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中国首次报道由肿胀链霉菌引起的马铃薯疮痂病

First Report of Potato Scab Caused by Streptomyces turgidiscabies in China.

作者信息

Zhao W Q, Liu D Q, Yu X M

机构信息

Biological Control Center of Plant Diseases and Plant Pests of Hebei Province, Department of Plant Pathology, Agricultural University of Hebei, Baoding 071001, China.

Biological Control Center of Plant Diseases and Plant Pests of Hebei Province, Department of Biochemistry and Cell Biology, Agricultural University of Hebei, Baoding 071001, China. This study was supported by the National Natural Science Foundation of China (No. 30700523) and the Hi-Tech Research and Development Program of China (No. 2006AA10A211).

出版信息

Plant Dis. 2008 Nov;92(11):1587. doi: 10.1094/PDIS-92-11-1587C.

DOI:10.1094/PDIS-92-11-1587C
PMID:30764453
Abstract

Potato scab, caused by several plant pathogenic Streptomyces species, is known to occur in potato-planting areas worldwide. Symptoms of disease on potato tubers are shallow, raised, or pitted corky lesions (2). In 1998, Streptomyces turgidiscabies was reported as a new potato scab pathogen from Hokkaido, Japan (3). Potato scab has been observed in many potato-cultivation areas in China and incidence of the disease was approximately 6 to 10% in some fields in 2006 (in our survey). To investigate the casual agent of scab disease, isolations were made from scabby potato tubers collected from different areas using oatmeal agar. Identification of an isolate from Shaanxi Province was based on morphological and physiological characterization followed by 16S rRNA confirmation. Characteristics were gray, aerial hypha, rectiflexuous spore chains, a smooth spore surface, and spores that were 0.5 to 0.7 × 1.0 to 1.2 μm. The strain did not produce melanin on tyrosine-peptone-yeast extract agar media, did not produce diffusible pigments, used all the International Streptomyces Project (ISP) sugars (4) as single carbon sources, used l-hydroxyproline, l-tyrosine, and l-histidine as single nitrogen sources but not l-methionine, grew at pH 4.5, was susceptible to streptomycin (20 μg ml), phenol (0.1%), and penicillin (10 IU ml) but not to crystal violet (0.5 μg ml), and produced HS. The identification was confirmed by comparison of its 16S rRNA sequence with the GenBank database using the BLAST program. The 16S rRNA sequence was amplified by PCR with primers S1: 5'-CATTCACGGAGAGTTTGATCC-3' and S2: 5'-AGAAAGGAGGTGATCCAGCC-3' and sequenced. BLASTn analysis of the sequence obtained showed the highest similarity (99.9%) with S. turgidiscabies type strain ATCC 700248 (GenBank Accession No. AB026221). The sequence was submitted to GenBank (Accession No. AM889495). Pathogenicity of the strain was tested in the greenhouse on potato tubers of cv. Favorita grown in pots (one plant per pot, three replicates). One hundred milliliters of inoculum (1 × 10 CFU ml) of the strain was mixed with sterile soil and vermiculite (1:1) in each pot. Potato plants were grown at 25°C and the soil was allowed to dry between waterings. The immature potato tubers were used to evaluate scab symptoms 10 weeks after planting. All tubers inoculated with the pathogen developed typical common scab symptoms consisting of erumpent, brown, corky lesions, which is different from the symptoms caused by S. reticuliscabiei (1). The noninoculated control tubers did not show scab symptoms. S. turgidiscabies was reisolated from lesions of diseased immature tubers. The pathogenicity test indicates that S. turgidiscabies caused scab disease on potato tubers. To our knowledge, this is the first report of S. turgidiscabies causing potato scab disease in China. References: (1) K. Bouchek-Mechiche et al. Int. J. Syst. Evol. Microbiol. 56:2771, 2006. (2) R. Loria et al. Plant Dis. 81:836, 1997. (3) K. Miyajima et al. Int. J. Syst. Bacteriol. 48:495, 1998. (4) E. B. Shirling and D. Gottlieb. Int. J. Syst. Bacteriol. 16:313, 1966.

摘要

马铃薯疮痂病由多种植物致病链霉菌引起,在全球马铃薯种植区均有发生。马铃薯块茎上的病害症状为浅的、凸起的或凹陷的木栓质病斑(2)。1998年,肿胀链霉菌被报道为来自日本北海道的一种新的马铃薯疮痂病病原菌(3)。在中国的许多马铃薯种植区都观察到了马铃薯疮痂病,2006年在我们的调查中,该病在一些田块中的发病率约为6%至10%。为了调查疮痂病的病原菌,使用燕麦片琼脂从不同地区采集的疮痂马铃薯块茎中进行分离。对来自陕西省的一个分离株的鉴定基于形态学和生理学特征,随后进行16S rRNA确认。其特征为灰色、气生菌丝、直形弯曲的孢子链、孢子表面光滑,孢子大小为0.5至0.7×1.0至1.2μm。该菌株在酪氨酸 - 蛋白胨 - 酵母提取物琼脂培养基上不产生黑色素,不产生可扩散色素,能利用所有国际链霉菌项目(ISP)糖类(4)作为单一碳源,能利用L - 羟脯氨酸、L - 酪氨酸和L - 组氨酸作为单一氮源,但不能利用L - 蛋氨酸,在pH 4.5下生长,对链霉素(20μg/ml)、苯酚(0.1%)和青霉素(10 IU/ml)敏感,但对结晶紫(0.5μg/ml)不敏感,并产生H2S。通过使用BLAST程序将其16S rRNA序列与GenBank数据库进行比较来确认鉴定结果。16S rRNA序列用引物S1:5'-CATTCACGGAGAGTTTGATCC-3'和S2:5'-AGAAAGGAGGTGATCCAGCC-3'通过PCR扩增并测序。对所得序列的BLASTn分析显示与肿胀链霉菌模式菌株ATCC 700248(GenBank登录号AB026221)的相似性最高(99.9%)。该序列已提交至GenBank(登录号AM889495)。在温室中对盆栽的费沃瑞它品种马铃薯块茎(每盆一株,三个重复)进行该菌株的致病性测试。将100毫升该菌株的接种物(1×10CFU/ml)与无菌土壤和蛭石(1:1)在每个盆中混合。马铃薯植株在25°C下生长,浇水之间让土壤干燥。种植10周后,使用未成熟的马铃薯块茎评估疮痂症状。所有接种病原菌的块茎都出现了典型的普通疮痂症状,包括隆起的、褐色的木栓质病斑,这与网状链霉菌引起的症状不同(1)。未接种的对照块茎没有出现疮痂症状。从患病未成熟块茎的病斑中重新分离出肿胀链霉菌。致病性测试表明肿胀链霉菌可引起马铃薯块茎的疮痂病。据我们所知,这是中国关于肿胀链霉菌引起马铃薯疮痂病的首次报道。参考文献:(1)K. Bouchek - Mechiche等人,《国际系统与进化微生物学杂志》56:2771,2006年。(2)R. Loria等人,《植物病害》81:836,1997年。(3)K. Miyajima等人,《国际系统细菌学杂志》48:495,1998年。(4)E. B. Shirling和D. Gottlieb,《国际系统细菌学杂志》16:313,1966年。