Plant Regeneration in Buckwheat (Fagopyrum esculentum Moench.) via Somatic Embryogenesis and Induction of Meristemoids in Abnormal Embryos

Abstract

An efficient in vitro regeneration protocol is reported for common buckwheat. A combination of 0.5 mg/l 2,4-D and 0.2 mg/l BAP with sucrose showed highest induction of somatic embryogenesis from cotyledon and hypocotyl explants. More than 35% of normal somatic embryos matured on MS. MS with 2% sucrose were found best for germination and conversion of somatic embryos to plantlets. In tissue culture, abnormal somatic embryos usually occur. In this report, abnormal embryos are also used to induce shoot organogenesis, adding to the number of final regenerants and ensuring full utilization of regenerative propagules. A treatment of 0.2 mg/l BAP induced meristemoids in 60% of underdeveloped embryos and a combination of 0.5 mg/l BAP and 0.5 mg/l AgNO3 led browning and senescence-free progression of shoot buds to well developed shoots, which were subsequently rooted in half strength MS containing 2% sucrose and 0.25 mg/l IBA. The regenerated plants survived acclimatization, flowered and set seeds.
 Plant Tissue Cult. & Biotech. 29(1): 33-47, 2019 (June)

Somatic Embryogenesis and Plant Regeneration from Cotyledon and Hypocotyl Explants of Fagopyrum esculentum Moench lpls Mutant

“…Moreover, abnormal somatic usually occurred in this experiment. Similar results were also observed in the tissue culture described by Saraswat and Kumar [16]. TDZ in combination with NAA was proved to be the most effective for direct somatic embryo formation and regeneration in Epipremnum aureum [29].…”

“…Moreover, TDZ and 6-BA in combination with NAA or potato puree could induce germination of buckwheat somatic embryos in our study. However, recent study suggested that a combination of 0.5 mgL −1 2,4-D and 0.2 mgL −1 BAP with sucrose showed highest induction of SE from cotyledon and hypocotyl explants of buckwheat [16]. In addition, embryogenic callus from cotyledon of buckwheat was found to be MS medium containing 0.2 mgL −1 KT, 2.0 mgL −1 BAP and 3% (w/v) sucrose [26].…”

“…Moreover, somatic embryogenesis from hypocotyl explants of buckwheat were reported on MS medium containing 2.0 mgL −1 IAA, 1.0 mgL −1 KT, 1.0 mgL −1 6-BA and 3% sucrose, and 2,4-D did not show any significant effect on callus induction or embryogenesis [15]. However, recent study suggested that a combination of 0.5 mgL −1 2,4-D and 0.2 mgL −1 BAP with sucrose showed obvious induction of somatic embryogenesis from cotyledon and hypocotyl explants of buckwheat [16]. Here, we developed a useful protocol for plant regeneration and SE from hypocotyl explants of F. esculentum, which provides a potential method for producing self-compatibility lines of buckwheat.…”

Buckwheat in Tissue Culture Research: Current Status and Future Perspectives

“…This was also observed by Gumerova et al [33], who noted that the regeneration from PECCs can be achieved in three ways: via the formation of vegetative buds, via the formation of somatic embryos on the PECCs' surface and via the whole PECC transformation in a sprouting embryo. The term embryogenic callus is also used by Saraswat and Kumar [48], where it was induced from hypocotyls and cotyledons. On the other hand, Valieva et al [49] describes an embryogenic callus, which can form embryoids, roots and buds, and a morphogenic callus, which can form vegetative buds and roots.…”

“…On the contrary, Woo et al [40] found that increased sucrose content did not have any effect on the callus induction or SE. In terms of callus induction and proliferation, Saraswat and Kumar [48] noticed that the hypocotyl explants responded better in media supplemented with glucose, while cotyledon explants responded better to the medium with sucrose as the carbohydrate source. Park et al [39] were able to regenerate plants from leaf-derived calli on MS medium devoid of hormones, but the regeneration rate was low and failed to regenerate plantlets from stem-derived calli.…”