Lebas B S M, Ochoa-Corona F M, Elliott D R, Tang Z, Alexander B J R
Plant Environmental Laboratory, Biosecurity New Zealand, Ministry of Agriculture and Forestry, P.O. Box 2095, Auckland 1015, New Zealand.
Plant Dis. 2005 Oct;89(10):1103-1108. doi: 10.1094/PD-89-1103.
High Plains virus (HPV) causes a potentially serious economic disease of cereals and is of quarantine importance for New Zealand. HPV is transmitted by the wheat curl mite Aceria tosichella, and neither the virus nor its vector is present in New Zealand. Cereal seeds imported to New Zealand are required to be certified HPV-free, as the virus is a regulated pest. A procedure was developed for inspecting plants and testing cereal seedlings in quarantine using reverse transcriptase polymerase chain reaction (RT-PCR) as a detection method. A sample of 50,655 sweet corn seeds was taken from an imported commercial line and germinated in containment. Symptomatic seedlings were collected at 3 and 4 ½ weeks after sowing. Eight out of 27 symptomatic samples tested HPV positive by RT-PCR and were confirmed by enzyme-linked immunosorbent assay (ELISA). Sequence analysis revealed that the HPV isolates had a 99.3 to 100% nucleotide identity and 99.0 to 100% amino acid similarity with the HPV USA isolate (GenBank accession no. U60141). HPV variants were detected by single stranded conformational polymorphism (SSCP) analysis but not by restriction fragment length polymorphism (RFLP).
高平原病毒（HPV）可引发一种对谷物具有潜在严重经济影响的疾病，且对新西兰具有检疫重要性。HPV由小麦曲叶螨传播，新西兰既不存在该病毒，也没有其传播媒介。由于该病毒属于受管制害虫，进口到新西兰的谷物种子必须经过认证，确保无HPV。已开发出一种程序，在检疫期间使用逆转录聚合酶链反应（RT-PCR）作为检测方法来检查植物并检测谷物幼苗。从一个进口商业品种中抽取了50655粒甜玉米种子样本，并在隔离环境中使其发芽。在播种后3周和4.5周收集出现症状的幼苗。27个出现症状的样本中有8个通过RT-PCR检测呈HPV阳性，并经酶联免疫吸附测定（ELISA）确认。序列分析显示，HPV分离株与美国HPV分离株（GenBank登录号U60141）的核苷酸同一性为99.3%至100%，氨基酸相似性为99.0%至100%。通过单链构象多态性（SSCP）分析检测到了HPV变体，但限制性片段长度多态性（RFLP）分析未检测到。
