利用煙草白肋種(White Burley)分離甜椒病毒

The use of white burley tobacco for isolaton of sweet pepper viruses

黃玉堂(Y. T Liu) ； 劉秀鈴(H. L. Liu) ； 盧耀村(Y. T. Lu)

《植物病理學會刊》 3卷2期 (1994/06) Pp. 91-100

https://doi.org/10.6649/PPB.199406_3(2).0003

摘要

本省甜椒的栽培，常遭受病毒的複合感染，造成生長後期皆出現嚴重的斑駁嵌紋病徵，而難以直接從後期罹病株分離鑑定爲害的病毒種類。從初發病出現較多的脈綠嵌紋病葉，以10倍量0.1 M磷酸緩衝液(pH 7.2)研磨，粗汁液機械接種至一系列判別植物，從接種的煙草white Burley (W. B.)所出現的病徵，加以單離後鑑定爲害病毒種類。首先從W. B.接種葉所出現的黃斑，經過單一病斑的分離，得到二個不同分離株；即只在接種葉出現黃斑(YS），及除接種葉外上位葉也會出現斑點(SYS)之兩個性質不同分離株。經過寄主範圍、血清反應及粒子形態觀察，獲知兩者是病原性有所不同的Chili veinal mottle virus，另外從煙草W. B.心葉所出現的嵌紋病徵，經過Sepharose CL-6B色層分析與病原性測定，在第一吸收峰(R1)及其肩狀凸起處(R2)所分離之病毒分離株，均能使甜椒出現退綠黃化的斑駁病徵。雖然兩分離株病毒，對胡瓜嵌紋病毒抗血清皆無spur的反應帶出現，但是經過單離後之R1病毒分離株爲30 nm見不到中心(core)的球狀病毒；而R2為28 nm,能見到core的球形病毒，即在粒子形態上有所不同。又在寄主範圍上，兩種分離株在煙草W. B.、N. glutinosa與萬桃花上呈現病徵也有所不同，由此推知兩者皆屬Cucumovirus群中的病毒，即R1爲胡瓜嵌紋病毒，而R2則屬花生矮化病毒中之一系統。

關鍵字

煙草White Burley ； 病毒分離 ； 甜椒病毒 ； 胡瓜嵌紋病毒群

並列摘要

Sweet pepper during the growing season were often affected with virus diseases. The diseased plants at the late growing stage usually showed severe mottle mosaic symptoms resulted from mixed infection; and it was difficult to isolate all causal viruses directly the mix-infected pepper plants. In an attempt to isolate viruses infecting pepper, the healthy pepper seedlings were planted in the field. From which the early infected leaves were collected and homogenized with 10 volumes of 0.1 M phosphate buffer (pH 7.2), and each of the saps was mechanically inoculated to several differential plants. Different types of symptoms were observed on inoculated leaves of tobacco ＂White Burley＂ (W. B.) plants, and they were used as a source to isolate the causal viruses. Two virus isolates were obtained from chlorotic spots by single lesion isolation; one isolate produced chlorotic spots only on the inoculated leaves, while the other induced chlorotic spots on the inoculated and upper leaves. According to host ranges, physical properties, serological tests and particle morphology, they were considered as two different pathogenic strains of chili veinal mottle virus. Virus isolation was also conducted from green island mosaic leaves of the tobacco. The infected leaves were subjected to partial purification by differential centrifugation and sepharose CL-6B molecular sieve chromatography. The elution samples that showed absorption at 254 nm were examined with an electron microscope (EM) and sap inoculated to pepper plants. The results showed that the infectivity appeared in the first fraction (RI) and the shoulder fraction (R2. The viruses in both fractions induced similar veinal mottle symptom on pepper leaves, but produced different symptoms on W. B. tobacco, Nicotiana glutinosa and Datura metel. EM examination revealed that the virions in R1 were isometric particles of 30 nm in diameter on which the virus core could not be observed; while those in R2 were isometric particles of 28 nm on which the virus core could be seen. Both viruses reacted with the cucumber mosaic virus antiserum in the double diffusion test. Based on the host reactions, virion morphology and scrological tests, the R1 virus was concluded as cucumber mosaic virus and the R2 virus possibly as a strain of peanut stunt virus of the same cucumovirus group

並列關鍵字

White Burley tobacco ； virus isolation ； sweet pepper virus ； cucumovirus group