WO2017219394A1 - 食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法 - Google Patents

食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法 Download PDF

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WO2017219394A1
WO2017219394A1 PCT/CN2016/089629 CN2016089629W WO2017219394A1 WO 2017219394 A1 WO2017219394 A1 WO 2017219394A1 CN 2016089629 W CN2016089629 W CN 2016089629W WO 2017219394 A1 WO2017219394 A1 WO 2017219394A1
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tremella
cultivar
edible fungus
production method
seed production
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PCT/CN2016/089629
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English (en)
French (fr)
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熊璐
顾世
肖奎
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四川省大真科技有限责任公司
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Publication of WO2017219394A1 publication Critical patent/WO2017219394A1/zh

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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
    • A01G18/50—Inoculation of spawn
    • 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
    • A01G18/20—Culture media, e.g. compost

Definitions

  • the invention relates to the technical field of edible fungi cultivation, in particular to a seed production method of edible fungus cultivars, a cultivar prepared thereby and a cultivation method of edible fungi.
  • Edible fungi are large fungi that are edible by humans and have fleshy or gummy fruiting bodies. China's edible fungus resources are very rich. In 2000, there were 938 edible fungi in China and more than 50 artificially cultivated species.
  • the edible part of the edible fungus is a fruiting body with sporulation structure, and the fruit protein content of the fruiting body is about 3-4% of fresh weight or 20-40% of dry weight, and the amino acid type is complete, and there are more nucleic acids and Various vitamins, including vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin PP (nicotinic acid), vitamin C (ascorbic acid) and vitamin D (ergosterol).
  • the mineral content is also rich, especially the phosphorus content is more, which is beneficial to the regulation of various physiological functions of the human body.
  • Tremella, fungus, and monkey head also have a variety of specific nourishing or medical uses.
  • Tremella or golden ear is a typical representative of such edible fungi.
  • Tremella or Golden Ear has a strict matching relationship with its accompanying bacteria, and the completion of its life history is inseparable from the synergistic effect of the accompanying bacteria.
  • the production of edible fungi begins with the mother species. After the original seed is expanded and the ear shape is observed, the transplanted expanded culture is selected as a cultivar. After a period of cultivation, the selective cultivar is excellent. The species that the average grower buys from the market is the cultivar.
  • the current production of edible fungi cultivars is mostly: the medium is compacted and the holes required for the original species are reserved at the center. The culture medium is sterilized and then inserted into the original species under aseptic conditions, and the cultivar can be obtained after a period of cultivation.
  • the seed production method of the above cultivars is convenient and easy, in edible fungi It is widely used in the preparation of cultivars. However, such a method has certain defects for edible fungi that need to cooperate with the accompanying bacteria to complete the life history.
  • Tremella although the mycelial growth range of its associated bacteria (Saccharomyces cerevisiae) can spread over the entire bacterial bag, the growth extension of the hyphae of the white fungus can only extend outward by about 2 cm at the inoculation point, and usually the bacteria bag or mushroom The diameter of the bottle is not less than 8 cm; this causes the hyphae of the accompanying bacteria to grow only on the side away from the center of the bag, but there is no hyphae or tallow hyphae.
  • inoculation is usually carried out by two methods, one is inoculation with clams, because it is not easy to distinguish between the fungus hyphae, the golden ear hyphae and their associated hyphae, which is prone to cavitation, ie There is only associated hyphae in the inoculation hole; one is to open the wood chips completely through a funnel-like tool, which is easy to cause hyphae damage, reduce the activity, and it is easy to appear only a small amount of white fungus in the inoculation hole or The phenomenon of hyphae.
  • the technical problem to be solved by the present invention is to provide a method for seeding edible mushroom cultivars, a cultivar prepared thereby, and a method for cultivating edible fungi.
  • the seed production method provided by the present invention can increase the yield of edible fungi.
  • the method for seeding the edible fungus cultivar provided by the present invention comprises: mixing the original seed, mixing with the culture medium, compacting, and then cultivating the edible fungus cultivar.
  • the seed production method of the cultivar provided by the present invention does not inoculate the original seed at the center of the culture medium, but pulverizes the original seed and uniformly mixes with the culture medium. Thereby, the hyphae of the edible fungi in the obtained cultivar can be uniformly distributed. In the past, it was considered that pulverizing the original species would destroy the integrity of the hyphae and make the growth of the hyphae in the cultured species poor.
  • the pulverized seed is the original species in which the ear group has appeared; the pulverization is specifically to break up the original seed.
  • the edible fungus is white fungus or golden ear.
  • Tremella fuciformis belongs to Tremellales, Termelaceae, Tremell. It is a moderate-temperature and aerobic fungus. The optimum temperature for growth of Tremella mycelium and fruiting bodies is 22-26 °C. The optimum pH value is Tremella fuciformis. 5.2 ⁇ 5.8, under the light conditions of 50 ⁇ 600Lx, the growth of the tremella fruit body is good. The completion of the life history of Tremella can not be separated from the synergy of the bacterium.
  • Tremella aurantialba Bandoni et Zang the same genus as Tremella, also known as yellow fungus, golden white fungus, yellow ear or brain ear.
  • the growth of Tremella mycelium is suitable for 23 ⁇ 25°C, the fruit body is 15 ⁇ 20°C, and the optimum pH is 5.8 ⁇ 6.2.
  • the hyphae growth of the golden ear does not require light, and the fruit body formation must have light induction.
  • the development of fruiting bodies requires adequate ventilation.
  • the poorly ventilated fruiting bodies are dull in color. Appropriate ventilation and sufficient oxygen can make the golden ear form natural orange and orange red pigment.
  • the natural growth and occurrence of the golden ear is inseparable from the fungus of the ear fungus.
  • the method for preparing cultivars provided by the present invention is suitable for common edible fungi. It is especially suitable for edible fungi that require associated bacteria such as white fungus and golden ear.
  • the method for seeding the Tremella cultivar is as follows: the Tremella species are pulverized, mixed with the culture medium, compacted, and cultured to obtain Tremella cultivars.
  • the original species is excessively pulverized, which would damage the hyphae excessively, which would reduce the activity of the original species. If the pulverization was insufficient, it could not be mixed well with the medium, and the distribution of hyphae in the obtained cultivar was uneven. .
  • the original species is pulverized to a diameter of 2 mm to 6 mm.
  • the culture medium provides a nitrogen source and a carbon source for the edible fungus, and the medium for cultivating the original species and the cultivar of the present invention is identical.
  • the medium includes 78 parts by mass of wood chips, 20 parts by mass of wheat bran, 1 part by mass of sucrose, and 1 part by mass of gypsum.
  • the sawdust is Cyclobalanopsis sinensis, and the sawdust is cut down within six months. Experiments have shown that freshly harvested sawdust has a better effect than long-term storage or fermentation of wood chips, and can improve the germination rate of the original or cultivar of Tremella.
  • the pH in the medium is 5.2 to 5.8.
  • the water content in the medium was 55 wt%.
  • the mass ratio of the original seed to the medium determines whether the original seed can be evenly mixed with the medium, and is heavier. It is only necessary that the hyphae can be evenly distributed in the culture species within a certain culture period at a suitable mass ratio. A too low mass ratio causes the hyphae to decay before it is filled with the medium, and an excessively high mass ratio does not further increase the growth of the hyphae in the medium, which in turn causes waste of resources.
  • the mass ratio of the original species to the medium is (50 to 150): (800 to 1000).
  • the mass ratio of stock to medium is 50:1000.
  • the mass ratio of stock to medium is 150:800.
  • the mass ratio of stock to medium is 100:900.
  • the stock of the present invention is prepared by inoculating the mother seed on a sterilized medium and culturing at 24 ° C for 25 days to obtain the original seed.
  • the Tremella species used in the present invention was purchased from the Sanming Fungi Research Institute; the strain number was 9903.
  • the mixed stock and culture medium are cultured under certain conditions, and the culture needs to reach a certain accumulated temperature to be a finished product of the cultivar.
  • the cultivar will grow healthily and rapidly at a suitable culture temperature. Excessive temperature makes the hyphae long, while the bacteria also proliferate rapidly, which tends to cause the hyphae to rot.
  • the too low temperature can not reach the accumulated temperature in a short time, prolonging the production cycle, and the edible fungus with a shorter growth cycle. It will undoubtedly cause waste of resources.
  • the culture temperature is 20 ° C ⁇ 25 ° C, the humidity does not exceed 70%; the culture time is 20 days ⁇ 25 days.
  • the humidity is air humidity.
  • the culture temperature was 22 ° C and the culture time was 25 days.
  • the temperature of the culture is 20 ° C and the culture time is 25 days.
  • the temperature of the culture is 25 ° C and the culture time is 20 days.
  • the woody pieces can be bonded together after 7 days of culture, and after 20 days of cultivation, white hair groups appear on the inner surface of the bacteria bag, and culture is cultivated after 25 days of cultivation.
  • kind of finished product
  • the medium needs to be rigorously sterilized before use to prevent the bacteria carried in the medium from affecting the development of the hyphae.
  • the method of autoclaving is more suitable for production practice.
  • the high temperature is 121 ° C, and the high pressure is 0.105 MPa to 0.15 MPa.
  • the culture medium is sterilized, and the sterilization temperature is 121 ° C and the pressure is 0.105. MPa ⁇ 0.15MPa, time is 2.5h.
  • the original seed and the culture medium can be better mixed by sufficient shaking, and then the mixed original seed and the medium are pressed.
  • the compaction described in the present invention is pressed to a density of from 0.42 g to 0.64 g/cm 3 .
  • the culture in the method provided by the present invention is carried out under aerobic conditions.
  • the culture of the present invention is carried out in a breathable bag.
  • the venting bag is made of polypropylene.
  • the form is an inner folded bag; it has a collar and a cover; the specification is 19cm ⁇ 45cm ⁇ 5s, and the opening length is 32 ⁇ 0.1cm. It has a 4 cm diameter vent.
  • the diameter of the ventilated bag is matched with the inner plug, and the collar is matched with the inner plug.
  • the collar has a flared shape with an inner diameter of 3.8 ⁇ 0.1 cm and a height of 3.2 ⁇ 0.1 cm.
  • Breathable bags, collars and inner plugs are required to be clean, non-destructive, smooth and burr-free.
  • the material of the collar and the inner plug is plastic, which can withstand high temperatures of 130 ° C and high pressure of 2.5 kg.
  • An edible fungus cultivar prepared by the seed production method of the present invention is an edible fungus cultivar prepared by the seed production method of the present invention.
  • the distribution of hyphae in the cultivars provided by the invention is more uniform, and the white fungus is taken as an example.
  • the hyphae of the white fungus are evenly distributed, and the ratio of the hyphae to the accompanying bacteria is suitable, and the cultivar can be used as a cultivar in production.
  • the present invention also provides a method for cultivating an edible fungus, which is inoculated with an edible fungus cultivar prepared by the preparation method of the present invention.
  • the present invention provides a method of cultivating a Tremella cultivar prepared by the method of the present invention.
  • the cultivation method is bagged white fungus or segmental white fungus.
  • the invention provides the effect of improving the ear output rate of the Tremella fuciformis cultivars prepared by the method More significant.
  • inoculation is inoculation or chopping.
  • the method for preparing the Tremella chinensis cultivar comprises: pulverizing the Tremella species and mixing with the medium in a mass ratio of (50-150): (800-1000), and compacting to a density of 0.42 g to 0.64. g/cm 3 , cultured at 20 ° C to 25 ° C for 20 days to 25 days under aerobic conditions to obtain Tremella cultivars.
  • the method for indoor cultivation of Tremella comprises: inoculating the Tremella cultivar prepared by the present invention into clinker or culture medium, and culturing for 50 days; the culture condition is 22 ° C, the humidity is 70%, and the air is ventilated 3 times a day. 30 minutes each time.
  • the clinker is a short section of 10 cm high in the lime water immersion section, and the electric drill is perforated, and the clinker is autoclaved.
  • the outdoor cultivation method of the white fungus provided by the present invention comprises: inocating the Tremella chinensis cultivar prepared by the present invention in the beginning of April and early April, and inoculation into the Cyclobalanopsis glauca L., and cultivating until mid-September.
  • the preparation method of the Cyclobalanopsis glauca L. is: cutting down the Cyclobalanopsis glauca tree, and drying it for 15 days to 25 days, and cutting into a section of 1 m of wood, drilling 2 to 3 cm deep.
  • the cutting time is after the Spring Festival, and the drying is to horror.
  • the invention provides a seed production method of an edible fungus cultivar, a prepared cultivar and an edible fungus cultivation method.
  • test materials, reagents and instruments used in the present invention are all common commercial products, and are commercially available.
  • Tremella species was purchased from Sanming Fungi Research Institute, number 9903.
  • the formula is 78% qinggang shavings, 20% wheat bran, 1% sucrose, 1% gypsum and 55% water content.
  • the Cyclobalanopsis glauca L. is the best tree species of the T. chinensis. The previous tests proved that cutting wood chips in the first half of the year is more conducive to the growth of Tremella compared to long-term storage, or fermenting wood chips.
  • the culture medium was placed in a mushroom bottle, compacted to a material height of 7 to 10 cm, and sealed with a cotton stopper.
  • the purchased mother seeds were picked into a sterilized and cooled medium, and cultured at 24 ° C for 25 days to obtain the original species.
  • the formula is 78% qinggang shavings, 20% wheat bran, 1% sucrose, 1% gypsum and 55% water content.
  • the Cyclobalanopsis glauca L. is the best tree species of the T. chinensis. The previous tests proved that cutting wood chips in the first half of the year is more conducive to the growth of Tremella compared to long-term storage, or fermenting wood chips.
  • the medium is placed in a ventilated bag, which is 0.8 kg to 1 kg per bag. Sterilize at 121 ° C, 0.105 MPa to 0.15 MPa for 2.5 hours, and use after cooling completely.
  • the stock prepared in Example 1 was broken up and dispersed to a diameter of about 2 mm to 6 mm.
  • the dispersed seed was inoculated into a breathable bag containing the medium in an amount of 100 g/bag, mixed, compacted, and compacted to a volume of 2000 cm 3 . Incubate at 22 ° C for 25 days, during which the air humidity does not exceed 70%.
  • Tremella mycelium After 7 days of cultivation, the growth of Tremella mycelium can bond the wood chips together. After 20 days of cultivation, white hair groups appear on the inner surface of the bacteria bag, and after 25 days of cultivation, they become cultivars.
  • the formula is 78% qinggang shavings, 20% wheat bran, 1% sucrose, 1% gypsum and 55% water content.
  • the Cyclobalanopsis glauca L. is the best tree species of the T. chinensis. The previous tests proved that cutting wood chips in the first half of the year is more conducive to the growth of Tremella compared to long-term storage, or fermenting wood chips.
  • the medium is placed in a ventilated bag, which is 0.8 kg to 1 kg per bag. Sterilize at 121 ° C, 0.105 MPa to 0.15 MPa for 2.5 hours, and use after cooling completely.
  • the stock prepared in Example 1 was broken up and dispersed to a diameter of about 2 mm to 6 mm.
  • the dispersed seed was inoculated into a ventilated bag containing the medium in an amount of 50 g/bag, mixed, compacted, and compacted to a volume of 2000 cm 3 . Incubate at 25 ° C for 20 days, during which the air humidity does not exceed 70%.
  • the Tremella mycelium grows to bond the wood chips together, and after 20 days of cultivation, it becomes a cultivar product.
  • the formula is 78% qinggang shavings, 20% wheat bran, 1% sucrose, 1% gypsum and 55% water content.
  • the Cyclobalanopsis glauca L. is the best tree species of the T. chinensis. The previous tests proved that cutting wood chips in the first half of the year is more conducive to the growth of Tremella compared to long-term storage, or fermenting wood chips.
  • the medium is placed in a ventilated bag, which is 0.8 kg to 1 kg per bag. Sterilize at 121 ° C, 0.105 MPa to 0.15 MPa for 2.5 hours, and use after cooling completely.
  • the stock prepared in Example 1 was broken up and dispersed to a diameter of about 2 mm to 6 mm.
  • the dispersed seed was inoculated into a breathable bag containing the medium at a dose of 150 g/bag, mixed, compacted, and compacted to a volume of 2000 cm 3 . Incubate at 20 ° C for 25 days, during which the air humidity does not exceed 70%.
  • the fungus filaments can grow the wood chips together. After 20 days of cultivation, yellow water appears on the inner surface of the bacteria bag, and after 25 days of cultivation, it becomes a cultivar product.
  • the formula is 78% qinggang shavings, 20% wheat bran, 1% sucrose, 1% gypsum and 55% water content.
  • the Cyclobalanopsis glauca L. is the best tree species of the T. chinensis. The previous tests proved that cutting wood chips in the first half of the year is more conducive to the growth of Tremella compared to long-term storage, or fermenting wood chips.
  • the culture medium is placed in a fungus bag, and each bag is filled with 0.8 kg to 1 kg.
  • the inoculation hole is compacted and reserved, and sterilized at 121 ° C, 0.105 MPa to 0.15 MPa for 2.5 hours, and completely cooled and used.
  • the stock pieces prepared in Example 1 were inserted into the inoculation wells in an inoculum amount of 100 g/bag; and cultured at 22 ° C for 25 days, during which the air humidity did not exceed 70%.
  • the fungus filaments can grow the wood chips together. After 20 days of cultivation, yellow water appears on the inner surface of the bacteria bag, and after 25 days of cultivation, it becomes a cultivar product.
  • the high-temperature sterilization clinker that is, the short-section clinker, is a test carrier for the ear white ear rate.
  • the cultivars prepared in Examples 2 to 4 were experimental subjects, and were inoculated with clams and stalks. This method was used to access the inoculation holes of short clinker to observe the ear rate and the indoor test of Tremella traits.
  • the cultivar prepared in Comparative Example 1 was used as a control, and was inserted into the inoculation hole of the short clinker in two ways, namely, inoculation with clams and mashing, to observe the recorded ear rate and the indoor test of the tremella trait.
  • the earing rate of the stalks inoculated with the cultivars of Examples 2 to 4 was increased by 30% compared with the cultivar of the cultivar of the cultivar of the cultivar of Example 2 to 4, and the white fungus grown by the ear sticks of the cultivars of Examples 2 to 4 was inoculated. It is tidy and tidy, and it is obvious that the growth is better than the control.
  • the collected tremella of different numbers were uniformly collected and collected, and the dry weight and the shape were determined. The results showed that the yield of the woods inoculated with the cultivars of Examples 2 to 4 was compared with that of the cultivar 1 The increase was 28.2%, and the first-grade product rate increased by 10.32%.
  • the method of improving the earing rate of the wood by using the cultivar seedlings of the Phyllostachys pubescens species is a highly efficient, stable and promising seed production technique, which can be applied to the cultivation of the wood fungus.
  • the application of this method is not limited to a certain strain, but can be applied to all Tremella strains suitable for planting on wood, providing technical support for the development of high quality segmental fungus industry. hold.
  • the application of the method can reduce the porosity of the ear stick, reduce the cost of strains and ear wood; improve the ease of use of growers; increase the economic value of production, promote the inheritance of traditional crafts and ensure the local economic development. Significance.

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Abstract

一种食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法。该制种方法制备的栽培种中菌丝的分布更加均匀。以银耳为例，其栽培种中的菌丝分布均匀，与伴生菌的比例适宜，以其作为生产中的栽培种，接种后能显著提高出耳率，减少耳棒空穴的现象，提高耳棒的生物转化率，增加种植户的收入。

Description

食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法
本申请要求于2016年06月24日提交中国专利局、申请号为201610478902.5、发明名称为“食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。
技术领域
本发明涉及食用菌栽培技术领域，尤其涉及食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法。
背景技术
食用菌是可供人类食用的，具有肉质或胶质子实体的大型真菌。中国食用菌资源十分丰富，2000年统计中国的食用菌达938种，人工栽培的50余种。食用菌可食用部分为具有产孢结构的子实体，其子实体的蛋白质含量约为鲜重的3～4％或干重的20～40％，含氨基酸种类齐全，还有较多的核酸和各种维生素，包括维生素B1(硫胺素)、维生素B2(核黄素)、维生素PP(烟酸)、维生素C(抗坏血酸)和维生素D元(麦角甾醇)等。矿物质的含量亦较丰富，尤其含磷质较多，有利于人体各项生理机能的调节，此外，银耳、木耳、猴头等还兼有多种特定的滋补或医疗用途。
食用菌的生产栽培中有一类必须与伴生真菌协同完成其生活史，其中，银耳或金耳为这类食用菌的典型代表。在生产中，银耳或金耳与其伴生菌有着严格的配对关系，其生活史的完成离不开伴生菌的协同作用。
食用菌的生产从母种开始，经过原种扩培和观察出耳形态后，选择生长优良的转接扩培成为栽培种，经过一段时间培养后，选择性状优良的成为成品栽培种。一般种植户从市场上购买的菌种即为栽培种。对于栽培种的制种方法，目前食用菌栽培种的制种多为：将培养基压实并在中心预留转接原种需要的孔洞。培养基经过灭菌后无菌条件下接入原种，经过一段时间的培养即可获得栽培种。上述栽培种的制种方法方便易行，在食用菌 栽培种的制备中广泛使用。但这样的方法对于需与伴生菌协同完成生活史的食用菌而言，却存在着一定的缺陷。
以银耳为例，虽然其伴生菌(香灰菌)的菌丝生长范围可以遍布整个菌袋，但是银耳的菌丝的生长延伸范围只能在接种点向外延伸2cm左右，而通常菌袋或蘑菇瓶直径为不低于8cm；这就导致了菌袋中远离中心的一侧仅生长了伴生菌的菌丝，却没有生长银耳菌丝或金耳菌丝。而对于食用菌而言，接种通常采用两种方法进行，一种是掰块接种，因不能很好的区别银耳菌丝、金耳菌丝与它们的伴生菌丝，极易出现空穴，即接种孔内只有伴生菌丝；一种是通过将木屑完全糅细后通过一个形似漏斗的工具敞栽，极易造成菌丝损伤，活性降低，同时极易出现接种孔内只是极少量的银耳或金耳菌丝的现象。如此之后，段木的出耳率将比预期的降低许多，即浪费了生产材料，也辜负了种植户的一番努力，阻碍段木银耳、段木金耳的产业发展，依然不能满足市场对高品质银耳的需求。当然种植户中也不乏有知悉银耳或金耳生物学特征的，那么他在使用市场上现有栽培种时，将扔弃掉部分只含纯伴生菌丝的木屑，成本明显增加。
因此，进一步改进食用菌栽培种的制种方式，以提高食用菌的品质和产量，减少资源的浪费十分必要。
发明内容
有鉴于此，本发明要解决的技术问题在于提供一种食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法。本发明提供的制种方法能够提高食用菌的产量。
本发明提供的食用菌栽培种的制种方法为：原种粉碎后与培养基混合、压实，然后培养获得食用菌栽培种。
本发明提供的栽培种的制种方法，不再在培养基的中心接种原种，而是将原种粉碎后与培养基均匀的混合。从而使所得栽培种中食用菌的菌丝能够均匀的分布。以往认为，粉碎原种会破坏菌丝的完整性，使培养种中菌丝的生长状况不良。本发明提供的方法中，粉碎原种采用的是已经出现耳基的原种；粉碎具体为将原种打散。
本发明中，食用菌为银耳或金耳。
银耳(Tremellafuciformis)属银耳目(Tremellales)，银耳科(Termelaceae)，银耳属(Tremell)，属中温、好气性真菌，银耳菌丝和子实体最适温度生长为22～26℃，最适pH值为5.2～5.8，在50～600Lx的光照条件下银耳子实体生长发育良好。银耳的生活史的完成离不开香灰菌的协同作用。
金耳(Tremella aurantialba Bandoni et Zang)，与银耳同属，又称黄木耳、金黄银耳、黄耳或脑耳。金耳菌丝体生长适温23～25℃，子实体为15～20℃，最适pH值为5.8～6.2，金耳的菌丝生长不需要光，子实体形成则必须有光诱导。子实体发育需要足够的通风，通风不良子实体色泽暗淡，适当的通风，充足的氧气才能使金耳形成自然的橙黄和橙红色素。金耳的自然生长和发生，都离不开其耳友菌粗毛硬革菌。
本发明提供的制备栽培种的方法适用于常见食用菌。特别适用于银耳、金耳等需要伴生菌的食用菌。
一些具体的实施例中，银耳栽培种的制种方法为：将银耳原种粉碎后与培养基混合、压实，经培养获得银耳栽培种。
以银耳为实验材料，证明原种粉碎过度，会过度损伤菌丝，使得原种的活性降低；而如果粉碎不足，则不能与培养基很好的混合，所得栽培种中菌丝的分布不均匀。本发明中，原种粉碎的至直径为2mm～6mm。
培养基为食用菌提供氮源和碳源，本发明培养原种和栽培种的培养基一致。
以银耳为例，培养基中包括木屑78质量份，麦麸20质量份，蔗糖1质量份，石膏1质量份。
所述木屑为青冈树木屑，且为半年内砍伐粉碎的木屑。实验表明，新鲜砍伐的木屑与长久储存或发酵的木屑相比，具有更好的效果，能够提高银耳的原种或栽培种的萌发率。
所述培养基中的pH值为5.2～5.8。
所述培养基中的含水量为55wt％。
原种与培养基的质量比决定原种是否能够与培养基混合均匀，更重 要的是只有在适宜的质量比下，在一定的培养时期内，菌丝才能够均匀的分布在培养种中。过低的质量比导致菌丝在布满培养基前就开始衰败，而过高的质量比也并不会进一步的提高菌丝在培养基中的生长状况，反而会造成资源的浪费。
以银耳为例，原种与培养基的质量比为(50～150)：(800～1000)。
一些实施例中，原种与培养基的质量比为50:1000。
一些实施例中，原种与培养基的质量比为150：800。
一些实施例中，原种与培养基的质量比为100:900。
本发明原种的制备方法为，将母种接种到经灭菌的培养基上，24℃培养25天，获得原种。
本发明采用的银耳母种购自三明真菌研究所；菌株编号为9903。
混合后的原种和培养基在一定条件下培养，培养需达到一定的积温才能够成为栽培种成品。栽培种在适宜的培养温度下，会健康快速的生长。过高的温度使菌丝徒长的同时，杂菌也迅速的繁殖，容易导致菌丝腐烂；而过低的温度在短时间内无法达到积温，延长了生产周期，对于生长周期较短的食用菌，无疑会造成资源的浪费。
以银耳为例，培养的温度为20℃～25℃，湿度不超过70％；培养的时间为20天～25天。
所述湿度为空气湿度。
一些实施例中，培养的温度为22℃，培养的时间为25天。
一些实施例中，培养的温度为20℃，培养的时间为25天。
一些实施例中，培养的温度为25℃，培养的时间为20天。
在本发明提供的培养条件下，经过7天培养后银耳菌丝生长可将木屑块粘结在一起，经过20天培养后可见菌袋内表面有白毛团出现，经过25天培养后成为栽培种成品。
培养基在使用前需要经过严格的灭菌，以避免培养基中所携带的杂菌影响使用菌丝的发育。高温高压灭菌的方式更适用于生产实践。所述高温为121℃，所述高压为0.105MPa～0.15MPa，。
以银耳为例，培养基经灭菌，所述灭菌的温度为121℃、压强为0.105 MPa～0.15MPa，，时间为2.5h。
对于好气性的真菌，不透气的材料不利于其菌丝的生长。以往的制种方法中，采用菌袋或玻璃瓶培养栽培种，透气性不佳，也是导致栽培种中菌丝分布不均的重要原因。并且，常规规制种培养是将接种块放入接种孔后，直接盖上盖子或系好套绳，放入适宜温度环境中即完成，其制成的菌丝无法均匀的分布在全部培养基中。本发明在制作栽培袋时未对基料进行压块，料较松散，在接种后通过充分的震荡可较好将原种与培养基混匀，再将混合好的原种与培养基的压平在透气袋无透气无纺布的一面，形成一个形似方形的木屑块，保证适宜的温度和充足的氧气。
本发明中所述的压实为压至密度为0.42g～0.64g/cm3。
本发明提供的方法中培养在有氧条件下进行。
本发明所述培养在透气袋中进行。
所述透气袋的材质为聚丙烯。形式为内折袋；具有套环和盖子；规格为19cm×45cm×5s，撑开长度32±0.1cm。其具有4cm直径透气孔。
透气袋直径与内塞配套，套环与内塞配套。套环呈喇叭形，上口内径3.8±0.1cm，高度3.2±0.1cm。
透气袋、套环、内塞皆要求干净、无破损，圆滑无毛刺。
套环与内塞的材质为塑料，能够耐130℃高温、2.5kg高压。
以本发明所述制种方法制备的食用菌栽培种。
本发明提供的栽培种中菌丝的分布更加均匀，以银耳为例，其栽培种中，银耳的菌丝分布均匀，与伴生菌的比例适宜，以其作为生产中的栽培种，接种后能显著提高出耳率，减少耳棒空穴的现象，提高耳棒的生物转化率，增加种植户的收入。
本发明还提供了一种食用菌栽培方法，其接种以本发明所述制备方法制备的食用菌栽培种。
以银耳为例，本发明提供的栽培方法为，接种以本发明提供方法制备的银耳栽培种。
栽培的方式为袋装银耳或段木银耳。
本发明提供方法制备的银耳栽培种对段木银耳的出耳率提高效果 更加显著。
所述接种的方式为掰块接种或揉碎敞栽。
具体的，本发明提供的制备银耳栽培种的方法为：将银耳原种粉碎后与培养基以质量比为(50～150)：(800～1000)混合、压实至密度为0.42g～0.64g/cm3，有氧条件下，20℃～25℃培养20天～25天，获得银耳栽培种。
本发明提供的银耳的室内栽培方法包括：将本发明制备的银耳栽培种接种于熟料或培养基，培养50天；所述培养的条件为22℃，湿度为70％，每天通风3次，每次30分钟。
优选的，培养20天后用棉沙布保湿，每天浇水3～5次。
所述熟料为经过石灰水浸泡的段高10cm、电钻打孔的短段，高压灭菌的熟料。
本发明提供的银耳的室外栽培方法包括：3月初4月初将本发明制备的银耳栽培种接种于青冈树段木，培养至9月中下旬。
所述青冈树段木的制备方法为：砍伐青冈树，晾晒15天～25天后截成1米的段木，钻孔2～3厘米深。
所述砍伐的时间为春节后，所述晾晒至惊蛰。
本发明提供的食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法，本发明提供方法制备的栽培种中菌丝的分布更加均匀，以银耳为例，其栽培种中，银耳的菌丝分布均匀，与伴生菌的比例适宜，以其作为生产中的栽培种，接种后能显著提高出耳率，减少耳棒空穴的现象，提高耳棒的生物转化率，增加种植户的收入。
具体实施方式
本发明提供了食用菌栽培种的制种方法及所制备的栽培种和食用菌栽培方法。本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内 对本文的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
本发明采用的试材、试剂、仪器皆为普通市售品，皆可于市场购得。
其中，银耳母种购自三明真菌研究所，编号为9903。
下面结合实施例，进一步阐述本发明：
实施例1原种的制备
1、配制培养基：
配方为青冈树木屑78％，麦麸20％，蔗糖1％，石膏1％，含水量55％。
青冈树为段木银耳最佳树种，前期试验证明半年内砍伐粉碎的木屑相对于长久储存，或发酵木屑更有利于银耳的生长。
培养基装入蘑菇瓶，压实至料高7～10cm，用棉花塞封口。
121℃，高压灭菌2.5小时，完全冷却后使用。
2、接种：
将购买的母种夹取一块放入灭菌、冷却后的培养基，24℃，培养25天后，获得原种。
实施例2栽培种的制备
1、配制培养基：
配方为青冈树木屑78％，麦麸20％，蔗糖1％，石膏1％，含水量55％。
青冈树为段木银耳最佳树种，前期试验证明半年内砍伐粉碎的木屑相对于长久储存，或发酵木屑更有利于银耳的生长。
培养基放入透气袋，每袋装0.8kg～1kg。121℃，0.105MPa～0.15MPa灭菌2.5小时，完全冷却后使用。
2、接种：
将实施例1制备的原种打散，打散至直径约2mm～6mm。
将打散的原种以100g/袋的量接种入装有培养基的透气袋，混匀后压 实，压实至体积为2000cm3。22℃培养25天，培养期间，空气湿度不超过70％。
经过7天培养后银耳菌丝生长可将木屑块粘结在一起，经过20天培养后可见菌袋内表面有白毛团出现，经过25天培养后成为栽培种成品。
实施例3栽培种的制备
1、配制培养基：
配方为青冈树木屑78％，麦麸20％，蔗糖1％，石膏1％，含水量55％。
青冈树为段木银耳最佳树种，前期试验证明半年内砍伐粉碎的木屑相对于长久储存，或发酵木屑更有利于银耳的生长。
培养基放入透气袋，每袋装0.8kg～1kg。121℃，0.105MPa～0.15MPa灭菌2.5小时，完全冷却后使用。
2、接种：
将实施例1制备的原种打散，打散至直径约2mm～6mm。
将打散的原种以50g/袋的量接种入装有培养基的透气袋，混匀后压实，压实至体积为2000cm3。25℃培养20天，培养期间，空气湿度不超过70％。
经过7天培养后银耳菌丝生长可将木屑块粘结在一起，经过20天培养成为栽培种成品。
实施例4栽培种的制备
1、配制培养基：
配方为青冈树木屑78％，麦麸20％，蔗糖1％，石膏1％，含水量55％。
青冈树为段木银耳最佳树种，前期试验证明半年内砍伐粉碎的木屑相对于长久储存，或发酵木屑更有利于银耳的生长。
培养基放入透气袋，每袋装0.8kg～1kg。121℃，0.105MPa～0.15MPa灭菌2.5小时，完全冷却后使用。
2、接种：
将实施例1制备的原种打散，打散至直径约2mm～6mm。
将打散的原种以150g/袋的量接种入装有培养基的透气袋，混匀后压实，压实至体积为2000cm3。20℃培养25天，培养期间，空气湿度不超过70％。
经过7天培养后银耳菌丝生长可将木屑块粘结在一起，经过20天培养后可见菌袋内表面有黄水出现，经过25天培养后成为栽培种成品。
对比例1栽培种的制备
1、配制培养基：
配方为青冈树木屑78％，麦麸20％，蔗糖1％，石膏1％，含水量55％。
青冈树为段木银耳最佳树种，前期试验证明半年内砍伐粉碎的木屑相对于长久储存，或发酵木屑更有利于银耳的生长。
培养基放入菌袋，每袋装0.8kg～1kg，压实并预留接种孔，121℃，0.105MPa～0.15MPa灭菌2.5小时，完全冷却后使用。
2、接种：
将实施例1制备的原种掰块接入接种孔，接种量为100g/袋；22℃培养25天，培养期间，空气湿度不超过70％。
经过7天培养后银耳菌丝生长可将木屑块粘结在一起，经过20天培养后可见菌袋内表面有黄水出现，经过25天培养后成为栽培种成品。
实施例5室内测试出耳率
主要以将青冈段木截断到更适宜大小，通过高压灭菌，既保留了青冈段木的有效成分，模拟自然生长；又缩短了段木出耳的生长周期，对验证新品种和新工艺的开发效果，具有快速有效的特点。以经过石灰水浸泡的段高10cm、电钻打孔的短段，高压灭菌的熟料，即短段熟料为银耳出耳率测试载体。
实施例2～4制备的栽培种为实验对象，以掰块接种和的糅细敞栽两 种方式接入短段熟料的接种孔中，以观察记录出耳率，和银耳性状的室内测试。
对比例1制备的栽培种为对照，以掰块接种和糅细敞栽两种方式接入短段熟料的接种孔中，以观察记录出耳率，和银耳性状的室内测试。
以上每个处理3个重复(即3袋栽培种)，每袋点种20个短段熟料。温室培养条件为恒温22℃，湿度为70％，每天通风3次，每次30分钟。培养20天后将熟料短段的塑料袋口打开，用棉沙布盖上袋口保湿，每天喷水3-5次。培养30天及30天起每5天观察记录出耳、污染情况，和采摘鲜耳称重至培养50天烘干测产量，并通过对比分析不同处理银耳的生长指标来评估其提高出耳率的能力和在短段熟料中的生长情况。结果如表1：
表1银耳的出耳情况
Figure PCTCN2016089629-appb-000001
结果表明(表1)，不论采用哪种接种方法，熟料短段在接种不同栽培种后出耳率差异显著。接种实施例2～4的栽培种的熟料短段的出耳率比对比例1制备的栽培种的出耳率增加30％以上，空穴率降低20％以上， 产量提高58.41％以上，该效果具有显著性(p<0.05)。因此，接种发明所述的栽培种能明显改善熟料短段的出耳效果，提高出耳率，降低污染率和空穴率，提高产量。
实施例6室外测试出耳率
以通江段木银耳主产区陈河乡砍伐的段木为出耳率室外测试。
以实施例2～4的栽培种和对比例1的栽培种为实验对象，接种方法为掰块接种。在陈河乡按照当地农事安排，春节后砍伐青冈树，晾晒20天左右，待惊蛰节气时截成1米的均匀段木。运回生产产地后钻孔2-3里面深，待接种后发菌40-60天，根据当年气候条件决定，5月底6月初进入事先准备好的出耳耳堂中进行出耳管理；根据银耳生长状况适时喷水防虫。9月中下旬，统计出耳情况。以上每个处理3个重复(即3袋栽培种)，每袋点种100斤耳棒。结果如表2：
表2银耳的出耳情况
生长指标(个) 出耳率 空穴率 一级品率 产量合计(g)
实施例2 70％ 30％ 80％ 256.4
实施例3 65％ 35％ 77％ 250.11
实施例4 70％ 30％ 80％ 255.10
对比例1 40％ 60％ 69％ 200.00
接种实施例2～4栽培种的段木的出耳率比块栽接种对比例1栽培种增加了30％，且接种实施例2～4栽培种的耳棒生长出的银耳，朵大呈菊花状而整齐，且明显看出长势好于对照。将采集的不同茬数的银耳烘干后统一收集齐，测定干重以及朵型，结果表明：接种实施例2～4栽培种的段木的产量与块栽接种对比例1栽培种相比，增加了28.2％，一级品率增加了10.32％。
上述结果表明：用改变段木银耳菌种栽培种制种方式，提高段木出耳率的方法是一种高效、稳定且应用前景很好的菌种制种技术，可应用于段木银耳栽培中。该种方法的应用不局限于某个菌株，而可以应用于所有适于种植在段木上的银耳菌株，为高质量的段木银耳产业发展提供技术支 持。同时，该方法的应用可减少耳棒的空穴率，降低菌种和耳木成本；提高种植户的易用性；提高产量带来经济价值，对促进传统工艺的传承以及保障当地经济发展有重要意义。
以上仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (10)

  1. 一种食用菌栽培种的制种方法，其特征在于，原种粉碎后与培养基混合、压实，然后培养获得食用菌栽培种。
  2. 根据权利要求1所述的制种方法，其特征在于，所述食用菌为银耳或金耳。
  3. 根据权利要求1所述的制种方法，其特征在于，所述培养基中包括木屑78质量份，麦麸20质量份，蔗糖1质量份，石膏1质量份。
  4. 根据权利要求1所述的制种方法，其特征在于，所述原种与培养基的质量比为(50～150)：(800～1000)。
  5. 根据权利要求1所述的制种方法，其特征在于，所述培养的温度为20℃～25℃，湿度不超过70％；培养的时间为20天～25天。
  6. 根据权利要求1所述的制种方法，其特征在于，所述培养的温度为22℃，培养的时间为25天。
  7. 根据权利要求1所述的制种方法，其特征在于，所述培养基经灭菌，所述灭菌的温度为121℃、压强为0.105MPa～0.15MPa，时间为2.5h。
  8. 根据权利要求1所述的制种方法，其特征在于，所述培养在有氧条件下进行。
  9. 权利要求1～8任一项所述制种方法制备的食用菌栽培种。
  10. 一种食用菌栽培方法，其特征在于，接种权利要求9所述的食用菌栽培种。
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CN112205242A (zh) * 2020-11-19 2021-01-12 中华全国供销合作总社昆明食用菌研究所 一种金耳液体菌种接耳栽培方法
CN112243798A (zh) * 2020-10-22 2021-01-22 山东同康农业开发有限公司 一种节能高效银耳菌株的选育及种植方法
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CN114711093A (zh) * 2022-03-30 2022-07-08 贵州省生物研究所 一种冬小包脚菇原种培养基及培养方法
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CN114982555A (zh) * 2022-07-07 2022-09-02 贵州美味鲜竹荪产业有限公司 适用于红托竹荪栽培种的培养基及使用生产方法

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CN110326485B (zh) * 2019-05-23 2023-07-07 福建农林大学 一种栽培血红栓菌栽培方法及应用
CN112042465A (zh) * 2020-09-10 2020-12-08 云南菌视界生物科技有限公司 一种通过金耳子实体与外源毛韧革菌组合选育金耳新品种的方法
CN112243798A (zh) * 2020-10-22 2021-01-22 山东同康农业开发有限公司 一种节能高效银耳菌株的选育及种植方法
CN112205242A (zh) * 2020-11-19 2021-01-12 中华全国供销合作总社昆明食用菌研究所 一种金耳液体菌种接耳栽培方法
CN112602531A (zh) * 2020-12-02 2021-04-06 镇江市菇满园生态农业有限公司 一种袋栽猴头菇大棚全智能化控制栽培的方法
CN113040002A (zh) * 2021-04-02 2021-06-29 甘肃省科学院生物研究所 一种羊肚菌菌种的栽培方法
CN114711093A (zh) * 2022-03-30 2022-07-08 贵州省生物研究所 一种冬小包脚菇原种培养基及培养方法
CN114847085A (zh) * 2022-05-26 2022-08-05 郴州市芝草农业科技开发有限公司 一种食用菌的无菌接种装置及方法
CN114847085B (zh) * 2022-05-26 2023-07-25 郴州市芝草农业科技开发有限公司 一种食用菌的无菌接种装置及方法
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