臺灣博碩士論文加值系統

台灣地區之煙草粉蝨種群 (Bemisia tabaci species complex) 及稻蝗類 (Oxya spp.) 以遺傳標示包括 RAPD-PCR、rDNA 及 mtDNA等方法分析粉蝨種群內關係及稻蝗類之種間關係。台灣地區現有之煙草粉蝨種群之基因組 DNA，可利用篩選出的逢機引子 OPA01、F12、91167 及 91074 進行 RAPD 擴增所得的圖譜可以區分出 A、B 品系，證實台灣確有煙草粉蝨 A、B 品系之存在。利用擴增的特異性片段序列設計出之引子組 Baf/Bar，對煙草粉蝨 B 品系具有專一性之鑑別力，增幅出約 500 bp 的片段，而 A 品系則無該片段產生。4 種台灣地區稻蝗類之基因組 DNA，以篩選出之逢機引子 UBC431、OPA01 及 OPA03 進行 RAPD 可增幅出具有辨識性之圖譜，據此分析稻蝗類之親緣關係，台灣稻蝗 (O. chinensis formosana) 與日本稻蝗 (O. japonica japonica) 最為相似，其次依序為台灣短翅稻蝗 (O. podisma) 和小稻蝗 (O. hyla intricata)。比對 4 種稻蝗 rDNA 之 ITS 區域核酸序列以及 mtDNA 之 COI 區域部分序列，兩者序列分析結果相同，台灣稻蝗及日本稻蝗亦為近緣種，但與其他兩種之親緣關係依序為台灣短翅稻蝗及小稻蝗。顯示台灣稻蝗與日本稻蝗無論在形態上和遺傳標示上，皆具有相當高之相似性，因此，此兩種稻蝗是屬親緣關係相當密切之種類。

The phylogenetic relationships among Bemisia tabaci species complex (A and B biotype) and four rice grasshoppers (Oxya spp.) in Taiwan were analyzed by molecular markers (RAPD-PCR, rDNA, and mtDNA). The A and B biotype of B. tabaci could be discriminated by RAPD-PCR with four selected primers (OPA01, F12, 91167, and 91074). The result confirmed that both A and B biotype of this whitefly really exist in Taiwan. The specific primer set (Baf/Bar) was designed based on the nucleotide sequence of the amplicon, amplified by RAPD-PCR with primer 91074. This primer set, with the genomic DNA of B biotype of the whitefly, by PCR yielded an amplicon, about 500 bp, while nothing with that of A biotype whitefly was found. It revealed that this primer set can be specifically used to discriminate B biotype of the whitefly from A biotype.
Four rice grasshoppers (O. chinensis formosana, O. japonica japonica, O. podisma, and O. hyla intricata) were examined by RAPD-PCR with three selected primers (UBC431, OPA01 and OPA03). These four species could be distinguished based on the amplicon’s patterns. The phylogenetic relationships as analyzed: showed that O. chinensis formosana and O. japonica japonica are most closely related, followed by O. podisma and then O. hyla intricata. Comparisons and analyses of the nucleotide sequences of the ITS region of rDNA and the COI region of mtDNA of the four Oxya species showed that O. chinensis formosana and O. japonica japonica are closely related, followed by O. podisma and then O. hyla intricata. Therefore, O. japonica japonica is similar to O. chinensis formosana not only in morphology, but also in molecular markers. We concluded that O. chinensis formosana maybe a sibling species of O. japonica japonica.

目錄
中文摘要
英文摘要
目錄i
表次iii
圖次iv
壹、前言1
貳、往昔研究3
一、粉蝨類3
二、稻蝗類5
三、分子標示8
參、材料與方法16
一、供試昆蟲16
二、染色體核型17
三、基因組 DNA 萃取17
四、DNA 片段之增幅18
五、RAPD-PCR19
六、DNA 純化20
七、勝任細胞之製備21
八、接合反應21
九、質體轉形21
十、重組質體鑑定22
十一、DNA 定序及序列分析23
十二、專一性 PCR23
十三、親緣關係推衍24
肆、結果25
一、粉蝨類25
二、稻蝗類26
伍、討論32
陸、參考文獻41
柒、誌謝52
捌、圖表53
玖、附錄82

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