A Temporary Immersion System Improves Regeneration of In Vitro Irradiated Recalcitrant Indica Rice (Oryza sativa L.) Embryogenic Calli

Plants

Jason Pérez¹  Ana Abdelnour-Esquivel¹  Rebeca Fait-Zúñiga¹  Alejandro Hernández-Soto²  Walter Vargas-Segura³  Randall Rojas-Vásquez⁴  Andrés Gatica-Arias⁴ 

[1] Biotechnology Research Center, Biology School, Costa Rica Institute of Technology, Cartago P.O. Box 159-7050, Costa Rica;Doctorado en Ciencias Naturales para el Desarrollo (DOCINADE), Instituto Tecnológico de Costa Rica, Universidad Nacional, Universidad Estatal a Distancia, Cartago P.O. Box 159-7050, Costa Rica;Gamma Irradiation Laboratory, School of Physics, Costa Rica Institute of Technology, Cartago P.O. Box 159-7050, Costa Rica;Plant Biotechnology Laboratory, School of Biology, University of Costa Rica, San José P.O. Box 2060, Costa Rica;

关键词: somatic embryogenesis;    Cobalt-60;    radiation-induced mutagenesis;    temporary immersion systems (TIS);   

DOI  :  10.3390/plants11030375

来源: DOAJ

【 摘 要 】

The development of gamma ray-mutated rice lines is a solution for introducing genetic variability in indica rice varieties already being used by farmers. In vitro gamma ray (⁶⁰Co) mutagenesis reduces chimeras and allows for a faster selection of desirable traits but requires the optimization of the laboratory procedure. The objectives of the present work were sequencing of matK and rbcL, the in vitro establishment of recalcitrant rice embryogenic calli, the determination of their sensitivity to gamma radiation, and optimization of the generation procedure. All sequenced genes matched perfectly with previously reported matK and rbcL O. sativa genes. Embryogenic calli induction improved using MS medium containing 2 mg L⁻¹ 2,4-D, and regeneration was achieved with MS medium with 3 mg L⁻¹ BA and 0.5 mg L⁻¹ NAA. The optimized radiation condition was 60 Gy, (LD20 = 64 Gy) with 83% regeneration. An immersion system (RITA^®, Saint-Mathieu-de-Tréviers, France) of either 60 or 120 s every 8 h allowed systematic and homogeneous total regeneration of the recalcitrant line. Other well-known recalcitrant cultivars, CR1821 and CR1113, also had improved regeneration in the immersion system. To our knowledge, this is the first study reporting the use of an immersion system to allow for the regeneration of gamma-ray mutants from recalcitrant indica rice materials.

【 授权许可】

Unknown