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丁香假单胞菌豌豆致病变种引起豌豆细菌性疫病在土耳其的首次报道。

First Report of Bacterial Blight Caused by Pseudomonas syringae pv. pisi on Pea in Turkey.

作者信息

Benlioglu K, Özyilmaz Ü, Ertan D

机构信息

Department of Plant Protection, Faculty of Agriculture, Adnan Menderes University, 09100 Aydin, Turkey.

出版信息

Plant Dis. 2010 Jul;94(7):923. doi: 10.1094/PDIS-94-7-0923A.

Abstract

In April of 2009, leaf blight symptoms were observed on field peas (Pisum sativum L.) grown in Söke, Torbali, and Ödemis counties in the Aegean Region of Turkey. Field inspections revealed disease incidence as high as 45% and the disease was found in 13 commercial fields. Initial symptoms consisted of small, dark green, water-soaked lesions on leaves, stipules, and stems near ground level. Lesions often enlarged and coalesced and turned chocolate brown with a water-soaked margin. Stem infections usually coalesced and girdled the stem spreading upward to stipules and leaflets forming a fan-like lesion on the stipule. A fluorescent, gram-negative bacterium was consistently isolated from diseased tissues onto King's B medium. Twelve strains (five from cv. Early Sweet, three from cv. Geneva, two from cv. Bolero, and two from cv. Carina) from thirteen pea fields were obtained. All strains metabolized glucose oxidatively, and their reactions in LOPAT tests were +, -, -, -, +, and thus classified as belonging to Pseudomonas syringae LOPAT group Ia (1). The 12 strains utilized homoserine, inositol, sorbitol, sucrose, mannitol, and mannose but did not utilize erythritol, trehalose, and L-tartarate. All showed ice nucleation activity but variable results were obtained for gelatin liquefaction and esculin hydrolysis. Identification of P. syringae pv. pisi was confirmed by sequencing the 16S rDNA with primers Univ-1390R (3) and 27F (2). Sequences of the three local strains (Bz2, Bz4, and Bz8) were 100% identical to a type culture strain. The nucleotide sequence of strain Bz4 was submitted to GenBank (Accession No. GU332546). Pathogenicity tests were performed on greenhouse-grown 2-week-old pea plants cv. Geneva as three replicates in 12-cm pots containing a steamed sand/peat/soil mixture. Plants were stab inoculated by puncturing the main stem at its junction with the stipules at the second node from the apical end with a 26-gauge needle through a 5-μl drop of 10 CFU/ml bacterial suspensions. Control plants were inoculated with sterile water. After 10 days of incubation in a growth chamber at 24 ± 1°C with a 14-h photoperiod, stems inoculated with pea isolates resulted in water-soaked tissue spreading from the site of inoculation along the veins on stipules and leaflets that were identical to symptoms seen in the field. Control plants remained symptomless. Isolates recovered from the symptomatic stems showed the same morphological and biochemical features of the original isolates. All physiological and biochemical tests as well as the pathogenicity assay were performed at least twice and the type strain of P. syringae pv. pisi (NCPPB 2585) was used as reference. On the basis of the physiological, biochemical, genetic, and pathological characteristics, all strains were identified as P. syringae pv. pisi. To our knowledge, this is the first report of P. syringae pv. pisi causing bacterial blight on pea in Turkey. Turkey currently produces approximately 93.000 t of peas annually and three-quarters of that is produced in Western Anatolia. The new disease may represent a limiting factor for future production. References: (1) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (2) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991. (3) D. Zeng et al. Appl. Environ. Microbiol. 62:4504, 1996.

摘要

2009年4月,在土耳其爱琴海地区的索凯、托尔巴利和厄代米斯县种植的豌豆(Pisum sativum L.)上观察到叶枯病症状。田间检查发现发病率高达45%,且在13个商业种植田发现了该病。最初症状表现为叶片、托叶和近地面茎上出现小的、深绿色、水渍状病斑。病斑常常扩大并融合,变成巧克力褐色,边缘有水渍状。茎部感染通常会融合并环绕茎部,向上蔓延至托叶和小叶,在托叶上形成扇形病斑。从病组织中持续分离出一种荧光、革兰氏阴性细菌,接种到金氏B培养基上。从13个豌豆田获得了12个菌株(5个来自早熟甜豌豆品种,3个来自日内瓦品种,2个来自博莱罗品种,2个来自卡丽娜品种)。所有菌株通过氧化代谢葡萄糖,它们在LOPAT试验中的反应为 +、-、-、-、+,因此被归类为丁香假单胞菌LOPAT Ia组(1)。这12个菌株利用高丝氨酸、肌醇、山梨醇、蔗糖、甘露醇和甘露糖,但不利用赤藓醇、海藻糖和L - 酒石酸盐。所有菌株均表现出冰核活性,但明胶液化和七叶苷水解的结果各不相同。通过用引物Univ - 1390R(3)和27F(2)对16S rDNA进行测序,确认了豌豆假单胞菌丁香假单胞菌致病变种(P. syringae pv. pisi)。三个本地菌株(Bz2、Bz4和Bz8)的序列与一个模式培养菌株100%相同。菌株Bz4的核苷酸序列已提交至GenBank(登录号GU332546)。对温室中生长2周的日内瓦品种豌豆植株进行致病性试验,在装有蒸汽消毒的沙子/泥炭/土壤混合物的12厘米花盆中进行三次重复。用26号针头通过5微升10 CFU/ml细菌悬浮液在主茎与顶端第二个节的托叶交界处穿刺进行刺伤接种。对照植株接种无菌水。在24 ± 1°C、14小时光周期条件下的生长室中培养十天后,接种豌豆分离株的茎出现水渍状组织,从接种部位沿托叶和小叶的叶脉蔓延,与田间观察到的症状相同。对照植株无症状。从有症状的茎中分离出的菌株表现出与原始分离株相同的形态和生化特征。所有生理生化试验以及致病性测定至少进行了两次,并以豌豆假单胞菌丁香假单胞菌致病变种(P. syringae pv. pisi)的模式菌株(NCPPB 2585)作为参考。根据生理、生化、遗传和病理特征,所有菌株均被鉴定为豌豆假单胞菌丁香假单胞菌致病变种。据我们所知,这是豌豆假单胞菌丁香假单胞菌致病变种在土耳其引起豌豆细菌性叶枯病的首次报道。土耳其目前每年生产约93000吨豌豆,其中四分之三产于安纳托利亚西部。这种新病害可能成为未来生产的限制因素。参考文献:(1)R. A. Lelliott等人,《应用细菌学杂志》29:470,1966年。(2)W. G. Weisburg等人,《细菌学杂志》173:697,1991年。(3)D. Zeng等人,《应用与环境微生物学》62:4504,1996年。

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