Avila-Quezada G, Silva-Rojas H V
Centro de Investigacion en Alimentacion y Desarrollo A.C., Ave 4 Sur 3820, Delicias, Chihuahua, Mexico 33089.
Colegio de Postgraduados, Campus Montecillo. Texcoco, Mexico, Mex. 56230.
Plant Dis. 2007 Sep;91(9):1200. doi: 10.1094/PDIS-91-9-1200A.
Mexico is a major avocado (Persea americana) producer in the world. Glomerella cingulata (anamorph Colletotrichum gloeosporioides) has been reported as a causal agent of anthracnose on avocado fruits worldwide (3), while G. acutata (anamorph Colletotrichum acutatum) has been identified as the cause of this disease only in New Zealand (2) and Australia (4). This study was done with the objective to determine the Glomerella spp. involved as the causal agents of avocado anthracnose in Mexico. From 2003 to 2006, avocado fruits cv. Hass with anthracnose symptoms appearing as brown-black lesions on the pericarp and soft rot in the mesocarp were collected in 10 counties in Michoacan, the leading avocado-producing Mexican state. Glomerella spp. were isolated on potato dextrose agar (PDA) for molecular and morphological identification. A phylogenetic analysis was done by amplifying the internal transcribed spacer region of rDNA for 28 of the isolates. Primers ITS5/NL4 was used and successfully amplified bands of approximately 1,000 bp. Each sequence corresponding to Glomerella spp. was compared with sequences deposited in the GenBank database using BLAST. The results from molecular approach indicated 86% of the isolates used in this study were G. cingulata and 14% were G. acutata. Sequences of both species were deposited in GenBank under Accession Nos. EF221828, EF221829, and EF221830 for G. cingulata and EF175780, EF221831, and EF221832 for G. acutata. Colonies of G. acutata that developed on PDA medium were pale gray, occasionally the lower surface was olive green, and the center was covered with orange-to-salmon pink masses of conidia and perithecia. Conidia grown in the same media were straight, fusiform, 8.2 to 16.5 μm long, and 2.7 to 4.0 μm wide (4). Pathogenicity tests of G. acutata were carried out by inoculating six healthy cv. Hass fruits (1) at three evenly spaced locations on the fruit surface with a needle dipped in a conidial mass from a 3-day-old monoconidial culture of G. acutata. Fruits were then incubated in a moist chamber for 3 days. Anthracnose symptoms were observed on healthy fruits inoculated with G. acutata, while control fruits inoculated with sterile water did not develop symptoms. The fungi were reisolated successfully to confirm the pathogen's identity using morphological key. To our knowledge, this is the first report of G. acutata causing anthracnose on avocado fruits in Mexico. References: (1) R. Guetsky et al. Phytopathology 95:1341, 2005. (2) W. F. T. Hartill. N. Z. J. Crop Hortic. Sci. 19:297, 1991. (3) D. Prusky. Annu. Rev. Phytopathol. 34:413, 1996. (4) J. H. Simmonds. Qld. J. Agric. Anim. Sci. 22:437, 1965.
墨西哥是世界上主要的鳄梨(油梨)生产国。围小丛壳菌(无性型为胶孢炭疽菌)在全球范围内被报道为鳄梨果实炭疽病的病原(3),而尖孢小丛壳菌(无性型为尖孢炭疽菌)仅在新西兰(2)和澳大利亚(4)被鉴定为该病的病原。本研究旨在确定在墨西哥作为鳄梨炭疽病病原的小丛壳菌种类。2003年至2006年,在墨西哥主要的鳄梨生产州米却肯州的10个县,采集了哈斯品种表现出炭疽病症状(果皮上出现棕黑色病斑,中果皮软腐)的鳄梨果实。将小丛壳菌分离在马铃薯葡萄糖琼脂(PDA)上进行分子和形态学鉴定。对28个分离株的核糖体DNA内部转录间隔区进行扩增,进行系统发育分析。使用引物ITS5/NL4并成功扩增出约1000bp的条带。将每个对应于小丛壳菌的序列与GenBank数据库中保存的序列进行BLAST比较。分子分析结果表明,本研究中使用的分离株86%为围小丛壳菌,14%为尖孢小丛壳菌。这两个种的序列已分别以围小丛壳菌的登录号EF221828、EF221829和EF221830以及尖孢小丛壳菌的登录号EF175780、EF221831和EF221832存入GenBank。在PDA培养基上生长的尖孢小丛壳菌菌落为浅灰色,偶尔下表面为橄榄绿色,中心覆盖着橙色至鲑鱼粉红色的分生孢子和子囊壳团块。在相同培养基上生长的分生孢子呈直形、梭形,长8.2至16.5μm,宽2.7至4.0μm(4)。通过用蘸有来自3日龄尖孢小丛壳菌单孢培养物分生孢子团的针在6个健康的哈斯品种果实(1)的果实表面三个等距位置接种,进行尖孢小丛壳菌的致病性测试。然后将果实置于保湿箱中培养3天。接种尖孢小丛壳菌的健康果实上观察到炭疽病症状,而接种无菌水的对照果实未出现症状。使用形态学特征成功重新分离出真菌以确认病原菌的身份。据我们所知,这是尖孢小丛壳菌在墨西哥导致鳄梨果实炭疽病的首次报道。参考文献:(1)R. Guetsky等人,《植物病理学》95:1341,2005年。(2)W. F. T. Hartill,《新西兰作物与园艺科学杂志》19:297,1991年。(3)D. Prusky,《植物病理学年度评论》3:413,1996年。(4)J. H. Simmonds,《昆士兰农业与动物科学杂志》22:437,1965年。