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加州首次报告黑曲霉引起酿酒葡萄(葡萄属)酸腐病

First Report of Aspergillus carbonarius Causing Sour Rot of Table Grapes (Vitis vinifera) in California.

作者信息

Rooney-Latham S, Janousek C N, Eskalen A, Gubler W D

机构信息

California Department of Food and Agriculture-Plant Pest Diagnostics, Sacramento, 95634.

Department of Plant Pathology, University of California, Davis 95616.

出版信息

Plant Dis. 2008 Apr;92(4):651. doi: 10.1094/PDIS-92-4-0651A.

Abstract

During the 2006 growing season, grape berries in several Red Globe vineyards in Kern County, California were found exhibiting black fungal sporulation and typical sour rot symptoms. Symptoms included berry cracking and leakage along with a pungent vinegar smell. In California, sour rot (also known as summer bunch rot) has been attributed to a complex of microorganisms that invade ripe berries following injury. Fungi typically associated with sour rot include Aspergillus niger, Alternaria tenuis, Cladosporium herbarum, Rhizopus arrhizus, and Penicillium spp. Various yeasts and bacteria have also been associated with the disease, especially Acetobacter bacteria, which give the grapes their pungent acetic acid smell (2). In June 2006, the two fungi most commonly isolated from affected berries (postveraison) were A. niger and A. carbonarius. Identification of the two species was confirmed by colony and spore morphology (1) and PCR analysis of the internal transcribed spacer (ITS) region of rDNA. Although A. niger was more common, A. carbonarius has not previously been reported as a pathogen of grape in California. Conidia of A. carbonarius were globose, dark with very rough walls, and could be distinguished from A. niger by their wall structure and larger size ([5.1] 6.0 to 7.6 [8.8] μm in diameter). Sequence analysis of the ITS region of isolates morphologically identified as A. carbonarius showed 100% similarity to known isolates of this species. To confirm pathogenicity, postveraison Red Globe berries on standing grapevines were wounded with a needle and inoculated by dipping entire clusters into A. carbonarius spore suspensions (10 conidia/ml) for 30 s. Sterile water was used as a control. Twenty berries on each cluster were wounded and 10 clusters were used for each treatment. The experiment was repeated in two different vineyards in Kern County in 2006 and 2007. After 48 h, water-soaked lesions could be seen on the wounding site of grapes inoculated with A. carbonarius. After 1 week, 100% of the inoculated grapes exhibited dark, black sporulation, and after 3 weeks, the clusters were almost completely rotted. Koch's postulates were completed by isolating A. carbonarius from the affected berries. A few (less than 5%) of the wounded control berries also exhibited black sporulation and rot after 3 weeks. These infections were probably the result of natural inoculum at the sites because spore traps placed in the vineyards also contained A. carbonarius and A. niger propagules. Furthermore, soil surveys in the same vineyards showed that both A. niger and A. carbonarius were present on plant debris on the vineyard floor and in the soil. To our knowledge, this is the first report of A. carbonarius causing sour rot of grapes in California. References: (1) M. A. Klich. Identification of Common Aspergillus Species. Centraalbureau voor Schimmelcultures, Urecht, The Netherlands, 2002. (2) J. J. Marois et al. Bunch rots: Miscellaneous secondary invaders and sour rot. Page 69 in: Grape Pest Management. 2nd ed. The Regents of the University of California, Division of Agriculture and Natural Resources, Oakland, 1992.

摘要

在2006年生长季,加利福尼亚州克恩县几个红地球葡萄园的葡萄浆果出现了黑色真菌孢子形成以及典型的酸腐症状。症状包括浆果开裂和渗漏,并伴有刺鼻的醋味。在加利福尼亚州,酸腐(也称为夏季穗腐病)被认为是由多种微生物引起的,这些微生物在果实受伤后侵入成熟的浆果。通常与酸腐相关的真菌包括黑曲霉、细链格孢、草本枝孢、少根根霉和青霉属。各种酵母和细菌也与该病有关,尤其是醋酸杆菌,它使葡萄产生刺鼻的醋酸味(2)。2006年6月,从受影响的浆果(转色期后)中最常分离出的两种真菌是黑曲霉和碳黑曲霉。通过菌落和孢子形态(1)以及对rDNA内转录间隔区(ITS)的PCR分析,确认了这两个物种的鉴定。虽然黑曲霉更为常见,但此前在加利福尼亚州尚未报道碳黑曲霉是葡萄的病原菌。碳黑曲霉的分生孢子呈球形,颜色深,壁非常粗糙,可通过其壁结构和较大的尺寸(直径[5.1]6.0至7.6[8.8]μm)与黑曲霉区分开来。对形态学鉴定为碳黑曲霉的分离株的ITS区域进行序列分析,结果显示与该物种的已知分离株100%相似。为了确认致病性,用针在直立葡萄藤上的转色期后红地球浆果上造成伤口,然后将整个果穗浸入碳黑曲霉菌孢子悬浮液(10个孢子/毫升)中30秒进行接种。用无菌水作为对照。每个果穗上有20个浆果被划伤,每种处理使用10个果穗。该实验于2006年和2007年在克恩县的两个不同葡萄园重复进行。48小时后,接种碳黑曲霉的葡萄伤口部位出现水渍状病斑。1周后,100%接种的葡萄出现黑色孢子形成,3周后,果穗几乎完全腐烂。通过从受影响的浆果中分离出碳黑曲霉,完成了柯赫氏法则验证。少数(不到5%)受伤的对照浆果在3周后也出现了黑色孢子形成和腐烂。这些感染可能是由于伤口处的自然接种物导致的,因为放置在葡萄园中的孢子捕捉器中也含有碳黑曲霉和黑曲霉的繁殖体。此外,在同一葡萄园进行的土壤调查表明,黑曲霉和碳黑曲霉都存在于葡萄园地面的植物残体和土壤中。据我们所知,这是碳黑曲霉在加利福尼亚州引起葡萄酸腐病的首次报道。参考文献:(1)M. A. Klich。常见曲霉属物种的鉴定。荷兰乌得勒支中央真菌培养中心,2002年。(2)J. J. Marois等人。穗腐病:其他次生侵染菌和酸腐病。载于:《葡萄病虫害管理》第2版。加利福尼亚大学董事会,农业与自然资源司,奥克兰,1992年,第69页。

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