一株野生金针菇菌种的分离、鉴定与生物学特性研究

doi:10.3969/j.issn.1004-1524.2023.05.12

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (5): 1088-1096.DOI: 10.3969/j.issn.1004-1524.2023.05.12

一株野生金针菇菌种的分离、鉴定与生物学特性研究

冯连荣¹(), 张妍¹, 赵鑫闻¹, 宋立志¹, 梁德军²^,^*()

1.辽宁省杨树研究所,辽宁盖州 115213
2.辽宁省经济林研究所,辽宁大连 116030

收稿日期:2022-06-17 出版日期:2023-05-25 发布日期:2023-06-01
作者简介:冯连荣(1983—),女,黑龙江齐齐哈尔人,硕士,正高级工程师,研究方向为杨树育种和林下食用菌栽培。E-mail:fenglianrong@163.com
通讯作者: *梁德军,Emai:liangdejun70@163.com
基金资助:
“十四五”国家重点研发计划(2021FYD2201200);“十三五”国家重点研发计划(2016YFD0600401)

Isolation, identification and biological characteristics of a wild Flammulina filiformis strain

FENG Lianrong¹(), ZHANG Yan¹, ZHAO Xinwen¹, SONG Lizhi¹, LIANG Dejun²^,^*()

1. Liaoning Provincial Institute of Poplar, Gaizhou 115213, Liaoning, China
2. Liaoning Institute of Economic Forestry, Dalian 116030, Liaoning, China

Received:2022-06-17 Online:2023-05-25 Published:2023-06-01

摘要/Abstract

摘要：

为给野生食用菌开发利用提供候选菌种资源,以采自杨树树桩上的野生大型真菌JC423为研究对象,通过形态学鉴定、出菇试验、ITS序列分析和系统发育分析,明确其分离地位。采用单因素试验,研究不同碳源、氮源、无机盐、维生素、pH值和温度对JC423菌丝生长的影响。结果表明,采集的野生食用菌依据子实体形态初步鉴定为金针菇。ITS序列分析获得773 bp目的片段;BLAST序列比对显示,目的片段与Flammulina filiformis strain F007相似性最高,为99.34%;下载部分相似性序列,用MEGA 7.0构建进化树,JC423与序列号为MN337400.1的 F. filiformis strain F007聚为一支,结合形态学特征将JC423鉴定为金针菇。生物学特性结果表明,JC423菌丝生长适宜碳源为果糖,适宜氮源为蛋白胨,适宜pH值范围为9~10,适宜生长温度为25 ℃,无机盐和维生素对菌丝生长影响不大。该研究明确了野生金针菇菌丝生长的营养需求和条件,研究结果可以为金针菇开发利用奠定基础。

关键词: 金针菇, 菌种分离, ITS序列, 生物学特性

Abstract:

The isolation, identification and biological characteristics of a wild edible fungus were studied to provide candidate strain resources for the development and utilization of wild edible fungi. The wild macrofungi JC423 collected from poplar stumps was taken as the research object, and its taxonomic status was clarified through morphological identification, fruiting test, ITS sequences analysis and phylogenetic analysis. The effects of different carbon sources, nitrogen sources, inorganic salts, vitamins, pH and temperature on the growth of JC423 hyphae were studied by single factor experiment. The results showed that the collected wild edible fungi were preliminarily identified as Flammulina velutiper according to the shape of the fruiting bodies. The 773 bp target fragment was obtained by ITS sequence analysis. BLAST sequence alignment showed that the highest similarity between this strain and F. filiformis strain F007 was 99.34%. Partially similar sequences were downloaded and an evolutionary tree was constructed by MEGA 7.0. JC423 and F. filiformis strain F007 with the serial number MN337400.1 were clustered together. Combined with morphological characteristics, JC423 was identified as F. filiformis. The results of biological characteristics showed that the suitable carbon source for the growth of JC423 mycelium was fructose, the suitable nitrogen source was peptone, the suitable pH range was 9-10, and the suitable growth temperature was 25 ℃. Inorganic salts and vitamins had little effect on mycelial growth. The study clarified the nutritional requirements and conditions for the growth of wild F. filiformis, and laid a foundation for the development and utilization of F. filiformis.

Key words: Flammulina filiformis, strain isolation, ITS sequence, biological characteristic

中图分类号:

S646.15

冯连荣, 张妍, 赵鑫闻, 宋立志, 梁德军. 一株野生金针菇菌种的分离、鉴定与生物学特性研究[J]. 浙江农业学报, 2023, 35(5): 1088-1096.

FENG Lianrong, ZHANG Yan, ZHAO Xinwen, SONG Lizhi, LIANG Dejun. Isolation, identification and biological characteristics of a wild Flammulina filiformis strain[J]. Acta Agriculturae Zhejiangensis, 2023, 35(5): 1088-1096.

图/表 7

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配方 Formula	菌丝长满袋时间 Time takes for hyphae to fill the bag/d	采收期 Harvest period/d	菇体整齐度 Mushroom uniformity	平均单袋产量 Average bag yield/g	袋数 Number of bags	总产量 Total yield/g	生物学效率 Biological efficiency/%
1	38	70	整齐Neat	330.15 aA	42	13 688.5	65.18
2	40	70	较整齐Relatively neat	298.00 aAB	42	12 516.0	59.60
3	40	68	整Neat	148.44 cC	42	6 232.48	29.68
4	41	71	较整齐Relatively neat	230.84 bB	42	9 695.28	46.17

配方 Formula	菌丝长满袋时间 Time takes for hyphae to fill the bag/d	采收期 Harvest period/d	菇体整齐度 Mushroom uniformity	平均单袋产量 Average bag yield/g	袋数 Number of bags	总产量 Total yield/g	生物学效率 Biological efficiency/%
1	38	70	整齐Neat	330.15 aA	42	13 688.5	65.18
2	40	70	较整齐Relatively neat	298.00 aAB	42	12 516.0	59.60
3	40	68	整Neat	148.44 cC	42	6 232.48	29.68
4	41	71	较整齐Relatively neat	230.84 bB	42	9 695.28	46.17

处理 Treatment		菌丝长势 Hyphae growth	菌丝密度 Hyphael density	生长速度 Speed of growth
碳源Carbon source	果糖Fructose	+++	浓密Dense mycelium	11.52±1.14 aA
	甘露醇 Mannitol	++	稀疏Sparse	9.41±1.30 bAB
	可溶性淀粉 Soluble starch	+++	浓密Dense mycelium	7.95±1.66 bcBC
	蔗糖 Sucrose	++	较密Relatively dense	7.10±1.30 cdBC
	葡萄糖 Glucose	++	较密Relatively dense	6.72±1.75 cdBC
	对照CK	+	稀疏Sparse	5.25±0.33 dC
氮源Nitrogen source	蛋白胨 Peptone	+++	浓密Dense mycelium	12.56±0.31 aA
	酵母膏Ysast extract	++	浓密Dense mycelium	12.44±0.14 aA
	硫酸铵Ammonium sumlphate	++	较密Relatively dense	10.91±1.16 bAB
	硝酸铵Ammonium nitrate	++	较密Relatively dense	10.35±1.41 bcB
	尿素Urea	+++	浓密Dense mycelium	9.00±0.84 cB
	对照CK	+	稀疏Sparse	9.87±1.12 bcB
无机盐Inorganic salt	氯化钠 Sodium chloride	+++	浓密Dense mycelium	12.45±0.26 aA
	硫酸钙 Calcium sulfate	+++	浓密Dense mycelium	12.43±0.85 aA
	硫酸镁 Magnesium sulfate	+++	浓密Dense mycelium	12.21±0.21 aA
	磷酸二氢钾Potassium dihydrogen phosphate	+++	较密Relatively dense	11.54±0.57 aA
	对照CK	+++	较密Relatively dense	11.75±0.58 aA
维生素Vitamin	V_B3	+++	浓密Dense mycelium	8.17±1.22 aA
	V_B1	+++	浓密Dense mycelium	8.06±0.66 aA
	V_B12	+++	浓密Dense mycelium	8.02±0.72 aA
	V_B2	++	浓密Dense mycelium	7.64±1.22 aA
	V_B6	++	较密Relatively dense	6.95±1.22 aA
	对照CK	++	较密Relatively dense	7.43±1.38 aA
pH	4	+++	浓密Dense mycelium	8.43±0.86 bcBCD
	5	+++	浓密Dense mycelium	8.96±0.48 abABC
	6	++	较密Relatively dense	8.00±0.72 cdCD
	7	++	较密Relatively dense	7.45±0.38 dDE
	8	++	较密Relatively dense	6.59±0.34 eE
	9	+++	浓密Dense mycelium	9.34±0.50 aAB
	10	+++	浓密Dense mycelium	9.64±0.22 aA
温度Temperature/℃	5	++	稀疏Sparse	2.32±0.21 eE
	10	++	较密Relatively dense	5.66±0.77 cC
	15	++	较密Relatively dense	4.39±0.21 dD
	20	+++	较密Relatively dense	9.70±0.44 bB
	25	+++	浓密Dense mycelium	11.37±0.09 aA
	30	+++	较密Relatively dense	9.70±0.60 bB
	35	-	-	0 fF

处理 Treatment		菌丝长势 Hyphae growth	菌丝密度 Hyphael density	生长速度 Speed of growth
碳源Carbon source	果糖Fructose	+++	浓密Dense mycelium	11.52±1.14 aA
	甘露醇 Mannitol	++	稀疏Sparse	9.41±1.30 bAB
	可溶性淀粉 Soluble starch	+++	浓密Dense mycelium	7.95±1.66 bcBC
	蔗糖 Sucrose	++	较密Relatively dense	7.10±1.30 cdBC
	葡萄糖 Glucose	++	较密Relatively dense	6.72±1.75 cdBC
	对照CK	+	稀疏Sparse	5.25±0.33 dC
氮源Nitrogen source	蛋白胨 Peptone	+++	浓密Dense mycelium	12.56±0.31 aA
	酵母膏Ysast extract	++	浓密Dense mycelium	12.44±0.14 aA
	硫酸铵Ammonium sumlphate	++	较密Relatively dense	10.91±1.16 bAB
	硝酸铵Ammonium nitrate	++	较密Relatively dense	10.35±1.41 bcB
	尿素Urea	+++	浓密Dense mycelium	9.00±0.84 cB
	对照CK	+	稀疏Sparse	9.87±1.12 bcB
无机盐Inorganic salt	氯化钠 Sodium chloride	+++	浓密Dense mycelium	12.45±0.26 aA
	硫酸钙 Calcium sulfate	+++	浓密Dense mycelium	12.43±0.85 aA
	硫酸镁 Magnesium sulfate	+++	浓密Dense mycelium	12.21±0.21 aA
	磷酸二氢钾Potassium dihydrogen phosphate	+++	较密Relatively dense	11.54±0.57 aA
	对照CK	+++	较密Relatively dense	11.75±0.58 aA
维生素Vitamin	V_B3	+++	浓密Dense mycelium	8.17±1.22 aA
	V_B1	+++	浓密Dense mycelium	8.06±0.66 aA
	V_B12	+++	浓密Dense mycelium	8.02±0.72 aA
	V_B2	++	浓密Dense mycelium	7.64±1.22 aA
	V_B6	++	较密Relatively dense	6.95±1.22 aA
	对照CK	++	较密Relatively dense	7.43±1.38 aA
pH	4	+++	浓密Dense mycelium	8.43±0.86 bcBCD
	5	+++	浓密Dense mycelium	8.96±0.48 abABC
	6	++	较密Relatively dense	8.00±0.72 cdCD
	7	++	较密Relatively dense	7.45±0.38 dDE
	8	++	较密Relatively dense	6.59±0.34 eE
	9	+++	浓密Dense mycelium	9.34±0.50 aAB
	10	+++	浓密Dense mycelium	9.64±0.22 aA
温度Temperature/℃	5	++	稀疏Sparse	2.32±0.21 eE
	10	++	较密Relatively dense	5.66±0.77 cC
	15	++	较密Relatively dense	4.39±0.21 dD
	20	+++	较密Relatively dense	9.70±0.44 bB
	25	+++	浓密Dense mycelium	11.37±0.09 aA
	30	+++	较密Relatively dense	9.70±0.60 bB
	35	-	-	0 fF

一株野生金针菇菌种的分离、鉴定与生物学特性研究

Isolation, identification and biological characteristics of a wild Flammulina filiformis strain

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