A functional genomics resource for Brassica napus : development of an EMS mutagenized population and discovery of FAE1 point mutations by TILLING

A - Papers appearing in refereed journals

Wang, N., Wang, Y., Tian, F., King, G. J., Zhang, C., Long, Y., Shi, L. and Meng, J. 2008. A functional genomics resource for Brassica napus : development of an EMS mutagenized population and discovery of FAE1 point mutations by TILLING. New Phytologist. 180 (4), pp. 751-765.

AuthorsWang, N., Wang, Y., Tian, F., King, G. J., Zhang, C., Long, Y., Shi, L. and Meng, J.
Abstract

Two ethylmethanesulfonate (EMS) mutant populations of the semi-winter rapeseed cv. Ningyou7 were constructed with high mutant load, to provide a TILLING platform for functional genomics in Brassica napus, and for introduction of novel allelic variation in rapeseed breeding. Forward genetic screening of mutants from the M2 populations resulted in identification of a large number of novel phenotypes. Reverse genetic screening focused on the potentially multi-paralogous gene FAE1 (fatty acid elongase1), which controls seed erucic acid synthesis in rapeseed. A B. napus BAC library was screened, and loci in a reference mapping population (TNDH) were mapped to conclude that there are two paralogous copies of FAE1, one on each of the B. napus A and C genomes. A new procedure is demonstrated to identify novel mutations in situations where two or more very similar paralogous gene copies exist in a genome. The procedure involves TILLING of single plants, using existing SNPs as a positive control, and is able to distinguish novel mutations based on primer pairs designed to amplify both FAE1 paralogues simultaneously. The procedure was applied to 1344 M2 plants, with 19 mutations identified, of which three were functionally compromised with reduced seed erucic acid content. New Phytologist (2008) 180: 751-765 (c) National Key Laboratory of Crop Genetic Improvement (2008). Journal compilation (c) New Phytologist (2008) doi: 10.1111/j.1469-8137.2008.02619.x.

KeywordsPlant Sciences
Year of Publication2008
JournalNew Phytologist
Journal citation180 (4), pp. 751-765
Digital Object Identifier (DOI)doi:10.1111/j.1469-8137.2008.02619.x
PubMed ID18811617
Open accessPublished as non-open access
FunderNational Basic Research Program of China
National Natural Science Foundation of China
Chinese Ministry of Education
Funder project or codeCentre for Crop Genetic Improvement (CGI)
Project: 4719
Project: 6029
ISSN0028646X
PublisherWiley
Grant ID2006CB101600
103175
30600373

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