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GrainGenes Reference Report: PRT-20-475

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Reference	PRT-20-475
Title	Enhancement of microspore culture efficiency of recalcitrant barley genotypes
Journal	Plant Cell Reports
Year	2001
Volume	20
Pages	475-481
Author	Li H Devaux P
Abstract	Summary: The culture response of isolated microspores of seven recalcitrant cultivars of barley has been largely improved by identifying an appropriate pretreatment and utilizing ovary co-cultivation. After comparison of three pretreatment media, medium B was shown to be most efficient for inducing microspore embryogenesis, while 0.3 M mannitol frequently used for the responsive cv. Igri was found to be ineffective for recalcitrant genotypes. A further significant improvement of embryogenesis was achieved by using ovary co-culture, which resulted in an overall 2.1-fold increase in embryo formation and 2.4-fold increase in green plant regeneration from all cultivars compared with the control. Optimal co-culture conditions were identified as 5 ovaries/ml medium kept over 20 days in induction culture. Microspore plating densities in cultures with and without co-culture were found to be optimal at 4x10(4)/ml and 8-12x10(4)/ml, respectively. The most effective and reproducible method for culturing microspores of recalcitrant genotypes appeared to be the combination of medium B pretreatment with ovary co-culture. By using this procedure, the genotypic difference in microspore embryogenesis could be reduced. It was found that medium B mainly enhanced percent live embryogenic microspores, and ovary co-culture subsequently improved cell division and embryogenic development. The method described here is important for the application of the microspore culture technique to barley breeding and biotechnology
Keyword	[ Hide all but 1 of 46 ] application barley barley breeding barley genotypes benzyladenine biochemical techniques biotechnology breeding cell cell culture cell division cell-division cultivar culture culture media density division embryo embryogenesis embryogenic microspores embryonic development genetic variation genotype genotypic difference green-plant-regeneration haploid hordeum vulgare improvement increase induction isolated microspores live mannitol media method microspore microspore culture microspore embryogenesis microspores ovaries ovary ovules plant embryos plant regeneration regeneration regenerative ability

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