Somatic embryogenesis of agriculturally important lupin species (Lupinus angustifolius, L. albus, L. mutabilis)

Abstract

Somatic embryos were obtained from immature cotyledons of Lupinus angustifolius, L. albus and L. mutabilis but not from L. luteus. Different kinds of basal media and plant growth regulators in primary and secondary culture were tested. The best induction media were based on BS'and were supplemented with 5 mg 1-1 2,4-D alone or with 0.25 mg 1-1 kinetin. Mature stage somatic embryos were obtained on media containing ABA (0.1-0.5 mg 1 -~) and a high NH4/NO 3 ratio. Embryo germination and plantlet development occurred on MS media supplemented with glutamine or GA 3.

Enhanced methionine levels and increased nutritive value of seeds of transgenic lupins ( Lupinus angustifolius L.) expressing a sunflower seed albumin gene

“…To introduce foreign genes into lupins, we first developed a robust regeneration protocol that could be used in combination with a gene delivery system such as A. tumefaciens. Although the regeneration of lupin explants from tissue culture has been reported (26)(27)(28), attempts to reproduce these results in our laboratory using Australian commercial cultivars were unsuccessful. After extensive testing of a range of tissue explants and media, we developed a reproducible regeneration system, based on organogenesis, using thinly sliced embryonic axes of maturing seeds.…”

Morphogenic Aspects of Somatic Embryogenesis

“…Among those legumes in which a callus phase is not evident prior to embryo formation, such as crownvetch [50], pea [51], and soybean [52,53], the frequency of embryogenesis is decreased when a cytokinin is used in conjunction with an auxin. In lupin species, addition of a cytokinin does not affect the embryogenic response if 2,4-0 is used as the inducing auxin, but does reduce it ifNAA is used as the inducing auxin [54]. The exact effect ofcytokinins during the induction process remains difficult to assess fully, given that studies only rarely include a cytokinin-free treatment whenever growth regulator studies are evaluated for their effectiveness in embryo induction.…”

Agrobacterium-mediated transformation of Lupinus mutabilis L. using shoot apical explants

“…Procedures were described for shoot induction from hypocotyl-derived callus (Sroga 1987) and of somatic embryogenesis from immature cotyledons (Nadolska-Orczyk 1992) of Lupinus, but other workers failed to reproduce these protocols (Phoplonker and Caligari 1992;Babaoglu, Power and Davey, unpublished). Cell suspensions of L. polyphyllus and L. hartwegii were transformed by oncogenic strains of Agrobacterium tumefaciens (Berlin et al 199 la,b), while tissues of L. angustifolius and L. luteus were inoculated with A. tumefaciens carrying the nptII, bar and gus genes (Somsap et al 1994).…”