金寨县桑黄的菌种鉴定与生长特性研究

doi:10.3969/j.issn.1004-1524.2021.12.03

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (12): 2234-2244.DOI: 10.3969/j.issn.1004-1524.2021.12.03

金寨县桑黄的菌种鉴定与生长特性研究

程安东(), 汪本勤^*()

六安职业技术学院,安徽六安 237158

收稿日期:2020-12-15 出版日期:2021-12-25 发布日期:2022-01-10
通讯作者: 汪本勤
作者简介:* 汪本勤,E-mail: 870772098@qq.com
程安东(1991—),男,安徽六安人,硕士,讲师,主要从事植物病理学研究。E-mail: 441683477@qq.com
基金资助:
安徽省教育厅自然科学重点研究项目(KJ2019A1062)

Identification and growth characteristics of Sanghuang in Jinzhai County

CHENG Andong(), WANG Benqin^*()

Lu’an Vocational Technical College, Lu’an 237158, China

Received:2020-12-15 Online:2021-12-25 Published:2022-01-10
Contact: WANG Benqin

摘要/Abstract

摘要：

针对皖西大别山金寨县永福桑黄菌种植合作社种植的桑黄菌开展研究,通过现代分子生物学技术对其18S rDNA 基因ITS区段进行克隆测序和序列特征分析,依据ITS序列分析结果将菌种鉴定为瓦宁木层孔菌(Sanghuangporus vaninii);对JS-1的基础培养基进行筛选,结果显示,PDA培养基最适合其生长;测定了JS-1菌株的生长特性,结果显示,温度在5~35 ℃区间内菌丝均可生长,最适生长温度为28 ℃;pH在4~10范围内菌丝均可生长,pH为7时最适合菌丝生长。以菌丝生长情况为指标筛选碳源和氮源,碳源为葡萄糖的培养基上菌丝生长速度最快且长势浓密,认为葡萄糖为其生长最适碳源;以马铃薯浸汁为氮源的培养基虽然生长速度略慢于蛋白胨和玉米粉,但菌落长势最为茂盛,且最早出现黄褐色软木状环纹,认为马铃薯浸汁为其生长最适氮源;在以葡萄糖为碳源,马铃薯浸汁为氮源的基础上,通过L₉(3⁴)正交试验得到JS-1菌株菌丝生长速度最快的培养条件为葡萄糖20 g·L^-1,马铃薯100 g·L^-1,pH为8,培养温度30 ℃;测定菌丝的致死温度为45 ℃。

关键词: 桑黄, 菌种鉴定, 培养基, 生长条件

Abstract:

In this study, researchers conducted a research on the strains of Sanghuang planted by Yongfu Plantation Cooperation of Jinzhai Country in Dabie Mountain Area and identified the strain as Sanghuang vaninii by cloning and sequencing the ITS region of its 18S rDNA gene using modern molecular biology techniques. Screening of the basal medium for JS-1 showed that PDA medium was the most suitable for the growth of JS-1. The growth characteristics of JS-1 strain were determined, and the results showed that the mycelium could grow in the temperature range of 5-35 ℃, and the optimum growth temperature was 28 ℃. The mycelium could grow in the pH range of 4-10, while the pH value of 7 was the optimal for the growth of mycelium. By taking the mycelium growth as an indicator to screen the carbon and nitrogen sources, it was found that the medium had the fastest and dense mycelium growth when glucose was used as the carbon source. Thus, glucose was considered the most suitable carbon source. When potato juice was used as the nitrogen source of the medium, despite of the slightly slower growth rate than peptone and corn flour, the colony growth was the most luxuriant, with the earliest appearance of yellow-brown cork-like ring. Thus, potato juice was considered the most suitable nitrogen source for its growth.On the basis of glucose as carbon source and potato extract as nitrogen source, L₉ (3 ⁴) orthogonal test showed that the optimal growth conditions of JS-1 strain were glucose 20 g·L^-1 and potato 100 g·L^-1, pH 8 and culture temperature 30 ℃. The mycelium lethal temperature was 45 ℃.

Key words: Sanghuang, species identification, medium, growth conditions

中图分类号:

S567.3⁺9

程安东, 汪本勤. 金寨县桑黄的菌种鉴定与生长特性研究[J]. 浙江农业学报, 2021, 33(12): 2234-2244.

CHENG Andong, WANG Benqin. Identification and growth characteristics of Sanghuang in Jinzhai County[J]. Acta Agriculturae Zhejiangensis, 2021, 33(12): 2234-2244.

图/表 14

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基础培养基 Medium	菌丝生长速度 Mycelium growth rate/ (mm·d^-1)	菌丝表面形态特征 Surface morphology of mycelial	菌丝长势 Mycelial growth vigor
木生菌标准培养基 Wood fungus standard medium	6.89±0.09 a	原基凸起一般,新生白色菌丝较多 Primordia protruding moderate. Large range of newly grown white mycelia	浓密 Thick
玉米粉综合培养基 Corn flour composite medium	6.32±0.07 b	原基凸起最小,新生白色色菌丝稀少 Primordia protruding the minimum Small range of newly grown white mycelia	稀疏 Thin
PDA培养基 Potato composite medium	6.14±0.12 b	原基凸起高,新生乳白色菌丝较多 Primordia protruding high Large range of newly grown white mycelia	浓密茂盛 Thick and lush

基础培养基 Medium	菌丝生长速度 Mycelium growth rate/ (mm·d^-1)	菌丝表面形态特征 Surface morphology of mycelial	菌丝长势 Mycelial growth vigor
木生菌标准培养基 Wood fungus standard medium	6.89±0.09 a	原基凸起一般,新生白色菌丝较多 Primordia protruding moderate. Large range of newly grown white mycelia	浓密 Thick
玉米粉综合培养基 Corn flour composite medium	6.32±0.07 b	原基凸起最小,新生白色色菌丝稀少 Primordia protruding the minimum Small range of newly grown white mycelia	稀疏 Thin
PDA培养基 Potato composite medium	6.14±0.12 b	原基凸起高,新生乳白色菌丝较多 Primordia protruding high Large range of newly grown white mycelia	浓密茂盛 Thick and lush

θ/℃	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
5	0.71±0.05 i	稀疏Thin
10	1.22±0.2 h	稀疏Thin
15	1.82±0.2 g	稀疏Thin
20	3.86±0.06 e	浓密Thick
25	4.12±0.09 d	浓密Thick
θ/℃	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
26	4.97±0.02 c	浓密茂盛Thick and lush
27	5.53±0.07 b	浓密茂盛Thick and lush
28	6.14±0.02 a	浓密茂盛Thick and lush
29	6.09±0.12 a	浓密茂盛Thick and lush
30	5.81±0.12 c	浓密Thick
35	2.42±0.05 f	稀疏Thin
40	-	死亡Death

θ/℃	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
5	0.71±0.05 i	稀疏Thin
10	1.22±0.2 h	稀疏Thin
15	1.82±0.2 g	稀疏Thin
20	3.86±0.06 e	浓密Thick
25	4.12±0.09 d	浓密Thick
θ/℃	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
26	4.97±0.02 c	浓密茂盛Thick and lush
27	5.53±0.07 b	浓密茂盛Thick and lush
28	6.14±0.02 a	浓密茂盛Thick and lush
29	6.09±0.12 a	浓密茂盛Thick and lush
30	5.81±0.12 c	浓密Thick
35	2.42±0.05 f	稀疏Thin
40	-	死亡Death

pH	菌丝生长速度 Mycelium growth rate/ (mm·d^-1)	菌丝长势 Mycelial growth vigor
3	-	死亡Death
4	3.26±0.18 e	稀疏Thin
5	5.42±0.21 d	浓密Thick
6	6.04±0.06 b	浓密茂盛Thick and lush
7	6.12±0.02 a	浓密茂盛Thick and lush
8	5.93±0.12 c	浓密茂盛Thick and lush
9	3.31±0.09 e	浓密Thick
10	1.71±0.02 f	稀疏Thin
11	-	死亡Death
12	-	死亡Death

金寨县桑黄的菌种鉴定与生长特性研究

Identification and growth characteristics of Sanghuang in Jinzhai County

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图/表 14

参考文献 23

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水平 Level	因素Factor
水平 Level	葡萄糖 Glucose/(g·L^-1)	马铃薯 Potato/(g·L^-1)	θ/℃	pH
1	10	100	26	6
2	20	200	28	7
3	30	300	30	8

碳源 Carbon sources	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
葡萄糖 Glucose	6.12±0.12 a	浓密茂盛 Thick and lushn
蔗糖 Sucrose	5.91±0.02 a	浓密茂盛 Thick and lush
半乳糖 Galactose	5.88±0.06 ab	浓密茂盛 Thick and lush
可溶性淀粉 Soluble starch	5.53±0.18 b	浓密茂盛 Thick and lushk
麦芽糖 Maltose	5.51±0.06 b	浓密茂盛 Thick and lushk
甘露糖Mannose	3.51±0.03 c	稀疏Thin
纤维乙糖 Fiber B suger	2.82±0.05 d	稀疏 Thin

氮源 Nitrogen sources	菌丝生长速度 Mycelium growth rate/(mm·d^-1)	菌丝长势 Mycelial growth vigor
蛋白胨Peptone	6.81±0.08 a	浓密Thick
玉米粉浸汁 Corn flour juice	6.32±0.02 b	稀疏 Thin
马铃薯浸汁 Potato juice	6.12±0.06 b	浓密茂盛 Thick and lush
牛肉浸膏Beef extract	5.15±0.14 c	浓密Thick
硝酸钠Sodium nitrate	5.03±0.11 c	稀疏Thin
酵母浸膏Yeast extract	4.13±0.26 d	稀疏Thin

编号 Number	(A)葡萄糖 Glucose/(g·L^-1)	(B)马铃薯 Potato/(g·L^-1)	(C) θ/℃	(D)pH	菌丝生长速度 Mycelia growth rate/(mm·d^-1)
1	1	1	1	1	4.81
2	1	2	2	2	5.27
3	1	3	3	3	5.61
4	2	1	3	2	6.05
5	2	2	1	3	5.47
6	2	3	2	1	4.43
7	3	1	2	3	5.17
8	3	2	3	1	5.09
9	3	3	1	2	4.59
K₁	5.23	5.34	4.96	4.78	—
K₂	5.32	5.28	4.96	5.30	—
K₃	4.95	4.88	5.58	5.42	—
R	0.37	0.46	0.62	0.64	—

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