同步接種 PRSV-DF 與 PRSV-SMN 的感染詴驗（DF＋SMN）中，台大八號體內 SMN 系統的第一週病毒量為 104 CNs，第二週上升至 106 CNs 且維持到第六週，第八週 時降至 5x105 CNs，第九週再增加至 106 CNs，第十與十二週降至 105 CNs；DF 系 統在接種後一週達到 5x105 CNs，第四週時上升至 106 CNs，第七週時增加至 107 CNs，
至十二週期間病毒量在 5x106~107 CNs 間變化（圖十二 A）。台農二號體內 SMN 系 統病毒量在第一週為 7x104 CNs，第二週上升至 5x105 CNs 並維持至第八週，第九 週後無法偵測到病毒量；DF 系統首週為 3x105 CNs，第二週增加至 106 CNs 且維持 至第五週，第六週增殖為 107 CNs 直至第十二週（圖十二 B）。紅妃體內 SMN 系統 病毒量在第一週為 4x103 CNs，第二週升至 106 CNs 後至第九週均維持在 5x105~106 CNs，第十週下降為 105 CNs，第十二週則偵測不到病毒量；DF 系統首週病毒量為 8x102 CNs，第二週升至 106 CNs，第八週增高為 107 CNs，第九週反降至 106 CNs，
第十週與第十二週再回到 107 CNs（圖十二 C）。同時接種兩病毒系統五週後，DF 系統病毒量開始領先 SMN 系統病毒量。
伍、討論
變（翁，1981；卲井，1986；林，1980; Lin et al, 1989；關，1990；廖，2004），十年前 與現今流行系統相差極大，危害程度不同。倘若要應用抗病育種來防治 PRSV，需探討是台農一號和紅妃早期低於 104 CNs 後期才爬升至 106 CNs，台農二號體內病毒量保持
量比單獨接種 SMN 高；紅妃體內 DF 系統與 SMN 系統有相同的變動情形，但 SMN 病 毒量低於 DF 一百倍。上述結果顯示，後進入的病毒系統難以取代已存在的病毒系統，
系統間沒有明顯的協力或拮抗的情形，這與前人研究同屬 Potyvirus 的 Plum pox virus 兩 系統的交互作用獲得相近之結果（Capote et al, 2006）。 病毒（Cucumber mosaic virus, CMV）、西瓜嵌紋病毒（Watermelon mosaic virus, WMV）、
矮南瓜黃化嵌紋病毒（Zucchini yellow mosaic virus, ZYMV），CMV 與 Potyvirus 間有協 力作用（Wang et al., 2002, Wang et al., 2004, Zeng et al., 2007），WMV、ZYMV、PRSV 同屬於 Potyvirus，彼此間交互作用尚不清楚，但可能會影響 PRSV 二系統之生態消長，
這方面值得未來做進一步之研究。
本研究以 real-time RT-PCR 作為分析病毒增殖之工具，綜合病徵發展與病毒增情形 二項資訊，在新耐（抗）病品系之選拔過程中，提供更詳實的評良參考。同步與非同步 的接種詴驗，探討不同病毒系統的交互作用關係，長時間觀察病徵與病毒量，測詴新品 種（如台大八號）耐病性之穩定，探討系統間之交互作用與族群趨勢，藉以預測田間病 毒系統之變化，研擬新防治策略並持續助益新耐病品種之開發。
陸、參考資料
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柒、圖與表
表 一、以反轉錄聚合酶連鎖反應(RT-PCR)與即時定量反轉錄聚合酶連鎖反應(real-time RT-PCR)偵測木瓜輪點病毒（PRSV）所 用之引子與探針。
Table 1. The nucleotides sequence of primers and probes used in the reverse transcription – polymerase chain reaction (RT-PCR) and real-time RT-PCR assays for the detected of Papaya ringspot virus.
Name Sequence
RT-PCR
Reverse cDNA F„5end 5‟-AAA TAA AAC ATC TCA ACA CAA CAC-3‟
R„829 5‟-TGC AAT CTC AGC TTC AAG TG-3‟
表 二、木瓜輪點病毒感染不同品種（系）木瓜寄主所產生的病徵級數鑑別
Table 2. Categorization of the symptom index induced by papaya ringspot virus in different cultivars after inoculation.
Symptom index Days post inoculation
Treatment Cultivar 14 28 42 56 70
＊值為帄均數±標準差。 Value are mean ± standard deviation.
病徵級數：分為 5 級：0 級，植株健康；1 級，30％系統葉出現輕微嵌紋、葉脈透化等現象；2 級，70％系統葉出現輕微嵌紋、
葉脈透化等現象；3 級，系統葉出現明顯嵌紋、葉脈透化及黃化現象；4 級，葉片出現嚴重嵌紋、壞疽斑、捲曲、畸形等病徵；
5 級，葉片出現嚴重嵌紋、葉脈透化、捲曲、畸形、嚴重黃化或植株死亡。
Symptom index：0、no symptom.；1、30% of systemic leaves showing mild mottling and vein clearing；2、70% of systemic leaves showing mild mottling and vein clearing；3、systemic leaves showing mottling, vein clearing and yellowing.4、system leaves showing severe mosaic, necrotic spots and deformation.；5、systemic leaves showing severe mosaic, vein-clearing, deformation, yellowing and decline.
圖 一、木瓜輪點病毒畸形系統(PRSV-DF)感染 6 種不同品種或品系（10A、10B、10F、TN1、TN2、RL）的木瓜在 28 天、56 天、70 天後之病徵表現。
圖 二、木瓜輪點病毒嚴重嵌紋壞疽系統(PRSV-SMN)感染 6 種不同品種或品系（10A、10B、10F、TN1、TN2、RL）的木瓜在 28 天、56 天、70 天後之病徵表現。
表 三、以反轉錄聚合酶鏈鎖反應(RT-PCR)偵測木瓜輪點病毒畸形系統(PRSV-DF)與嚴重嵌紋系統(PRSV-SMN)於不同品種中之 增殖情形。
Table 3. Detection of the replication of PRSV-DF and PRSV-SMN individually infecting the different papaya cultivars(lines) by the RT-PCR assays
-, 單位像素密度值 (AVG) < 500;+, 500 ~ < 5000;++, 5000 ~ < 10000;+++, 10000 ~ < 15000;++++, > 15000。單位像素密度值等於單 位面積內亮度值。黑色為 0; 白色為 66535
-, AVG(average value of pixel) < 500;+, 500 ~ < 5000;++, 5000 ~ < 10000;+++, 10000 ~ < 15000;++++, > 15000. (Black is 0; White is Days Post Inoculation
Treatment Cultivar 7 14 21 28 35 42 49 56 63 70
圖 三、以即時定量反轉錄聚合酶連鎖反應(real-time RT-PCR)偵測木瓜輪點病毒畸形系 統(PRSV-DF)在不同的木瓜品種(台農一號、台農二號與紅妃)中之增殖動態。
Figure 3. The monitoring of multiplicative dynamic of PRSV-DF in the different papaya cultivars (TN 1, TN 2 and Red-Lady) using the real-time RT-PCR assays.
圖 四、以即時定量反轉錄聚合酶連鎖反應(real-time RT-PCR)偵測木瓜輪點病毒嚴重嵌 紋壞疽系統(PRSV-SMN)在不同的木瓜品種(台農一號、台農二號與紅妃)中之增殖動 態。
Figure 4. The monitoring of multiplicative dynamic of PRSV-SMN in the different papaya cultivars (TN 1, TN 2 and Red-Lady) using the real-time RT-PCR assays.
圖 五、木瓜輪點病毒畸形系統(PRSV-DF)接種在不同木瓜品種（NTU8、TN2、RL）木瓜後 14 天、28 天、56 天與 70 天後之病 徵表現。
Figure 5. The symptom-expression of different papaya cultivars (NTU 8, TN 2 and RL) infected by PRSV-DF 14, 28, 56, and 70 days