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Analysis of MS2Bnap genomic DNA homologous to MS2 gene from Arabidopsis thaliana in two dominant digenic male sterile accessions of oilseed rape (Brassica napus L.)

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Abstract

PCR technique was employed to isolate gene homologous to the MS2Bnap (X99922.1) from two rapeseed (Brassica napus L.) dominant digenic male sterile lines, namely 220A (male sterile) and 220B (male fertile), 6A (male sterile) and 6C (male fertile). The isolated 2,581 bp sequences from 220A (named 220A-gDNA, GenBank accession number AY288778), 220B (220B-gDNA, AY257490), 6A (6A-gDNA, DQ060318) and 6C (6C-gDNA, DQ060319) all contained six introns. Forty-one single nucleotide polymorphism (SNP) sites were detected by alignment of these four sequences, seven of them dispersed in the exon regions. Two SNPs (1247, 1656) were detected between 220A-gDNA and 220B-gDNA, and the one at nucleotide 1247 of 220A-gDNA with A replaced by C was a missense mutation, which may be the putative male sterility site in 220A. All eight SNPs identified between 6A-gDNA and 6C-gDNA were located in the third intron, so the proteins encoded by them are the same. The one SNP between 6A-/6C-gDNA and 220A-/220B-gDNA at nucleotide 2474 of 220A-/220B-gDNA with C replaced by G was a missense mutation. Mutation site of BNMS2PROT (CAA68190.1) encoded by MS2Bnap in 220A(254) and 6A/6C(584) is different, which indicated dominant digenic male sterile line 220AB and 6CA have some difference in the molecular level. Comparison of structure of MS2Bnap in B. napus with that of MS2 in Arabidopsis thaliana revealed that the similarity of exons between these two genes is higher than that of introns.

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Acknowledgements

This work was partly supported by China Postdoctoral Science Foundation (2004036075), Chinese Chunhui Project (Z2004-1-61007), Scientific Research Project of Key laboratory of Oil Crop Genetics and Improvement of Chinese Ministry of Agriculture (Wuhan, China), The Project for the Excellent Young Scientists of Northwest A&F University (04ZR006) and The Program for Excellent Young Talents in Northwest A&F University. Rapeseed DDGMS line 6CA was kindly provided by Mr Shulin Li at Shanghai Academy of Agricultural Sciences, Shanghai, China.

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Authors and Affiliations

  1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, Hubei, 430062, People’s Republic of China

    S. W. Hu, Y. F. Fan, X. L. Guo, C. H. Dong, S. Y. Liu & H. Z. Wang

  2. College of Agronomy, Northwest A&F University, Yangling, Shaanxi, 712100, People’s Republic of China

    S. W. Hu, Y. F. Fan & C. Y. Yu

  3. College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, 712100, People’s Republic of China

    H. X. Zhao

  4. Department of Biology, Saint Mary’s University, B3H 3C3, Halifax, Nova Scotia, Canada

    G. L. Sun

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  1. S. W. Hu
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Correspondence to H. Z. Wang.

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Communicated by H. C. Becker

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Hu, S.W., Fan, Y.F., Zhao, H.X. et al. Analysis of MS2Bnap genomic DNA homologous to MS2 gene from Arabidopsis thaliana in two dominant digenic male sterile accessions of oilseed rape (Brassica napus L.). Theor Appl Genet 113, 397–406 (2006). https://doi.org/10.1007/s00122-006-0303-5

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