不同保藏方法对金针菇“上研1号”栽培特性的影响
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摘要
针对金针菇(Flammulina velutipes)菌种在保藏中难以保持遗传性能相对稳定的问题,以金针菇新品种"上研1号"为供试菌株,采用继代培养法(C)、液体石蜡法(P)、液氮法(N)和蒸馏水法(W)保藏两年后,观测活化菌丝的生长、液体菌种性能及工厂化栽培性状,比较不同保藏方法对金针菇栽培特性的影响。研究结果表明:液氮法保藏处理和继代培养保藏处理中,活化菌丝生长速度快,菌落浓白、均一;液体菌种成球度、菌球大小和密度等方面均表现优异,液体菌种终止pH在5.9～6.0之间;菌丝分支能力强,锁状联合明显且数量较多。在工厂化栽培中,液氮法保藏处理子实体整齐度好、产量高(每瓶372.67 g);液氮法和继代培养法保藏处理的A级商品菇比例相近,商品性状优良。液氮法与继代培养法保藏处理在两年内均可维持金针菇新品种的栽培特性,但液氮保藏法操作更为便捷,是保藏金针菇品种的稳定、可靠、省力的方法。
The genetic stability of Flammulina velutipes strains is difficult to be maintained during storage.Aiming at this issue,the new cultivar "Shangyan 1" was used as the test strain and 4 preservation methods were assessed: the subculture preservation method(C), liquid paraffin preservation method(P), liquid nitrogen preservation method(N) and distilled water preservation method(W). Cultivation characteristics including the growth rate of activated strain, indexes of liquid spawn and industrial cultivation performance were observed after 2 years, and the effects of different preservation methods on the cultivation characteristics were compared. The results showed that after C and N treatments, the mycelia grew fast, the colonies were white and uniform, the indexes of liquid spawn such as sphericity, size and density were excellent, and the termination pH values were between 5. 9 and 6.0. The microscopic examination revealed that mycelial growth direction was clear with many branches and obvious clamp cells. Among the industrial cultivation indexes, the best uniformity and the highest average yield(372.67 g/bottle) were observed after N treatment. Both N and C treatments maintained the cultivation characteristics of the new F. velutipes cultivar within 2 years, by providing similar ratios of A-grade products with outstanding commercial characters after reactivation. However, liquid nitrogen preservation is easier to operate, and hence recommended as a stable, reliable and labor-saving method for the preservation of F. velutipes cultivar.
The genetic stability of Flammulina velutipes strains is difficult to be maintained during storage.Aiming at this issue,the new cultivar "Shangyan 1" was used as the test strain and 4 preservation methods were assessed: the subculture preservation method(C), liquid paraffin preservation method(P), liquid nitrogen preservation method(N) and distilled water preservation method(W). Cultivation characteristics including the growth rate of activated strain, indexes of liquid spawn and industrial cultivation performance were observed after 2 years, and the effects of different preservation methods on the cultivation characteristics were compared. The results showed that after C and N treatments, the mycelia grew fast, the colonies were white and uniform, the indexes of liquid spawn such as sphericity, size and density were excellent, and the termination pH values were between 5. 9 and 6.0. The microscopic examination revealed that mycelial growth direction was clear with many branches and obvious clamp cells. Among the industrial cultivation indexes, the best uniformity and the highest average yield(372.67 g/bottle) were observed after N treatment. Both N and C treatments maintained the cultivation characteristics of the new F. velutipes cultivar within 2 years, by providing similar ratios of A-grade products with outstanding commercial characters after reactivation. However, liquid nitrogen preservation is easier to operate, and hence recommended as a stable, reliable and labor-saving method for the preservation of F. velutipes cultivar.
引文
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[3]中国食用菌协会关于印发全国食用菌2016年度产量、产值和出口统计调查结果的函,中食菌协函[2017]11号,http://www.cefa.org.cn/2017/10/24/10250.html
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[12]张丹,王瑞娟,徐珍,等.金针菇新品种'上研1号'[J].园艺学报,2017,44(S2):2701-2702.
[13]唐木田郁夫.中国金针菇工厂化生产中的问题[J].食药用菌,2018,26(1):23-25.
[14]李洪荣.双孢蘑菇液氮保藏菌株的遗传稳定性分析[J].中国农学通报,2011,27(22):276-280.
[15]王世东,周学政.食用菌菌种的质量检测处理[J].食用菌,2003,25(3):20-21.
[16]吕和平,吴红宇,邢仁昌,等.食用菌菌株的长期保藏处理[J].中国食用菌,2007,26(2):29-30.
[17] DRAKE JW, CHARLESWORTH B, CHARLES WORTH D, et al. Rates of spontaneous mutation[J].Genetics, 1998, 148(4):1667-1686.
[18]韩芹芹,王金宁,李国庆,等.液氮保藏菌种对平菇菌丝体和子实体的影响[J].安徽农业大学学报,2012,39(6):1003-1007.
[19]顾金刚,李世贵,姜瑞波,等.真菌保藏技术研究进展[J].菌物学报,2007,26(2):316-320.
[20] MATA G, PEREZ-MERLO R. Spawn viability in edible mushrooms after freezing in liquid nitrogen without a cryoprotectant[J]. Cryobiology,2003,47(1):14-20.
[21] HOMOLKA L, LISA L, NERUD F. Basidiomycete cryopreservation on perlite:evaluation of a new method[J].Cryobiology, 2006,52(3):446-453.
[2]刘德云,徐珍.平菇金针菇安全生产技术指南[M].北京:中国农业出版社,2012:95-97.
[3]中国食用菌协会关于印发全国食用菌2016年度产量、产值和出口统计调查结果的函,中食菌协函[2017]11号,http://www.cefa.org.cn/2017/10/24/10250.html
[4]张丹,朱爱莲,赵静,等.采用木屑培养基传代的金针菇菌种稳定性[J].食用菌学报,2018,25(2):29-34.
[5]杨新美.中国食用菌栽培学[M].北京:农业出版社,1988:206-212.
[6] LITTLE GN, GORDON MA. Suivival of fungus cultures maintained under mineral oil for twelve years[J].Mycologia, 1967, 59(4):733-736.
[7] HWANG SW. Long-term preservation of fungus cultures with liquid nitrogen refrigeration[J]. Applied Microbiology, 1966, 14(5):784-788.
[8]杨慧,尚晓冬,王瑞霞,等.蒸馏水与液氮保藏金针菇菌种特性的比较[J].食用菌学报,2010,17(4):23-25.
[9] BORMAN A,SZEKELY A, CAMPBELL C, et al. Evaluation of the viability of pathogenic filamentous fungi after prolonged storage in sterile water and review of recent published studies on storage methods[J].Mycopahologia, 2006, 161(6):361-368.
[10]张瑞颖,胡丹丹,左雪梅,等.食用菌菌种保藏技术研究进展[J].食用菌学报,2010,17(4):84-88.
[11]孔维丽,袁瑞奇,孔维威,等.食用菌菌种保藏历史、现状及研究展望概述[J].中国食用菌,2015,34(5):1-5.
[12]张丹,王瑞娟,徐珍,等.金针菇新品种'上研1号'[J].园艺学报,2017,44(S2):2701-2702.
[13]唐木田郁夫.中国金针菇工厂化生产中的问题[J].食药用菌,2018,26(1):23-25.
[14]李洪荣.双孢蘑菇液氮保藏菌株的遗传稳定性分析[J].中国农学通报,2011,27(22):276-280.
[15]王世东,周学政.食用菌菌种的质量检测处理[J].食用菌,2003,25(3):20-21.
[16]吕和平,吴红宇,邢仁昌,等.食用菌菌株的长期保藏处理[J].中国食用菌,2007,26(2):29-30.
[17] DRAKE JW, CHARLESWORTH B, CHARLES WORTH D, et al. Rates of spontaneous mutation[J].Genetics, 1998, 148(4):1667-1686.
[18]韩芹芹,王金宁,李国庆,等.液氮保藏菌种对平菇菌丝体和子实体的影响[J].安徽农业大学学报,2012,39(6):1003-1007.
[19]顾金刚,李世贵,姜瑞波,等.真菌保藏技术研究进展[J].菌物学报,2007,26(2):316-320.
[20] MATA G, PEREZ-MERLO R. Spawn viability in edible mushrooms after freezing in liquid nitrogen without a cryoprotectant[J]. Cryobiology,2003,47(1):14-20.
[21] HOMOLKA L, LISA L, NERUD F. Basidiomycete cryopreservation on perlite:evaluation of a new method[J].Cryobiology, 2006,52(3):446-453.
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