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Title of Journal: Mol Biol Rep

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Abbravation: Molecular Biology Reports

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Springer Netherlands

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DOI

10.1002/jez.1400650204

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ISSN

1573-4978

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Development of molecular markers specific to petal

Authors: Xiaolin Yu Yapei Liu Yanxia Lv Zhenning Liu Zhujun Chen Gang Lu Jiashu Cao
Publish Date: 2014/01/03
Volume: 41, Issue: 2, Pages: 769-778
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Abstract

To establish a simple and rapid method for isolating mitochondrial DNA mtDNA from Brassica vegetables the effects of different factors on mtDNA extraction were investigated firstly A new protocol includes five steps organelle isolation deoxyribonuclease treatment lysis RNase treatment and deproteinization Results indicate that a 15 minlysis time can achieve higher mtDNA yields from etiolated seedlings Moreover it is found that the inflorescence of the cytoplasmic male sterile CMS line is unfit for the isolation of mtDNA The mtDNA isolated using this method is intact and pure and can be used for further molecular analysis Subsequently the genomic and transcriptional differences of atps and coxs genes on the mitochondria between the petaloidtype CMS line and its maintainer line have been identified RFLP analysis revealed that out of the five atps and three coxs genes except of atp4 and cox3 the others mtDNA protein coding genes exhibited polymorphisms respectively This results suggest that atps and coxs genes are located in a long mtDNA fragment and the mtDNA evolves rapidly in structure between the CMS line and its maintainer line in tuber muster Northern blot analysis showed that the expression level of these genes in flower bud is higher than that of leaf and flower and that alternative splicing have been found among the atp6 atp8 and cox3 genes respectively Our results modified a efficient protocol for isolating the mtDNA and provided some novel molecular markers indicating the CMS trait in tuber mustard The comparative analysis presented in this study allows a more comprehensive understanding of the molecular mechanism on CMS in Brassica cropsThis research was partially supported by the Breeding Project of SciTech Foundation of Zhejiang Province 2012C1290361 and the Nature Science Foundation of Zhejiang Province Y3090294 The authors gratefully acknowledge Dr Gergely Gulyas and Dr Gen Hattori for their helpful advice and we particularly thank Dr Johnson Cameron for the stimulating discussions and for critically reading the manuscript

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