北苍术根腐病病原鉴定及生物学特性研究

doi:10.13304/j.nykjdb.2021.0468

中国农业科技导报 ›› 2022, Vol. 24 ›› Issue (5): 137-144.DOI: 10.13304/j.nykjdb.2021.0468

• 动植物健康 • 上一篇

北苍术根腐病病原鉴定及生物学特性研究

霍佳欢¹(), 温晓蕾¹^,²(), 李双民³, 冯丽娜¹, 兰淑慧¹, 董立新⁴, 郭思柔¹, 栗佳宁¹, 王建华¹, 齐慧霞¹()

^1.河北科技师范学院农学与生物科技学院, 河北省作物逆境生物学重点实验室, 河北秦皇岛 066600
^2.河北农业大学植物保护学院, 河北保定 071002
^3.河北省昌黎县职业技术教育中心, 河北秦皇岛 066600
^4.秦皇岛市植保植检站, 河北秦皇岛 066000

收稿日期:2021-06-08 接受日期:2021-09-22 出版日期:2022-05-15 发布日期:2022-06-06
通讯作者: 齐慧霞
作者简介:霍佳欢E-mail：2469634717@qq.com
温晓蕾E-mail：xiaoleiwen@sina.com第一联系人：霍佳欢和温晓蕾为共同第一作者。

Identification and Biological Characteristics of the Pathogen Causing Root Rot of Atractylodes chinensis

Jiahuan HUO¹(), Xiaolei WEN¹^,²(), Shuangmin LI³, Lina FENG¹, Shuhui LAN¹, Lixin DONG⁴, Sirou GUO¹, Jianing LI¹, Jianhua WANG¹, Huixia QI¹()

^1.Hebei Key Laboratory of Crop Stress Biology，College of Agronomy and Biotechnology，Hebei Normal University of Science ＆ Technology，Hebei Qinhuangdao 066600，China
^2.College of Plant Protection，Hebei Agricultural University，Hebei Baoding 071001，China
^3.Hebei Changli vocational and Technical Education Center，Hebei Changli 066600，China
^4.Qinhuangdao Plant Protection and Inspection Station，Hebei Qinhuangdao 066600，China

Received:2021-06-08 Accepted:2021-09-22 Online:2022-05-15 Published:2022-06-06
Contact: Huixia QI

摘要/Abstract

摘要：

根腐病严重影响北苍术生产，育苗期至成株期均有发生。为明确引起河北省秦皇岛地区北苍术根腐病病原菌，采用组织分离法获得致病菌菌株，并通过形态学及分子生物学相结合的方法对其进行鉴定，同时分析温度、氮源、碳源对病原菌生长的影响以及菌丝的致死温度。结果表明，引起秦皇岛地区北苍术根腐病的病原菌为尖孢镰刀菌Fusarium oxysporum （MK849925），该病原菌在PDA培养基上菌落近圆形，呈辐射状生长，似绒毛，粉紫色，较干燥。菌丝侧生分支，有隔膜；分生孢子均无色，大型分生孢子呈渐尖型镰刀状，有1~5个分隔，小型分生孢子多为卵形，具1~2个分隔，厚垣孢子圆形、无色。其致病力较强，接种后10 d病部呈深褐色、失水干枯。该病原菌较适宜的生长条件为25 ℃、以酵母浸粉为基础氮源的培养基，培养5 d，菌落直径可达7.17 cm；在不同碳源中，对葡萄糖和蔗糖的利用率较高，而对于可溶性淀粉的利用率较低。菌丝在75 ℃恒温条件下处理10 min后失去生命力。

关键词: 北苍术, 根腐病, 尖孢镰刀菌, 病原鉴定, 生物学特性

Abstract:

Root rot seriously affects the production of Atractylodes chinensi，which occurrs from seedling to mature periods. In order to clarify the pathogen causing Atractylodis chinensi root rot in Qinhuangdao area of Hebei Province， tissue isolation， morphological and molecular biological identification were used to isolate and identify the pathogen， and the pathogenicity was identified. At the same time， the effects of temperature， nitrogen source and carbon source on the growth of the pathogen and the lethal temperature of the mycelium were analyzed. The results showed that the pathogen causing root rot of Atractylodis chinensi in Qinhuangdao area was Fusarium oxysporum （MK849925）. The colony of the pathogen on PDA medium was nearly round， radial， villous， pink purple and dry. Mycelium lateral branched with septum， the conidia were all colorless and the large conidia were acuminate sickle with 1~5 septations， the small conidia were mostly oval with 1~2 septations. The chlamydospores were round and colorless. Its pathogenicity was strong， and the disease showed dark brown dehydration dry after inoculation for 10 d. The pathogen grew better at 25 ℃ and on the medium with yeast extract powder as the nitrogen source. After 5 d culture， the colony diameter reached 7.17 cm. Among different carbon sources， the utilization rates of glucose and sucrose were high， while the utilization rate of soluble starch was low. The mycelium lost its vitality after treated at 75 ℃ for 10 min.

Key words: Atractylodes chinensis, root rot, Fusarium oxysporum, pathogen identification, biological characteristic

中图分类号:

S432

霍佳欢, 温晓蕾, 李双民, 冯丽娜, 兰淑慧, 董立新, 郭思柔, 栗佳宁, 王建华, 齐慧霞. 北苍术根腐病病原鉴定及生物学特性研究[J]. 中国农业科技导报, 2022, 24(5): 137-144.

Jiahuan HUO, Xiaolei WEN, Shuangmin LI, Lina FENG, Shuhui LAN, Lixin DONG, Sirou GUO, Jianing LI, Jianhua WANG, Huixia QI. Identification and Biological Characteristics of the Pathogen Causing Root Rot of Atractylodes chinensis[J]. Journal of Agricultural Science and Technology, 2022, 24(5): 137-144.

图/表 10

图1 北苍术根腐病症状

Fig. 1 Symptoms of Atractylodes chinensi root rot

图2 北苍术根腐病病原菌形态特征

Fig. 2 Morphological characteristics of root rot pathogens of Atractylodes chinensi

图3 CZG1菌株ITS序列系统发育树

Fig. 3 Phylogenetic tree of CZG1 strain based on ITS sequence

图4 北苍术根腐病室内接种发病症状

Fig 4 Symptoms of Atractylodes chinensi Root Rot by indoor inoculation

图5 温度对菌落生长的影响注：图中不同小写字母表示在P<0.05水平差异显著。

Fig. 5 Effect of temperature on colony growthNote：Different lowercase letters in the figure indicate significant differences at P<0.05 level.

表1 碳源对菌落直径生长的影响

Table 1 Influence of carbon source on the growth of colony diameter

碳源 Carbon source	菌落直径Colony diameter/cm					平均生长速率 Average growth rate/（cm·d^-1）
碳源 Carbon source	1 d	2 d	3 d	4 d	5 d	平均生长速率 Average growth rate/（cm·d^-1）
葡萄糖Glucose	0.85±0.05 bc	2.60±0.00 a	4.17±0.06 b	5.60±0.00 a	6.75±0.00 a	1.350
蔗糖Sucrose	0.97±0.06 a	2.45±0.00 b	3.97±0.06 c	5.30±0.00 b	6.70±0.05 a	1.340
果糖 Fructose	0.82±0.03 c	2.40±0.00 c	3.75±0.05 c	4.93±0.03 c	6.32±0.03 b	1.264
肌醇 Inositol	0.92±0.03 ab	2.63±0.03 a	4.20±0.00 a	5.57±0.06 a	6.28±0.03 b	1.256
甘露醇 Mannitol	0.87±0.03 bc	2.13±0.03 d	3.50±0.00 d	4.87±0.00 d	6.28±0.08 b	1.256
木糖Xxylose	0.72±0.06 d	2.10±0.00 de	3.50±0.00 de	4.78±0.03 e	6.03±0.03 c	1.206
乳糖Lactose	0.90±0.00 ab	2.60±0.00 a	3.77±0.08 b	4.68±0.03 f	5.47±0.06 d	1.094
可溶性淀粉 Soluble starch	0.53±0.03 e	2.07±0.06 e	3.23±0.10 e	4.72±0.03 f	5.38±0.14 d	1.076

图6 不同碳源对菌落生长的影响注：A~H分别葡萄糖、蔗糖、果糖、肌醇、甘露醇、木糖、乳糖、可溶性淀粉。

Fig. 6 Effect of different carbon sources on colony growthNote： A~H are glucose， sucrose， fructose， inositol， mannitol， xylose， lactose， soluble starch， respectively.

表2 氮源对菌落直径生长的影响

Table 2 Influence of nitrogen source on the growth of colony diameter

氮源 Nitrogen source	菌落直径Colony diameter/cm					平均生长速率 Average growth rate/（cm·d^-1）
氮源 Nitrogen source	1 d	2 d	3 d	4 d	5 d	平均生长速率 Average growth rate/（cm·d^-1）
酵母浸粉 Yeast extract	0.77±0.33 d	2.60±0.00 b	4.47±0.08 a	5.67±0.23 a	7.17±0.08 a	1.434
硝酸钠 Sodium nitrate	0.98±0.03 a	2.62±0.03 ab	4.22±0.03 c	5.53±0.03 ab	6.80±0.00 b	1.360
牛肉膏 Beef extract	0.83±0.03 c	2.65±0.00 a	4.35±0.05 b	5.40±0.05 bc	6.77±0.03 b	1.354
硝酸钾 Potassium nitrate	0.77±0.03 d	2.5±0.00 c	4.02±0.03 d	5.35±0.05 c	6.75±0.05 b	1.350
蛋白胨 Peptone	0.62±0.03 e	2.35±0.00 d	3.97±0.03 d	5.40±0.00 bc	6.70±0.00 b	1.340
尿素 Urea	0.63±0.03 e	2.02±0.06 f	3.33±0.10 f	4.67±0.08 d	6.15±0.00 c	1.230
氯化铵 Ammonium chloride	0.95±0.00 ab	2.37±0.03 d	3.63±0.08 e	3.77±0.10 e	3.68±0.23 d	0.736
硫酸铵 Ammonium sulfate	0.93±0.03 b	3.05±0.00 e	3.10±0.00 g	3.37±0.03 f	3.53±0.03 d	0.706

图7 不同氮源对菌落生长的影响注：A~H分别为酵母浸粉、硝酸钠、牛肉膏、硝酸钾、蛋白胨、尿素、氯化铵、硫酸铵。

Fig. 7 Effect of different nitrogen sources on colony growthNote： A~H are yeast extract， sodium nitrate， beef extract， potassium nitrate， peptone， urea，ammonium chloride， ammonium sulfate， respectively.

图8 北苍术根腐病菌致死温度注：A~J分别为40、45、50、55、60、65、70、75、80、85℃。

Fig. 8 Lethal temperature of Atractylodes chinensi root rot pathogenNote： A~J are 40， 45， 50， 55， 60， 65， 70， 75， 80， 85℃， respectively.

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北苍术根腐病病原鉴定及生物学特性研究

Identification and Biological Characteristics of the Pathogen Causing Root Rot of Atractylodes chinensis

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