Isolated microspore culture of oat (Avena sativa L.) for the production of doubled haploids : effect of pre-culture and post-culture conditions

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DOIResolve DOI: http://doi.org/10.1007/s11240-013-0385-0
AuthorSearch for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titlePlant Cell, Tissue and Organ Culture
ISSN0167-6857
Volume116
Issue1
Pages8996; # of pages: 8
SubjectAvena sativa; Culture conditions; Doubled haploid; Media composition; Microspore culture; Microspore embryogenesis; Oat; Plant breeding programs; Cell culture; Plants (botany); Seed
AbstractThe production of doubled haploid (DH) plants from microspores is an important technique used in plant breeding programs and basic research. Although doubled haploidy efficiencies in wheat and barley are sufficient for breeding purposes, oat (Avena sativa L.) is considered recalcitrant. The objective of this project was to develop a protocol for the production of microspore-derived embryos of oat and further develop these embryos into fertile DH plants. A number of experiments were conducted evaluating the factors influencing microspore embryogenesis, i.e. donor plant conditions, pretreatments, media composition, and culture conditions. The initial studies yielded little response, and it was not until high microspore densities (106 microspores/mL and greater) were used that embryogenesis was achieved. Depending on the treatment, yields of over 5,000 embryos/106 microspores were obtained for breeding line 2000QiON43. The doubled haploidy protocol includes: a 0.3 M mannitol pretreatment of the tillers for 7 days, culture in W14 basal medium with a pH of 6.5-7.5, a microspore density of 106 microspores/mL, and continuous incubation at 28 °C incubation. The resulting embryos observed after 28 days were plated onto solidified W14 medium with 0.8 or 1.0 g/L activated charcoal. A colchicine treatment of 0.2 % colchicine for 4 h resulted in conversion of 80 % of the plants from haploid to DH. This protocol was successful for the production of oat microspore-derived embryos and DH green plants with minimal albinism. DH seed was produced and planted for evaluation in a field nursery.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; Aquatic and Crop Resource Development
Peer reviewedYes
NRC number54686
NPARC number21270793
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Record identifier5a9526df-e02a-4e92-903a-a59bfa9ee24a
Record created2014-02-17
Record modified2016-05-09
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