Li J, Gao J M, Han Y H, Sun Y X, Huang Q
College of Plant Protection, Yunnan Agricultural University, Kunming, 650201, Yunnan, China. This work was supported by National Natural Science Foundation of China (No. 31270067).
Plant Dis. 2014 Sep;98(9):1270. doi: 10.1094/PDIS-12-13-1290-PDN.
The loquat (Eriobotrya japonica (Thunb.) Lindl.), an important perennial evergreen fruit crop that originated in China and is also cultivated as an ornamental plant, is well known all over the world for high nutritional, medicinal, economic, and ecological values (4). So far, it has been grown in more than 30 countries (1) and is becoming an important industry in China, Spain, Japan, India, Pakistan, and Turkey (2). During an investigation from May to August of 2013, severely withered loquat plants were observed in Kunming city of Yunnan Province (25°02' N; 102°42' E), Southwest China. Initial symptoms were brown lesions of leaves and canker on one to several branches, leaves of the whole tree turned wilted. Cross sections of diseased plants revealed irregularly shaped brown discoloration in the xylem of the trunk. A fungus was consistently isolated from the leaf when diseased leaves were incubated between two slices of fresh carrot root. Spore masses were picked from the apices of perithecia and transferred to malt extract agar medium (MEA) and incubated at 25°C. After perithecium formation, observed perithecia were black, globose (176.0 to 303.2 × 186.0 to 274.3 μm) and showed a long black neck (634.2 to 809.9 μm). Ascospore had the typical format of a "hat" with dimensions of 4.6 to 6.3 × 3.3 to 4.8 μm. Cylindrical endoconidia (7.1 to 36.1 × 2.9 to 6.0 μm) were found. Chlamydospores were ovoid or obpyriform and smooth (8.6 to 12.1 × 6.9 to 12.1 μm). PCR amplification was carried out for one isolate, YT2. The ITS region of rDNA was sequenced using the procedures of Thorpe et al (3). Analysis of ITS sequence data (GenBank Accession Nos. KF963101 and KF963102) showed that the isolates were 99% homologous to the isolate of Ceratocystis fimbriata from diseased Colocasia esculenta in Cuba, China, and Hawaii (AY526304 to 06) by BLAST analysis. Therefore, the fungus was identified as C. fimbriata based on morphological and molecular characteristics. Pathogenicity of the six isolates from this study was tested by inoculation of 1-year-old pot grown seedlings of loquat. The soil of six plants was inoculated by drenching with 40 ml spore suspension (10 spores/ml). Control plants were inoculated with 40 ml of sterile distilled water. The plants were maintained in a controlled greenhouse at 25°C and watered weekly. After inoculation for 1 week, all plants produced wilt symptoms; as the disease progressed, leaves withered and died after 3 weeks of inoculation while control plants remained symptomless. C. fimbriata was successfully re-isolated from the infected trees and no fungal growth was observed in the controls. The pathogenicity assay showed that C. fimbriata was pathogenic to loquat. To our knowledge, this is the first report of C. fimbriata causing wilt of loquat in China. Since C. fimbriata is one of the most aggressive plant pathogens on a wide variety of perennial as well as agronomic crop plants worldwide, and the infection can pose a significant threat to the production of loquat, it is critical to deploy appropriate management strategies to limit the fungus spread. References: (1) J. J. Feng et al. Acta Hort. 750:117, 2007. (2) J. Janick. Acta Hort. 750:27, 2007. (3) D. J. Thorpe et al. Phytopathology 95:316, 2005. (4) J. Yan et al. Pak. J. Bot. 44:1215, 2012.
枇杷(Eriobotrya japonica (Thunb.) Lindl.)是一种重要的多年生常绿果树作物,原产于中国,也作为观赏植物种植,因其具有高营养、药用、经济和生态价值而闻名于世(4)。到目前为止,它已在30多个国家种植(1),并在中国、西班牙、日本、印度、巴基斯坦和土耳其成为一项重要产业(2)。在2013年5月至8月的一次调查中,在中国西南部云南省昆明市(北纬25°02′;东经102°42′)观察到枇杷植株严重枯萎。最初症状是叶片出现褐色病斑,一至多个枝条上出现溃疡,整棵树的叶子枯萎。患病植株的横切面显示树干木质部有不规则形状的褐色变色。当将病叶在两片新鲜胡萝卜根之间培养时,始终能从叶片中分离出一种真菌。从子囊壳顶端采集孢子团,转移到麦芽提取物琼脂培养基(MEA)上,在25°C下培养。子囊壳形成后,观察到的子囊壳为黑色,球形(176.0至303.2×186.0至274.3μm),并有一个长的黑色颈部(634.2至809.9μm)。子囊孢子具有典型的“帽子”形状,尺寸为4.6至6.3×3.3至4.8μm。发现圆柱形内生分生孢子(7.1至36.1×2.9至6.0μm)。厚垣孢子呈卵形或倒梨形,表面光滑(8.6至12.1×6.9至12.1μm)。对一个分离株YT2进行了PCR扩增。使用Thorpe等人(3)的方法对rDNA的ITS区域进行了测序。ITS序列数据分析(GenBank登录号KF963101和KF963102)表明,通过BLAST分析,这些分离株与来自古巴、中国和夏威夷患病芋艿(Colocasia esculenta)的Fimbriata炭疽菌分离株(AY526304至06)有99%的同源性。因此,根据形态和分子特征,该真菌被鉴定为Fimbriata炭疽菌。通过接种1年生盆栽枇杷幼苗测试了本研究中六个分离株的致病性。用40毫升孢子悬浮液(10个孢子/毫升)浇灌六株植物的土壤。对照植株接种40毫升无菌蒸馏水。将植株置于25°C的可控温室中,每周浇水一次。接种1周后,所有植株都出现了枯萎症状;随着病情发展,接种3周后叶片枯萎死亡,而对照植株无症状。成功地从受感染的树木中重新分离出Fimbriata炭疽菌,对照中未观察到真菌生长。致病性测定表明Fimbriata炭疽菌对枇杷具有致病性。据我们所知,这是中国关于Fimbriata炭疽菌引起枇杷枯萎的首次报道。由于Fimbriata炭疽菌是全球多种多年生以及农艺作物上最具侵袭性 的植物病原体之一,且这种感染会对枇杷生产构成重大威胁,因此部署适当的管理策略以限制真菌传播至关重要。参考文献:(1)J. J. Feng等人,《园艺学报》750:117,2007。(2)J. Janick,《园艺学报》750:27,2007。(3)D. J. Thorpe等人,《植物病理学》95:316,2005。(4)J. Yan等人,《巴基斯坦植物学杂志》44:1215,2012。