CN103947454A - Artificial culturing method of lepista sordida mycelium and culturing medium thereof - Google Patents

Artificial culturing method of lepista sordida mycelium and culturing medium thereof Download PDF

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CN103947454A
CN103947454A CN201410192113.6A CN201410192113A CN103947454A CN 103947454 A CN103947454 A CN 103947454A CN 201410192113 A CN201410192113 A CN 201410192113A CN 103947454 A CN103947454 A CN 103947454A
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medium
mycelium
culture medium
glucose
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刘宇
王建瑞
姜德凯
图力古尔
程显好
董洪新
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Ludong University
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Abstract

本发明公开了一种花脸香蘑菌丝体的人工培养方法及其培养基,培养基配方组分按重量份为:松针30-50份、蛋白胨5-12份、酵母浸粉6-12份、葡萄糖2.5-10份、琼脂15-20份;其人工培养方法为:将松针清洗放入水中煮沸20-30分钟,过滤取汁;将松汁定容至1000ml,然后依次加入蛋白胨、酵母浸粉、葡萄糖、琼脂,煮沸混匀;将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟;后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种;将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管后冷藏,本发明操作简单,缩短培养周期短,无污染无废弃物,菌丝体产量高。 The invention discloses a method for artificially cultivating the mycelium of the variegated mushroom and its culture medium. The formula components of the culture medium are: 30-50 parts of pine needles, 5-12 parts of peptone, and 6-12 parts of yeast extract powder , 2.5-10 parts of glucose, 15-20 parts of agar; the artificial culture method is: wash the pine needles, put them in water and boil for 20-30 minutes, filter the juice; dilute the pine juice to 1000ml, then add peptone, yeast extract powder, glucose, agar, boil and mix well; divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes; finally take out the medium and let it cool naturally to 20 ℃-30℃, make a slant culture medium, insert the purified mushroom strains under aseptic conditions; put the inoculated culture medium in a constant temperature incubator, and cultivate it at 20-30℃ for 5-10 The test tube is refrigerated until the mycelium is overgrown. The invention has the advantages of simple operation, shortened culture period, no pollution and waste, and high mycelium output.

Description

一种花脸香蘑菌丝体的人工培养方法及其培养基A kind of artificial cultivation method and culture medium of the mushroom mycelia

  the

技术领域: Technical field:

本发明涉及一种花脸香蘑菌丝体的人工培养方法及其培养基,属于属于生药学技术领域。  The invention relates to a method for artificially cultivating the mycelia of A. variegata and its culture medium, and belongs to the technical field of pharmacognosy. the

背景技术:     Background technique:

花脸香蘑属于真菌界 Fungi,担子菌门 Basidiomycota,伞菌纲Agaricomycetes、伞菌目Agaricales、口蘑科Tricholomataceae、香蘑属Lepista的大型真菌。其鲜嫩甜美, 香味浓郁,口感极好,素有“一家食其味, 十家闻其香”的美誉。在欧洲,花脸香蘑的受欢迎程度能够与松茸和牛肝菌齐名, 在法国被视为上等的食材,具有很高的食用价值。 Flower face mushroom belongs to Fungi Kingdom Fungi, Basidiomycota Basidiomycota, Agaricomycetes, Agaricales Agaricales, Tricholomataceae Tricholomataceae, Lepista large fungi. It is tender and sweet, with strong fragrance and excellent taste. It is known as "one family eats its taste, and ten families smell its fragrance". In Europe, mushrooms are as popular as matsutake and porcini mushrooms. In France, they are regarded as high-quality ingredients with high food value.

此外,花脸香蘑不仅味道鲜美, 而且含有许多人体必需的维生素和氨基酸以及丰富的蛋白质,例如苏氨酸、丙氨酸、谷氨酰胺、丝氨酸、谷氨酸、缬氨酸、亮氨酸、天冬酰胺酸、天门冬氨酸和甘氨酸[8]。此外花脸香蘑的子实体还含有甘露醇、阿拉伯糖醇、葡萄糖、甘油、果糖和海藻糖,其中海藻糖的含量是最高的。 In addition, the flower face mushroom is not only delicious, but also contains many essential vitamins and amino acids and rich protein, such as threonine, alanine, glutamine, serine, glutamic acid, valine, leucine, Asparagine, aspartic acid and glycine [8] . In addition, the fruiting body of the mushroom also contains mannitol, arabitol, glucose, glycerin, fructose and trehalose, among which the content of trehalose is the highest.

在药用价值方面,据元朝的《日用草本》和清代黄官绣的《本草术经》记载,长期食用花脸香蘑能够有效地防止血管硬化, 可以治疗肝硬化、糖尿病、肾脏病、胆结石等。到目前为止,国外关于花脸香蘑的药用价值的研究主要还是集中在它的抗氧化活性和抗菌活性两方面上。花脸香蘑的子实体还含有丰富的维生素B1,能够调节生物体正常的糖代谢, 促进神经传导,对预防脚气有很好的效果。花脸香蘑子实体的热水提取物具有显著的抗癌活性,其药理试验证明花脸香蘑子实体提取物对小白鼠艾氏瘤 和肉瘤S180的抑制率分别为100%和90%。而子实体煎煮的汁能够帮助治疗脓肿、砂眼和伤口。 In terms of medicinal value, according to the "Daily Herbs" of the Yuan Dynasty and the "Compendium of Materia Medica" written by Huang Guanxiu of the Qing Dynasty, long-term consumption of mushrooms can effectively prevent vascular sclerosis, and can treat liver cirrhosis, diabetes, kidney disease, Gallstones etc. So far, foreign researches on the medicinal value of Amanita chinensis mainly focus on its antioxidant activity and antibacterial activity. The fruiting body of the mushroom is also rich in vitamin B1, which can regulate the normal glucose metabolism of the organism, promote nerve conduction, and have a good effect on preventing athlete's foot. The hot water extract of the fruiting body of the flower face mushroom has significant anticancer activity, and its pharmacological test proves that the inhibition rate of the fruiting body extract of the flower face mushroom on the mouse Ehrlich tumor and sarcoma S180 is 100% and 90%, respectively. The decoction of the fruiting body can help treat abscesses, trachoma and wounds.

近年来,花脸香蘑逐渐被世人所认识了解,对它的市场需求不断增加,从而导致野生资源逐年减少,很难再满足市场的需求,对花脸香蘑进行人工驯化栽培势在必行,花脸香蘑菌丝体培养过程中生长速度慢,菌丝稀疏,不健壮,是制约花脸香蘑人工驯化栽培的一个技术难题。      目前,香蘑类菌丝体的人工培养大都采用马铃薯葡萄糖琼脂培养基( PDA ) ;马铃薯综合培养基( CPDA) ;麸皮葡萄糖琼脂培养基(麸皮50-70 g、葡萄糖20 g、琼脂18-20 g);玉米粉综合培养基(玉米粉20-30 g、葡萄糖20 g、磷酸二氢钾2 g、硫酸镁0. 5 g、琼脂18-20 g);堆肥浸出液培养基【堆肥(干) 100 g、葡萄糖20 g、琼脂18-20 g】等,这些培养基用于花脸香蘑菌丝培养过程中,生长速度慢,菌丝末端产生分支菌丝较多,稀疏。 In recent years, the flower-faced mushroom has gradually been recognized by the world, and the market demand for it has been increasing, which has led to a decrease in wild resources year by year, and it is difficult to meet the market demand. It is imperative to artificially domesticate and cultivate the flower-faced mushroom. The growth rate of the mycelium of the mushroom is slow, and the mycelium is sparse and not strong, which is a technical problem restricting the artificial domestication and cultivation of the mushroom. At present, most of the artificial cultivation of mushroom mycelium adopts potato dextrose agar medium (PDA); potato comprehensive medium (CPDA); bran glucose agar medium (bran 50-70 g, glucose 20 g, agar 18 -20 g); corn flour comprehensive medium (corn flour 20-30 g, glucose 20 g, potassium dihydrogen phosphate 2 g, magnesium sulfate 0.5 g, agar 18-20 g); compost leach medium [compost ( Dry) 100 g, glucose 20 g, agar 18-20 g], etc., these media are used in the process of cultivating the mycelia of the mushroom, the growth rate is slow, and there are many branched mycelia at the end of the mycelium, which are sparse.

发明内容 : Invention content:

本发明的目的在于克服上述已有技术的不足而提供一种操作简单,缩短培养周期短,无污染无废弃物,菌丝体产量高的花脸香蘑菌丝体的人工培养方法及其培养基。 The purpose of the present invention is to overcome the above-mentioned deficiencies in the prior art and provide a kind of simple operation, shorten the cultivation period, no pollution and no waste, the artificial culture method and culture medium of the mycelium of the mycelium with high mycelium yield .

本发明的目的可以通过如下措施来达到:一种用于培养花脸香蘑菌丝体的培养基,其特征在于其配方组分按重量份为:松针30-50份、蛋白胨5-12份、酵母浸粉6-12份、葡萄糖2.5-10份、琼脂:15-20份。 The object of the present invention can be achieved through the following measures: a kind of medium for cultivating the mycelia of the fragrant mushroom, is characterized in that its formula components are by weight: 30-50 parts of pine needles, 5-12 parts of peptone, Yeast extract powder 6-12 parts, glucose 2.5-10 parts, agar: 15-20 parts.

为了进一步实现本发明的目的,其配方组分按重量份为:松针36份、蛋白胨8份、酵母浸粉6份、葡萄糖4份、琼脂:18份。 In order to further realize the object of the present invention, its formula components by weight are: 36 parts of pine needles, 8 parts of peptone, 6 parts of yeast extract powder, 4 parts of glucose, 18 parts of agar.

一种花脸香蘑菌丝体的人工培养方法,其特征在于具体工艺步骤为: A method for artificially cultivating the mycelium of the fenugreek mycelium, characterized in that the specific process steps are:

(1)松针前处理: (1) Pretreatment of pine needles:

将收集到的松针进行清洗,放入1000ml水中,煮沸20-30分钟,过滤取汁; Wash the collected pine needles, put them into 1000ml water, boil for 20-30 minutes, filter to get the juice;

(2)培养基制备: (2) Culture medium preparation:

将松汁定容至1000ml,然后依次加入蛋白胨、酵母浸粉、葡萄糖、琼脂,煮沸混匀,然后进行分装; Dilute the pine juice to 1000ml, then add peptone, yeast extract powder, glucose, agar in turn, boil and mix well, and then pack;

(3)培养基分装,灭菌: (3) Subpackage and sterilize the medium:

 将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟; Divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes;

(4)冷却,接种: (4) cooling, inoculation:

 灭完菌后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种; After sterilization, take out the culture medium, cool it naturally to 20°C-30°C, make a slant medium, and insert the purified mushroom strains under aseptic conditions;

(5)培养: (5) Cultivate:

 将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管; Place the inoculated culture medium in a constant temperature incubator, and incubate at 20-30°C for 5-10 days until the mycelium is full of the test tube;

(6)菌种保藏:将培养好的菌种放于4℃的冰箱中进行保藏。 (6) Preservation of strains: Store the cultured strains in a refrigerator at 4°C.

本发明同已有技术相比可产生如下积极效果:花脸香蘑是一种珍稀、优质野生菌种,在野生状态下,数量少,产量低、但其色泽优美,气味浓香,味道鲜嫩,营养价值和药用价值极高,且价格昂贵。目前,适合花脸香蘑菌种优良培养基研究甚少,本发明培养基所使用的原料无污染,不会产生任何垃圾,简单实用,缩短培养周期,一般缩短4天左右,并且较其他培养基菌丝体长稠密、粗壮,呈淡紫色,生活力旺盛,可促使花脸香蘑菌丝体快速生长,为花脸香蘑实现大规模工业化生产及医药事业的发展都具有非常重要的意义。 Compared with the prior art, the present invention can produce the following positive effects: the mushroom is a rare and high-quality wild strain. In the wild state, the quantity is small and the yield is low, but its color is beautiful, the smell is strong, and the taste is fresh and tender. The nutritional value and medicinal value are extremely high, and the price is expensive. At present, there are very few researches on the excellent culture medium suitable for the strains of A. variegata. The raw materials used in the culture medium of the present invention are non-polluting, will not produce any garbage, are simple and practical, and shorten the cultivation period, generally about 4 days, and compared with other culture mediums. The mycelium is dense, strong, lavender in color, and has strong vitality, which can promote the rapid growth of the mycelium of the flower face mushroom, which is of great significance for the large-scale industrial production of the flower face mushroom and the development of the medical industry.

具体实施方式 : Detailed ways :

    下面对本发明的具体实施方式作详细说明: Below the specific embodiment of the present invention is described in detail:

实施例1: Example 1:

(1)松针前处理: (1) Pretreatment of pine needles:

将收集到的30g松针进行清洗,放入1000ml水中,煮沸20-30分钟,过滤取汁; Wash the collected 30g pine needles, put them into 1000ml water, boil for 20-30 minutes, filter to get the juice;

(2)培养基制备: (2) Culture medium preparation:

将松汁定容至1000ml,然后依次加入5g蛋白胨、6g酵母浸粉、2.5g葡萄糖、15g琼脂,煮沸混匀,然后进行分装;  Dilute the pine juice to 1000ml, then add 5g peptone, 6g yeast extract powder, 2.5g glucose, 15g agar in turn, boil and mix well, and then pack;

(3)培养基分装,灭菌: (3) Subpackage and sterilize the medium:

 将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟; Divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes;

(4)冷却,接种: (4) cooling, inoculation:

 灭完菌后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种; After sterilization, take out the culture medium, cool it naturally to 20°C-30°C, make a slant medium, and insert the purified mushroom strains under aseptic conditions;

(5)培养: (5) Cultivate:

 将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管; Place the inoculated culture medium in a constant temperature incubator, and incubate at 20-30°C for 5-10 days until the mycelium is full of the test tube;

(6)菌种保藏:将培养好的菌种放于4℃的冰箱中进行保藏。 (6) Preservation of strains: Store the cultured strains in a refrigerator at 4°C.

实施例2: Example 2:

(1)松针前处理: (1) Pretreatment of pine needles:

将收集到的36g松针进行清洗,放入1000ml水中,煮沸20-30分钟,过滤取汁; Wash the collected 36g pine needles, put them into 1000ml water, boil for 20-30 minutes, filter to get the juice;

(2)培养基制备: (2) Culture medium preparation:

将松汁定容至1000ml,然后依次加入8g蛋白胨、6g酵母浸粉、4g葡萄糖、18g琼脂,煮沸混匀,然后进行分装;  Dilute the pine juice to 1000ml, then add 8g peptone, 6g yeast extract powder, 4g glucose, 18g agar in turn, boil and mix well, and then pack;

(3)培养基分装,灭菌: (3) Subpackage and sterilize the medium:

 将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟; Divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes;

(4)冷却,接种: (4) cooling, inoculation:

 灭完菌后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种; After sterilization, take out the culture medium, cool it naturally to 20°C-30°C, make a slant medium, and insert the purified mushroom strains under aseptic conditions;

(5)培养: (5) Cultivate:

 将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管; Place the inoculated culture medium in a constant temperature incubator, and incubate at 20-30°C for 5-10 days until the mycelium is full of the test tube;

(6)菌种保藏:将培养好的菌种放于4℃的冰箱中进行保藏。 (6) Preservation of strains: Store the cultured strains in a refrigerator at 4°C.

实施例3: Example 3:

(1)松针前处理: (1) Pretreatment of pine needles:

将收集到的50g松针进行清洗,放入1000ml水中,煮沸20-30分钟,过滤取汁; Wash the collected 50g pine needles, put them into 1000ml water, boil for 20-30 minutes, filter to get the juice;

(2)培养基制备: (2) Culture medium preparation:

将松汁定容至1000ml,然后依次加入12g蛋白胨、12g酵母浸粉、10g葡萄糖、20g琼脂,煮沸混匀,然后进行分装;  Dilute the pine juice to 1000ml, then add 12g peptone, 12g yeast extract powder, 10g glucose, 20g agar in turn, boil and mix well, and then pack;

(3)培养基分装,灭菌: (3) Subpackage and sterilize the medium:

 将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟; Divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes;

(4)冷却,接种: (4) cooling, inoculation:

 灭完菌后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种; After sterilization, take out the culture medium, cool it naturally to 20°C-30°C, make a slant medium, and insert the purified mushroom strains under aseptic conditions;

(5)培养: (5) Cultivate:

 将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管; Place the inoculated culture medium in a constant temperature incubator, and incubate at 20-30°C for 5-10 days until the mycelium is full of the test tube;

(6)菌种保藏:将培养好的菌种放于4℃的冰箱中进行保藏。 (6) Preservation of strains: Store the cultured strains in a refrigerator at 4°C.

以上的实施例仅仅是对本发明的优选实施方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通工程技术人员对本发明的技术方案作出的各种变形和改进,均应落入本发明的权利要求书确定的保护范围内。 The above embodiment is only a description of the preferred implementation of the present invention, and is not intended to limit the scope of the present invention. On the premise of not departing from the design spirit of the present invention, ordinary engineers and technicians in the field can make various modifications to the technical solution of the present invention. and improvements, all should fall within the scope of protection determined by the claims of the present invention.

Claims (3)

1.一种用于培养花脸香蘑菌丝体的培养基,其特征在于其配方组分按重量份为:松针30-50份、蛋白胨5-12份、酵母浸粉6-12份、葡萄糖2.5-10份、琼脂:15-20份。 1. A culture medium for cultivating the mycelia of the fenugreek mushroom, characterized in that its formula components are in parts by weight: 30-50 parts of pine needles, 5-12 parts of peptone, 6-12 parts of yeast extract powder, glucose 2.5-10 parts, agar: 15-20 parts. 2.根据权利要求1所述的一种用于培养花脸香蘑菌丝体的培养基,其特征在于其配方组分按重量份为:松针36份、蛋白胨8份、酵母浸粉6份、葡萄糖4份、琼脂:18份。 2. A kind of medium for cultivating the mycelium of A. versicolor according to claim 1 is characterized in that its formula components are by weight: 36 parts of pine needles, 8 parts of peptone, 6 parts of yeast extract powder, 4 parts of glucose, 18 parts of agar. 3.一种花脸香蘑菌丝体的人工培养方法,其特征在于具体工艺步骤为: 3. a method for artificially cultivating the mycelium of the fragrant mushroom mycelium, characterized in that the specific process steps are: (1)松针前处理: (1) Pretreatment of pine needles: 将收集到的松针进行清洗,放入1000ml水中,煮沸20-30分钟,过滤取汁; Wash the collected pine needles, put them into 1000ml water, boil for 20-30 minutes, filter to get the juice; (2)培养基制备: (2) Culture medium preparation: 将松汁定容至1000ml,然后依次加入蛋白胨、酵母浸粉、葡萄糖、琼脂,煮沸混匀,然后进行分装; Dilute the pine juice to 1000ml, then add peptone, yeast extract powder, glucose, agar in turn, boil and mix well, and then pack; (3)培养基分装,灭菌: (3) Subpackage and sterilize the medium:  将配制好的培养基分装于试管中,盖上胶塞,在120-130℃条件下灭菌20-30分钟; Divide the prepared medium into test tubes, cover with rubber stoppers, and sterilize at 120-130°C for 20-30 minutes; (4)冷却,接种: (4) cooling, inoculation:  灭完菌后取出培养基,自然冷却至20℃-30℃,制成斜面培养基,在无菌条件下,接入经过纯化的花脸香蘑菌种; After sterilization, take out the culture medium, cool it naturally to 20°C-30°C, make a slant medium, and insert the purified mushroom strains under aseptic conditions; (5)培养: (5) Cultivate:  将接好种的培养基置于恒温培养箱里,20-30℃培养5-10天,直至菌丝长满试管; Place the inoculated culture medium in a constant temperature incubator, and incubate at 20-30°C for 5-10 days until the mycelium is full of the test tube; (6)菌种保藏:将培养好的菌种放于4℃的冰箱中进行保藏。 (6) Preservation of strains: Store the cultured strains in a refrigerator at 4°C.
CN201410192113.6A 2014-05-08 2014-05-08 Artificial culturing method of lepista sordida mycelium and culturing medium thereof Pending CN103947454A (en)

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CN104584862A (en) * 2015-01-12 2015-05-06 高佩金 Method for producing lepista sordida in field commercial way
CN105766368A (en) * 2016-03-01 2016-07-20 鲁东大学 Method for cultivating lepista sordida (Schum.: Fr.) Sing.with miscanthus sacchariflorus and culture medium of method
CN106854093A (en) * 2016-12-30 2017-06-16 高佩金 The artificial method for preparing Lepista sordida culture medium and cultivating Lepista sordida
CN108048335A (en) * 2017-12-25 2018-05-18 内蒙古自治区农牧业科学院 Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection
CN108048335B (en) * 2017-12-25 2021-02-05 内蒙古自治区农牧业科学院 Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof
CN108293599A (en) * 2018-02-28 2018-07-20 中华全国供销合作总社昆明食用菌研究所 A kind of Lepista sordida and its separation expanding propagation method and soil covering culture method
CN108293599B (en) * 2018-02-28 2020-05-19 中华全国供销合作总社昆明食用菌研究所 Lepista sordida, and separation propagation method and soil-covering cultivation method thereof
CN111066574A (en) * 2020-01-15 2020-04-28 青岛农业大学 Method for preparing Lepista sordida cultivars by using mushroom dregs
CN113862159A (en) * 2021-10-12 2021-12-31 天津春发生物科技集团有限公司 Method for preparing tricholoma matsutake mushroom seasoning by fungus liquid culture method
CN113862159B (en) * 2021-10-12 2024-04-30 天津春发生物科技集团有限公司 Method for preparing tricholoma matsutake seasoning by fungus liquid culture method
CN115058347A (en) * 2022-06-07 2022-09-16 青岛农业大学 Lepista sordida, preparation method of non-saccharide extract of Lepista sordida and application of non-saccharide extract

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