木瓜輪點病毒鞘蛋白轉基因木瓜之抗病性
Other Title
Resistance Evaluation of the Transgenic Papaya Lines Expressing the Coat Protein Gene of Papaya Ringspot Virus
Journal
植物保護新科技研討會專刊
Part Of
農業試驗所特刊第57號
Pages
57-64
ISBN
957-00-7487-6
Date Issued
1996-06
Date
1996-06
Author(s)
Abstract
為了利用植物遺傳工程方式將木瓜輪點病毒(PRV)鞘蛋白(coat protein, CP)基因轉移至木瓜以產生抗病性狀，木瓜轉型作用之進行係以金剛砂製造傷口，經由Agrobacterium tumcfaciens媒介，將GUS或PRV 5’前導序列表現的PRV 鞘蛋白基因導入木瓜幼胚分化組織中，並再生成轉基因植株。經由PCR 及西方漬染法分析，確定CP 基因存在轉型木瓜株系中，並可表現產生PRV的鞘蛋白，計獲得包含GUS 前導序列的10個株系及PRV 5’前導序列的3 個株系。先以組培養技術繁殖其R0植株，經由發根、馴化，使其在溫室生長兩個月後，進行挑戰接種，以測定其抗病性狀。實驗中以來自臺灣、夏威夷及泰國三個不同來源的PRV YK、HA 及TH 系統供試“接種後在溫室中觀察其結果，不表現病徵之植株再以酵素連結免疫呈色法（E LlsA ）及生物分析偵測是否有病毒增殖。抗病篩選得到四個抗同源PRV YK的株系：GCP 16-0、GCP 17-0、GCP 17- 1及GCP 17-3，在此四株系中，GCP 17-1及GCP 17-3對異源的PRV HA 及TH 系統仍有高度抗性，GCP 16-0則對異源病毒只有中度抗性，而GCP 17-0的抗病能力為系統專一性的抗病現象，只對同源病毒系統有抗病力。目前此四抗病毒株系仍繼續於隔離溫室中培養，其生長情形與一般木瓜一致。但此四株系均為雌株，以日昇之花粉授粉，果實生長情形與外觀形狀也都正常。目前GCP 17-1之R1 種子已發芽，並於溫室中栽培，再次進行抗病檢定中。由於GCP 17-1及GCP 17-3對不同來源的三個病毒系統都有極高的抗病力，其未來可望不受地區限制而能廣泛應用以防治木瓜輪點病毒。
The coat protein (CP) of a local mosaic strain of papaya ringspot vrius (PRV YK) was previously constructed in the Ti-vector for generation of transgenic papaya resistant to PRV infection. The CP gene with a GUS or the PRV leader sequence was transferred to embryogenic tissues derived from immature embryos of papaya via Agrobacterium -mediated transformation assisted by carborundumwounding treatment. The presence and the expression of the transgene in the putative transgenic lines regenerated were confirmed by PCR detection and Southern blotting. R0 plants of ten CP-transgenic lines with the GUS leader and three lines with the virus leader were established by micropropagatio n The plants were challenged with three different strains of PRV originated from Taiwan (YK), Hawaii (HA), and Thailand (TH). The infection of PRV was determined by symptom observation, ELISA and bioassay. Four lines of GCP 16-0, 17-0, 17-1, and 17-3 were found to be resistant to the homologous YK strain. Among them, GCP 17-1 and 17- 3 were highly resistant to the heterologous HA and TH strains, GCP 16-0 were moderately resistant to the heterologous strain HA and GCP 17-0 was specifically resistant only to the homologous YK strain. All four lines were female and their horticultural properties were similar to the untransformed papaya. The R1 plants of GCP 17-1 crossed with Sunrise papaya have been obtained and their resistance to PRV infection are currently evaluated under greenhouse conditions. The broad resistance in GCP 17-1 and 17-3 indicated that they have a great potential to be applied in different areas for control of the notorious PRV.
Event(s)
植物保護新科技研討會
Proceeding of A Symposium on New Techniques of Plant Protection
Subjects
木瓜輪點病毒
鞘蛋白
木瓜
papaya ringspot virus
coat protein
papaya
Publisher
農業試驗所
Type
conference paper
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