TÁI SINH CÂY ĐẬU TƯƠNG CHUYỂN GENE MANG CẤU TRÚC pZY102::GmDREB7A THÔNG QUA CHỌN LỌC BẰNG PHOSPHINOTHRICIN

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Tác giả: Hoàng Mậu Chu, Thu Hà Dương, Đức Hùng Nguyễn, Thị Hải Yến Nguyễn, Thị Hồng Trần

Ngôn ngữ: vie

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Thông tin xuất bản: Tạp chí Khoa học và Công nghệ - Đại học Thái Nguyên, 2023

Mô tả vật lý: tr.377-383

Bộ sưu tập: Metadata

ID: 340142

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Đậu tương là loại cây trồng mẫm cảm với hạn và mặn, do vậy nghiên cứu biện pháp tăng cường khả năng chống chịu hạn và mặn của cây đậu tương là vấn đề đang được quan tâm. Protein DREB là nhân tố phiên mã làm tăng cường biểu hiện các gene liên quan đến tính chịu hạn và mặn của đậu tương, tuy nhiên đến nay còn nhiều gene DREB ở đậu tương còn chưa rõ chức năng. Nghiên cứu này nhằm tạo cây đậu tương chuyển gene phục vụ đánh giá vai trò của gene GmDREB7 đối với các gene liên quan đến tính chống chịu hạn và mặn của cây đậu tương. Các phương pháp biến nạp di truyền, tái sinh, chọn lọc in vitro và phân tích cây chuyển gene đã được sử dụng để tạo cây đậu tương chuyển gene phục vụ nghiên cứu chức năng của gene GmDREB7. Từ 150 mẫu được biến nạp cấu trúc pZY102::GmDREB7A đã thu được 447 chồi và qua 3 lần sàng lọc bằng phosphonothricin (ppt) đã thu được 17 cây in vitro được chuyển gene GmDREB7A. Nhân dòng in vitro và chọn lọc các cây T0 bằng ppt thu được 8 dòng đậu tương, ký hiệu là T0-1, T0-2, T0-3, T0-4, T0-5, T0-6, T07, T0-8. Kết quả phân tích PCR 8 dòng đậu tương kháng ppt trong nhà lưới thu được 7 dòng dương tính với PCR (T0-1, T0-2, T0-3, T0-4, T0-5, T0-6, T07, T0-8). Các dòng đậu tương chuyển gene GmDREB7A tiếp tục được theo dõi và phân tích ở các thế hệ tiếp theo.Soybean is a crop sensitive to drought and salinity stress, so research on measures to improve drought and salinity tolerance of soybean is a matter of current concern. The DREB protein is a transcription factor that enhances the expression of genes related to drought and salinity tolerance in soybean
however, there are many DREB genes in soybean whose function is still unknown. This study aimed to create transgenic soybean plants to evaluate the role of the GmDREB7 gene on genes involved in the abiotic stress tolerance of soybean. The methods of genetic transformation, regeneration, in vitro selection and analysis of transgenic plants were used to create transgenic soybean plants for the functional study of the GmDREB7 gene. From 150 samples transformed with pZY102:GmDREB7A construct, 447 shoots were obtained, and 17 in vitro GmDREB7A transgenic plants were obtained after three screenings with phosphinothricin (ppt). In vitro cloning and selection of T0 plants by ppt obtained 8 soybean lines, denoted T0-1, T0-2, T0-3, T0-4, T0-5, T0-6, T07, T0- 8. Analysis of 8 lines of ppt-resistant soybeans in greenhouses by PCR resulted in 7 PCR-positive transgenic soybean lines (T0-1, T0-2, T0-3, T0-4, T0-5, T0-6, T07, T0-8). The GmDREB7A transgenic soybean lines continued to be monitored and analyzed in subsequent generations.

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