设施韭菜根腐病致病镰孢菌鉴定及生物学特性研究

doi:10.16420/j.issn.0513-353x.2021-0051

摘要/Abstract

摘要：

韭菜根腐病是设施连作多年生韭菜的重要病害之一。为明确辽宁省设施韭菜根腐病的病原菌种类,从韭菜感病根系分离纯化获得45株镰孢菌,结合形态学特征及EF-1a、Histone3、IGS基因片段序列分析结果,鉴定为尖孢镰孢菌（Fusarium oxysporum）、层出镰孢菌（F. proliferatum）和腐皮镰孢菌（F. solani）。经柯赫氏法则证病,确定均可引起韭菜根腐病。对病原菌的生物学特性研究结果表明,3种镰孢菌最适生长温度为30 ℃左右;最适pH 7 ~ 8;黑暗条件有利于病原菌菌丝生长;菌丝的致死温度为75 ℃10 min,分生孢子致死温度为56 ℃10 min。病原菌对碳源鼠李糖以及氮源硝酸钠、甘氨酸利用率最高。综上,尖孢镰孢菌、层出镰孢菌和腐皮镰孢菌是辽宁省设施韭菜根腐病的主要病原菌,其中尖孢镰孢菌分布最广,致病力最强,属于优势致病菌。

关键词: 韭菜, 根腐病, 镰孢菌, 鉴定, 生物学特性

Abstract:

Chinese chive（Allium tuberosum）root rot is one of the main diseases of continuous cropping chive in greenhouse. In order to explore the main pathogens of chive root rot disease in Liaoning Province,45 isolates of the genus Fusarium were generated from diseased chive roots and finally identified as F. oxysporum,F. proliferatum and F. solani based on their morphological characteristics and EF-1a,Histone3,rDNA-IGS sequences analysis. These three Fusarium species were confirmed to be the pathogenic agents of the disease according to Koch’s rule. Studies on the biological characteristics of the three pathogens showed that the most suitable growth temperature is about 30 ℃;the optimal growth pH value are at pH 7-8;full dark is conducive to the mycelial growth of the three species;their mycelia can be killed at 75 ℃ for 10 min;their conidia can not germinate at 56 ℃ for 10 min. These pathogens have the highest utilization rate of rhamnose as carbon source and sodium nitrate and glycine as nitrogen source. In summary,F. oxysporum,F. proliferatum and F. solani are the principal pathogenic fungi causing root rot of A. tuberosum,of which F. oxysporum is the most dominant pathogenic species because of its wide distribution and stronger pathogenicity.

Key words: Chinese chive, root rot, Fusarium, identification, biological characteristics

中图分类号:

S633.3

高晓梅, 吕国忠, 李杨, 敖静, 刘晓辉, 孙玉禄. 设施韭菜根腐病致病镰孢菌鉴定及生物学特性研究[J]. 园艺学报, 2022, 49(4): 893-906.

GAO Xiaomei, LÜ Guozhong, LI Yang, AO Jing, LIU Xiaohui, SUN Yulu. Identification of the Pathogens Causing Root Rot on Allium tuberosum and Study on Their Biological Characteristics[J]. Acta Horticulturae Sinica, 2022, 49(4): 893-906.

图/表 11

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病原菌Pathogens	发病率/% Morbidity	病情指数Disease index
尖孢镰孢菌F. oxysporum	96.88 ± 1.46 ab	87.01 ± 1.79 a
层出镰孢菌F. proliferatum	95.71 ± 1.21 b	80.83 ± 2.08 b
腐皮镰孢菌F. solani	98.90 ± 0.33 a	86.18 ± 1.51 a

病原菌Pathogens	发病率/% Morbidity	病情指数Disease index
尖孢镰孢菌F. oxysporum	96.88 ± 1.46 ab	87.01 ± 1.79 a
层出镰孢菌F. proliferatum	95.71 ± 1.21 b	80.83 ± 2.08 b
腐皮镰孢菌F. solani	98.90 ± 0.33 a	86.18 ± 1.51 a

试验 Test	培养条件 Culture condition	菌落直径/mm Colony diameter
试验 Test	培养条件 Culture condition	尖孢镰孢菌 F. oxysporum	层出镰孢菌 F. proliferatum	腐皮镰孢菌 F. solani
碳源Carbon source	蔗糖Sucrose	83.94 ± 0.50 a	69.86 ± 1.78 c	57.75 ± 0.40 c
	菊糖Inulin	83.85 ± 1.20 a	76.21 ± 0.53 b	58.07 ± 0.22 c
	葡萄糖Glucose	84.64 ± 2.39 a	67.48 ± 1.03 c	56.66 ± 0.15 c
	山梨糖Sorbose	68.96 ± 0.76 c	53.79 ± 1.98 e	51.26 ± 0.75 d
	木糖Xylose	68.71 ± 1.60 c	62.91 ± 1.77 d	57.72 ± 1.02 c
	乳糖Lactose	83.74 ± 0.50 a	76.65 ± 1.23 b	60.61 ± 1.18 b
	果糖Fructose	75.96 ± 1.28 b	66.64 ± 3.28 c	57.08 ± 2.88 c
	鼠李糖Rhamnose	82.53 ± 0.87 a	82.02 ± 2.36 a	65.21 ± 1.21 a
	甘露糖Mannose	78.38 ± 3.62 b	74.52 ± 1.07 b	58.47 ± 0.66 c
氮源Nitrogen source	NH₄Cl	77.01 ± 1.71 a	58.51 ± 1.55 c	51.01 ± 1.80 d
	NaNO₃	74.32 ± 0.76 a	62.74 ± 2.55 b	59.68 ± 0.49 b
	KNO₃	68.42 ± 2.47 bcd	66.70 ± 1.39 a	54.82 ± 2.74 c
	酒石酸铵Ammonium tartrate	58.06 ± 3.62 e	48.73 ± 1.55 d	55.22 ± 1.40 c
	甘氨酸Glycine	72.57 ± 1.97 ab	56.06 ± 0.28 c	63.22 ± 2.90 a
	胱氨酸Cystine	65.15 ± 3.84 d	65.64 ± 0.81 ab	59.86 ± 1.54 b
	丙氨酸Alanine	66.22 ± 4.71 cd	56.94 ± 1.32 c	61.47 ± 2.09 a
	CK（PDA）	71.67 ± 2.99 abc	56.92 ± 4.16 c	61.40 ± 0.32 a
温度/℃ Temperature	10	15.19 ± 0.35 e	16.04 ± 0.37 f	12.29 ± 0.32 e
	15	42.66 ± 0.77 d	43.54 ± 0.83 e	29.66 ± 1.09 d
	20	57.31 ± 2.04 c	55.81 ± 0.41 d	43.04 ± 1.56 c
	25	72.46 ± 2.10 b	71.26 ± 2.07 b	52.25 ± 1.17 b
	30	78.39 ± 3.83 a	74.82 ± 1.61 a	56.86 ± 1.06 a
	35	71.74 ± 1.83 b	70.66 ± 1.75 b	54.47 ± 1.83 ab
	40	58.16 ± 3.70 c	67.06 ± 1.41 c	45.09 ± 2.41 c
试验 Test	培养条件 Culture condition	菌落直径/mm Colony diameter
试验 Test	培养条件 Culture condition	尖孢镰孢菌 F. oxysporum	层出镰孢菌 F. proliferatum	腐皮镰孢菌 F. solani
pH	4	41.91 ± 0.91 c	44.32 ± 0.26 e	30.95 ± 3.11 e
	5	56.01 ± 0.32 b	55.27 ± 1.54 d	47.47 ± 1.24 cd
	6	61.13 ± 0.24 a	72.11 ± 0.28 b	49.40 ± 0.30 bcd
	7	62.56 ± 2.27 a	76.07 ± 2.72 a	51.52 ± 0.78 ab
	8	60.81 ± 4.95 a	68.36 ± 1.30 c	54.80 ± 2.09 a
	9	60.75 ± 3.20 a	67.35 ± 0.13 c	52.27 ± 1.76 ab
	10	53.74 ± 0.73 b	65.87 ± 1.08 c	50.74 ± 1.67 bc
	11	52.46 ± 0.34 c	44.31 ± 2.18 e	46.59 ± 3.68 d
光照Illumination	全光照 Full illumination	70.28 ± 1.44 c	67.42 ± 3.54 b	50.17 ± 3.30 b
	全黑暗 Full darkness	78.34 ± 0.84 a	75.27 ± 0.81 a	60.02 ± 1.36 a
	12 h光照/12 h黑暗 12 h illumination/12 h darkness	73.28 ± 1.34 b	70.54 ± 1.51 ab	56.86 ± 0.86 a

试验 Test	培养条件 Culture condition	菌落直径/mm Colony diameter
试验 Test	培养条件 Culture condition	尖孢镰孢菌 F. oxysporum	层出镰孢菌 F. proliferatum	腐皮镰孢菌 F. solani
碳源Carbon source	蔗糖Sucrose	83.94 ± 0.50 a	69.86 ± 1.78 c	57.75 ± 0.40 c
	菊糖Inulin	83.85 ± 1.20 a	76.21 ± 0.53 b	58.07 ± 0.22 c
	葡萄糖Glucose	84.64 ± 2.39 a	67.48 ± 1.03 c	56.66 ± 0.15 c
	山梨糖Sorbose	68.96 ± 0.76 c	53.79 ± 1.98 e	51.26 ± 0.75 d
	木糖Xylose	68.71 ± 1.60 c	62.91 ± 1.77 d	57.72 ± 1.02 c
	乳糖Lactose	83.74 ± 0.50 a	76.65 ± 1.23 b	60.61 ± 1.18 b
	果糖Fructose	75.96 ± 1.28 b	66.64 ± 3.28 c	57.08 ± 2.88 c
	鼠李糖Rhamnose	82.53 ± 0.87 a	82.02 ± 2.36 a	65.21 ± 1.21 a
	甘露糖Mannose	78.38 ± 3.62 b	74.52 ± 1.07 b	58.47 ± 0.66 c
氮源Nitrogen source	NH₄Cl	77.01 ± 1.71 a	58.51 ± 1.55 c	51.01 ± 1.80 d
	NaNO₃	74.32 ± 0.76 a	62.74 ± 2.55 b	59.68 ± 0.49 b
	KNO₃	68.42 ± 2.47 bcd	66.70 ± 1.39 a	54.82 ± 2.74 c
	酒石酸铵Ammonium tartrate	58.06 ± 3.62 e	48.73 ± 1.55 d	55.22 ± 1.40 c
	甘氨酸Glycine	72.57 ± 1.97 ab	56.06 ± 0.28 c	63.22 ± 2.90 a
	胱氨酸Cystine	65.15 ± 3.84 d	65.64 ± 0.81 ab	59.86 ± 1.54 b
	丙氨酸Alanine	66.22 ± 4.71 cd	56.94 ± 1.32 c	61.47 ± 2.09 a
	CK（PDA）	71.67 ± 2.99 abc	56.92 ± 4.16 c	61.40 ± 0.32 a
温度/℃ Temperature	10	15.19 ± 0.35 e	16.04 ± 0.37 f	12.29 ± 0.32 e
	15	42.66 ± 0.77 d	43.54 ± 0.83 e	29.66 ± 1.09 d
	20	57.31 ± 2.04 c	55.81 ± 0.41 d	43.04 ± 1.56 c
	25	72.46 ± 2.10 b	71.26 ± 2.07 b	52.25 ± 1.17 b
	30	78.39 ± 3.83 a	74.82 ± 1.61 a	56.86 ± 1.06 a
	35	71.74 ± 1.83 b	70.66 ± 1.75 b	54.47 ± 1.83 ab
	40	58.16 ± 3.70 c	67.06 ± 1.41 c	45.09 ± 2.41 c
试验 Test	培养条件 Culture condition	菌落直径/mm Colony diameter
试验 Test	培养条件 Culture condition	尖孢镰孢菌 F. oxysporum	层出镰孢菌 F. proliferatum	腐皮镰孢菌 F. solani
pH	4	41.91 ± 0.91 c	44.32 ± 0.26 e	30.95 ± 3.11 e
	5	56.01 ± 0.32 b	55.27 ± 1.54 d	47.47 ± 1.24 cd
	6	61.13 ± 0.24 a	72.11 ± 0.28 b	49.40 ± 0.30 bcd
	7	62.56 ± 2.27 a	76.07 ± 2.72 a	51.52 ± 0.78 ab
	8	60.81 ± 4.95 a	68.36 ± 1.30 c	54.80 ± 2.09 a
	9	60.75 ± 3.20 a	67.35 ± 0.13 c	52.27 ± 1.76 ab
	10	53.74 ± 0.73 b	65.87 ± 1.08 c	50.74 ± 1.67 bc
	11	52.46 ± 0.34 c	44.31 ± 2.18 e	46.59 ± 3.68 d
光照Illumination	全光照 Full illumination	70.28 ± 1.44 c	67.42 ± 3.54 b	50.17 ± 3.30 b
	全黑暗 Full darkness	78.34 ± 0.84 a	75.27 ± 0.81 a	60.02 ± 1.36 a
	12 h光照/12 h黑暗 12 h illumination/12 h darkness	73.28 ± 1.34 b	70.54 ± 1.51 ab	56.86 ± 0.86 a