矮南瓜黃化嵌紋病毒（ZYMV）經瓜類種子傳播的特性及其檢測

比較生物分析(bioassay)、間接法酶聯抗體免疫分析(indirect ELISA)、西方墨漬法(western blot)及反轉錄聚合酶連鎖反應(RT-PCR)等檢定方法之靈敏度，據以研擬瓜類種子帶矮南瓜黃化嵌紋病毒(ZYMV)的標準檢測流程，其中包括群體測試的採樣及統計分析、供試樣品製備、適合的檢驗方法及檢測結果的表示等。以出口種子檢測來測試標準檢測流程，其中1個南瓜種子批號有ZYMV檢出；以＂Seedhealth＂程式計算，病毒檢出率為0.47% ± 0.46%，其中ELISA陽性反應的檢體經RT-PCR檢測，明確顯示ZYMV的存在，核酸解序其RT-PCR產物，進一步確定檢測結果；但以出芽試驗檢測同批種子，未能在子代苗株發現病株。檢測市售胡瓜種子，所有9個供試品種都呈陰性反應。ZYMV接種胡瓜，部分抗病品種的果實與種子以ELISA檢驗均無ZYMV檢出，取感病品種的罹病株果實採樣，新鮮種子中可測得26%帶有ZYMV，種子乾燥15 d後，全數供試種子均無ZYMV檢出。西瓜罹病株果實之果蒂、果底、果皮、外果肉、內果肉及果中柱均有ZYMV分佈，未成熟的種子則比成熟的種子帶有較多病毒，通常西瓜「甜美人」比「黑美人」的種子較容易測出ZYMV。另置於10℃冰箱保存1年的「甜美人」西瓜種子，仍有60%可檢出ZYMV，而出芽試驗所檢測的種子傳染率仍為0。

關鍵字

標準檢測流程；病毒檢測；胡瓜；西瓜；南瓜

並列摘要

Zucchini yellow mosaic virus (ZYMV) had been comparatively detected by bioassay, indirect enzyme-linked immunosorbent assay (ELISA), western blot, and reverse transcription-PCR (RT-PCR). Accordingly, a standard operating procedure for detection of ZYMV carried by cucurbit seeds was set. The procedure includes sampling technique and statistical analysis mode for group test, specimen preparation for testing, acceptable methods for ZYMV detection, and formal expression of tested results. To practice the procedure, a seed lot of Cucurbit sp. for quarantine test was tested by ELISA and the proportion of positive was 0.47% ± 0.46% calculated by ‘Seedhealth’ program. ZYMV involved in these positively detected specimens were confirmed by the analysis of RT-PCR and the nucleotide sequencing of RT-PCR products. However, no seedlings from the same seed lot were infected by ZYMV when tested by growing-out test. All tested seeds collected from 9 local commercial cucumber cultivars were detected to be completely negative. Plants of various cucumber cultivars were inoculated with ZYMV, no virus was detected in fruit or seed derived from some resistant cultivars; whereas, ZYMV-infected fruits were harvested from symptomatic plants of susceptible cultivars. The incidence of ZYMV on fresh seeds in infected fruits was estimated to be 26%; even so, the virus became undetectable after dry preservation of the seeds for 15 days. The distribution of ZYMV on fruit of watermelon was observed, and the virus was found in fresh pedicle, fruit base, fruit peel, outer pulp, inner pulp and innermost pulp. Immature seeds contained more virus than that of mature seeds. Regularly, it’s easier to detect ZYMV in seeds of watermelon cv. 'Tien-Mei-Zen' than that of 'Dark Belle.' The virus was still detectable in 60% of the 'Tien-Mei-Zen' seeds stored at 10℃ for 1 year, but the seed transmission rate detected by growing out test was kept 0.

並列關鍵字

Standard operating procedure ； Virus indexing ； Cucumber ； Watermelon ； Pumpkin

國際替代計量

矮南瓜黃化嵌紋病毒（ZYMV）經瓜類種子傳播的特性及其檢測

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