宁夏地区酿酒葡萄溃疡病病原菌Lasiodiplodia theobromae鉴定、致病力分析及生物学特性研究

doi:10.16420/j.issn.0513-353x.2025-0382

摘要/Abstract

摘要：

为明确宁夏地区酿酒葡萄溃疡病的病原种类、致病力及生物学特性，采集银川、永宁、青铜峡等5个县（市）罹病的‘赤霞珠’‘西拉’‘贵人香’‘霞多丽’‘美乐’‘马瑟兰’和‘维欧尼’等7个酿酒葡萄品种植株样品，采用组织分离法进行病原分离并纯化，用烫伤法和灌根法对所分离的纯化菌株进行致病力测定。通过形态学观察和多基因（ITS-TEF1-TUB2）联合系统发育分析，确定病原菌种类。通过测定病原菌菌丝生长和产孢量，确定病原菌最优碳源、氮源、pH值及致死温度。研究结果表明，引起宁夏酿酒葡萄溃疡病的菌株均为可可毛色二孢（Lasiodiplodia theobromae），其中致病力最强的菌株为FR2。致病菌株对酿酒葡萄枝条、果梗和根系均具有致病力。生物学特性研究显示，该菌在以草酸铵和乙酸铵为氮源、葡萄糖和蔗糖为碳源、pH 5 ~ 7时生长和产孢较好，致死温度为55 ℃。

关键词: 葡萄, 葡萄溃疡病, 可可毛色二孢, 致病力, 生物学特性

Abstract:

To clarify the pathogen species，pathogenicity and biological characteristics of grapevine canker in Ningxia，samples of diseased Cabernet Sauvignon and Syrah wine grape plants from five counties and cities including Yinchuan，Yongning and Qingtongxia were collected. Diseased plant samples from five counties/cities，including Yinchuan，Yongning，and Qingtongxia，covering seven wine grape cultivars：‘Cabernet Sauvignon’‘Syrah’‘Italian Riesling’‘Chardonnay’‘Merlot’‘Marselan’and ‘Viognier’. Pathogen isolation and purification were conducted using tissue isolation method. Pathogenicity of the isolated and purified strains was determined by scalding method and root irrigation method. The pathogen species were identified through morphological observation and multi-gene （ITS-TEF1-TUB2）combined phylogenetic analysis. The optimal carbon and nitrogen sources，pH value， and lethal temperature of the pathogen were determined by measuring mycelial growth and spore production. The results showed that all the strains causing the canker disease of wine grapes in Ningxia were Lasiodiplodia theobromae，among which FR2 had the strongest pathogenicity. The pathogenic strains were pathogenic to grape branches，pedicels and roots. According to the study on biological characteristics，the fungus grew and sporulated well when ammonium oxalate and ammonium acetate are used as nitrogen sources，glucose and sucrose as carbon sources，and the pH value is 5-7，with a lethal temperature of 55  ℃.

Key words: grapevine, grape canker disease, Lasiodiplodia theobromae, pathogenicity, biological characteristics

蒲占悦, 赵楠, 王一冬, 辛颖, 顾沛雯. 宁夏地区酿酒葡萄溃疡病病原菌Lasiodiplodia theobromae鉴定、致病力分析及生物学特性研究[J]. 园艺学报, 2026, 53(8): 2454-2468.

PU Zhanyue, ZHAO Nan, WANG Yidong, XIN Ying, GU Peiwen. Identification，Pathogenicity Analysis，and Biological Characterization of Lasiodiplodia theobromae Causing Grapevine Canker in Ningxia Wine Grape Region[J]. Acta Horticulturae Sinica, 2026, 53(8): 2454-2468.

图/表 11

图1 田间葡萄溃疡病的发病症状（赤霞珠） a：发病后葡萄根茎部溃烂增生；b：葡萄根茎部和根系坏死、腐烂；c：罹病葡萄地上部叶片边缘呈焦灼状坏死病斑

Fig. 1 Symptoms of grapevine canker disease in the field（Cabernet Sauvignon） a：Hyperplasia and ulceration at the rootstock base；b：Necrosis and rot in both the rootstock base and root system；c：Scorch-like necrotic lesions along the leaf margins in the aboveground parts of diseased plants

表1 酿酒葡萄溃疡病疑似病原菌的来源及分离菌株数

Table 1 Sources and number of isolated strains of suspected pathogens of canker disease in wine grapes

分离部位Tissue of isolation	属 Genus	赤霞珠 Cabernet Sauvignon	西拉 Syrah	马瑟兰 Marselan	维欧妮 Viognier	贵人香 Italian Riesling	霞多丽 Chardonnay	美乐 Merlot	总数 Total number
根茎 Rhizome	镰刀菌属 Fusarium	6	6	6	6	4	6	4	38
	毛色二孢属 Lasiodiplodia	13	3	0	0	2	1	0	19
	茎点霉属 Phoma	1	0	0	0	0	0	0	1
	丝核菌属 Rhizoctonia	1	0	0	0	0	0	0	1
	角担菌属 Certatohasidium	1	0	0	0	0	0	0	1
	丛赤壳属 Dactylonectria	3	2	0	1	2	1	1	10
枝条 Branch	镰刀菌属 Fusarium	3	4	3	3	3	3	4	23
	毛色二孢属 Lasiodiplodia	0	0	0	0	0	0	0	0
	茎点霉属 Phoma	0	0	1	0	0	0	0	1
	丝核菌属 Rhizoctonia	0	0	0	0	0	0	0	0
	角担菌属 Certatohasidium	0	0	0	0	0	0	0	0
	丛赤壳属 Dactylonectria	1	0	0	0	0	0	0	1
果梗Stalk	镰刀菌属 Fusarium	3	3	3	2	2	2	3	18
	毛色二孢属 Lasiodiplodia	1	0	0	0	0	0	0	1
	茎点霉属 Phoma	0	0	0	0	1	0	0	1
	丝核菌属 Rhizoctonia	0	0	0	0	0	0	0	0
	角担菌属 Certatohasidium	0	0	0	0	0	0	0	0
	丛赤壳属 Dactylonectria	0	0	0	0	0	0	0	0
	总数Total number	33	18	13	12	14	13	12	115

图2 7株代表性疑似菌株离体枝条致病力验证（赤霞珠） A：枝条上病斑扩展症状；B：枝条上病斑面积在接种3 d和5 d时扩展大小（n = 4）。不同小写字母表示差异显著（P < 0.05）。下同

Fig. 2 Pathogenicity validation of detached branches from seven representative suspected strains（Cabernet Sauvignon） A：Symptomatic lesion expansion on grapevine shoots；B：Lesion area measurements at 3 d and 5 d post-inoculation（n = 4）. Different lowercase letters indicate significant differences（P < 0.05）. The same below

图3 酿酒葡萄品种离体枝条的致病力验证 A：每个品种从左到右依次为对照，接种3和5 d的枝条；B：枝条上病斑面积

Fig. 3 Pathogenicity assay of detached shoots from wine grape cultivars A：From left to right，each variety is the control，and the branches inoculated for 3 and 5 days，respectively；B：Area of diseased spots on branches

图4 酿酒葡萄果穗致病力验证（赤霞珠）

Fig. 4 Pathogenicity verification of wine grape clusters（Cabernet Sauvignon）

图5 盆栽试验的致病力验证（贵人香） a：盆栽试验病株症状；b、c：FR2引发的根部表皮腐烂并伴有灰白色霉层；d：体式显微镜下FR2引发侧根坏死腐烂；e：根部横截面显示FR2引发的木质部坏死；f：横截面显示FR2引发的根部腐烂坏死；g ~ i：横纵切面显示FR2引发的梭形溃疡斑；j：葡萄根茎部分离的病原菌菌株

Fig. 5 Pot experiment of strain FR2（Italian Riesling） a：Symptoms of diseased plants in pot experiments；b，c：Epidermal rot of roots caused by FR2 accompanied by grayish-white mold layer；d：Necrosis and rot of lateral roots caused by FR2 under stereomicroscope；e：Cross-section of roots showing necrosis of xylem caused by FR2；f：Cross-section showing root rot and necrosis caused by FR2；g-i：Longitudinal and transverse sections showing fusiform ulcers caused by FR2；j：Pathogenic strains isolated from the root and stem of grape

图6 Lasiodiplodia theobromae形态特征（A）及ITS-EF1α-TUB联合序列NJ系统发育树（B） a ~ c：菌落形态；d：PDA分生孢子器；e、f：分生孢子器产生分生孢子；g：成熟的分生孢子；h：分生孢子萌发；i、j：成熟分生孢子和分生孢子萌发；k：侧丝（很长）；l：分生孢子器；m：分生孢子梗（很短）

Fig. 6 Morphological characteristics of Lasiodiplodia theobromae（A）and NJ phylogenetic tree based on combined ITS-EF1α-TUB sequences（B） a-c：Colony morphology；d：Pycnidia formed on PDA medium；e，f：Conidia production within pycnidia；g：Mature conidia；h：Conidial germination；i，j：Mature conidia and conidial germination；k：Paraphyses（notably elongated）；l：Pycnidium；m：Conidiophores（markedly short）

图7 不同氮源下FR2菌株菌落生长及产孢情况

Fig. 7 Colony growth and spore production of FR2 strain under different nitrogen sources

图8 不同碳源下病菌FR2菌株菌落生长及产孢情况

Fig. 8 Colony growth and spore production of FR2 strain under different carbon sources

图9 不同 pH 下FR2菌株菌落生长及产孢情况

Fig. 9 Colony growth and spore production of FR2 strain at different pH values

图10 FR2菌株致死温度测定

Fig. 10 Determination of lethal temperature for FR2 strain

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