Tzeng S J, Sun E J, Hsiao W W
Department of Plant Pathology and Microbiology, National Taiwan University, Taipei, Taiwan, R. O. C.
Experimental Forest of College of Bio-Resources and Agriculture, National Taiwan University, Nantou, Taiwan, R. O. C.
Plant Dis. 2010 Apr;94(4):487. doi: 10.1094/PDIS-94-4-0487C.
Since 2005, coconut fruits (Cocos nucifera L.) harvested from trees in southern Taiwan have shown symptoms of a fruit rot disease. The disease occurs only on harvested coconut fruits and is more serious during warmer seasons. Principal symptoms are blackening of the exocarps, mesocarps, and then the endocarps of coconut fruits from pedicel end or wounds on fruits. A fungus was consistently recovered from diseased fruits. Colonies on potato dextrose agar (PDA) were white, becoming black 1 to 2 days later, with a strong fruity aroma. The fungus produced two asexual spores, including phialospore-type conidia, which were hyaline to mid brown, cylindrical to somewhat oval and thick walled when mature, 8.5 to 16.0 × 4.7 to 6.7 μm, released from phialides measuring 80.0 to 155.0 × 5.0 to 7.8 μm, and chlamydospores, which were oval, black, 13.4 to 25.0 × 8.9 to 12.5 μm, and formed in short chains from specialized hyphal tips. Perithecia were produced after 2 months by mating isolates on coconut exocarp placed on PDA. Perithecia were brown, globose, 280 μm in diameter, with a 1,100-μm long neck. Ascospores generally were ellipsoid, but some were unequally bent, 12 × 3 μm. The temperature for hyphal growth of this fungus was 25 to 30°C, with a growth rate of 3.3 cm per day. It did not grow at temperatures lower than 10°C or higher than 35°C. On the basis of these characteristics, this fungus was identified as Ceratocystis paradoxa (Dade) C. Moreau (anamorph Thielaviopsis paradoxa (de Seynes) Höhn) (1). The internal transcribed spacer (ITS) region of mycelial DNA of two isolates was amplified with ITS5 and ITS4 primers and the PCR products were sequenced (GenBank Accession Nos. GU358207 and GU358206). BLAST analysis of both sequences showed 99% sequence similarity with C. paradoxa strain WIN(M) 925 (Accession No. DQ318203). Two of the isolates (BCRC 34425 and BCRC 34426) were deposited in the Bioresource Collection and Research Center, Hsinchu, Taiwan. Twenty-four detached healthy coconut fruits were inoculated by placing spore suspensions (1 × 10 spores/ml) of eight wild type isolates onto wounds created by removing the calyx. Symptoms similar to those observed in the field developed on inoculated fruits after incubating at room temperature for 10 days, but did not develop on six fruits that were wounded but not inoculated. The same pathogen was reisolated from the inoculated fruits but not the control fruits. C. paradoxa has been reported as causing disease of coconut fruits in Brazil (2), but to our knowledge, this is the first report of the disease in Taiwan. References: (1) G. Morgan-Jones. No. 143 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Surrey, England, 1967. (2) V. Rossetti. "Crostas Pretas" das folhas e "Podridão Basal" dos frutos de coqueiro. O Biológico 21:54, 1955.
自2005年以来,从台湾南部树木上采收的椰子果实(Cocos nucifera L.)出现了果实腐烂病症状。该病仅发生在采收后的椰子果实上,在温暖季节更为严重。主要症状是椰子果实从果柄端或果实伤口处开始,外果皮、中果皮然后是内果皮变黑。从患病果实中一直分离出一种真菌。在马铃薯葡萄糖琼脂(PDA)上的菌落为白色,1至2天后变黑,有强烈的果香气味。该真菌产生两种无性孢子,包括瓶梗孢子型分生孢子,成熟时透明至中褐色,圆柱形至略呈椭圆形,壁厚,8.5至16.0×4.7至6.7μm,从80.0至155.0×5.0至7.8μm的瓶梗中释放,以及厚垣孢子,椭圆形,黑色,13.4至25.0×8.9至12.5μm,由特化的菌丝顶端形成短链。通过将分离物在置于PDA上的椰子外果皮上配对培养2个月后产生子囊壳。子囊壳褐色,球形,直径280μm,有一个1100μm长的颈部。子囊孢子通常为椭圆形,但有些弯曲不均,12×3μm。该真菌菌丝生长的温度为25至30°C,生长速率为每天3.3cm。在低于10°C或高于35°C的温度下不生长。基于这些特征,该真菌被鉴定为奇异长喙壳菌(Ceratocystis paradoxa (Dade) C. Moreau)(无性型为奇异拟茎点霉(Thielaviopsis paradoxa (de Seynes) Höhn))(1)。用ITS5和ITS4引物扩增了两个分离物菌丝体DNA的内部转录间隔区(ITS),并对PCR产物进行了测序(GenBank登录号:GU358207和GU-358206)。对这两个序列的BLAST分析显示与奇异长喙壳菌菌株WIN(M) 925(登录号:DQ318203)的序列相似性为99%。其中两个分离物(BCRC 34425和BCRC 34426)保藏于台湾新竹的生物资源保存及研究中心。通过将8个野生型分离物的孢子悬浮液(1×10孢子/ml)接种到去除花萼造成的伤口上,对24个离体健康椰子果实进行接种。在室温下培养10天后,接种果实上出现了与田间观察到的症状相似的症状,但6个仅受伤未接种的果实未出现症状。从接种果实中重新分离出了相同的病原菌,而对照果实中未分离到。奇异长喙壳菌已被报道在巴西引起椰子果实病害(2),但据我们所知,这是该病害在台湾的首次报道。参考文献:(1)G. Morgan-Jones。载于《英联邦真菌研究所病原真菌和细菌描述》第143号。英联邦真菌研究所,英国萨里,1967年。(2)V. Rossetti。椰子叶的“黑斑病”和果实的“基部腐烂病”。《生物学》21:54,1955年。