Inácio C A, Pereira-Carvalho R C, Morgado F G A, Fonseca M E N, Boiteux L S
Universidade de Brasília (UnB) Departamento de Fitopatologia, 70910-900, Brasília-DF, Brazil.
Embrapa Hortaliças (CNPH), CP 218, 70359-970, Brasília-DF, Brazil.
Plant Dis. 2011 Oct;95(10):1318. doi: 10.1094/PDIS-06-11-0464.
Fruit rots caused by distinct fungal pathogens are commonly observed on tomatoes (Solanum lycopersicum L.) throughout all major production areas in Brazil. Samples of fruits displaying white mycelial growth associated with a profuse salmon-color sporulation were collected in greenhouse-grown tomatoes in Brasília-DF in February 2011. The isolated fungus displayed pink-to-white colonies containing several conidiophores with conidia. Mycelia displayed hyaline hyphae as much as 4 μm in diameter; conidiophores were simple or branched, 112 to 300 (360) μm long, and 2 to 4 μm wide. Conidia were produced in basipetal chains (frequently clustered), were ellipsoidal to pyriform with oblique and prominent truncate basal scars, two-celled, hyaline, and (14-) 16 to 26 (-28) × (6-) 7 to 10 (-12) μm. These characteristics allocated the specimen to Trichothecium roseum (Pers.). Koch's postulates were fulfilled for one fungal isolate by either spraying 10 intact fruits or by placing a drop of a spore suspension (adjusted to 10 conidia/ml) into three to five wounds created on 10 mature fruits of each of two tomato cultivars (Santa Clara and Dominador) by puncturing each fruit with a sterile needle. Five fruits of each cultivar were treated with sterile water as the mock-inoculated control treatment. Identical symptoms to those of the original fruit were observed only in the T. roseum-inoculated samples 5 to 7 days after using both inoculation procedures. Total DNA was extracted from a pure colony of the fungus growing on potato dextrose agar medium and used as template in PCR assays with the internal transcribed spacer (ITS)-4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS-5 (5'-GGAAGTAAAAGTCGTAACAAGG-3') primer pair (2). A single amplicon of approximately 630 bp was observed and directly sequenced. Sequence analysis of the Brazilian isolate (GenBank No. JN081877) indicated identity levels of 99% with T. roseum isolates reported on Leucadendron xanthoconus in South Africa (GenBank No. EU552162) and isolates from strawberry fruits in South Korea (GenBank No. HM355750). However, phylogenetic analysis was unable to discriminate isolates of T. roseum from Passalora (GenBank No. EF432764) and Fusarium (GenBank No. GU183369) isolates, confirming the low genetic variability of the ITS region in Hypocreales (3). T. roseum has been reported to be infecting greenhouse tomatoes in the United States (4) and causing postharvest disease of tomatoes in Argentina (1). To our knowledge, this is the first report of T. roseum infecting greenhouse tomatoes in Brazil. References: (1) G. Dal Bello. Australas. Plant Dis. Notes 3:103, 2008. (2) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (3) L. Lombard et al. Stud. Mycol. 66:31, 2010. (4) A. W. Welch, Jr. et al. Plant Dis. Rep. 59:255, 1975.
在巴西所有主要产区的番茄(Solanum lycopersicum L.)上,由不同真菌病原体引起的果实腐烂现象普遍可见。2011年2月,在巴西利亚联邦区温室种植的番茄中,采集到了一些果实样本,这些果实上有白色菌丝体生长,并伴有大量鲑鱼色的孢子形成。分离出的真菌呈现出粉红色至白色的菌落,包含几个带有分生孢子的分生孢子梗。菌丝体的透明菌丝直径可达4μm;分生孢子梗简单或有分支,长112至300(360)μm,宽2至4μm。分生孢子以向基链的形式产生(常成簇),呈椭圆形至梨形,基部有倾斜且明显的截断疤痕,有两个细胞,透明,大小为(14-)16至26(-28)×(6-)7至10(-12)μm。这些特征表明该标本属于粉红单端孢(Trichothecium roseum (Pers.))。通过对两种番茄品种(圣克拉拉和多米纳多)的10个成熟果实进行穿刺造成三到五个伤口,然后向其中喷洒10个完整果实,或向每个伤口滴一滴孢子悬浮液(调整至10个分生孢子/ml),对一种真菌分离株进行了柯赫氏法则验证。每个品种的五个果实用无菌水进行处理,作为模拟接种的对照处理。在两种接种方法实施5至7天后,仅在接种粉红单端孢的样本中观察到了与原始果实相同的症状。从在马铃薯葡萄糖琼脂培养基上生长的真菌纯菌落中提取总DNA,并将其用作PCR分析的模板,使用内部转录间隔区(ITS)-4(5'-TCCTCCGCTTATTGATATGC-3')和ITS-5(5'-GGAAGTAAAAGTCGTAACAAGG-3')引物对(2)。观察到一个约630bp的单一扩增子,并直接进行了测序。对巴西分离株(GenBank登录号:JN081877)的序列分析表明,其与南非黄锥花树(Leucadendron xanthoconus)上报道的粉红单端孢分离株(GenBank登录号:EU552162)以及韩国草莓果实上的分离株(GenBank登录号:HM355750)的同一性水平为99%。然而,系统发育分析无法区分粉红单端孢与帕萨洛拉属(Passalora,GenBank登录号:EF432764)和镰刀菌属(Fusarium,GenBank登录号:GU183369)的分离株,这证实了肉座菌目(Hypocreales)中ITS区域的低遗传变异性(3)。据报道,粉红单端孢在美国感染温室番茄(4),在阿根廷导致番茄采后病害(1)。据我们所知,这是粉红单端孢感染巴西温室番茄的首次报道。参考文献:(1)G. Dal Bello。Australas. Plant Dis. Notes 3:103, 2008。(2)N. L. Glass和G. C. Donaldson。Appl. Environ. Microbiol. 61:1323, 1995。(3)L. Lombard等人。Stud. Mycol. 第66卷:31页,2010。(4)A. W. Welch, Jr.等人。Plant Dis. Rep. 59:255, 1975。