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巴西巴伊亚州茄腐镰刀菌胡椒专化型有性繁殖的首次报道。

First Report of Sexual Reproduction of Fusarium solani f. sp. piperis in Bahia, Brazil.

作者信息

Vaz A B, Elizei V G, Costa S S, Pfenning L H, Ventura J A

机构信息

Departamento de Fitopatologia, Universidade Federal de Lavras, 37200-000 Lavras MG, Brazil.

Instituto Capixaba de Pesquisa, Assistência Técnica e Extensãdo Rural, 29052-010, Vitória ES, Brazil.

出版信息

Plant Dis. 2012 Oct;96(10):1581. doi: 10.1094/PDIS-05-12-0469-PDN.

DOI:10.1094/PDIS-05-12-0469-PDN
PMID:30727344
Abstract

Black pepper (Piper nigrum L.) is a popular spice native of India, and Brazil is one of its most important producing countries. The main disease of black pepper in Brazil is fusariosis, caused by Fusarium solani f. sp. piperis. Symptoms include leaf chlorosis and defoliation, blight of stems or stem cuttings, and root and foot decay. During surveys conducted in the south of the state of Bahia, municipalities of Taperoá (13°34'S, 39°10'W) and Valencia (13°20'S, 39°14'W), stems of diseased plants covered with red or salmon-colored perithecia were observed, while twigs showed leaf chlorosis, leading to early death of the plants. Ascomata were solitary or in groups, mostly superficial or surrounded by mycelia, globose, subglobose, ovoid, and 122 to 400 μm diameter. Microscopic examination revealed unitunicate, cylindric asci, 60 to 90 × 8.5 to 16 μm, thin-walled, containing eight ascospores arranged obliquely in two rows. Ascospores are hyaline, elliptical to oblong, one-septate, constricted at the central septum, 10 to 16 × 4 to 6.5 μm (means ± S.D.: 13.1 ± 1.4 × 5.1 ± 0.6 μm), length/width (L/W) 1.9 to 3.7. Single-spored cultures were transferred to SNA medium (incubated at 20°C for 7 days with 12-h photoperiod) and on potato dextrose agar (25°C in dark) for characterization. The anamorph is characterized by the presence of chlamydospores, canoe-shaped sporodochial macroconidia with three to four septae, and microconidia formed on long monophialidic conidiophores. Based on morphological markers, isolates were identified as F. solani. The partial fragment of the TEF-1α gene of single-spored isolates (CML 2186, 2187, 2188, 2189, 2190, and 2191) were sequenced. BLAST analysis of the sequence resulted in 94 to 99% identity with a reference strain of F. solani f. sp. piperis (NRRL 22570, CML 1888). For pathogenicity tests, cv. Bragantina was used and two isolates were inoculated as 5-mm diameter mycelial plugs on the stem of four plants each. Four control plants were treated only with sterile culture medium. Plants were maintained in the greenhouse at 25°C and 75 to 85% relative humidity under 70% shade. All inoculated plants showed initial symptoms of stem necrosis in inoculated branches 7 days after inoculation. Symptoms were not observed on stems of control plants. Isolates were successfully reisolated and identified as F. solani f. sp. piperis, fulfilling Koch's postulates. Representative isolates were deposited at the Coleção Micológica de Lavras (CML) at Universidade Federal de Lavras, Brazil. Production of perithecia of the pathogen has been previously reported only in Pará and Espírito Santo States (1,3). It is not yet confirmed if this taxon is homothallic or heterothallic. To our knowledge, this is the first report of the associated teleomorph of F. solani f. sp. piperis infecting and causing black pepper fusariosis in Bahia, Brazil. The results suggest that the spread of ascospores from perithecia is likely to be one of the main inoculum sources of the disease on adjacent vines. There is evidence that this special form of F. solani actually represents a distinct species pathogenic to black pepper (2). References: (1) F. C. Albuquerque and S. Ferraz. Experientiae 22:133, 1976. (2) K. O'Donnell. Mycologia 92:919, 2000. (3) J. A. Ventura et al. Fitopatol. Bras. 11:361, 1986.

摘要

黑胡椒(Piper nigrum L.)是一种原产于印度的受欢迎的香料,巴西是其最重要的生产国之一。巴西黑胡椒的主要病害是镰刀菌病,由茄腐镰刀菌胡椒专化型(Fusarium solani f. sp. piperis)引起。症状包括叶片黄化和落叶、茎或茎插条枯萎以及根和基部腐烂。在巴伊亚州南部的塔佩罗阿(南纬13°34′,西经39°10′)和巴伦西亚(南纬13°20′,西经39°14′)市进行调查时,观察到患病植株的茎上覆盖着红色或鲑鱼色的子囊壳,而嫩枝出现叶片黄化,导致植株过早死亡。子囊果单个或成群,大多表生或被菌丝包围,球形、近球形、卵形,直径122至400μm。显微镜检查显示,子囊单囊壁、圆柱形,60至90×8.5至16μm,薄壁,含有八个呈两排斜向排列的子囊孢子。子囊孢子无色,椭圆形至长圆形,具一个隔膜,在中央隔膜处缢缩,10至16×4至6.5μm(平均值±标准差:13.1±1.4×5.1±0.6μm),长宽比(L/W)为1.9至3.7。将单孢子培养物转移至SNA培养基(在20°C、12小时光周期下培养7天)和马铃薯葡萄糖琼脂(在黑暗中25°C培养)上进行鉴定。无性型的特征是存在厚垣孢子、具三至四个隔膜的独木舟形分生孢子座大型分生孢子以及在长的单瓶梗分生孢子梗上形成的小型分生孢子。基于形态学标记,分离物被鉴定为茄腐镰刀菌。对单孢子分离物(CML 2186、2187、2188、2189、2190和2191)的TEF-1α基因的部分片段进行了测序。序列的BLAST分析结果显示与茄腐镰刀菌胡椒专化型的参考菌株(NRRL 22570,CML 1888)的同一性为94%至99%。进行致病性测试时,使用了Bragantina品种,将两个分离物以直径5mm的菌丝块接种到四株植物的茎上。四株对照植物仅用无菌培养基处理。将植物置于温室中,温度为25°C,相对湿度为75%至85%,遮荫70%。所有接种的植物在接种后7天,接种枝条的茎上出现了茎坏死的初始症状。对照植物的茎上未观察到症状。成功从接种植物上重新分离出分离物,并鉴定为茄腐镰刀菌胡椒专化型,符合柯赫氏法则。代表性分离物保藏于巴西拉夫拉斯联邦大学的拉夫拉斯真菌保藏中心(CML)。此前仅在帕拉州和圣埃斯皮里图州报道过该病原菌子囊果的产生(参考文献1,3)。目前尚未证实该分类单元是同宗配合还是异宗配合。据我们所知,这是首次报道茄腐镰刀菌胡椒专化型的相关有性型在巴西巴伊亚州感染并引起黑胡椒镰刀菌病。结果表明,来自子囊果的子囊孢子传播可能是该病在相邻藤蔓上的主要接种源之一。有证据表明,这种特殊形式的茄腐镰刀菌实际上代表了一种对黑胡椒致病的独特物种(参考文献2)。参考文献:(1) F. C. Albuquerque和S. Ferraz。Experientiae 22:133, 1976。(2) K. O'Donnell。Mycologia 92:919, 2000。(3) J. A. Ventura等人。Fitopatol. Bras. 11:361, 1986。

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