Non-destructive in vitro selection of microspore-derived embryos with the fertility restorer gene for CMS Ogu-INRA in winter oilseed rape (Brassica napus L.)

Abstract

We have bred low glucosinolate (GSL) winter oilseed rape lines carrying the fertility restorer for the CMS Ogu-INRA system. The original restorer line BO20 contained 31μmol/g GSL in seeds, but by crossing this line with various low GSL CMS lines, followed by repeated selection of fertile segregants, we were able to obtain fertile lines with a mean GSL content in seeds of 11.8 μmol/g. This result confirmed that the gene(s) controlling the GSL content are not closely linked to the fertility restorer gene. The results confirm, that the SCAR marker SG34 is closely associated with the fertility restoring allele, and facilitates so the selection of fertile segregants; however, the marker is unable to distinguish between the homozygous RfRf and the heterozygous Rfrf genotypes.

Early in vitro selection of winter oilseed rape (Brassica napus L.) plants with the fertility restorer gene for CMS Shaan 2A via non-destructive molecular analysis of microspore-derived embryos

“…Rf gene was amplified in 104 (51.7%) accessions, which corresponds to the assumed 1:1 segregation ratio for a single copy gene. It differs from our previous results for Ogu-INRA restorers (Havlíčková et al 2015) where the occurrence of genotypes with Rf gene was substantially lower (37%). In total, 172 (85.6%) plants were regenerated from the above-mentioned embryos to the generative stage.…”

“…A microspore culture technique has been developed for doubled haploid (DH) production in most important Brassica species. Non-destructive markerassisted selection may provide early identification of all genotypes carrying the desired gene in the selected material, in just three weeks of microspore culture under optimal conditions (Havlíčková et al 2015). For hybrid breeding of oilseed rape (B. napus L.), several cytoplasmic male sterility (CMS) systems have been widely used to develop commercial cultivars.…”

“…Surprisingly, DNA isolation even from smaller embryos (2.0-3.0 mm) provided quite a sufficient amount of material for DNA extraction. However, it may be difficult to obtain such a small sample always in a non-destructive way, without the risk of damage to the embryo tip or loss of the sample during routine manipulation (Havlíčková et al 2014(Havlíčková et al , 2015. The length of the amplified fragment was approx.…”

Cytological and Molecular Characterization of a New Ogura Cytoplasmic Male Sterility Restorer of Brassica napus L.

“…On the manipulation of genetic improvement, heterosis utilization is effective in enhancing rapeseed yield [ 1 , 2 ]. Among heterosis utilization systems, the cytoplasmic male sterility (CMS) system is the major type, which mainly includes Polima CMS and Ogura CMS [ 3 , 4 ]. Currently, more than 80% of rapeseed varieties belong to hybrids including Polima CMS, recessive genic male sterility, and chemical hybridization in China.…”