The Current Status of Geminiviruses Occurred on Passion Fruit in Taiwan

在台灣感染百香果的病毒計有胡瓜嵌紋病毒 (Cucumber mosaic virus, CMV)、東亞百香果病毒(East Asian passiflora virus, EAPV)、百香果斑駁病毒(Passion fruit mottle virus, PaMV)、百香果漣葉病毒(Passion fruit crinkle virus, PCV) 4 種。2011 年3 月在埔里地區採集到黃綠不均且葉片變形百香果樣本，利用ELISA 檢測未受到上述4 種病毒感染，利用滾輪法(rolling circle amplification, RCA) 卻測到疑似雙生病毒感染。RCA 產物以BamHI 處理，得到約3.0 Kb 之產物，選殖至載體，選殖株以EcoRI 酵素處理確認時出現2 類不同切位DNA 片段。定序分析後分別有2745 與2732 個核苷酸，與GenBank 登錄之序列比對結果，2745 個核苷酸者(Accession No. KC161185) 與一品紅捲葉病毒(Euphorbia leaf curl virus, EuLCV) DNA-A 有98.5%的相同度，2732 個核苷酸者(Accession No. KC161184) 與番木瓜捲葉廣東病毒(Papaya leaf curl virus, PaLCuGDV) DNA-A 有91.7%的相同度。根據2 病毒序列分析結果設計PCR 引子對，利用電泳分析調查2 種病毒發生情形。此2 病毒應已普遍發生於百香果台農1 號多時，於宜蘭、花蓮、苗栗及南投地區等百香果栽培區均有發現，因為台農1 號百香果需經由嫁接苗繁殖，因此該2 種病毒隨著嫁接苗普遍發生。病毒感染後造成百香果葉片斑駁、葉面不平整，各百香果園觀察到的病徵嚴重程度不一，但進入春夏季之後氣溫回升，葉片變形情況趨於輕微至觀察不到病徵，百香果生長回復正常，而造成忽略。 The main variety of passion fruit (Passiflora edulis x Passiflora edulis f. flavicarpa) cultivated in Taiwan is ‘Tainung No. 1 ’which is a hybrid and propagated only by grafting. In the spring of 2011, plants with severe systemic mosaic and malformation on leaves were found in some orchards located in Puli, Nantou in the middle of Taiwan. However, after growing up to three months, most of these diseased plants became symptomless when the weather became warmer. In March of 2011, 2 leaf samples exhibiting mosaic and 3 samples showing malformation were collectec and tested by DAS-ELISA; none were positively reacted with antibodies against Cucumber mosaic virus (CMV), East Asian passiflora virus (EAPV), Passion fruit mottle virus (PaMV) or Passion fruit crinkle virus (PCV) that have been occurred in Taiwan. The rolling circle amplification (RCA) with hexamer primers was adopted to analyze the potential begomoviruses which were prevalent on the other crops in Taiwan. The RCA amplified products were digested with BamHI and separated on 1.2% agarose by gel electrophoresis. A fragment, about 3 kb, was purified from each gel and cloned into the respective site of pBluescript SK(-) individually. Clones were screened by EcoRI digestion and two types of restriction fragement length patterns were found among them. One type clone containing 2745nucleotides (Accession No. KC161185) with 98.5% identity to Euphorbia leaf curl virus (EuLCV) and the other type clone containing 2732 nucleotide (Accession No. KC161184) with 91.7 identity to Papaya leaf curl Guangdong virus (PaLCuGDV) were revealed by nucleotide comparisons of their DNA-A in the GenBank. Accordingly, we confirmed the existence of passiflora isolates of EuLCV and PaLCuGDV. In a brief survey, allpassion fruit leaf samples were detected EuLCV and/or PaLCuGDV infectionTo our knowledge, this is the first report of begomoviruses associated passion fruit in Taiwan and in the Asia