Purification and Serology of an Isolate of Zucchini Yellow Mosaic Virus

矮南瓜黃化嵌紋病毒之純化與血清學特性之研究

黃秋雄(C. H. Huang) ； 許秀惠(S. H. Hseu) ； 趙玉珍(Y. J. Chao)

《中華農業研究》 35卷4期 (1986/12) Pp. 495-503

https://doi.org/10.29951/JARC.198612.0009

摘要

將Gonsalves及Ishii純化木瓜輪點病毒所使用之等密度平衡離心法(Zucchini yellow mosaic virus, ZYMV)略加修改，可成功地純化矮南瓜黃化嵌紋病毒(zucchini yellow mosaic virus, ZYMV)。以接種ZYMV-No.7分離株之矮南瓜爲純化材料，估計每百克病葉可獲得22毫克之純化病毒。紫外光吸收比值A260/280約在1.27至1.31之間。所純化之病毒懸浮液適量混合輔助劑(adjvuant)後曲于入小白兔使產生抗血清。利用界面環沈澱法，將抗血清與本病毒之粗汁液及純化之病毒反應，測得血清力價分別爲2048及4096。
本省九個ZYMV分離株及一個美國佛州ZYMV-FL分離株，以SDS-瓊脂免疫擴散法來探討各分離株血清類綠關係，結果發現所有分離株均與ZYMV-No.7抗血清形成明顯沈澱帶且互爲融合，證明各分離株均有相同血清類緣關係，但與WMV-1或WMV-2（佛州分離株）無關。進一步試驗，發現本省之ZYMV分離株與一個來自佛州大學之WMV-2抗血清有血清類緣關係。由是可知，本省之ZYMV分離株異於佛州一個WMV-2分離株，但與WMV-2部份分離株有血清類綠關係。

關鍵字

病毒純化 ； 血清學研究 ； 矮南瓜黃化嵌紋病毒

並列摘要

An isolate of zucchini yellow mosaic virus, ZYMV-7, was purified and its antiserum produced. Systemic leaves from inoculated zucchini plants taken 9-10 days after inoculation had the highest virus concentration, and were used in virus purification. Yields of purified virus estimated by spectrophotometer were 18-22 mg/100g leaf tissues. The O. D. ratio of A260/280 from several separate preparations ranged from 1.27 to 1.31. The purified virus preparation obtained from the first isopycnic centrifugation in cesium sulfate was found contaminated with host proteins, however, the contaminated materials could be removed by a second isopycnic centrifugation. The reciprocal titers of antiserum against homologous antigens in crude leaf extracts and purified virus preparations were 2048 and 4096, respectively, in ring interface precipitin tests. The titer of this antiserum against crude sap antigens was 16 in SDS-immunodiffusion tests. Antisera collected at several bleedings reacted with host constituents weakly, but the addition of 2% bovine serum albumin to the antisera or to agar medium completely removed the non-specific reactions in SDS-gel diffusion tests. Antiserum to ZYMV-7 produced precipitin bands with all 10 ZYMV isolates, i. e., ZYMV-1 to ZYMV-9 from Taiwan, and ZYMV-FL from Florida. There were no serological relationships of ZYMV-7 with WMV 1 and WMV 2 (Florida isolate). However, an antiserum to WMV-2, obtained from University of Florida, reacted with ZYMV isolates strongly but formed definite spurs with WMV-2 in SDS-gel diffusion tests, indicating that ZYMV isolates from Taiwan are different from WMV-2 Florida isolate but have some serological relations to certain types of WMV-2.

並列關鍵字

purification ； serology ； zucchini yellow mosaic virus