is dry, spray water to moisten the soil.

(6) Lei Zhi management: Lei Zhi can appear 10 ~ 15 days after burial. Pay attention to drainage in rainy days. In addition, a circle of toxic soil mixed with mirex should be scattered around the breeding field to trap and kill termites. Measures such as spraying water, ventilation, shading and heat preservation were taken to control the temperature in the shed at 24 ~ 28℃, the relative humidity at 85% ~ 90%, the light intensity at 300 ~ 1000 LX, the air was fresh and the soil was loose and moist.

(3) Pests and diseases and their control

Most of them are infected by Penicillium, Mucor and Rhizopus.

Prevention and treatment methods: ① Strict aseptic operation during inoculation; ② Thoroughly disinfect the culture; ③ Reduce humidity through proper ventilation; (4) Scrape off the miscellaneous bacteria and surrounding bark with a charred blade, and then apply thick lime milk for prevention and control, or fill the holes with absorbent cotton dipped in 75% ethanol solution, so as to eliminate serious polluters in time.

Third, harvest.

1. Timely harvest

It takes 40 ~ 50 days from the appearance of mycelium to the stop of mycelium growth. The color of mycelium has changed from light yellow to reddish brown, the color of cap handle is consistent, from soft to hard, spore powder is ejected, and mycelium is mature. You can harvest at this time.

2. Harvest method

Cut Ganoderma lucidum from the stem base with a sharp knife or branch scissors, leaving a stem pedicle of 0.5 ~ 1 cm. After the incision is healed, it will form a cap primordium and a new fruiting body. According to the different volume of wood, it can be cut continuously for 2 ~ 3 years. For example, if spore powder is collected by bottle culture or bag culture, the fruiting body differentiation can last 1 month. The collection method is to cover the fungus cover with a paper bag and collect.