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Characterization of BcMF23a and BcMF23b, two putative pectin methylesterase genes related to pollen development in Brassica campestris ssp. chinensis

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Abstract

Two homologous genes, Brassica campestris Male Fertility 23a (BcMF23a) and Brassica campestris Male Fertility 23b (BcMF23b), encoding putative pectin methylesterases (PMEs) were isolated from Brassica campestris ssp. chinensis (syn. Brassica rapa ssp. chinensis). These two genes sharing high sequence identity with each other were highly expressed in the fertile flower buds but silenced in the sterile ones of genic male sterile line system (‘Bcajh97-01A/B’). Results of RT-PCR and in situ hybridization suggested that BcMF23a and BcMF23b were pollen-expressed genes, whose transcripts were first detected at the binucleate pollen and maintained throughout to the mature pollen grains. Western blot indicated that both of the putative BcMF23a and BcMF23b proteins are approximately 40 kDa, which exhibited extracellular localization revealed by transient expression analysis in the onion epidermal cells. The promoter of BcMF23a was active specifically in pollen during the late pollen developmental stages, while, in addition to the pollen, BcMF23b promoter drove an extra gene expression in the valve margins, abscission layer at the base of the first true leaves, taproot and lateral roots in seedlings.

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Abbreviations

BcMF23 :

Brassica campestris Male Fertility 23

PME:

Pectin methylesterase

ORF:

Open reading frame

HAP:

Hours after pollination

RT-PCR:

Reverse transcript polymerase chain reaction

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Acknowledgements

This research was supported by the National Program on Key Basic Research Projects (No. 2012CB113900), the Natural Science Foundation of China (No. 31272176), and the Key Sci-Technology Project of Zhejiang Province (No. 2010C12004).

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Authors and Affiliations

  1. Laboratory of Cell and Molecular Biology, Institute of Vegetable Science, Zhejiang University, Hangzhou, 310058, China

    Sue Lin, Li Huang, Xiaolin Yu, Xingpeng Xiong, Xiaoyan Yue, Tingting Liu, Ying Liang, Meiling Lv & Jiashu Cao

  2. Key Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Ministry of Agriculture, Hangzhou, 310058, China

    Sue Lin, Li Huang, Xiaolin Yu, Xingpeng Xiong, Xiaoyan Yue, Tingting Liu, Ying Liang, Meiling Lv & Jiashu Cao

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  1. Sue Lin
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Correspondence to Jiashu Cao.

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Lin, S., Huang, L., Yu, X. et al. Characterization of BcMF23a and BcMF23b, two putative pectin methylesterase genes related to pollen development in Brassica campestris ssp. chinensis . Mol Biol Rep 44, 139–148 (2017). https://doi.org/10.1007/s11033-016-4090-z

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