广西杀虫性大型有毒真菌筛选及菌丝体人工培养的初步研究
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摘要
本文总结了广西杀虫性大型有毒真菌筛选及菌丝体人工培养的初步研究结果。从广西不同地区采集到28种大型有毒真菌,这些大型有毒真菌分属13科,其中鹅膏菌科(Amanitaceae)5种,白蘑科(Tricholomataceae)4种,蘑菇科(Agaricacea)、鬼伞科(Coprinaceae)、牛肝菌科(Boletaceae)各3种。将这28种大型有毒真菌的新鲜子实体和培养物,分别选用水、乙醇、石油醚作为溶剂进行提取、获得粗提取物,然后以斜纹夜蛾(Prodeniaa litura)和小菜蛾(Pultella xylostella)作为供试害虫,进行杀虫活性的测定试验,结果发现:有24种大型有毒真菌新鲜子实体粗提取物对斜纹夜蛾幼虫具有杀虫活性,有23种对小菜蛾幼虫具有杀虫活性。其中角鳞白鹅膏菌(Amanita solitaria)、角鳞灰鹅膏菌(A.spissacea)、残斑托鹅膏菌(A.sychnopyramis)的蒸馏水粗提取物对斜纹夜蛾和小菜蛾幼虫的杀虫作用非常明显,其3 d校正死亡率均为100%。除硬黄皮马勃(Sclerderma flavidum)、橘黄裸伞(Gymnopilus spectabilis)、残斑托鹅膏菌外其余大型有毒真菌的杀虫活性在国内均为首次报道。墨汁鬼伞(Coprinus atramentarius)和绿褐裸伞(Gymnopilus aeraginosus)的固体培养菌丝体的粗提取物对斜纹夜蛾具有杀虫活性,灰鹅膏菌(A.vaginata),暗褐毒鹅膏菌(A.brunnescens)等4种大型有毒真菌液体培养产物对斜纹夜蛾和小菜蛾幼虫有杀虫活性。
对这28种大型有毒真菌进行分离、人工培养的试验结果表明,暗褐毒鹅膏菌、灰鹅膏菌、粪生光盖伞(Psilocybe coprophila)等10种大型有毒真菌能够分离、培养得到菌丝体。其中暗褐毒鹅膏菌、灰鹅膏菌和绿褐裸伞菌丝体的分离和培养在国内尚属首次。结合杀虫活性试验结果,选择既具有一定杀虫活性又能人工分离培养的墨汁鬼伞、绿褐裸伞、暗褐毒鹅膏菌和灰鹅膏菌这4种大型有毒真菌进行人工培养特性的试验研究,结果如下,墨汁鬼伞菌丝体的适宜固体培养基为PDA+牛粪浸液培养基,以乳糖和蛋白胨为适宜的碳氮源,适宜培养温度为28～32℃,适宜pH为7.0;绿褐裸伞菌丝体的适宜固体培养基为PDA+树叶浸出液培养基,碳源为葡萄糖、氮源为干酪素;适宜培养温度为30℃,适宜pH为6.0左右;暗褐毒鹅膏菌、灰鹅膏菌的菌丝体适宜固体培养基均为PDM培养基,适宜碳源分别麦芽糖和甘露醇;适宜氮源分别为酵母粉和干酪素;适宜培养温度分别为30℃、28℃,适宜pH分别为5.0和6.0。
为了给规模化人工培养提供依据,对较有研究和开发价值的墨汁鬼伞进行了菌丝体液体培养试验研究,结果显示,墨汁鬼伞菌丝体的适宜液体培养基组成为葡萄糖2.0%,脱脂豆粉0.5%,K_2HPO_4 0.3%,MgSO_4·5H_2O 0.3%,10%牛粪浸液;适宜培养温度为28℃,适宜pH为7.0,适宜装液量为90 ml(250 ml锥形瓶),接种量以体积分数10%为好。
This paper was a preliminary study on screening toadstools with insecticidal activity and their mycelia culture in Guangxi.Twenty-eight species of toadstools were collected from different regions of Guangxi.They were under 13 families.The family with most toadstool was Amanitaceae,5 species;the second one was Tricholomataceae,4 species;and the Agaricacea, Coprinaceae and Boletaceae were 3 species.The crude of the 28 toadstools were extracted from fresh fruitbodies and cultured mixture using distilled water,ethanol and petroleum ether as menstruum respectively.Then the crude extracts were used to detect whether they had insecticidal activity against Prodeniaa litura and Pultella xylostella.The results showed that during 3 days,the crude extracts from 24 species of toadstools had insecticidal activity against Prodeniaa litura,and from 23 species of toadstools had insecticidal activity against Pultella xylostella.The crude extracts with distilled water about Amanita solitaria,A.spissacea and A.sychnopyramis were toxin to Prodeniaa litura and Pultella xylostella,which the corrected mortalities were 100%.The insecticidal activity of the toadstools was first reported except Sclerderma flavidum,Gymnopilus spectabilis and A.sychnopyramis.The crude extracts from solid culture mycelia of Coprinus atramentarius and Gymnopilus aeraginosus were toxic to Prodeniaa litura.And the crude extracts from liquid culture mycelia of 4 species of toadstools such as Amanita vaginata and A.brunnescens were toxic to Prodeniaa litura and Pultella xylostella.
Isolation and artificial culture of 28 species of toadstools were conducted. The results showed that only 10 species of toadstools such as A.brunnescens、A.vaginata,Psilocybe coprophila could be artificial cultured.The isolation and artificial culture of A.brunnescens,A.vaginata and G.aeraginosus were first reported.The optimal artificial culture conditions of 4 toadstool species which not only had insecticidal activity but also could be isolated as below respectively:
The optimal solid media for C.atramentarius mycelia was PDA plus extraction from cattle manure;the optimal carbon and nitrogen sources were Lactose and peptone;the optimal temperature for the mycelium growth was 28～32℃;and the optimal pH was about 7.0.The optimal solid media for G. aeraginosus mycelia was PDA plus extraction from tree leaves;the optimal carbon and nitrogen sources were glucose and casein;the optimal temperature for the mycelia growth was 30℃;and the optimal pH was about 6.0.The optimal solid media for A.brunnescens and A.vaginata were PDM;the optimal carbon sources were maltose and mannite;the optimal nitrogen sources were yeast extract and casein respectively;the optimal temperature for the mycelia growth was 30℃and 28℃,the optimal pH was 5.0 and 6.0.
The optimal liquid media for mycelia growth of C.atramentarius were 2.0%glucose,0.5%defatted soybean powder,0.3%K_2HPO_4,0.3% MgSO4_4·5H_2O and 10%cattle manure.The optimal growth conditions for mycelia of C.atramentarius were:culture temperature 28℃,pH 7.0,90 ml substrata in 250 ml shake flask and inoculating amount 10%.The results of liquid culture provide scientific basis for the mass culture of the toadstools.
对这28种大型有毒真菌进行分离、人工培养的试验结果表明,暗褐毒鹅膏菌、灰鹅膏菌、粪生光盖伞(Psilocybe coprophila)等10种大型有毒真菌能够分离、培养得到菌丝体。其中暗褐毒鹅膏菌、灰鹅膏菌和绿褐裸伞菌丝体的分离和培养在国内尚属首次。结合杀虫活性试验结果,选择既具有一定杀虫活性又能人工分离培养的墨汁鬼伞、绿褐裸伞、暗褐毒鹅膏菌和灰鹅膏菌这4种大型有毒真菌进行人工培养特性的试验研究,结果如下,墨汁鬼伞菌丝体的适宜固体培养基为PDA+牛粪浸液培养基,以乳糖和蛋白胨为适宜的碳氮源,适宜培养温度为28～32℃,适宜pH为7.0;绿褐裸伞菌丝体的适宜固体培养基为PDA+树叶浸出液培养基,碳源为葡萄糖、氮源为干酪素;适宜培养温度为30℃,适宜pH为6.0左右;暗褐毒鹅膏菌、灰鹅膏菌的菌丝体适宜固体培养基均为PDM培养基,适宜碳源分别麦芽糖和甘露醇;适宜氮源分别为酵母粉和干酪素;适宜培养温度分别为30℃、28℃,适宜pH分别为5.0和6.0。
为了给规模化人工培养提供依据,对较有研究和开发价值的墨汁鬼伞进行了菌丝体液体培养试验研究,结果显示,墨汁鬼伞菌丝体的适宜液体培养基组成为葡萄糖2.0%,脱脂豆粉0.5%,K_2HPO_4 0.3%,MgSO_4·5H_2O 0.3%,10%牛粪浸液;适宜培养温度为28℃,适宜pH为7.0,适宜装液量为90 ml(250 ml锥形瓶),接种量以体积分数10%为好。
This paper was a preliminary study on screening toadstools with insecticidal activity and their mycelia culture in Guangxi.Twenty-eight species of toadstools were collected from different regions of Guangxi.They were under 13 families.The family with most toadstool was Amanitaceae,5 species;the second one was Tricholomataceae,4 species;and the Agaricacea, Coprinaceae and Boletaceae were 3 species.The crude of the 28 toadstools were extracted from fresh fruitbodies and cultured mixture using distilled water,ethanol and petroleum ether as menstruum respectively.Then the crude extracts were used to detect whether they had insecticidal activity against Prodeniaa litura and Pultella xylostella.The results showed that during 3 days,the crude extracts from 24 species of toadstools had insecticidal activity against Prodeniaa litura,and from 23 species of toadstools had insecticidal activity against Pultella xylostella.The crude extracts with distilled water about Amanita solitaria,A.spissacea and A.sychnopyramis were toxin to Prodeniaa litura and Pultella xylostella,which the corrected mortalities were 100%.The insecticidal activity of the toadstools was first reported except Sclerderma flavidum,Gymnopilus spectabilis and A.sychnopyramis.The crude extracts from solid culture mycelia of Coprinus atramentarius and Gymnopilus aeraginosus were toxic to Prodeniaa litura.And the crude extracts from liquid culture mycelia of 4 species of toadstools such as Amanita vaginata and A.brunnescens were toxic to Prodeniaa litura and Pultella xylostella.
Isolation and artificial culture of 28 species of toadstools were conducted. The results showed that only 10 species of toadstools such as A.brunnescens、A.vaginata,Psilocybe coprophila could be artificial cultured.The isolation and artificial culture of A.brunnescens,A.vaginata and G.aeraginosus were first reported.The optimal artificial culture conditions of 4 toadstool species which not only had insecticidal activity but also could be isolated as below respectively:
The optimal solid media for C.atramentarius mycelia was PDA plus extraction from cattle manure;the optimal carbon and nitrogen sources were Lactose and peptone;the optimal temperature for the mycelium growth was 28～32℃;and the optimal pH was about 7.0.The optimal solid media for G. aeraginosus mycelia was PDA plus extraction from tree leaves;the optimal carbon and nitrogen sources were glucose and casein;the optimal temperature for the mycelia growth was 30℃;and the optimal pH was about 6.0.The optimal solid media for A.brunnescens and A.vaginata were PDM;the optimal carbon sources were maltose and mannite;the optimal nitrogen sources were yeast extract and casein respectively;the optimal temperature for the mycelia growth was 30℃and 28℃,the optimal pH was 5.0 and 6.0.
The optimal liquid media for mycelia growth of C.atramentarius were 2.0%glucose,0.5%defatted soybean powder,0.3%K_2HPO_4,0.3% MgSO4_4·5H_2O and 10%cattle manure.The optimal growth conditions for mycelia of C.atramentarius were:culture temperature 28℃,pH 7.0,90 ml substrata in 250 ml shake flask and inoculating amount 10%.The results of liquid culture provide scientific basis for the mass culture of the toadstools.
引文
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