應用農桿菌進行台中亞蔬四號番茄之轉殖

Transformation of Tomato Variety 'Taichung Asveg No.4' Mediated by Agrobacterium

本試驗係以子葉或下胚軸為培植體，建立臺灣栽培種番茄台中亞蔬四號品種之農桿菌媒介法轉殖系統。番茄之子葉或下胚軸與含質體pBI121的農桿菌LBA 4404共培養後，於含有100mg/l kanamycin的培養基上進行篩選及培養，可獲得擬轉殖枝條。部分具有kanamycin抗性的技條，可表現GUS活性。擬轉殖枝條培養於含有100mg/l kanamycin之發根培養基，待發根形成植株後，移至黑軟盆中種植。藉由GUS活性分析、聚合酶連鎖反應與南方氏核酸雜交分析，證實外來基因已穩定轉殖於番茄植株中並可表現。

A transformation system using cotyledon and hypocotyl segment as explant was developed for the tomato variety ”Taichung Asveg No.4”. Cotyledon and hypocotyl segments of tomato were co-cultivated with Agrobacterium tumefaciens LBA 4404 harboring the binary plasmid pBI 121 (conferring β-glucuronidase activity and resistance to kanamycin) and cultured on the medium supplemented with 100 mg/l kanamycin to form shoots and select putatively transformants. The plants showing kanamycin-resistant and GUS (β-glucuronidase) activity were rooted on the medium containing 100 mg/l kanamycin, and transferred to puts. Stable integration and expression of the transgenes were confirmed by GUS assay, polymerase chain reaction (PCR), and Southern hybridization analysis.