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題名 | 東方果實蠅(雙翅目:果實蠅科)性別決定相關基因之表現分析及變形基因之功能研究=Expression Analyses of Sex-Related Genes and Functional Studies of the Transformer Gene in Bactrocera dorsalis (Diptera: Tephritidae) |
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作者 | 李嘉欣; 張誠; 戴淑美; | 書刊名 | 臺灣昆蟲 |
卷期 | 37:2 2017.09[民106.09] |
頁次 | 頁38-50 |
分類號 | 387.795 |
關鍵詞 | 東方果實蠅; 性別決定機制; 變形基因; RNA干擾; Bactrocera dorsalis; Transformer gene; Sex determination; RNA interference; |
語文 | 中文(Chinese) |
中文摘要 | 東方果實蠅 (Bactrocera dorsalis Hendel) 是臺灣果樹最重要的害蟲之一,其危害主要肇因 於雌蟲在果實上產卵,導致果實損傷與產量降低。由於果實受損程度與田間雌蟲數量相關,若能全 面性了解東方果實蠅的性別決定機制,將有助於新的防治策略擬定,以降低田間雌成蟲的數量。根 據文獻指出變形基因 (transformer, tra) 是果實蠅性別決定的開關,控制 tra 的表現即可影響其性 別。因此,本研究從不同時期的東方果實蠅萃取 RNA,再以反轉錄 (reversed transcription) 與 聚合酶連鎖反應 (polymerase chain reaction) 分析 tra、tra-2、雙性基因 (doublesex, dsx) 及卵 黃蛋白基因 (yolk protein 1, yp1) 等性別決定相關基因的表現模式,試圖了解這些基因的表現與 雌、雄個體發展的關係。結果顯示卵期的 Bdtraf 相對表現量高,證實母體會透過卵子給予胚胎 traf mRNA,作為胚胎性別決定的基礎;然而幼蟲及蛹期因處於性別特徵不明顯的階段,各性別相關 基因的表現量皆不高。羽化之後,雌成蟲的 Bdtraf 、Bddsxf 及 Bdyp1 表現量與雄成蟲的 Bddsxm 表現量皆隨著日齡漸增,此結果應與成蟲需啟動下游大量性別專一性基因有關。Bdtra-2 雖為雌雄 皆有的輔助性基因,但於幼蟲、蛹、成蟲時期均呈現穩定表現,推測可能參與其他未知的生理作用。 其次,以 Bdtra 的雙股 RNA (double stranded RNA, dsRNA) 干擾東方果實蠅的蛹期發育,在 68 個注射 Bdtra dsRNA 的蛹中分別有 16 個與 19 個羽化為雄成蟲及雌成蟲,性別比例接近 1:1。 存活的雌成蟲中有 9 隻產卵器變形,9 隻中又有 7 隻雌成蟲不產卵及 1 隻產出無效卵。雄成蟲大部 分皆具有正常的外生殖器,但部分個體無子代產生。推測在蛹期默化 tra 之表現,可能間接經由干 擾下游 dsxf 造成雌蟲產卵器變形與無法產卵,但不會影響東方果實蠅之性別決定。 |
英文摘要 | The oriental fruit fly, Bactrocera dorsalis (Hendel), is one of the most economically detrimental insect pests in Taiwan. The females puncturing the skin of fruit to deposit eggs and the larvae feeding in fruit cause severe damage to fruits and lead to huge economic losses. Because the number of female flies in the field is correlated with the degree of loss, more knowledge on the sex-determination mechanism would be beneficial to developing efficient control strategies for the pest. In tephritid flies, the transformer gene (tra) is a key switch in sex determination. To learn more about this process, the relative expression patterns of sex-related genes, including tra, transformer-2 (tra-2), doublesex (dsx), and yolk protein gene (Bdyp1), in Bactrocera dorsalis were examined at different developmental stages using reverse transcription polymerase chain reaction. High levels of Bdtraf observed in both unfertilized and fertilized eggs were consistent with previous observations that the female deposits of traf mRNA into eggs. During larval and pupal stages, many gender characteristics are still indefinite and expression levels of these sex-related genes are relatively low compared with those of adults. After eclosion, the expressions of Bdtraf , Bddsxf , and Bdyp1 in females and Bddsxm in males increase gradually with age. The phenomena indicate these products are functionally required and critical in downstream sex differential regulation. Because TRA-2 is present in males and females, its constant expression in all stages implies it might have other physiological functions in addition to forming the TRA/TRA-2 complex. For functional analysis, Bdtra double-stranded RNA (dsRNA) covering exon 2 to exon 4 was injected into 68 early pupae. A total of 16 male and 19 female flies eclosed successfully, with a sex ratio close to 1:1 and none of the expected intersex or XX-reversed males detected. However, eight dsRNA-treated females had deformed ovipositors, and six of these females laid no eggs. Most of the male adults possessed normal external genitalia, but a few individuals had no offspring. These results suggested that timing for conducting RNAi of Bdtra is critical for XX-reversed male formation and that Bdtra silencing in the pupal stage was unable to affect the sex determination of B. dorsalis. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。