NDLI: A high throughput Brassica napus microspore culture system: influence of percoll gradient separation and bud selection on embryogenesis

Content Provider	Springer Nature Link
Author	Bhowmik, Pankaj Dirpaul, Joan Polowick, Patricia Ferrie, Alison M. R.
Copyright Year	2011
Abstract	Microspore culture for the purpose of developing doubled haploid plants is routine for numerous plant species; however, the embryo yield is still very low compared with the total available microspore population. The ability to select and isolate highly embryogenic microspores would be desirable for high embryo yield in microspore culture. To maximize the efficiency of canola microspore culture, a combination of bud size selection and microspore fractionation using a Percoll gradient was followed. This approach has consistently given high embryo yields and uniform embryo development. Microspores isolated from buds 1.5 to 4.4 mm in length of Brassica napus genotypes Topas 4079, DH12075, Westar and 0025 formed embryos at different frequencies. The most embryogenic bud size range varied with each cultivar: Topas 4079 3.5–3.9 mm, DH12075 2.0–2.4 mm, and Westar and 0025 2.5–2.9 mm. When the microspores from 2.0 to 2.4 mm buds of DH12075 were carefully layered on top of a discontinuous Percoll gradient of 10, 20 and 40%, and subsequently spun through the Percoll layers by centrifugation, bands were formed containing populations of microspores of uniform developmental stage. The middle layer of the gradient contained the late uninucleate and early binucleate microspores that were the most embryogenic. In addition, the relationship between the bud size, developmental stage of isolated microspores, Percoll gradient concentration and the embryogenic frequency of each cultivar were studied. Optimization of these factors is required for each genotype evaluated.
Starting Page	359
Ending Page	362
Page Count	4
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	106
Issue Number	2
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2011-01-07
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Brassica napus Microspore Embryogenesis Percoll Plant Pathology Plant Physiology Plant Sciences Plant Genetics & Genomics
Content Type	Text
Resource Type	Article
Subject	Horticulture

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