Plantlet Regeneration via Somatic Embryogenesis from Hypocotyls of Common Buckwheat (Fagopyrum esculentum Moench.)

Abstract

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) 2.0 mg · L ). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

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

“…Later, shoot organogenesis was established and reported with different explants such as cotyledon (Luthar and Marchetti 1994, Berbec and Doroszewska 1999, Klcova and Gubisova 2008, Lee et al 2009, hypocotyl (Berbec and Doroszewska 1999, Jin et al 2002, Klcova and Gubisova 2008, Hou et al 2014, Adachi et al 1989, Lachman and Adachi 1990, leaf (Woo et al 2004), leaf petiole (Slawinska et al 2009) and nodal segment (Kachonpadungkitti et al 2001, Klcova and Gubisova 2008, Chen et al 2012, Majid et al 2015, utilizing various media regimens and obtaining varying frequencies. Regeneration ability via somatic embryogenesis has been published in a few reports using immature zygotic embryo (Neskovic et al 1987), leaf and stem (Park et al 1999), cotyledon (Woo et al 2000, Wang et al 2016, hypocotyl (Gumerova et al 2003, Han et al 2011, Wang et al 2016, Kwon et al 2013). However, no reports are available on the investigation of the best carbon source and explant type for somatic embryogenesis based in vitro regeneration in common buckwheat.…”

Buckwheat in Tissue Culture Research: Current Status and Future Perspectives

“…Protocols for buckwheat regeneration from several explants such as hypocotyls [14,20,42,46,47,[54][55][56], immature inflorescence [17], nodal segments [57], immature embryos [34,58], anthers [44,59], leaf petioles [60] and cotyledons [40,54,61,62] were developed. In Table 2, the optimal conditions for the plantlet regeneration from different explants, F. esculentum, F. tataricum and F. cymosum, are summarised.…”

“…Saraswat and Kumar [48] speculated that low maturity of somatic embryos supplemented with KNO 3 is due to the buckwheat being a salt-sensitive crop. Hypocotyl derived protoplast MS 0.1 mg/L NAA+ 0.5 mg/L 6-BA + 0.1 mg/L GA 3 [19] Leaf and stem MS 0.2 mg/L IAA + 2.0 mg/L 6-BA [16] Anther MS 2.5 mg/L 6-BA + 0.5 mg/L IAA [44] Cotyledon MS + B5 vits 0.5 mg/L IAA + 0.25 mg/L IBA [61] Hypocotyl and cotyledon LS 0.05-0.1 mg/L TDZ + 0.5 mg/L IAA [63] Leaf and stem MS Hormone free [39] Cotyledon MS 2.0 mg/L 6-BA + 0.2 mg/L KT [40] Nodal segment MS 1.0 mg/L KT [67] Anther B5 1.0 mg/I NAA + 1.0-2.0 mg/L 6-BA [18] Hypocotyl MS 2.0 mg/L 6-BA + 1.0 mg/L KT [43] Hypocotyl B5 2.23 mg/L 6-BA + 0.17 mg/L IAA [33,47] Leaf MS 0.2 mg/L KT + 2.0-3.0 mg/L 6-BA [41] Cotyledon and hypocotyl MS + B5 vits 1.0 mg/L 6-BA [57] Nodal segment and shoot apex MS + B5 vits 2.0 mg/L 6-BA + 0.2 mg/L IAA [57] Cotyledon MS 4.0 mg/L 6-BA + 7.0 mg/L AgNO 3 [68] Leaf petiole MS 1.0 mg/L 6-BA + 1.0 mg/L 2iP + 1.0 mg/L TIBA [60] Hypocotyl MS 1.0 mg/L KT + 1.0 mg/L 6-BA + 2.0 mg/L IAA [42] Hypocotyl MS 1.0 mg/L NAA + 1.0 mg/L 6-BA [55] Hypocotyl and cotyledon MS 0.2 mg/L 6-BA + 0.5 mg/L AgNO 3 [48] F. tataricum…”

Biotechnological Methods for Buckwheat Breeding

“…Kostyukova and Rumyantseva [ 125 ] studied the formation of embryoidogenic callus, connected with certain competent cells in procambial and subepidermal layers from cotyledons of immature embryos of Tartary buckwheat. Somatic embryogenesis from hypocotyls of Tartary and common buckwheat inducing callus formation followed by plantlet regeneration with the use of a complex combination of growth regulators has been developed [ 115 , 126 ].…”