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Upregulation of the AT-hook DNA binding gene BoMF2 in OguCMS anthers of Brassica oleracea suggests that it encodes a transcriptional regulatory factor for anther development

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Abstract

Ogura cytoplasmic male sterility (OguCMS) is the most important CMS system used for F1 hybrid cabbage production worldwide. The anther abortion and defective pollen development exhibited in OguCMS are coordinately regulated by the mitochondrial male sterile gene orf138 and many nuclear transcriptional regulatory factors. AT-hook DNA binding proteins regulate cell-specific gene expression. In this study, we cloned the gene encoding the AT-hook DNA binding protein BoMF2 using the cDNA-AFLP TDF sequence, which was upregulated in OguCMS cabbage flower buds, as a querying probe. BoMF2 contains a 783-nt continuous complete open reading frame encoding a 260 amino-acid polypeptide. In vivo transient expression assays using GFP fusions showed that BoMF2 protein was located in the nucleus. BoMF2 was preferentially expressed in cabbage stamens, with a short expression window at anther development stage 7–8. However, in OguCMS flowers, BoMF2 expression continued into the mature pollen stage and was concomitant with the continued proliferation of tapetum cells exhibited in this mutant. Arabidopsis plants overexpressing BoMF2 showed significantly shorter siliques than the wild type, as well as decrease of pollen viability. These results suggest that BoMF2, a transcriptional regulatory factor, might regulate tapetum proliferation during anther development.

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Abbreviations

BoMF2:

Brassica oleracea male fertility 2

OguCMS:

Ogura cytoplasmic male sterility

qRT-PCR:

Quantitative real time RT-PCR

AHL:

AT-hook nuclear localized protein

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Acknowledgments

This work was supported by Grants from the Natural Science Foundation of China (31372065), the National High Technology R&D Program of China (2012AA100105, 2012AA020103) and the Key Projects in the National Science & Technology Pillar Program during the Twelfth Five-Year Plan Period (2012BAD02B01, 2012BAD50G01).

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Authors and Affiliations

  1. Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing, 100097, China

    Jungen Kang, Yingying Guo, Yujuan Chen, Hailong Li, Lei Zhang & Haixia Liu

  2. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China), Ministry of Agriculture, Beijing, 100097, China

    Jungen Kang, Yingying Guo, Yujuan Chen, Hailong Li, Lei Zhang & Haixia Liu

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  1. Jungen Kang
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Correspondence to Jungen Kang.

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Kang, J., Guo, Y., Chen, Y. et al. Upregulation of the AT-hook DNA binding gene BoMF2 in OguCMS anthers of Brassica oleracea suggests that it encodes a transcriptional regulatory factor for anther development. Mol Biol Rep 41, 2005–2014 (2014). https://doi.org/10.1007/s11033-014-3048-2

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