Secor G A, Lee I-M, Bottner K D, Rivera-Varas V, Gudmestad N C
Department of Plant Pathology, North Dakota State University, Fargo 58105.
Molecular Plant Pathology Laboratory, USDA, ARS, Beltsville, MD 20705.
Plant Dis. 2006 Mar;90(3):377. doi: 10.1094/PD-90-0377B.
An outbreak of a new potato disease occurred in Texas and Nebraska causing a serious defect in potato chips produced from commercial processing potatoes. The defect consists of patchy brown discoloration of chips and can be a cause for rejection of contracted potatoes by the processor. Infected potato plants exhibit symptoms of the purple top wilt syndrome similar to those of the purple top disease in processing potatoes caused by clover proliferation phytoplasma recently found in Washington and Oregon (3). Foliar symptoms include stunting, chlorosis, slight purple coloration of new growth, swollen nodes, proliferated axillary buds, and aerial tubers. Tuber symptoms include mild vascular discoloration and brown flecking of medullary rays. Seed potatoes from affected plants produce hair sprouts. Total nucleic acid was extracted from leaf and stolon tissue of symptomatic plants in the field and from tuber samples exhibiting the defect from commercial storages. Nested polymerase chain reactions (PCR) were performed using phytoplasma-universal 16SrDNA-based primers (P1/P7 and R16F2n/R16R2) (2) to detect the presence of phytoplasmas in these samples. A negative control, devoid of DNA templates in the reaction mix, was included in all PCR assays. In 2004, 13 foliar samples tested positive for phytoplasmas using PCR. None of the apparently symptomless plants or tubers tested positive. Restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified 16S rDNA using enzymes AluI, MseI, HhaI, BfaI, and Tsp509I indicated that four samples are associated with a phytoplasma belonging to subgroup A (16SrI-A) of the "Candidatus Phytoplasma asteris" (aster yellows phytoplasma) group (16SrI), and nine plant samples were associated with a new phytoplasma related to, but distinct from, the stolbur phytoplasma group (16SrXII). Nucleotide sequence analysis of cloned 16S rDNAs (GenBank Accession Nos. DQ174114-DQ174123) confirmed the results on the basis of RFLP analyses. Sequences of cloned 16S rDNAs were analyzed with previously described phytoplasma strains available in GenBank using DNAStar's (Madison, WI) Lasergene software MegAlign program. The new phytoplasma is only distantly related to the stolbur phytoplasma, sharing 96.6% sequence homology. In 2005, 14 defective tuber samples from storage and 16 symptomatic plants from the field tested positive for the new phytoplasma. In Texas and Nebraska, it appears that at least two distinct phytoplasmas seem to be involved in the disease complex contributing to the defects of processed products produced from infected potatoes. Previous reports have suggested a similar defect of chipping potatoes, but the phytoplasma associated with the disease was not identified (1). To our knowledgek, this the first report of this new phytoplasma associated with disease and defects of potato and the first report of this phytoplasma in the United States. References: (1) E. E. Bantarri et al. Trans. ASAE 33:221, 1990. (2) I.-M. Lee et al. Int. J. Sys. Bacteriol. 48:1153, 1998. (3) I.-M. Lee et al. Plant Dis. 88:429, 2004.
德克萨斯州和内布拉斯加州爆发了一种新的马铃薯疾病,导致商用加工马铃薯制成的薯片出现严重缺陷。这种缺陷表现为薯片上有斑驳的褐色变色,可能导致加工商拒收合同规定的马铃薯。受感染的马铃薯植株表现出紫顶萎蔫综合征的症状,类似于最近在华盛顿州和俄勒冈州发现的由三叶草增殖植原体引起的加工马铃薯紫顶病(3)。叶片症状包括生长迟缓、黄化、新梢轻微发紫、节肿大、腋芽增殖和气生块茎。块茎症状包括轻度维管束变色和髓射线褐色斑点。受影响植株的种薯会长出毛状芽。从田间有症状植株的叶片和匍匐茎组织以及商业储存中出现缺陷的块茎样品中提取了总核酸。使用基于植原体通用16SrDNA的引物(P1/P7和R16F2n/R16R2)(2)进行巢式聚合酶链反应(PCR),以检测这些样品中是否存在植原体。所有PCR检测均包括一个反应混合物中不含DNA模板的阴性对照。2004年,13个叶片样品经PCR检测植原体呈阳性。所有明显无症状的植株或块茎检测均为阴性。使用AluI、MseI、HhaI、BfaI和Tsp509I酶对PCR扩增的16S rDNA进行限制性片段长度多态性(RFLP)分析表明,4个样品与属于“星状植原体”(翠菊黄化植原体)组(16SrI)A亚组(16SrI-A)的植原体相关,9个植株样品与一种与 stolbur 植原体组(16SrXII)相关但不同的新植原体相关。克隆的16S rDNA的核苷酸序列分析(GenBank登录号:DQ174114-DQ174123)在RFLP分析的基础上证实了结果。使用DNAStar公司(威斯康星州麦迪逊)的Lasergene软件MegAlign程序,将克隆的16S rDNA序列与GenBank中先前描述的植原体菌株进行分析。这种新植原体与stolbur植原体关系较远,序列同源性为96.6%。2005年,14个储存中的有缺陷块茎样品和16个田间有症状植株经检测,这种新植原体呈阳性。在德克萨斯州和内布拉斯加州,似乎至少有两种不同的植原体参与了导致受感染马铃薯加工产品出现缺陷的病害复合体。以前的报告曾提到薯片马铃薯有类似的缺陷,但未鉴定出与该病相关的植原体(1)。据我们所知,这是关于这种与马铃薯病害和缺陷相关的新植原体的首次报告,也是该植原体在美国的首次报告。参考文献:(1)E. E. Bantarri等人,《美国农业工程师学会会刊》33:221,1990年。(2)I.-M. Lee等人,《国际系统细菌学杂志》48:1153,1998年。(3)I.-M. Lee等人,《植物病害》88:429,2004年。