Abstract
Cytoplasmic male sterility (CMS) is an important factor to observe heterosis in Brassica rapa. Although several studies have documented the rearrangements of mitochondrial DNA and dysfunction in the mitochondria have been observed in most types of CMS, the basis of the molecular mechanisms involved in these processes and other effects on CMS remain unclear. In this study, suppression subtractive hybridization was performed in the flowers of an alloplasmic Polima CMS system from B. rapa ssp. chinensis to identify genes that are differentially expressed between fertile and sterile plants. A total of 443 clones were isolated (156 were upregulated in fertile buds, and 287 were upregulated in sterile ones). Real-time RT-PCR further demonstrated the credibility of SSH. Among these genes, many membrane protein genes (LTP12, PIP2A, and GRP14) were inhibited in the sterile male line. Mitochondrial membrane potential (MMP) assay was then performed. Results showed that the sterile MMP was unstable and failed to create a potential difference; thus, mitochondrial dysfunction occurred. Moreover, abnormal microtubules and photosynthetic pathways were found in sterile male cells. Unstable MMP, nutritional deficiency, and abnormal microtubules were the causes of Polima CMS in Brassica campestris. H2O2, MDA, and O2–, accumulated as byproducts of energy metabolism disorder in sterile male cells.
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This work was supported by the National Natural Science Foundation of China (31272173, 31301782), the National High Technology Research and Development Program of China (863 Program, 2012AA100101, 2012AA100202), the Natural Science Foundation of Jiangsu Province (BK2011643, BK20130673), and the Science and Technology Innovation Fund of Nanjing Agricultural University (KYZ201111, KJ2013015).
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Ying Li and Tongkun Liu contributed equally to this work.
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Supplementary Fig. S1
Analysis of subtraction efficiency by PCR. (DOC 163 kb)
Supplementary Table S1
Putative identities of cDNA clones from forward SSH library. (XLS 68 kb)
Supplementary Table S2
Putative identities of cDNA clones from reverse SSH library. (XLS 44 kb)
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Li, Y., Liu, T., Duan, W. et al. Instability in mitochondrial membranes in Polima cytoplasmic male sterility of Brassica rapa ssp. chinensis . Funct Integr Genomics 14, 441–451 (2014). https://doi.org/10.1007/s10142-014-0368-1
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DOI: https://doi.org/10.1007/s10142-014-0368-1