Chen C-C, Ko W-F, Lin C-Y, Jan F-J, Hsu H T
Taichung District Agriculture Improvement Station, Changhua 515, Taiwan.
Department of Plant Pathology, National Chung Hsing University, Taichung 402, Taiwan.
Plant Dis. 2003 Dec;87(12):1539. doi: 10.1094/PDIS.2003.87.12.1539C.
Calla lilies are ornamental plants of major economic importance in Taiwan. They are grown in the central and northern areas of the island, and ≈3 million stems are shipped annually. Calla lilies are susceptible to several viruses (1). Infections by Cucumber mosaic virus, Dasheen mosaic virus, Turnip mosaic virus, and Watermelon silver mottle virus were reported in Taiwan. Recently, virus-like symptoms including yellow mottling, light yellow spot, yellow ringspot, and mosaic were observed on leaves of field-grown calla lilies from Changhua County, located in central Taiwan. In March 2001, a virus culture was isolated from diseased calla lilies and established in Chenopodium quinoa Willd. and Nicotiana benthamiana Domin. When inoculated with the virus, healthy calla lilies developed chlorotic spots that enlarged and fused to form large, yellow patches on inoculated leaves. Symptoms were similar to those on the naturally infected plants observed in the fields. The virus induced chlorotic local lesions on C. quinoa, C. ficifolium Sm., C. amaranticolor Coste & Reyn, Cucurbita moschata Duchesne ex Poir, Lisianthus russellianum (Don.) Griseb, Phaseolus angularis Wight, Vigna angularis Willd., and V. radiata (L.) Wilczek. In addition to the localized chlorotic spots on inoculated leaves, systemic invasion of the virus was also observed 8 to 10 days postinoculation in Dianthus caryophyllus L., D. chinensis L., and Glycine max Merr. In N. benthamiana, the only symptom observed was systemic wilting. Examination of 2% of uranyl-acetate-stained samples using electron microscopy revealed the presence of spherical particles ≈34 to 35 nm in diameter in crude extracts of leaves of diseased calla lilies, or infected C. quinoa. Similar particles were also observed in the cytoplasm but not in the nuclei in ultrathin sections of virus-infected leaf tissues of C. quinoa and N. benthamiana. Differential centrifugation followed by sucrose density gradient centrifugation of tissue extracts of infected C. quinoa yielded virions with similar size. Sodium dodesyl sulfate polyacrylamide gel electrophoresis of the purified virus showed a single structural polypeptide ith a M of 41.6 kDa. The viral antigen reacted positively with its homologous antiserum and an antiserum against Carnation mottle virus (CarMV; Agdia, Inc., Elkhart, IN) in double antibody sandwich enzyme-linked immunosorbent assay. Using primers 5'-CTCCATGGTCATGGAA(A/G)ATAAA GGAGAA and 3'-CAACAAATATCCTACACTGTCCTAGGTG specific to the coat protein (CP) gene of CarMV, an expected viral CP gene product of 1.05 kb was amplified by reverse transcription-polymerase chain reaction from total RNA isolated from infected N. benthamiana. Comparisons of the 1,047-nucleotide CP gene with those of 15 CarMV isolates available in GenBank showed 94.6 to 98.2% nucleotide identity and 94.8 to 96.8% amino acid identity. Results from current studies indicate that the virus infecting calla lilies is an isolate of CarMV. To our knowledge, this is the first report of CarMV infection in calla lilies. The occurrence of CarMV in calla lilies has direct implication for the economically important nursery and floral industry in Taiwan. Reference: (1) F. W. Zettler and R. D. Hartman. Dieffenbachia, Caladium, and Zantedeschia. Pages 464-470 in: Virus and Virus-Like Diseases of Bulb and Flower Crops. G. Loebenstein, R. H. Lawson, and A. A. Brunt, eds. John Wiley and Sons, West Sussex, U.K., 1995.
马蹄莲是台湾具有重要经济价值的观赏植物。它们生长在该岛的中部和北部地区,每年约有300万根茎被运送。马蹄莲易受多种病毒感染(1)。台湾曾报道过黄瓜花叶病毒、芋花叶病毒、芜菁花叶病毒和西瓜银斑驳病毒的感染。最近,在位于台湾中部彰化县的田间种植的马蹄莲叶片上观察到了病毒样症状,包括黄色斑驳、浅黄色斑点、黄色环斑和花叶。2001年3月,从患病马蹄莲中分离出一种病毒培养物,并在藜和本氏烟中建立。用该病毒接种健康马蹄莲后,接种叶片上出现褪绿斑点,这些斑点扩大并融合形成大的黄色斑块。症状与田间自然感染的植物相似。该病毒在藜、 ficifolium、 amaranticolor、南瓜、洋桔梗、赤小豆、绿豆和豇豆上诱导产生褪绿局部病斑。除了接种叶片上的局部褪绿斑点外,在接种后8至10天,还观察到该病毒在石竹、中国石竹和大豆中发生系统侵染。在本氏烟中,观察到的唯一症状是系统萎蔫。使用电子显微镜检查2%醋酸铀染色的样品,发现在患病马蹄莲或受感染藜的叶片粗提物中存在直径约34至35 nm的球形颗粒。在藜和本氏烟病毒感染叶片组织的超薄切片的细胞质中也观察到类似颗粒,但在细胞核中未观察到。对受感染藜的组织提取物进行差速离心,然后进行蔗糖密度梯度离心,得到了大小相似的病毒粒子。纯化病毒的十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示有一条分子量为41.6 kDa的单一结构多肽。在双抗体夹心酶联免疫吸附试验中,病毒抗原与其同源抗血清以及抗康乃馨斑驳病毒(CarMV;Agdia公司,美国印第安纳州埃尔克哈特)的抗血清反应呈阳性。使用针对CarMV外壳蛋白(CP)基因的引物5'-CTCCATGGTCATGGAA(A/G)ATAAA GGAGAA和3'-CAACAAATATCCTACACTGTCCTAGGTG,通过逆转录-聚合酶链反应从感染本氏烟中分离的总RNA中扩增出预期的1.05 kb病毒CP基因产物。将1047个核苷酸的CP基因与GenBank中可获得的15个CarMV分离株的CP基因进行比较,核苷酸同一性为94.6%至98.2%,氨基酸同一性为94.8%至96.8%。目前的研究结果表明,感染马蹄莲的病毒是CarMV的一个分离株。据我们所知,这是关于CarMV感染马蹄莲的首次报道。CarMV在马蹄莲中的出现对台湾经济上重要的苗圃和花卉产业有直接影响。参考文献:(1)F. W. Zettler和R. D. Hartman。花叶万年青、五彩芋和马蹄莲。载于:球根花卉和花卉作物的病毒及病毒样病害。G. Loebenstein、R. H. Lawson和A. A. Brunt编。John Wiley and Sons,英国西苏塞克斯,1995年,第464 - 470页。