THIẾT LẬP ĐIỀU KIỆN TẠO VÀ NUÔI CẤY CALLUS IN VITRO CHO NGHIÊN CỨU STRESS MẶN Ở GIỐNG LÚA KHANG DÂN 18

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Tác giả: Thị Sen Đào, Thị Mai Hoa Điêu, Thu Trang Hà, Văn Quyền Nguyễn, Thị Dung Vũ, Thị Quỳnh Hương Vương

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Thông tin xuất bản: Tạp chí Khoa học - Trường Đại học Sư phạm Hà Nội: Khoa học Tự nhiên, 2020

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Giống Khang Dân 18 là một trong những giống lúa được trồng phổ biến ở Việt Nam, nơi đang bị đe dọa bởi sự xâm nhập mặn. Nghiên cứu này được tiến hành nhằm thiết lập điều kiện nuôi cấy callus và đánh giá khả năng chịu stress mặn in vitro ở giống lúa Khang Dân 18. Đầu tiên, các điều kiện khác nhau bao gồm hàm lượng chất điều hòa sinh trưởng (chất ĐHST) bổ sung, thời gian nuôi cấy và điều kiện chiếu sáng được khảo sát nhằm tạo và nuôi cấy được callus in vitro phù hợp. Môi trường thích hợp tạo để callus là môi trường ¼-MS bổ sung 10 - 40 µM 2,4-Dichlorophenoxyacetic acid (2,4-D). Khảo sát các khoảng thời gian nuôi cấy callus 1 tuần, 2 tuần và 4 tuần cho thấy khoảng thời gian nuôi cấy ảnh hưởng rõ rệt đến hình thái và chất lượng callus. Callus được tạo ra từ hạt sau 1 tuần nuôi cấy có tính chất và hình thái thích hợp để nghiên cứu tính chống chịu stress mặn hơn callus sau 1 tháng. Điều kiện chiếu sáng 10 h\ngày và điều kiện tối hoàn toàn là điều kiện thích hợp để tạo callus. Môi trường bổ sung 10 µM 2,4-D cho chất lượng callus tốt hơn các môi trường bổ sung cả 1-Naphthylacetic acid (NAA) và 2,4-D. Tỉ lệ tạo rễ và chồi của callus ở ba điều kiện chiếu sáng 10 h\ngày, 24 h\ngày và tối hoàn toàn có sự sai khác có ý nghĩa thống kê. Điều kiện nuôi cấy callus in vitro được sử dụng cho nghiên cứu về khả năng chịu mặn của callus. Sự sinh trưởng của callus suy giảm đáng kể khi nồng độ NaCl tăng lên 3 - 6‰ (P = 0,05), đặc biệt điều kiện 9‰ NaCl gây suy giảm mạnh về khối lượng và thay đổi rõ nét về mặt hình thái callus. Điều kiện nuôi cấy tối ưu ở nghiên cứu này có thể được áp dụng cho các nghiên cứu in vitro về stress và khả năng chịu mặn của giống lúa Khang Dân 18 và các giống lúa tương tự.The Khang Dan 18 is one of the rice cultivars widely cultivated in Vietnam, which is threatened by saline intrusion. This study is conducted to establish an in vitro culture system for evaluate salt tolerance of the rice cultivar Khang Dan 18. First, different culture conditions, including plant growth regulator concentration, culture period, light regime, were optimized to induce and maintain callus in vitro. Culture media supplemented with 10 - 40 µM 2,4- Dichlorophenoxyacetic acid (2,4-D) are suitable for inducing callus. Different culture periods (1, 2 and 4 weeks) significantly affected the callus morphology and nature. The 10-hour light photoperiod is the most effective condition for callus induction and multiplication. Callus cultured in the medium supplemented exclusively with 2,4-D showed higher quality compared to those in media supplemented with 1-Naphthylacetic acid (NAA) and 2,4-D. Different photoperiods also affected callus differentiation in terms of root\shoot induction. The established culture system was then employed to evaluate salt tolerance of the callus. The growth of callus was significantly decreased when the concentrations of NaCl increased (3, 6 and 9‰, P = 0.05), especially at the concentration of 9‰ NaCl caused a strong decrease in callus weight and an obvious change in callus morphology. This culture system may be applied for in vitro salt stress studies on the cultivar Khang Dan 18 and similar rice cultivars.

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