Hà Trần Minh Dũng * , Dương Ngọc Kiều Thi , Lê Tấn Đức Nguyễn Hữu Hổ

* Tác giả liên hệHà Trần Minh Dũng

Abstract

The suitable regenerating medium for 10-day old cotyledon of tomato was MS containing  0.5 mg/l BA, 0.5 mg/l kinetin, 0.1 mg/l IAA and 8.4 g/l agar. pH was adjusted  at 5.8.The lethal dose of kanamycin on cotyledonary segments (control) was 100 mg/l.The introduction of plasmid pCAMBIA 2301 ? Cry1Ab into Agrobacterium tumefaciens train  LBA 4404 and the presence of Cry1Ab gene were tested by PCR technique. The OD600nm = 1 and 100 àM acetosyringone resulted in the highest transient GUS activity (45.6 ± 5.1%) on transgenic cotyledonary fragments. The presence of Cry1Ab gene in 2-week old cotyledonary fragments and 3-month old transgenic tomato in vitro plants was evidenced by PCR.

Keywords: Agrobacterium tumefaciens LBA 4404, Hong Chau tomato variety, Lycopersicon esculentum Mill.

Tóm tắt

Môi trường tái sinh chồi cây cà chua in vitro từ lá mầm là MS có bổ sung 0,5 mg/l BA, 0,5 mg/l kinetin, 0,1 mg/l IAA, 8,4 g/l agar và pH 5,8; ngưỡng gây chết của kanamycin sulphate đối với lá mầm đối chứng là 100 mg/l. Việc biến nạp plasmid pCAMBIA 2301 ? Cry1Ab vào Agrobacterium tumafaciens dòng LBA 4404 và kiểm tra gen Cry1Ab đã được thực hiện thành công với sản phẩm khuếch đại của gen Cry1Ab là 559 bp. Mật độ OD600nm = 1 và 100 àM acetosyringone cho tỷ lệ biểu hiện GUS cao nhất với 45,6 ± 5,1%.  Kết quả PCR cho thấy có sự hiện diện của gen Cry1Ab ở một số lá mầm chuyển gen 2 tuần tuổi và lá cà chua 3 tháng tuổi. 
Từ khóa: Gen Cry1Ab, Agrobacterium tumefaciens LBA 4404, cà chua Hồng Châu, Lycopersicon esculentum Mill.

Article Details

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