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短小新壳梭孢引起中国刺葡萄穗枯病和叶斑病的首次报道

First Report of Neofusicoccum parvum Causing Panicle Blight and Leaf Spot on Vitis heyneana in China.

作者信息

Wu D D, Fu G, Ye Y F, Hu F Y, Mou H F, Qin L L, Jiang N

机构信息

Biotechnology Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, P. R. China.

Microbiology Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, P. R. China.

出版信息

Plant Dis. 2015 Mar;99(3):417. doi: 10.1094/PDIS-05-14-0515-PDN.

Abstract

The climbing vine, Vitis heyneana Roem. & Schult, is a member of the grape family endemic to Asia. Its fruits are used in wine production, and its roots, stems, and leaves can be used in medicinal materials. This plant is grown in Southwest China, as well as in India, Bhutan, and Nepal. Mulao Autonomous County in Guangxi Province is the only artificial cultivation area in China. During the summer of 2013, a panicle blight and leaf spot were detected on V. heyneana on four farms in Mulao Autonomous County. The symptoms were observed from the onset of florescence through fruit harvest. Brown lesions initially appeared at the base of a panicle and then extended to the whole panicle, finally causing the panicle to die and fruit to drop. When the disease developed on leaves, the symptom initially appeared as small dark brown circular spots, later enlarging into irregular spots (average diameter 6 mm) with a light brown center and dark brown rim. With severe disease, some individual leaves were affected by numerous spots, leading to premature senescence. Small sections of diseased tissue excised from 10 panicle and 10 leaf samples were plated on potato dextrose agar (PDA) and incubated at 28°C. Fungal colonies developed, initially with abundant white aerial mycelium, which turned olivaceous gray after 5 days and formed black pycnidia after 25 days. The conidia were hyaline, ellipsoidal to fusiform, externally smooth, thin-walled, and nonseptate. Thirty conidia were measured; the dimensions were 12.0 to 17.5 × 4.0 to 6.0 μm. Morphological characteristics of the isolates were similar to the descriptions of Neofusicoccum parvum (3). The isolate MPT-1 was selected as a representative for molecular identification. Genomic DNA was extracted and used for PCR to amplify the internal transcribed spacer (ITS) region and partial translation elongation factor 1-alpha (EF1-α) gene, using primers ITS1/ITS4 and EF1-728F/EF1-986R, respectively. The obtained ITS sequence (GenBank Accession No. KJ599627) and EF1-α sequence (KM921768) showed >99% homology with several GenBank sequences of N. parvum. Morphological and molecular results confirmed the isolate as N. parvum. For pathogenicity tests, detached, young healthy panicles and leaves of V. heyneana were surface-sterilized, wounded by sterile needle, and inoculated with mycelial plugs (3 mm in diameter) of four N. parvum isolates. Ten panicles and 10 leaves were used for every isolate. Control panicles and leaves were treated with sterile PDA plugs. All the samples were placed in a humid chamber (RH 90%, 28°C, 12 h of light) for 3 days. Symptoms similar to those observed in the field developed on all panicles and leaves inoculated with N. parvum isolates. N. parvum was reisolated from all inoculated, symptomatic tissues. The controls remained symptomless. N. parvum has been reported to cause trunk canker on V. vinifera (2), dieback on Cupressus funebris (3), and a leaf spot on Myristica fragrans (1). To our knowledge, this is the first report of N. parvum causing panicle blight and leaf spot on V. heyneana in China. Panicle blight caused a large number of fruits to drop and reduced the yield seriously. Some effective measures should be taken to control this disease. References: (1) V. Jayakumar et al. New Dis. Rep. 23:19, 2011. (2) J. Kaliternam et al. Plant Dis. 97:1656, 2013. (3) S. B. Li et al. Plant Dis. 94:641, 2010.

摘要

藤本植物毛葡萄(Vitis heyneana Roem. & Schult)是葡萄科的一种植物,为亚洲特有。其果实用于酿酒,根、茎、叶可入药。该植物在中国西南部以及印度、不丹和尼泊尔均有生长。广西壮族自治区的仫佬族自治县是中国唯一的人工种植区。2013年夏季,在仫佬族自治县的4个农场中,发现毛葡萄出现了穗枯病和叶斑病。从花期开始到果实收获期都观察到了这些症状。褐色病斑最初出现在穗基部,然后扩展到整个果穗,最终导致果穗死亡、果实脱落。当叶片发病时,症状最初表现为小的深褐色圆形斑点,随后扩大为不规则斑点(平均直径6毫米),中心浅褐色,边缘深褐色。病情严重时,部分叶片上有大量病斑,导致叶片提前衰老。从10个果穗和10片叶片样本上切取的小块患病组织接种到马铃薯葡萄糖琼脂(PDA)培养基上,于28°C培养。真菌菌落生长,最初有大量白色气生菌丝,5天后变为橄榄灰色,25天后形成黑色分生孢子器。分生孢子无色,椭圆形至梭形,表面光滑,薄壁,无隔膜。测量了30个分生孢子;大小为12.0至17.5×4.0至6.0微米。分离菌株的形态特征与葡萄小穴壳菌(Neofusicoccum parvum)的描述相似。选择分离菌株MPT - 1作为分子鉴定的代表。提取基因组DNA,分别使用引物ITS1/ITS4和EF1 - 728F/EF1 - 986R进行PCR扩增内部转录间隔区(ITS)和部分翻译延伸因子1 - α(EF1 - α)基因。获得的ITS序列(GenBank登录号:KJ599627)和EF1 - α序列(KM921768)与GenBank中多个葡萄小穴壳菌序列的同源性>99%。形态学和分子生物学结果证实该分离菌株为葡萄小穴壳菌。为进行致病性测试,将毛葡萄健康的幼嫩果穗和叶片摘下,进行表面消毒,用无菌针穿刺致伤,然后接种4个葡萄小穴壳菌分离菌株的菌丝块(直径3毫米)。每个分离菌株接种10个果穗和10片叶片。对照果穗和叶片接种无菌PDA块。所有样品置于湿度箱(相对湿度90%,28°C,光照12小时)中3天。接种葡萄小穴壳菌分离菌株的所有果穗和叶片均出现了与田间观察到的症状相似的症状。从所有接种且出现症状的组织中重新分离出了葡萄小穴壳菌。对照仍无症状。据报道,葡萄小穴壳菌可引起酿酒葡萄的树干溃疡病(2)、柏木的枝枯病(3)以及肉豆蔻的叶斑病(1)。据我们所知,这是葡萄小穴壳菌引起中国毛葡萄穗枯病和叶斑病的首次报道。穗枯病导致大量果实脱落,严重降低了产量。应采取一些有效措施来防治这种病害。参考文献:(1)V. Jayakumar等人,《新病害报道》23:19,2011年。(2)J. Kaliternam等人,《植物病害》97:1656,2013年。(3)S. B. Li等人,《植物病害》94:641,2010年。

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