CMV 共 34 株（單獨感染有 29 株），BBTV 有 7 株（單獨感染有 6 株），BSV 共有 2 株（皆為單獨染病）。BBrMV 與 CMV 複合感染有 3 株，BBrMV 與 BBTV 複合感染有 0 株，CMV 與 BBTV 複合感染 有1 株。未發現其他三種病毒與 BSV 複合感染，也未發現三種以上 病毒複合感染。
六、 即時反轉錄聚合酶連鎖反應 ( Real-time RT-PCR ) 與單步反轉錄聚合酶連鎖反應 one step RT-PCR 敏感 度測試比較
利用所設計出的Taqman 系統之 BBrMV 142 assay 建立標準曲線，
得出回歸曲線公式Ct = -3.527 log (Qty) + 43.224，利用該公式可推算出 copy numbers（圖十三 A、B）。藉由 10 倍序列稀釋原始總核酸濃度 104 ng/ uL 之核酸，得到 8 個濃度（包含原始濃度），每 microliter ( uL ) 分 別為104 ng、10.4 ng、1.04 ng、104 pg、10.4 pg、1.04 pg、104 fg 與 10.4 fg。將上述濃度分別以 real-time RT-PCR 與 one step RT-PCR 進行偵 測後可得到圖十四A 與 B，根據兩圖相互對照之結果可知 real-time RT-PCR 可測到最小濃度為 10.4 pg/ uL，比起傳統 one-step RT-RT-PCR 敏感約 100 倍（圖十四 A、B），且對健康香蕉核酸並不會反應。
將田間採集到帶有BBrMV 的核酸樣本以 BBrMV 142 assay 進行偵 測，偵測之五個樣本中F2 ( TC 5 from Pingtung )、F16 ( Oolong from Yunlin )、F35 ( TC 7 from TBRI ) 、F53 ( L-PC from Kaohsiung )、F92 ( Saba from TBRI ) 僅有 F53 無法被 BBrMV 142 assay 偵測到（但可被 one step RT-PCR 偵測到）。
伍、 討論
( leaf blotch, Victoria blight ) 之感病性中有觀察到。帶有抗 Pucciniacoronata var. avenae 抗性基因 Pc2 gene 的燕麥，會對 Cochliobolus victoriae 造成的葉枯病 “ Victoria blight ” 感病；而帶有抗 Victoria blight 抗性基因Vb gene 的植株，則對 crown rust 感病 ( Rines and Luke 1985;
Litzenberger 1949; Wolpert et al. 2002 ) 。番茄對鐮胞菌萎凋病第三型 ( tomato fusarium wilt race 3 ) 之抗病性與細菌性斑點病第四型之感病性 ( tomato bacterial spot T4 ) 的關係中也有觀察到類似現象。帶有抗 F.
oxysporum f.sp. lycopersici race 3 之 I-3 gene 的番茄植株同時也會對 Xanthomonas perforans T4 造成的細菌性斑點病更為感病 ( Hutton et al.
2014; Li et al. 2018 )。先前有兩篇報導指出寶島蕉 ( GCTCV-218 ) 之耐 病性可能是與該品系的NPR1-like gene 或根部之 cell wall-strengthening gene 相關 ( VAN DEN Berg et al. 2007; Endah et al. 2008 )，而 Swarupa et 中，阿拉伯芥被Oilseed rape mosaic virus 感染後隨著各個葉片的生長，
病徵會達到一個極大值，接著新長出之葉子便會recover ( Kørner et al.
2018 )。這樣的現象在許多其他的植物－病毒間的交互關係報導中也有 觀察到此現象，該批研究團隊發現在阿拉伯芥中雖然有病毒coat protein 的累積，但病毒的RNA silencing suppressor 的功能卻降低。而 Ghoshal and Sanfaçon 在 2015 年的 review 統整了幾派說法，認為這種現象常常 伴隨著RNA silencing 的參與 ( PTGS ) 或是寄主抑制病毒蛋白的轉譯 ( TGS ) 的機制，而 RNA silencing 也常常會調控寄主的基因表現來抑制 病徵的表現 ( Ghoshal and Sanfaçon 2015 )。或許在香蕉與 BBrMV 的交
互作用中也存在著這樣的情形，未來值得探討。
Zhang 等人所得結果相符合 ( Zhang et al. 2016 )；而 BBrMV 雖然能夠 長時間存在姑婆芋體內，但分布情況或許不平均或是病毒量很少，導致 雖然能夠利用RT-PCR 偵測病毒的存在，但卻不穩定，姑婆芋六株植株 內僅有兩棵到六個月時還能偵測到病毒的存在。過去在研究香蕉萎縮病
時，確定BBTV 的中間寄主包含了月桃、美人蕉、野薑花及芋（曹，
1998; Pinili et al. 2013; Ram and Summanwar 1984）。研究香蕉萎縮病的經 驗中，可知道蕉蚜具有食性上的選擇，原先在香蕉上的蕉蚜可能會不喜
BBTV 感染芋 ( Colocasia esculenta )，沒病徵且維持在低濃度的例子相 似 ( Pinili et. al. 2013; Ram and Summanwar 1984 )。田間採樣時也於香蕉
即BBrMV 在蕉區分布仍然很少，而目前抗黃葉病之台蕉五號、台蕉七
Rodoni et al. 1999 )。先前研究顯示，BBrMV coat protein 的 N-terminal 端 變異度較大 ( Rodoni et al. 1999 )，而 2013 年於厄瓜多有報導 Cavendish 蕉株感染BBrMV，以 RT-PCR 偵測時使用不同的 primer，有些確定已染 病的BBrMV 樣本竟無法成功增幅，該篇研究推測可能是因為 isolate 不 real time PCR 與 one step PCR 之敏感度測試結果，real time RT-PCR 比起 one step RT-RT-PCR 靈敏 100 倍左右。於是進一步比對設計出之 探針序列，該樣本在探針的地方有一個nucleotide 的不同，推測可能是 由於taqman 系統之專一性十分靈敏而使得本次設計之 BBrMV 142 assay
無法用於該分離株。以上述結果，臺灣可能存在不同BBrMV 的
strains，亟待未來加以探討。而本次設計出之 BBrMV 142 assay 或許在 未來能應用於系統的鑑別。
田間採集得到的樣本同時也偵測了其他三種重要香蕉病毒
（BBTV、CMV、BSV）的存在，發現田間葉部有嵌紋狀病斑的蕉株大 部分仍是感染CMV，而 BBrMV 及 BSV 並不多。BBrMV 在臺灣田間有 與CMV 複合感染的案例，不過尚未發現 BBrMV 與 BBTV 或與 BSV 複 合感染的情形。
距離馮（2006）上次的調查經過十二年，BBrMV 在臺灣的疫情雖 稍有擴大，但仍是零星幾個案例。可能由於臺灣的農民在耕作上與病蟲
害的防治有一定水準，而BBrMV 在田間最主要傳播的媒介是蕉蚜，過
去因為BBTV 在田間危害嚴重，農民早已將蕉蚜視為重要防治項目之
一，蕉蚜密度低而傳染源（vector）少讓 BBrMV 在田間仍未大流行。
BBrMV 目前在臺灣僅有零星案例，但產香蕉大宗的一些國家如菲
律賓、厄瓜多、哥倫比亞、印度等地皆已經有BBrMV 發生普遍的報
導。臺灣是黃葉病Foc 4 首個發生地以及抗黃葉病華蕉品系研發之重 鎮，不應忽視該病毒病害的問題。臺灣對於香蕉黃葉病的抗病品種的選 種愈來愈多，當外銷到其他國家時，必定需經過檢疫。菲律賓、印度等 地皆有對BBrMV 的減產評估，損失皆在 40 % 左右 ( Diekmann and Putter 1996; EFSA 2008; Thomas et al. 2000; Selvarajan and Singh, 1997 )，
黃葉病目前雖是世界上香蕉產業的頭號病害，但若忽略了對BBrMV 病
毒病害的感病性問題，而將BBrMV 感病的品種販售至疫情嚴重的區
域，可能會對當地產業造成危害。
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柒、 表
表一、實驗中RT-PCR 所使用引子對及 real-time PCR 所使用之引子對/探針序列。
Table 1. Primer pairs and real-time PCR(Taqman) primer/ probe sequences used in this study.
Reference or source BBrMV324-F 5’-AACGCTCAGCCTACTTTTCG-3’ 洪挺軒
（實驗室常備）
BBrMV324-R 5’-CATATCACGCTTCACATCTTCA-3’
BBrMV902-F 5'-ATCTGGAACGGAGTCAACC-3'
（馮，2006）
BBrMV902-R 5’-ATTCATGTTTCACCCCAAGC-3’
BBrMV142-F 5'-GGGAGCACTTAGAGGATTGAATGA-3'
（本人）
BBrMV142-R 5'-CCGAGTGTTTGATCCACGAA-3' BBrMV142-P FAM-CTAGAGAAGCACACACGC-MGB
CMV 245-F 5’-TCCCACGGCGATAAAGGAC-3’ 吳柏辰
（未發表）
CMV 245-R 5’- CAATCGAGAGTTTCACACAAGC-3’
BBTV -536-F 5'-CGCACTAACTCGGGACATCTG-3' 吳柏辰
（未發表）
BBTV -536-R 5'-GGCAGAGGTATTGTGGAAGACAG-3'
BBTV -536-R 5'-GGCAGAGGTATTGTGGAAGACAG-3'