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Isolation and characterization of three duplicated PISTILLATA genes in Brassica napus

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Abstract

Three coding region cDNAs of duplicated PISTILLATA-like (PI-like) MADS-box genes, BnPI-1, BnPI-2 and BnPI-3, were isolated from B. napus by RT-PCR. The sequence analysis showed that the three PI cDNAs possessed 627, 627 and 625 nucleotides, respectively, and their nucleotide sequences had 96.49–98.72% similarity. Due to a deletion of two nucleotides, the protein sequence in the downstream of the frameshift site was altered in BnPI-3. Therefore, there were only 171 amino acids coded by BnPI-3, while there were 208 ones coded by BnPI-1 or BnPI-2. The deduced amino acid identity between BnPI-1 and BnPI-2 was 97.6% and the amino acid sequence of BnPI-1 and BnPI-2 shared 72.6% identity with BnPI-3. The deduced amino acid sequences of the coded proteins indicated high homology with the members of the PI family of MADS-box proteins. RT-PCR analysis showed that BnPI transcription was only detectable in petals and stamens. The yeast two-hybrid assays results showed that the three BnPI proteins exhibited different dimerization affinities with three BnAP3. BnPI-1 and BnPI-2 could form strong heterodimers with BnAP3. The dimerization affinity of BnPI-1 with BnAP3-4 is the strongest in all the combinations, while the affinity of BnPI-3 with BnAP3-4 is the weakest. The dimerization affinity to BnAP3-4 of BnPI-1 is 3.5 times of that of BnPI-3. The distinguished weak interaction to AP3 of BnPI-3 is probably due to the loss of the PI motif. The divergences of sequence and affinity of protein interaction might reflect some functional divergence of the three PI genes in B. napus.

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Acknowledgement

This work was supported by the National Natural Science Foundation of China (Grant Nos. 30571174 and 30871540).

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Authors and Affiliations

  1. Key Laboratory of Ministry of Education for Bio-resources and Eco-environment, School of Life Sciences, Sichuan University, Chengdu, 610064, People’s Republic of China

    Wei Deng, Lin Zhou, Yuntao Zhou, Yujia Wang, Maolin Wang & Yun Zhao

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  1. Wei Deng
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  2. Lin Zhou
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  3. Yuntao Zhou
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  4. Yujia Wang
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  5. Maolin Wang
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  6. Yun Zhao
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Correspondence to Yun Zhao.

Additional information

The sequences reported in this paper have been deposited in GenBank with the accession numbers, EU159431, EU159432 and EU159433, respectively.

Wei Deng and Lin Zhou are committed as the first author.

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Deng, W., Zhou, L., Zhou, Y. et al. Isolation and characterization of three duplicated PISTILLATA genes in Brassica napus . Mol Biol Rep 38, 3113–3120 (2011). https://doi.org/10.1007/s11033-010-9981-9

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  • DOI: https://doi.org/10.1007/s11033-010-9981-9

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