Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production |
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Authors: | A M R Ferrie K L Caswell |
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Institution: | (1) Plant Biotechnology Institute, National Research Council, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada |
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Abstract: | An isolated microspore culture provides an excellent system for the study of microspore induction and embryogenesis, provides
a platform for an ever-increasing array of molecular studies, and can produce doubled haploid (DH) plants, which are used
to accelerate plant-breeding programs. Moreover, isolated microspore cultures have several advantages over anther culture,
wherein presence of the anther walls can lead to the development of diploid, somatic calli and plants. Although protocols
for isolated microspore culture vary from laboratory to laboratory, the basic steps of growing donor plants, harvesting floral
organs, isolating microspores, culturing and inducing microspores, regenerating embryos, and doubling the chromosomes, remain
the same. Over the past few years, a large proportion of the research reports on isolated microspore culture have focused
on cereal and Brassica species. For some of these species, isolated microspore culture protocols are well established and routinely used in laboratories
around the world for developing new varieties, as well as for basic research in areas such as genomics, gene expression, and
genetic mapping. Although these species are considered highly responsive to microspore culture, improvements in efficiency
are still being made. However, with many species, isolated microspore culture is simply not yet efficient enough at producing
DH plants to be cost-effective for breeding programs. There has been a recent resurgence of haploidy research with response
being reported in some species once considered recalcitrant. Future research programs aimed at elucidating pathways involved
in microspore induction and embryogenesis will be of benefit, as will novel approaches to improve the efficiency of microspore
culture for DH production. With many species, anther culture has proven to be more effective than isolated microspore culture,
necessitating more research to clarify the contribution of the anther wall to embryogenesis. The development of molecular
markers for use in determining the gametic origin of regenerated plants, irrespective of their ploidy, would also be beneficial.
In this review, we aim to provide an overview of the basic isolated microspore culture protocol with an emphasis on recent
progress in several crop species. |
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