Abstract
Most aspects of microspore culture protocol have the capacity to cause stress to microspores, hence, less stressful treatments might be required to avoid deleterious effects. In stressed plants, polyamines and trehalose can act as compatible solutes or osmoprotectants by stabilizing proteins and biological membranes. To improve green plant regeneration in wheat microspore culture, this study assessed the effects of polyamines (putrecine, spermidine, spermine) and trehalose on androgenic response namely embryogenesis, green plant regeneration and ploidy of green plants regenerated in three spring wheat genotypes. Microspores of the genotypes produced significant numbers of embryos and green plants among polyamine treatments but trehalose had no effect (P ≤ 0.05). Polyamine treatments for 30 min generally produced more green plants per 100 microspores than the 60 min treatments in all three genotypes. At least three out of twelve polyamine treatments in each genotype improved the production of double haploid plants and seed setting in regenerants. Wheat genotype, concentration and duration of polyamine treatment had significant impact on embryogenesis and regeneration of green plants in this study. The study also showed that polyamines could be used to accelerate cultivar development in wheat breeding.
Highlights
Microspores are haploid spores that develop into male gametophytes in heterosporous plants
The objective of this work was to investigate a protocol for efficient wheat microspore culture by the application of polyamines and trehalose, to wheat microspores to improve or increase the number of microspore-derived double haploid green plants
Two PA treatments, 1.0 mM Put and 1.0 mM Spm for 30 min produced the highest number of Green regenerated plants (GRPs)/100 embryoids in ICARDA39 whiles in DH83Z, the 0.5 mM combination treatments of (Put + Spm) and (Spm + Spd) yielded the highest GRPs/100 embryoids (Figure 5(a))
Summary
Microspores are haploid spores that develop into male gametophytes in heterosporous plants. Microspore culture (androgenesis) is currently used to produce double haploid plants for breeding and genetic research to obtain homozygous lines [1,2,3,4]. Wheat double haploid lines obtained through maize pollination and anther culture methods are being cultivated [7,8]. The application of androgenesis in a number of crops including wheat still requires improvement because many lines are recalcitrant, have a very low response to the protocol and/or often produce a high percent of albinos. Exogenous application of polyamines (PAs) to wheat anthers improved the green plant regeneration but overall effect was dependent on duration of the pretreatment and on the genotype [10]
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