Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not has a worldwide distribution infecting species from over 80 genera of plants (1). Apart from being an important pathogen of apple trees in many countries, B. dothidea can cause pre- and postharvest decay on apple fruit (2). It has been known to cause canker and dieback of forest trees in Serbia (3), but has not been recorded either on apple trees or apple fruit. In December 2010, apple fruit cv. Idared (Malus × domestica Borkh.) with symptoms of white rot were collected from one storage in the area of Svilajnac in Serbia. The incidence of the disease was low but the symptoms were severe. Affected fruit were brown, soft, and almost completely decayed, while the internal decayed tissue appeared watery and brown. A fungus was isolated from symptomatic tissue of one fruit after surface sterilization with 70% ethanol (without rinsing) and aseptic removal of the skin. Small fragments of decayed tissue were placed on potato dextrose agar (PDA) and incubated in a chamber at 22°C under alternating light and dark conditions (12/12 h). Fungal colonies were initially whitish, but started turning dark gray to black after 5 to 6 days. Pycnidia were produced after 20 to 25 days of incubation at 22°C and contained one-celled, elliptical, hyaline conidia. Conidia were 17.19 to 23.74 μm (mean 18.93) × 3.72 to 4.93 μm (mean 4.45) (n = 50). These morphological characteristics are in accordance with those described for the fungus B. dothidea (4). Genomic DNA was isolated from the fungus and internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The nucleotide sequence has been assigned to GenBank Accession No. KC994640. BLAST analysis of the 528-bp segment showed a 100% similarity with several sequences of B. dothidea deposited in NCBI GenBank, which confirmed morphological identification. Pathogenicity was tested by wound inoculation of five surface-sterilized, mature apple fruit cv. Idared with mycelium plugs (5 mm in diameter) of the isolate grown on PDA. Five control fruit were inoculated with sterile PDA plugs. After 5 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22°C, typical symptoms of white rot developed on inoculated fruit, while wounded, uninoculated, control fruit remained symptomless. The isolate recovered from symptomatic fruit showed the same morphological features as original isolate. To the best of our knowledge, this is the first report of B. dothidea on apple fruit in Serbia. Apple is widely grown in Serbia and it is important to further investigate the presence of this pathogen in apple storage, as well as in orchards since B. dothidea may cause rapid disease outbreaks that result in severe losses. References: (1) G. H. Hapting Agriculture Handbook 386, USDA, Forest Service, 1971. (2) A. L. Jones and H. S. Aldwinckle Compendium of Apple and Pear Diseases. APS Press, St. Paul, MN, 1990. (3) D. Karadžic et al. Glasnik Šumarskog Fakulteta 83:87, 2000. (4) B. Slippers et al. Mycologia 96:83, 2004.

译文

:Botryosphaeria dothidea(Moug .: Fr.)Ces。 &De Not在全球范围内感染80多种植物的物种(1)。除了在许多国家/地区是苹果树的重要病原菌外,多虫双歧杆菌还可以导致苹果果实收获前和收获后的腐烂(2)。在塞尔维亚(3)已知会引起森林树木的萎缩和枯萎(3),但在苹果树或苹果果实上均未见记录。 2010年12月,苹果果实简历。从塞尔维亚的Svilajnac地区的一个仓库中收集了具有白腐病症状的Idared(Malus×domestica Borkh。)。该病的发生率低,但症状很严重。受影响的果实为棕色,柔软,几乎完全腐烂,而内部腐烂的组织则呈水状和褐色。在用70%乙醇(无需冲洗)进行表面灭菌并无菌去除皮肤后,从一种水果的症状组织中分离出一种真菌。将腐烂组织的小片段放在马铃薯葡萄糖琼脂(PDA)上,并在22°C的暗室中于明暗条件下交替孵育(12/12 h)。真菌菌落最初呈白色,但在5至6天后开始从深灰色变成黑色。在22°C下孵育20至25天后,产生了碧萝cn,并含有单细胞,椭圆形,透明分生孢子。分生孢子为17.19至23.74μm(平均18.93)×3.72至4.93μm(平均4.45)(n = 50)。这些形态特征与描述真菌B. dothidea(4)的特征一致。从真菌中分离出基因组DNA,并用引物ITS1 / ITS4扩增rDNA的内部转录间隔区(ITS)。该核苷酸序列已分配给GenBank登录号KC994640。 528 bp片段的BLAST分析显示,其与NCBI GenBank中存放的多个B. dothidea序列具有100%的相似性,这证实了形态学鉴定。通过伤口接种五种经表面灭菌的成熟苹果果实简历来测试其致病性。厌倦了在PDA上生长的菌株的菌丝体塞(直径5毫米)。用无菌PDA塞接种了五个对照水果。在塑料容器中孵育5天后,在22°C的高湿度(RH 90至95%)下,接种的果实会出现典型的白腐症状,而受伤,未接种的对照果实仍无症状。从有症状果实中分离出的分离株表现出与原始分离株相同的形态特征。据我们所知,这是B. dothidea对塞尔维亚苹果果实的首次报道。苹果在塞尔维亚广泛种植,重要的是要进一步调查这种病原体在苹果储藏室以及果园中的存在,因为多头芽孢杆菌可能引起快速的疾病暴发,并造成严重的损失。参考文献:(1)G. H. Hapting农业手册386,美国农业部,林业局,1971年。(2)A. L. Jones和H. S. Aldwinckle《苹果和梨病简编》。 APS出版社,明尼苏达州圣保罗,1990年。(3)D.Karadžic等。 GlasnikŠumarskogFakulteta 83:87,2000。(4)B. Slippers等。 Mycologia 96:83,2004年。

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