ISSN 2167-0447
International Journal of Agricultural Sciences ISSN 2167-0447 Vol. 7 (1), pp. 1228-1233, January, 2017. ©International Scholars Journals
Full Length Research Paper
Developing an efficient protocol for plant regeneration of manihot esculenta crantz via somatic embryogenesis induction from immature leaf and apical shoots
1Do Hai Lan*, 2Do Xuan Dong, 2Le Van Son, 2Le Tran Binh, and 2Chu Hoang Ha
1Faculty of Biochemistry, Tay Bac University, Quyet Tam District - Sơn La City,
2Institute of Biotechnology, National Key Laboratory of Gene Technology.
Received 12 September, 2016; Revised 03 January, 2017; Accepted 16 January, 2017 and Published 16 January, 2017
Email: [email protected]
Abstract
In this study, we developed a method for the induction of somatic embryogenesis in two cassava genotypes from Vietnam is described. The explants used were immature leaves and apical shoots isolated from in vitro 2-3-week plants. Somatic embryogenesis was achieved at high frequencies by the addition in the induction medium of the auxin picloram over a wide range of concentrations. Our results show that the highest rate of somatic embryos formation for both cultivars was obtained by using 12 mg/l picloram supplemented to Murashigh - Skoog (MS) media. For immature leaves: the number of embryos per explant was similar between two cultivars for 4 weeks, the KM94 cultivar gave a higher rate (81,4 ±1,7%) than the KM140 cultivar (70,4 ± 2,9%). For apical shoots: the number of embryos per explant was similar between two cultivars for 4 weeks, the KM94 cultivar gave a higher rate (82 ±1.7%) than the KM140 cultivar (59.6 ± 2.9%). Somatic embryos were subsequently transferred onto media (MS supplemented with 0.3 mg/l BAP) for the highest frequency of plantlet regeneration (KM140 - 82,1 ± 3,1% and KM94 - 79,2 ± 2,3%), for immature leaves and (KM140 - 81 ±3.2% and KM94 - 79 ± 2.3%) for apical shoots. Shoots with the length of about 1.0-1.5 cm were transferred onto free - hormone MS media for 100% of rooting for 2 weeks. Complete plantlets were cultivated on a mixture of rice husk and sand-soil under ratio 4:6 in a greenhouse. This protocol required 16 to 18 weeks and is entirely appropriate for mass production of various cassava genotypes and further genetic transformation experiments.
Keywords: Cassava (Manihot esculentum Crantz), Callus culture, immature leaf, apical shoots, somatic embryos, regeneration.