混合兩種百香果病毒 East Asian Passiflora virus 多元抗體以提升百香果不同馬鈴薯Y屬病毒之廣效性檢測

Abstract

罹染病毒的百香果若出現果實木質化病徵，造成畸型變小、果汁率低、風味變差，對品質與產量影響甚大。在台灣，East Asian Passiflora virus (EAPV) AO 與IB 系統及Telosma mosaic virus (TeMV) 為可感染百香果之3 種主要的馬鈴薯Y 屬 (Potyvirus) 病毒。本研究以細菌生合成系統分別誘導表現EAPV-AO 和EAPV-IB 鞘蛋白 (coat protein; CP) 作為抗原，經兔免疫注射後製備出具有廣效檢測19種不同馬鈴薯Y屬病毒之多元抗體，分別稱之為Anti-EAPV-AOCP 和Anti-EAPV-IBCP，並於間接式-酵素連結免疫吸附反應 (indirect enzymelinked immunosorbent assay; indirect ELISA) 檢測中發現以此二抗體等比例混合 (稱之為Anti-EAPVmix) 進行檢測，也可用於檢出台灣及泰國的百香果TeMV，並提升對EAPV-AO、EAPV-IB 和TeMV 等3 種病毒的檢出率。應用Anti-EAPVmix 檢測病毒在百香果植株枝條上之分布顯示，以近新芽之枝條上段部位葉片所測得之病毒檢出率較高，適合作為提升檢出病毒之採樣部位。本研究顯示混合兩種EAPV 的多元抗體，一次檢測即可達到監測至少3 種台灣主要的百香果Potyvirus 屬病毒，並可擴及其他不同的potyvirus，且可節省檢測所需耗材和降低成本之效益，可強化對田間百香果病毒發生的監測及健康種苗的病毒檢測內控機制。 When the virus-infected passion fruits showed woodiness symptoms on fruits, the fruits were deformed and smaller resulting in a low juice rate, poor flavor and quality, which could cause a great impact on yield. Two East Asian Passiflora viruses (EAPV-AO and EAPV-IB) and Telosma mosaic virus (TeMV) are the three main potyviruses infecting passion fruits in Taiwan. In this study, the polyclonal antibodies against the coat proteins (CP) of EAPV-AO and EAPV-IB were respectively prepared from immunizing rabbits with the bacterial expressed viral CPs, revealing that both antibodies have a wide-spectrum response with 19 tested potyviruses. In indirect enzyme-linked immunosorbent assay (indirect ELISA), the equivalent mixture of polyclonal antibodies against the CPs of EAPV-AO and EAPV-IB, denoted Anti-EAPVmix, could also be used to detect passionfruit isolates of TeMV from Taiwan and Thailand, and to improve the detection rate of EAPV-AO, EAPV-IB, and TeMV. The distribution of viruses on plant branches was detected by Anti-EAPVmix. The virus was detected at a high rate from the upper part near new leaves, which is recommended for sampling. Our result showed that the mixture of EAPV polyclonal antibodies can be used to detect not only the three major passiflora viruses in Taiwan but also other potyviruses to save the detection time and costs. The detection method can strengthen the field survey of passiflora viruses and the internal control of healthy seedlings.