Sequence analysis of the gene correlated with cytoplasmic male sterility (CMS) in rape-seed (Brassica napus) Polima and Shaan 2A

Abstract

orf224 is a CMS-related mitochondrial gene discovered in Polima cytoplasm. Shaan 2A CMS line is the parent of the first rapeseed hybrid cultivar Qinyou No. 2 that has been grown in many regions of China. In this work, genomic DNA of Polima CMS line and Shaan 2A CMS line were used as templates, two primers of specific oligonucleotides at 5′ and 3′ ends were used, PCR was performed, the amplification fragments were cloned into pGEM-T Easy vectors and DNA sequences were determined. The CMS-associated gene, orf224-1 present in Shaan 2A CMS line, has a sequence highly homologous to the orf224 of the Polima CMS line, except for one nucleotide at position +398. There were only one base (AAC→AGC) and one amino acid (Asn →Ser) differences between the two. The homologies of the two sequences in nucleotide and amino acid were 99.9% and 99.6%, respectively. It is concluded that orf224 in Polima CMS line and orf224-1 of Shaan 2A CMS line are the allele at the same locus in mitochondria.