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Tái sinh thực vật từ đầu chồi Rosa được cryoprotect bằng phương pháp vitrification giọt kết hợp
Tóm tắt
Các đầu chồi thu được từ cây Rosa nuôi cấy in vitro (ba giống) đã được bảo quản đông lạnh thành công bằng phương pháp vitrification giọt kết hợp và sau đó đã tái sinh được các chồi. Các đầu chồi bị cắt (dài 1–4 mm) được ủ trong môi trường lỏng MS bổ sung 2.5 mg l−1 thiamine, 0.2 mg l−1 biotin, 0.2 mg l−1 pyridoxine, 0.25 mg l−1 6-benzylaminopurine (BAP), 0.5 mg l−1 axit gibberellic (GA3) và 0.08 M sucrose trong 24 giờ. Sau đợt ủ, các đầu chồi được tiền nuôi cấy trong môi trường MS này chứa 0.1 đến 1.0 M sucrose trong 24 và 48 giờ tương ứng. Các đầu chồi đã tiền nuôi cấy được làm khô với dung dịch bảo vệ cryo PVS2 nồng độ cao trong 10–30 phút ở nhiệt độ thường, trước khi nhúng trực tiếp vào nitơ lỏng. Sau khi làm nóng trở lại nhanh chóng trong môi trường lỏng đã đề cập, các đầu chồi được cấy trên môi trường MS đã điều chỉnh (5 g l−1 agar) có bổ sung vitamin và các chất điều hòa sinh trưởng thực vật như đã nêu trên để tái sinh. Các đầu chồi được bảo quản đông lạnh đã bắt đầu phát triển lại trong vòng 10 ngày và tái sinh chồi trong vòng 3 tuần. Số lượng đầu chồi phát triển lại cao nhất là 64.44% cho giống Kardinal, 67.73% cho giống Fairy và 57.57% cho giống Maidy.
Từ khóa
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