泰山云芝TN-1生物学特性及驯化栽培研究
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摘要
药用真菌是指能治疗疾病、具有药用价值的一类真菌,即对人体有保健作用,对疾病有预防、抑制或治疗作用的真菌。云芝属药用真菌中的一种木腐性药用真菌,其有效成分云芝多糖对治疗慢性肝炎、乙型肝炎有显著疗效,对提高人体免疫力,抗癌治癌也较有效,具有较高的经济价值和较好开发前景。泰山优越的气候条件使其蕴藏着丰富野生经济药用真菌资源,泰山云芝久负盛名,野生资源丰富。近年来,由于生态环境及季节等自然条件的限制,加之人为破坏和掠取,导致泰山野生云芝资源不断减少,亟需通过人工开发利用泰山云芝的野生资源,达到保护泰山云芝野生资源和开发野生药用菌资源为人所用的双重目的。关于泰山云芝的生物学特性及驯化栽培研究尚未见有研究报道。
2006-2009年在泰山区域内对野生云芝资源的调查采集,共分离保存菌株23株,经拮抗试验和回接出菇试验,筛选出TN-1、TN-2、TN-3三个农艺性状良好的菌株,其中以TN-1菌丝浓密、发菌快、子实体生长整齐,产量高,作为重点研究。经野生菌形态特征、菌丝培养特征和显微观察,初步鉴定泰山云芝TN-1菌株为云芝Coriolus versicolor (L.:Fr.) Quél.,别名杂色云芝。
生物学特性研究表明,适宜泰山云芝TN-1菌丝体生长的碳源是麦芽糖、蔗糖,氮源为蛋白胨、酵母膏,影响其生长速度因子主次顺序为麦芽糖、蛋白胨、KH2PO4、MgSO4;泰山云芝TN-1菌丝体在5℃～35℃时均能生长,最适生长温度在25～28℃,泰山云芝TN-1在黑暗条件下菌丝生长较快,在pH5.5～6.5时菌丝生长速度最快。
液体培养泰山云芝TN-1菌丝最佳条件筛选试验,初步确定最佳液体培养条件为:温度控制在25℃～30℃,以28℃为最适培养温度,最适液体培养基pH值为5～6,500mL三角瓶装液量150mL,摇瓶转速最佳转速在120～150r/min,培养6d可得最大菌丝生长量。
本研究通过泰山云芝人工驯化栽培研究,筛选出适于泰山云芝生长的各级菌种培养基,确定了影响泰山云芝出菇效果的最佳条件,如出菇方法、培养基含水量、装料量、出菇温度、湿度。结合试验研究结果,初步建立了泰山云芝人工驯化栽培技术体系。
Medicinal fungi is a class of fungi which are possessed of medicinal value and could cure diseases.Versicolor is a kind of wood-rotting medicinal fungi. The active ingredient PSK of Versicolor not only have significantly curative effect on the chronic hepatitis and hepatitis B, but also could prevent cancers and improve the human immunity. Thus it has a high economic value and good development prospect. The climatic conditions of Taishan is suitable for the medicinal growth and there have many resources. In recent years, the resources are declining due to the ecological environment , natural conditions, and man-made destruction and plunde. Development and utilization the wild resources of versicolor in Taishan could not only protect the wild resources,but also improve the exploit value of medicinal fungi. So far, the biological characteristics of Taishan versicolor and domestication cultivation has yet been reported.
From 2006 to 2009, the wild resources of versicolor in Taishan were detected and collected. Twenty-three strains were collected and three ones ( TN-1, TN-2, TN-3 ) that prossessing good traits were isolated and preserved by antagonistic test and fruiting tests. Our result showed that TN-1 strain was the best one, so we research it deeply. We identified the TN-1 strain is Coriolus versicolor (L.:Fr.) Quél. from the characteristics of morphological, mycelium culture and microscopic observation.
Biological characteristics research showed that the optimal carbon source is maltose and sucrose, and the optimal nitrogen source is peptone and yeast extract. The primary and secondary order of mycelial growth factors is maltose, peptone, KH2PO4 and MgSO4. The mycelium of TN-1 strain can grow from 5℃to 35℃and the optimal growth tempreture was found to be 25℃-28℃. TN-1 grows faster under the dark condition . The optimal pH of mycelial growth was found to be 5.5 - 6.5.
The optimal conditions in liquid culture of TN-1 were screened and the results are as followed.The temperature should be controlled at 25℃-30℃and the optimal temperture was found to be 28℃. The optimum pH value was 5-6. It could get the largest mycelium growth amount under the optimal tempreture and pH in 500mL bottle that filling 150mL liquid culture after being cultured six days .The optimal shake-flask speed is 120-150r/min.
In this study, we obtain one versicolor strain by artificial domestication cultivation, The suitable medium for mycelial growth in different stages were screened. We also determined the the best fruiting condition such as fruiting method, the moisture of medium, capacity of loading feeding, fruiting temperature and humidity. In the conclusion, the artificial domestication cultivation technology system of versicolor was initially established .
2006-2009年在泰山区域内对野生云芝资源的调查采集,共分离保存菌株23株,经拮抗试验和回接出菇试验,筛选出TN-1、TN-2、TN-3三个农艺性状良好的菌株,其中以TN-1菌丝浓密、发菌快、子实体生长整齐,产量高,作为重点研究。经野生菌形态特征、菌丝培养特征和显微观察,初步鉴定泰山云芝TN-1菌株为云芝Coriolus versicolor (L.:Fr.) Quél.,别名杂色云芝。
生物学特性研究表明,适宜泰山云芝TN-1菌丝体生长的碳源是麦芽糖、蔗糖,氮源为蛋白胨、酵母膏,影响其生长速度因子主次顺序为麦芽糖、蛋白胨、KH2PO4、MgSO4;泰山云芝TN-1菌丝体在5℃～35℃时均能生长,最适生长温度在25～28℃,泰山云芝TN-1在黑暗条件下菌丝生长较快,在pH5.5～6.5时菌丝生长速度最快。
液体培养泰山云芝TN-1菌丝最佳条件筛选试验,初步确定最佳液体培养条件为:温度控制在25℃～30℃,以28℃为最适培养温度,最适液体培养基pH值为5～6,500mL三角瓶装液量150mL,摇瓶转速最佳转速在120～150r/min,培养6d可得最大菌丝生长量。
本研究通过泰山云芝人工驯化栽培研究,筛选出适于泰山云芝生长的各级菌种培养基,确定了影响泰山云芝出菇效果的最佳条件,如出菇方法、培养基含水量、装料量、出菇温度、湿度。结合试验研究结果,初步建立了泰山云芝人工驯化栽培技术体系。
Medicinal fungi is a class of fungi which are possessed of medicinal value and could cure diseases.Versicolor is a kind of wood-rotting medicinal fungi. The active ingredient PSK of Versicolor not only have significantly curative effect on the chronic hepatitis and hepatitis B, but also could prevent cancers and improve the human immunity. Thus it has a high economic value and good development prospect. The climatic conditions of Taishan is suitable for the medicinal growth and there have many resources. In recent years, the resources are declining due to the ecological environment , natural conditions, and man-made destruction and plunde. Development and utilization the wild resources of versicolor in Taishan could not only protect the wild resources,but also improve the exploit value of medicinal fungi. So far, the biological characteristics of Taishan versicolor and domestication cultivation has yet been reported.
From 2006 to 2009, the wild resources of versicolor in Taishan were detected and collected. Twenty-three strains were collected and three ones ( TN-1, TN-2, TN-3 ) that prossessing good traits were isolated and preserved by antagonistic test and fruiting tests. Our result showed that TN-1 strain was the best one, so we research it deeply. We identified the TN-1 strain is Coriolus versicolor (L.:Fr.) Quél. from the characteristics of morphological, mycelium culture and microscopic observation.
Biological characteristics research showed that the optimal carbon source is maltose and sucrose, and the optimal nitrogen source is peptone and yeast extract. The primary and secondary order of mycelial growth factors is maltose, peptone, KH2PO4 and MgSO4. The mycelium of TN-1 strain can grow from 5℃to 35℃and the optimal growth tempreture was found to be 25℃-28℃. TN-1 grows faster under the dark condition . The optimal pH of mycelial growth was found to be 5.5 - 6.5.
The optimal conditions in liquid culture of TN-1 were screened and the results are as followed.The temperature should be controlled at 25℃-30℃and the optimal temperture was found to be 28℃. The optimum pH value was 5-6. It could get the largest mycelium growth amount under the optimal tempreture and pH in 500mL bottle that filling 150mL liquid culture after being cultured six days .The optimal shake-flask speed is 120-150r/min.
In this study, we obtain one versicolor strain by artificial domestication cultivation, The suitable medium for mycelial growth in different stages were screened. We also determined the the best fruiting condition such as fruiting method, the moisture of medium, capacity of loading feeding, fruiting temperature and humidity. In the conclusion, the artificial domestication cultivation technology system of versicolor was initially established .
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