Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique

Abstract

Pea and lupin plants have been potentially rich sources of protein for human and animal diets. Currently, in Poland the main source of protein is soya cake, imported to Poland in amount of 2 mln tons per year. For substitution of soya with domestic protein plants breeding methods for higher and stable yield of grain legumes should be developed. About 12 generations are needed to develop a new cultivar and under central or north European climate conditions only one generation per year is feasible in the field. 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]). Pisum and Lupinus species are known to be recalcitrant to in vitro culture [4][5][6][7]. In spite of numerous studies focused on the production of doubled haploids (DH), no important results have been noted to date [8][9][10][11]. 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. Currently, SSD populations are frequently used as alternatives to DH populations in genetic and genomic studies (see, for example, [12,13]). 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. 4 months (including verbalization) [14]. 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.The aim of the present studies was to establish in vitro conditions for the culture of pea, and narrow-leafed and yellow lupin embryos as the first step in research aimed at shortening generation cycles in the SSD technique used to accelerate the development of homozygous populations. Material and methodsMaterial for the studies consisted of two pea cultivars (Pisum sativum L.) -Cysterski and Akord, two narrow-leafed lupin cultivars (Lupinus angustifolius L.) -Kadryl and Sonet, and two yellow lupin cultivars (L. luteus L.) -Mister and Taper. Pea and lupin cultivar seeds were sown in the experimental field at Wiatrowo near Poznań. Pods from each pea and lupin cultivar were detached 21-28 days after flowering, when seeds were still-green but fully developed. Seeds taken out of pods were sterilized by successive dips into ethanol 70% (3 min) and AbstractThe 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 a...

QTLs for earliness and yield-forming traits in the Lubuski × CamB barley RIL population under various water regimes

“…The parent Cam/B1/CI08887//CI05761 (hereafter referred as CamB) is the Syrian breeding line supplied to Dr. A. Górny by Drs S. Grando and S. Ceccarelli from ICARDA in Aleppo and Lubuski is an old Polish cultivar derived from a Heines-Haisa/Skrzeszowicki hybrid. The examined population was developed by means of the single seed descent (SSD) technique (up to F 8 ) (Goulden 1939) associated with in vitro culture of immature embryos (Surma et al 2013). Out of 150 developed RILs 100 were randomly chosen for the present experiments.…”

Grain Legumes

“…Therefore, effective methods to shorten multiyear breeding cycles are not available. Surma et al (2013) tried to elaborate the single-seed descent (SSD) technique using in vitro cultures of lupin embryos ( L. luteus and L. angustifolius). It appeared that lupin roots and shoots grow well in vitro, but further plantlet acclimatization ex vitro is rather difficult.…”

“…Moreover, some of the regenerated plants have survived the transfer to greenhouse conditions where they flowered and set pods/seeds. These results constitute a very optimistic step towards the rapid attainment of succeeding generations via the SSD technique (Surma et al 2013).…”

“…A combination of the SSD method and in vitro culture of embryos was recently proposed for lupins (Surma et al 2013). As a first step towards that goal, culture conditions for L. angustifolius and L. luteus embryos dissected from fully developed, green seeds have been established.…”

In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik)

“…Modified SSD systems involving a first step of immature seeds in vitro culture for germination and a second step ex vitro for full development have been proposed to significantly shorten the breeding cycles in grain legumes such as pea and lupin (Ochatt et al 2002;Surma et al 2013). Recently, in lentil, the length of the generation cycle has been reduced using an in vitro only strategy (Mobini et al 2014) and an in vivo only strategy ).…”