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A functional genomics resource for Brassica napus : development of an EMS mutagenized population and discovery of FAE1 point mutations by TILLING
Author(s) -
Wang Nian,
Wang Yajie,
Tian Fang,
King Graham J.,
Zhang Chunyu,
Long Yan,
Shi Lei,
Meng Jinling
Publication year - 2008
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/j.1469-8137.2008.02619.x
Subject(s) - tilling , biology , rapeseed , genetics , brassica , genome , erucic acid , mutant , population , functional genomics , computational biology , arabidopsis , gene , genomics , mutation , botany , demography , sociology
Summary• 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.