Abstract
The aim of the studies was to establish in vitro conditions for the culture of pea and lupin embryos as the first step in the development of an in vitro assisted single seed descent technique for the attainment of homozygous populations. Materials for the study included of pea, and narrow-leafed and yellow lupin cultivars. Embryos dissected from mature but still-green seeds were cultured in vitro on two modified MS media and under three temperature regimes. Shoot and root lengths of regenerated plants were measured after 7, 14 and 21 days of culture. For pea plants full-strength MS medium with 4 g l<sup>−1</sup> agar and temperature 22/ 20°C (day/night) appeared to be the most conducive to shoot and root development, whereas for lupin plants lower temperatures were more propitious: 12°C in the dark for narrow-leafed lupin and 16/ 12°C (day/night) for yellow lupin. Almost all the cultured embryos developed into plants, but not all the regenerated plants survived acclimation to ex vitro conditions.
Highlights
Pea and lupin plants have been potentially rich sources of protein for human and animal diets
The present results demonstrate that pea and lupin embryos cultured in vitro develop in plants, which are ready for transfer into pots after ca. 4 weeks
Stafford and Davies [22], who cultured in vitro immature pea embryos on MS medium, showed that growth of embryos was dependent on sucrose concentrations
Summary
Pea and lupin plants have been potentially rich sources of protein for human and animal diets. For that reason the development of new tools is needed to accelerate obtaining homozygosity in breeding of pea and lupins. Homozygosity can be attained by selfing successive generations or by hapoidization of hybrids using different in vitro techniques and doubling chromosomes in haploid plants (for review see, e.g., [1,2,3]). For shortening the breeding cycle, the single seed descent technique (SSD) in combination with in vitro culture of immature embryos may be an alternative to the DH system. The SSD technique connected with embryo in vitro culture allows the attainment of homozygous lines in a relatively short time. Such an approach applied to the development of winter barley SSD lines enabled the shortening of the generation cycle to ca. Ochatt et al [15] proposed the reduction of pea generation cycles based on in vitro culture of embryos and a greenhouse strategy, but no information was found on embryo culture for accelerating breeding process in lupins
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