新疆万寿菊枯萎病病原菌鉴定及生物学特性研究

doi:10.16420/j.issn.0513-353x.2022-0999

园艺学报 ›› 2023, Vol. 50 ›› Issue (12): 2723-2734.doi: 10.16420/j.issn.0513-353x.2022-0999

新疆万寿菊枯萎病病原菌鉴定及生物学特性研究

努尔斯曼姑丽·艾海提¹, 阿依加玛丽·阿卜杜扎伊尔¹, 孙永民², 韩文杰³, 郭庆元¹^,^*(), 王丽丽¹^,^*()

¹ 新疆农业大学农学院，农业农村部西北荒漠绿洲农林外来入侵生物防控重点实验室，农林有害生物监测与安全防控重点实验室，乌鲁木齐 830052
² 新疆维吾尔自治区花卉业管理中心，乌鲁木齐 830052
³ 晨光生物科技集团莎车有限公司，新疆莎车 844700

收稿日期:2023-04-13 修回日期:2023-10-15 出版日期:2023-12-25 发布日期:2023-12-29
通讯作者:
*（E-mail：guoqingyuan3009@sina.com，
1136862740@qq.com）
基金资助:
新疆维吾尔自治区花卉协会横向课题(2019年); 新疆农业大学大学生创新项目(dxscx2022008)

Identification and Biological Characteristics of the Pathogens Causing Marigold Wilt in Xinjiang

Nursimangul·Aihaiti¹, Aigamarie·Abdulzaire¹, SUN Yongmin², HAN Wenjie³, GUO Qingyuan¹^,^*(), WANG Lili¹^,^*()

¹ Key Laboratory of Prevention and Control of Invasive Alien Species in Agriculture & Forestry of the North-western Desert Oasis（Co-construction by Ministry and Province），Ministry of Agriculture and Rural Affairs，Key Laboratory of the Pest Monitoring and Safety Control of Crops and Forests，Agricultural College，Xinjiang Agricultural University，Urumqi 830052，China
² Flower Industry Management Center of Xinjiang Uygur Autonomous Region，Urumqi 830052，China
³ Chenguang Biotechnology Group Shache Co., Ltd.，Shache，Xinjiang 844700，China

Received:2023-04-13 Revised:2023-10-15 Published:2023-12-25 Online:2023-12-29

摘要/Abstract

摘要：

为明确新疆万寿菊枯萎病病菌种类和生物学特性，对其开展组织分离、单孢纯化、致病性测定和形态学结合系统发育学（ITS和EF-1α序列）的菌种鉴定以及生物学特性测定。结果表明共有4种病菌，藤仓镰刀菌（Fusarium fujikuroi）、茄腐镰刀菌（Fusarium solani）、木贼镰刀菌（Fusarium equiseti）和黄色镰刀菌（Fusarium culmorum）。菌种最适生长温度均为25 ℃;最适pH为6 ~ 7。藤仓镰刀菌在全黑暗条件下，其余病原菌在全光照条件下利于菌丝生长。藤仓镰刀菌在碳源乳糖，氮源硝酸钾或PDA和PSA培养基;木贼镰刀菌在碳源乳糖、氮源酵母浸粉或PDA培养基;茄腐镰刀菌在碳源乳糖、氮源蛋白胨或察氏培养基;黄色镰刀菌在碳源可溶性淀粉、氮源酵母浸粉和氯化铵或PSA、PDA和CA培养基的条件下，最利于各菌丝生长。致死温度分别是64、63、63和52 ℃。其中，茄腐镰刀菌分离频率最高为68%，是新疆万寿菊枯萎病的主要病原菌。

关键词: 新疆, 万寿菊枯萎病, 镰刀菌, 鉴定, 生物学特性

Abstract:

The purpose of the study was to clarify the species and biological characteristics of the pathogens of marigold wilt in Xinjiang Uygur Autonomous Region. A series of experiments were conducted through tissue isolation，single spore purification，pathogenicity determination，morphological Characteristics，phylogenetic analysis of ITS and EF-1α sequences and identification of biological characteristics. The results showed that four pathogens were identified as Fusarium fujikuroi，F. solani，F. equiseti and F. culmorum. The optimum temperature for the growth of all Fusarium species was 25 ℃ and the optimum pH range was 6-7. Full-darkness was conducive to the mycelial growth of F. fujikuroi，but full-light was suitable for the rest species. They differed in their optimum growing conditions. The mycelial of F. fujikuroi grew better on media of PDA and PSA，and was in favor of lactose and potassium nitrate as the optimal carbon and nitrogen sources. For F. equiseti，the optimal medium was PDA，and the optimal carbon and niteogen sources were lactose and yeast extract. For F. solani，the best medium was Czapek medium，and the best carbon and nitrogen sources were lactose and peptone. For F. culmorum，the beneficial media were PSA，PDA and CA，and the beneficial carbon and nitrogen sources were soluble starch，yeast extract and ammonium chloride. Their mycelial lethal temperature were at 64 ℃，63 ℃，63 ℃，52 ℃ for 10 min respectively. Among of them，F. solani is the most dominant pathogenic species causing marigold wilt in Xinjiang because of highest separation frequency of 68%.

Key words: Xinjiang, marigold wilt, Fusarium, identification, biological characteristics

努尔斯曼姑丽·艾海提, 阿依加玛丽·阿卜杜扎伊尔, 孙永民, 韩文杰, 郭庆元, 王丽丽. 新疆万寿菊枯萎病病原菌鉴定及生物学特性研究[J]. 园艺学报, 2023, 50(12): 2723-2734.

Nursimangul·Aihaiti, Aigamarie·Abdulzaire, SUN Yongmin, HAN Wenjie, GUO Qingyuan, WANG Lili. Identification and Biological Characteristics of the Pathogens Causing Marigold Wilt in Xinjiang[J]. Acta Horticulturae Sinica, 2023, 50(12): 2723-2734.

图/表 12

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培养基 Media	菌落直径/mm Colony diameter
培养基 Media	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
玉米粉琼脂 Corn meal agar（CMA）	48.50 ± 0.84 g	41.83 ± 0.41 b	46.17 ± 0.41 f	55.17 ± 0.41 c
燕麦琼脂 Oat meal agar（OA）	60.33 ± 0.52 d	53.33 ± 0.52 ab	40.67 ± 1.03 h	58.33 ± 0.52 b
胡萝卜葡萄糖琼脂 Carrot dextrose agar（CA）	57.83 ± 0.41 e	46.67 ± 0.52 ab	48.33 ± 0.52 e	84.83 ± 0.41 a
察氏Czapek（Czapek）	68.83 ± 0.41 b	55.83 ± 0.41 a	54.33 ± 0.52 d	54.17 ± 0.98 d
水琼脂Water agar（WA）	52.67 ± 0.52 f	40.17 ± 0.41 b	44.83 ± 0.41 g	22.00 ± 0.63 f
马铃薯蔗糖琼脂 Potato saccharose agar（PSA）	70.17 ± 0.41 a	53.67 ± 0.52 ab	56.67 ± 0.52 b	84.83 ± 0.41 a
马铃薯葡萄糖琼脂 Potato D-gluose agar（PDA）	70.33 ± 0.52 a	51.83 ± 0.41 ab	57.67 ± 0.52 a	84.67 ± 0.52 a
马铃薯胡萝卜琼脂 Potato carrot agar（PCA）	64.33 ± 0.52 c	54.33 ± 0.52 ab	55.67 ± 0.52 c	49.17 ± 0.75 e

培养基 Media	菌落直径/mm Colony diameter
培养基 Media	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
玉米粉琼脂 Corn meal agar（CMA）	48.50 ± 0.84 g	41.83 ± 0.41 b	46.17 ± 0.41 f	55.17 ± 0.41 c
燕麦琼脂 Oat meal agar（OA）	60.33 ± 0.52 d	53.33 ± 0.52 ab	40.67 ± 1.03 h	58.33 ± 0.52 b
胡萝卜葡萄糖琼脂 Carrot dextrose agar（CA）	57.83 ± 0.41 e	46.67 ± 0.52 ab	48.33 ± 0.52 e	84.83 ± 0.41 a
察氏Czapek（Czapek）	68.83 ± 0.41 b	55.83 ± 0.41 a	54.33 ± 0.52 d	54.17 ± 0.98 d
水琼脂Water agar（WA）	52.67 ± 0.52 f	40.17 ± 0.41 b	44.83 ± 0.41 g	22.00 ± 0.63 f
马铃薯蔗糖琼脂 Potato saccharose agar（PSA）	70.17 ± 0.41 a	53.67 ± 0.52 ab	56.67 ± 0.52 b	84.83 ± 0.41 a
马铃薯葡萄糖琼脂 Potato D-gluose agar（PDA）	70.33 ± 0.52 a	51.83 ± 0.41 ab	57.67 ± 0.52 a	84.67 ± 0.52 a
马铃薯胡萝卜琼脂 Potato carrot agar（PCA）	64.33 ± 0.52 c	54.33 ± 0.52 ab	55.67 ± 0.52 c	49.17 ± 0.75 e

温度/℃ Temperature	菌落直径/mm Colony diameter
温度/℃ Temperature	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
5	11.17 ± 0.41 g	11.33 ± 0.52 g	12.17 ± 0.41 g	9.83 ± 0.41 g
10	23.00 ± 0.63 f	14.17 ± 0.41 f	16.33 ± 0.52 f	22.33 ± 0.52 f
15	31.33 ± 0.52 e	18.50 ± 0.55 e	20.83 ± 0.41 e	52.33 ± 0.52 c
20	42.17 ± 0.41 c	31.83 ± 0.41 d	36.17 ± 0.41 d	70.83 ± 0.98 b
25	75.17 ± 0.41 a	56.83 ± 0.41 a	63.83 ± 0.41 a	84.83 ± 0.41 a
30	62.33 ± 0.52 b	53.00 ± 0.63 b	58.67 ± 0.52 b	41.67 ± 0.52 d
35	32.67 ± 0.52 d	33.67 ± 0.82 c	42.17 ± 0.41 c	23.33 ± 0.52 e

温度/℃ Temperature	菌落直径/mm Colony diameter
温度/℃ Temperature	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
5	11.17 ± 0.41 g	11.33 ± 0.52 g	12.17 ± 0.41 g	9.83 ± 0.41 g
10	23.00 ± 0.63 f	14.17 ± 0.41 f	16.33 ± 0.52 f	22.33 ± 0.52 f
15	31.33 ± 0.52 e	18.50 ± 0.55 e	20.83 ± 0.41 e	52.33 ± 0.52 c
20	42.17 ± 0.41 c	31.83 ± 0.41 d	36.17 ± 0.41 d	70.83 ± 0.98 b
25	75.17 ± 0.41 a	56.83 ± 0.41 a	63.83 ± 0.41 a	84.83 ± 0.41 a
30	62.33 ± 0.52 b	53.00 ± 0.63 b	58.67 ± 0.52 b	41.67 ± 0.52 d
35	32.67 ± 0.52 d	33.67 ± 0.82 c	42.17 ± 0.41 c	23.33 ± 0.52 e

pH	菌落直径/mm Colony diameter
pH	藤仓镰刀菌F. fujikuroi	茄腐镰刀菌F. solani	木贼镰刀菌F. equiseti	黄色镰刀菌F. culmorum
5	60.33 ± 0.52 f	50.33 ± 0.52 f	50.33 ± 0.52 f	30.17 ± 5.81 f
6	71.83 ± 0.41 a	54.00 ± 0.63 b	56.83 ± 0.41 a	70.33 ± 0.52 ab
7	68.17 ± 0.41 b	54.83 ± 0.41 a	55.67 ± 0.52 b	72.83 ± 0.41 a
8	64.00 ± 0.63 d	53.17 ± 0.41 c	56.17 ± 0.41 b	67.33 ± 0.52 b
9	66.50 ± 0.84 c	51.33 ± 0.52 de	54.83 ± 0.41 c	42.17 ± 4.83 d
10	64.50 ± 0.84 d	51.83 ± 0.41 d	53.17 ± 0.41 d	47.33 ± 0.52 c
11	60.17 ± 0.41 e	51.00 ± 0.63 e	51.50 ± 0.84 e	40.00 ± 0.89 de
12	57.00 ± 0.63 g	48.83 ± 0.41 g	48.83 ± 0.41 g	37.00 ± 0.89 e

新疆万寿菊枯萎病病原菌鉴定及生物学特性研究

Identification and Biological Characteristics of the Pathogens Causing Marigold Wilt in Xinjiang

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碳源 Carbon source	菌落直径/mm Colony diameter
碳源 Carbon source	藤仓镰刀菌F. fujikuroi	茄腐镰刀菌F. solani	木贼镰刀菌F. equiseti	黄色镰刀菌F. culmorum
无碳 Carbonless	59.67 ± 0.52 ab	46.33 ± 0.82 e	46.50 ± 0.84 de	18.00 ± 1.26 cd
蔗糖Sucrose	66.83 ± 0.41 a	51.50 ± 0.55 b	50.33 ± 0.52 bc	15.17 ± 0.41 e
山梨醇 Sorbitol	64.50 ± 0.55 ab	52.17 ± 0.41 b	49.83 ± 0.41 c	18.50 ± 1.22 c
乳糖 Lactose	70.00 ± 0.63 a	54.67 ± 0.52 a	51.67 ± 0.52 a	17.17 ± 0.75 d
葡萄糖 Glucose	66.33 ± 0.52 a	52.67 ± 0.52 b	50.67 ± 1.03 b	15.83 ± 0.98 e
木糖醇 Xylitol	64.33 ± 0.52 ab	49.00 ± 3.95 c	46.33 ± 0.52 de	21.17 ± 0.41 b
麦芽糖 Maltose	55.00 ± 24.49 b	51.83 ± 0.41 b	49.83 ± 0.41 c	19.00 ± 0.89 c
可溶性淀粉 Soluble starch	59.83 ± 0.41 ab	46.83 ± 0.41 de	46.83 ± 0.41 d	44.67 ± 0.52 a
甘露醇 Mannitol	66.50 ± 0.55 a	51.50 ± 0.55 b	50.17 ± 0.41 bc	15.33 ± 0.52 e
D-果糖 D- fructose	63.83 ± 0.41 ab	48.17 ± 0.41 cd	45.83 ± 0.41 e	16.17 ± 0.75 e

氮源 Nitrogen source	菌落直径/mm Colony diameter
氮源 Nitrogen source	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
无氮 Nitrogen free	59.50 ± 0.55 e	48.17 ± 4.02 e	44.67 ± 0.52 f	10.17 ± 0.41 f
硝酸钠 Sdium nitrate	65.50 ± 0.55 b	52.33 ± 0.52 c	51.50 ± 0.55 c	21.67 ± 0.52 e
硝酸钾 Potassium nitrate	67.33 ± 0.52 a	51.83 ± 0.41 cd	49.83 ± 0.41 d	22.33 ± 0.52 e
尿素 Urea	25.50 ± 0.55 g	33.33 ± 0.52 f	26.83 ± 0.75 g	22.17 ± 1.33 e
氯化铵 Ammonium chloride	64.33 ± 0.82 c	51.83 ± 0.41 cd	49.67 ± 0.52 d	64.33 ± 0.52 a
硫酸铵 Ammonia sulfate	64.83 ± 0.41 c	50.17 ± 0.41 d	47.67 ± 0.52 e	46.33 ± 0.52 d
酵母浸粉 Yeast extract	60.33 ± 0.52 e	55.33 ± 0.52 b	55.33 ± 0.52 a	64.33 ± 0.52 a
酵母膏 Yeast cream	60.00 ± 0.63 ef	56.67 ± 0.52 ab	53.00 ± 0.89 b	51.50 ± 1.22 c
蛋白胨 Peptone	63.17 ± 0.41 d	57.67 ± 0.52 a	54.83 ± 0.41 a	61.33 ± 1.03 b

光照 Light	菌落直径/mm Colony diameter
光照 Light	藤仓镰刀菌 F. fujikuroi	茄腐镰刀菌 F. solani	木贼镰刀菌 F. equiseti	黄色镰刀菌 F. culmorum
全光照 All light	70.4 ± 0.55 b	78.20 ± 0.45 a	80.80 ± 0.45 a	84.60 ± 0.55 a
光暗交替 Dark/ Light	61.4 ± 1.14 c	67.80 ± 0.45 b	72.40 ± 0.55 b	74.60 ± 0.89 b
全黑暗 All dark	80.8 ± 0.45 a	58.60 ± 0.55 c	61.80 ± 0.84 c	62.00 ± 1.00 c

种类 Species	温度/℃ Temperature
种类 Species	45	50	51	52	53	54	55	60	61	62	63	64	65
藤仓镰刀菌 F. fujikuroi	+	+	+	+	+	+	+	+	+	+	+	-	-
茄腐镰刀菌 F. solani	+	+	+	+	+	+	+	+	+	+	-	-	-
木贼镰刀菌 F. equiseti	+	+	+	+	+	+	+	+	+	+	-	-	-
黄色镰刀菌 F. culmorum	+	+	+	-	-	-	-	-	-	-	-	-	-

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新疆万寿菊枯萎病病原菌鉴定及生物学特性研究

Identification and Biological Characteristics of the Pathogens Causing Marigold Wilt in Xinjiang

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