Abstract
Buckwheat (Fagopyrum esculentum, Family Polygonaceae) is an annual pseudo-cereal crop with healing benefits. However, the genetic improvement of common buckwheat has achieved only limited success, mainly due to buckwheat’s dimorphic flowers and heteromorphic self-incompatibility. Here, we develop a useful protocol for indirect somatic embryogenesis and subsequent plant regeneration from hypocotyl explants of F. esculentum. Firstly, the initial calli of hypocotyl explants were induced on Murashige and Skoog (MS) basal medium containing 2.0 mgL−1 2,4-D and 1.5 mgL−1 6-BA for 30 days culture, and then the yellowish white friable embryogenic calli were developed when the initial calli were transferred to fresh MS basal medium supplemented with 1.0 mgL−1 6-BA and 0.5 mgL−1 thidiazuron (TDZ)two to three times subculture at 40–60 days intervals. Subsequently, the somatic embryos were able to germinate from embryogenic callus sub-cultured on MS basal medium containing 1.0 mgL−1 6-BA and 0.5 mgL−1 TDZ with 15% potato puree for 20 days subculture. Finally, maximum mean percentage (75.75%) of somatic embryo-derived plants were obtained when the mature somatic embryos were transferred to MS basal medium without growth regulators for 40 days culture. Our result provides a useful protocol for plant regeneration and SE from hypocotyl explants of F. esculentum.
Highlights
Buckwheat (Fagopyrum esculentum, Family Polygonaceae) is an annual pseudo-cereal crop originated from southern China [1]
4.0 mgL−1 ) of 2,4-D in combination with different levels (0.5, 1.0, or 1.5 mgL−1 ) of 6-BA or KT, the initial calli were developed at cotyledon edges and hypocotyl basal end, and the frequency of cotyledon and hypocotyl explants with calli produced was evaluated (Supplementary Table S1)
Indirect somatic embryogenesis from immature embryos of buckwheat occurred on B5 medium sucrose when 5.0 mgL−1 2,4-D and 0.1 mgL−1 KT were supplied at the beginning of the culture period with 6% sucrose when 5.0 mgL−1 2,4-D and 0.1 mgL−1 KT were supplied at the beginning of the culture for 5 days, and transferred to B5 medium plus 2.2 mgL−1 BAP, 0.17 mgL−1 Indole-3-acetic acid (IAA) for germination of period for 5 days, and transferred to B5 medium plus 2.2 mgL−1 BAP, 0.17 mgL−1 IAA for somatic embryos [14]
Summary
Buckwheat (Fagopyrum esculentum, Family Polygonaceae) is an annual pseudo-cereal crop originated from southern China [1]. Buckwheat seeds (strictly achenes) contain proteins with healing benefits rich in lysine, dietary fiber, antioxidants, phenolic compounds and rutin [1,2,3]. The entomophilous flowers are open and fertile for only one Agronomy 2019, 9, x FOR PEER REVIEW day [9]. All these factors accounting for the low seed set and grain yields have made Buckwheat fall out offertile favorfor as only cropsone forday culture despite nutritious and healing benefits
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