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南京地区早竹褐色秆条斑病由芦竹节菱孢引起的首次报道

First Report of Brown Culm Streak of Phyllostachys praecox Caused by Arthrinium arundinis in Nanjing, China.

作者信息

Chen K, Wu X-Q, Huang M-X, Han Y-Y

机构信息

College of Forest Resources and Environment, Nanjing Forestry University, Nanjing, Jiangsu 210037, China.

出版信息

Plant Dis. 2014 Sep;98(9):1274. doi: 10.1094/PDIS-02-14-0165-PDN.

Abstract

Early spring shoot bamboo (Phyllostachys praecox) is profitable for the production of bamboo shoots and is widely cultivated in Jiangsu and Zhejiang provinces in China. In recent years, a new disease named brown culm streak has commonly occurred in bamboo plantations. During initial stages, small yellow or sandy beige spots (1 to 2 mm diameter) develop on the culm. Spots expand and develop into fine brown streaks (10 to 15 × 1 to 2 mm), and the quantity of spots increases greatly at the mid-stage of the disease. The streaks continue to expand (10 to 30 × 2 to 3 mm) and may coalesce into brown patches that cover 70% of the culm surface in late stages of disease. Color of the internal bamboo cavity gradually changes from white to brown without an alcoholic odor, and plants eventually die. Diseased tissues (1 × 1.5 cm) were taken from the edges of brown streaks, surface disinfected in 0.1% HgCl for 3 min, rinsed in distilled water three times, and incubated on potato dextrose agar (PDA) at 25°C. Based on colony and conidial morphology, six different fungi (ZBY01, ZBY02, ZBY03, ZBY04, ZBY05, and ZBY06) were isolated. A suspension of hyphal fragments (10 pieces/ml) of each of the fungi was used to inoculate plants in the field. Healthy bamboo culms were stabbed with alcohol-sterilized needles after surface disinfection and wounds of each bamboo were inoculated with 3 ml hypha fragment suspensions by paintbrush, then covered with a piece of sterile plastic film. A control treatment was inoculated with sterile water. Each treatment had two replicates. Twenty days after inoculation, only fungus ZBY01 caused similar symptoms with the diseased bamboo in the field and was re-isolated from necrotic tissue of inoculated plants, thus fulfilling Koch's postulates. Mycelium of fungus ZBY01 covered entire PDA plates (90 mm diameter) after incubating for 6 days at 25°C and was white, flat, and floccose with moderate aerial mycelium. Mycelium consisted of smooth, hyaline, branched, septate hyphae, 2 to 3 μm diameter. Conidiophores were erect, septate, pale brown, smooth, and reduced to conidiogenous cells. Conidiogenous cells were pale brown, smooth, ampulliform, 6 to 9 μm long; the apical neck was 1 to 3 μm long, basal part 5 to 6 μm long. Conidia were 1-celled, dark brown to black, smooth, lenticular, 5 to 7 μm in diameter, 2 to 4 μm wide, lemon-shaped in side view, and spherical from top view with an equatorial ring. Based on morphological characteristics, it was identified as Arthrinium arundinis (Corda) Dyko & B. Sutton (1). The internal transcribed spacer (ITS) region from the isolate ZBY01 was amplified with PCR using ITS1-ITS4 primer pairs and the amplicon sequenced. Size of the sequenced region was 540 bp and had 99% identity with A. arundinis, which was in accord with morphological identification. The sequence was deposited in Gen Bank under accession number KF850624. To our knowledge, brown culm streak of P. praecox is a new disease of bamboo that has not been reported in other countries; however, A. arundinis has been reported as a pathogen on barley in Fairfield, Montana. Reference: (1) P. W. Crous and J. Z. Groenewald. A phylogenetic re-evaluation of Arthrinium. IMA Fungus. 4:133, 2013.

摘要

早竹(Phyllostachys praecox)笋用效益显著,在中国江苏和浙江广泛种植。近年来,竹林中普遍出现一种名为竹秆褐条斑病的新病害。发病初期,竹秆上出现小黄斑或沙米色斑点(直径1至2毫米)。斑点扩大并发展成细褐色条纹(10至15×1至2毫米),病害中期斑点数量大幅增加。条纹继续扩大(10至30×2至3毫米),后期可能融合成褐色斑块,覆盖竹秆表面的70%。竹腔内颜色逐渐从白色变为褐色,无酒精气味,植株最终死亡。从褐色条纹边缘取患病组织(1×1.5厘米),在0.1% HgCl中表面消毒3分钟,用蒸馏水冲洗3次,然后在25°C下于马铃薯葡萄糖琼脂(PDA)上培养。根据菌落和分生孢子形态,分离出六种不同真菌(ZBY01、ZBY02、ZBY03、ZBY04、ZBY05和ZBY06)。将每种真菌的菌丝片段悬浮液(10片/毫升)用于田间接种植株。健康竹秆经表面消毒后用酒精消毒的针刺伤,用毛笔将3毫升菌丝片段悬浮液接种到每根竹子的伤口上,然后用一块无菌塑料薄膜覆盖。对照处理接种无菌水。每个处理有两个重复。接种20天后,只有真菌ZBY01引起与田间患病竹子相似的症状,并从接种植株的坏死组织中再次分离得到,从而满足柯赫氏法则。真菌ZBY01的菌丝体在25°C下培养6天后覆盖整个PDA平板(直径90毫米),呈白色、扁平、絮状,气生菌丝适中。菌丝体由光滑、透明、分枝、有隔膜的菌丝组成,直径2至3微米。分生孢子梗直立、有隔膜、浅褐色、光滑,退化为产孢细胞。产孢细胞浅褐色、光滑、瓶形,长6至9微米;顶端颈部1至3微米长,基部5至6微米长。分生孢子单细胞,深褐色至黑色,光滑,透镜形,直径5至7微米,宽2至4微米,侧面观为柠檬形,顶面观为球形,有赤道环。根据形态特征,将其鉴定为燕麦节菱孢(Arthrinium arundinis (Corda) Dyko & B. Sutton (1))。使用ITS1 - ITS4引物对通过PCR扩增分离物ZBY01的内转录间隔区(ITS)区域,并对扩增产物进行测序。测序区域大小为540 bp,与燕麦节菱孢的同一性为99%,与形态学鉴定结果一致。该序列已存入Gen Bank,登录号为KF850624。据我们所知,早竹的竹秆褐条斑病是一种竹子新病害,在其他国家尚未见报道;然而,燕麦节菱孢已被报道为蒙大拿州费尔菲尔德大麦上的病原菌。参考文献:(1) P. W. Crous和J. Z. Groenewald。节菱孢属的系统发育重新评估。IMA真菌。4:133,2013年。

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