Chen X R, Xing Y P, Zhang T X, Zheng J T, Xu J Y, Wang Z R, Tong Y H
College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, China.
Ningbo Technology Extension Center for Forestry and Specialty Forest Products, Ningbo 315010, China.
Plant Dis. 2012 May;96(5):764. doi: 10.1094/PDIS-01-12-0065-PDN.
Red bayberry (Myrica rubra Seib. & Zucc.) has great economic importance in eastern and southern China. However, increasing cultivation of red bayberry has resulted in an increase in diseases such as leaf necrosis. In April 2011, a survey was conducted to identify the causal agents of leaf necrosis of red bayberry (cv. Biqi) in Cixi City, Zhejiang Province. Symptoms began with oval and pale brown lesions (2 mm in diameter) that developed into a round to irregular shape (4 to 12 mm in diameter) with pale brown centers and dark brown borders. After approximately 4 months, necrotic lesions expanded to the leaf tips or margins. Black acervuli developed on lesions at later stages. Leaf tissues were surface sterilized with 0.5% sodium hypochlorite for 3 min and rinsed in sterile water before plating onto potato dextrose agar (PDA). Seven isolates were obtained from four samples from four fields on PDA at 25°C. The colonies were cottony white with filiform edges and produced a honey yellow color into the agar at 7 days. Conidia were produced in ink-like fruiting bodies at 4 days at 25°C on PDA. Conidia were straight or slightly curved, fusiform, and five celled with constrictions at the septa. Conidia ranged from 18.7 to 25.8 × 6.2 to 7.7 μm with hyaline apical and basal cells. Thirteen percent of the apical cells had two and the rest had three hyaline appendages ranging from 11.2 to 26.0 μm long. Basal appendages were hyaline, straight, and varied from 3.6 to 5.8 μm long. The color of three median cells was light to dark brown and demonstrated versicolorous. These morphological characteristics matched those of Pestalotiopsis sydowiana (Bresadola) Sutton (1). The morphological identification of the fungus was confirmed by nucleotide blast analysis of the 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA regions (GenBank Accession No. JQ322999), which revealed 100% with those of other P. sydowiana isolates (e.g., GenBank Accession No. FJ478105). Koch's postulates were confirmed with 20 healthy leaves of the same size on three branches of three plants in the field. Leaves were wounded by pressing slightly with sterile needles. Mycelial plugs (5 mm in diameter) obtained from the periphery of 7-day-old cultures were placed onto the wounds and covered with sterile-water-saturated cotton. Wounded leaves treated with sterile agar plugs served as controls. The inoculated leaves were sealed in moist plastic bags for 24 h to establish high humid conditions at 21 to 30°C. After 23 days, symptoms on all inoculated leaves were identical to those described above, whereas noninoculated control leaves did not show any symptoms. The fungus was consistently reisolated from the lesions. To our knowledge, this is the first report of P. sydowiana causing leaf necrosis of M. rubra in China. Results can help to better understand the diseases threatening red bayberry trees and develop effective control strategies for better fruit production. Reference: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961.
杨梅(Myrica rubra Seib. & Zucc.)在中国东部和南部具有重要的经济价值。然而,杨梅种植面积的不断增加导致了诸如叶片坏死等病害的增多。2011年4月,对浙江省慈溪市荸荠种杨梅叶片坏死的病原菌进行了调查。症状始于椭圆形、浅褐色病斑(直径2毫米),之后发展成圆形至不规则形(直径4至12毫米),病斑中心浅褐色,边缘深褐色。约4个月后,坏死病斑扩展至叶尖或叶缘。后期病斑上产生黑色分生孢子盘。叶片组织用0.5%次氯酸钠表面消毒3分钟,并用无菌水冲洗后,接种到马铃薯葡萄糖琼脂(PDA)培养基上。在25℃条件下,从四个果园的四个样本中在PDA培养基上获得了7个分离株。菌落呈棉絮状白色,边缘丝状,7天后培养基中产生蜜黄色。在25℃条件下,接种在PDA培养基上4天后,分生孢子在墨汁状子实体中产生。分生孢子直或稍弯,梭形,具5个细胞,隔膜处缢缩。分生孢子大小为18.7至25.8×6.2至7.7微米,顶端和基部细胞透明。13%的顶端细胞有2根附属丝,其余顶端细胞有3根透明附属丝,长度在11.2至26.0微米之间。基部附属丝透明、直,长度在3.6至5.8微米之间。中间3个细胞的颜色从浅褐色到深褐色,呈现杂色。这些形态特征与聚生拟盘多毛孢(Pestalotiopsis sydowiana (Bresadola) Sutton)相符(1)。通过对核糖体DNA区域的5.8S亚基及侧翼内转录间隔区(ITS1和ITS2)进行核苷酸比对分析(GenBank登录号:JQ322999),进一步证实了该真菌的形态学鉴定结果,其与其他聚生拟盘多毛孢分离株(如GenBank登录号:FJ478105)的序列相似度达100%。通过对田间三株植物三个枝条上20片相同大小的健康叶片进行柯赫氏法则验证。用无菌针轻轻刺伤叶片。从7日龄培养物边缘获取直径5毫米的菌丝块,放置在伤口上,并用饱和无菌水的棉花覆盖。用无菌琼脂块处理的受伤叶片作为对照。接种后的叶片密封在潮湿的塑料袋中24小时,以在21至30℃营造高湿度条件。23天后,所有接种叶片上的症状与上述描述一致,而未接种的对照叶片未出现任何症状。该真菌能持续从病斑中重新分离得到。据我们所知,这是聚生拟盘多毛孢引起中国杨梅叶片坏死的首次报道。研究结果有助于更好地了解威胁杨梅树的病害,并制定有效的防治策略以提高果实产量。参考文献:(1)E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961.