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Cloning and characterization of boron transporters in Brassica napus

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Abstract

Six full-length cDNA encoding boron transporters (BOR) were isolated from Brassica napus (AACC) by rapid amplification of cDNA ends (RACE). The phylogenic analysis revealed that the six BORs were the orthologues of AtBOR1, which formed companying with the triplication and allotetra-ploidization process of B. napus, and were divided into three groups in B. napus. Each group was comprised of two members, one of which was originated from Brassica rapa (AA) and the other from Brassica oleracea (CC). Based on the phylogenetic relationships, the six genes were named as BnBOR1;1a, BnBOR1;1c, BnBOR1;2a, BnBOR1;2c, BnBOR1;3a and BnBOR1;3c, respectively. The deduced BnBOR1 s had extensive similarity with other plant BORs, with the identity of 74–96.8% in amino acid sequence. The BnBOR1;3a and BnBOR1;3c resembled AtBOR1 in number and positions of the 11 introns, but the others only have 9 introns. After the gene duplication, there was evidence of purifying selection under a divergent selective pressure. The expression patterns of the six BnBOR1 s were detected by semi-quantitative RT–PCR. The BnBOR1;3a and BnBOR1;3c showed a ubiquitous expression in all of the investigated tissues, whereas the other four genes showed similar tissue-specific expression profile. Unlike the non-transcriptional regulation of AtBOR1, the expression of BnBOR1;1c and BnBOR1;2a were obviously induced by boron deficiency. This study suggested that the BOR1 s had undergone a divergent expression pattern in the genome of B. napus after that the B. napus diverged from Arabidopsis thaliana.

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Abbreviations

B:

Boron

MIPs:

Major intrinsic proteins

RACE:

Rapid amplification of cDNA ends

MYA:

Million years ago

CDS:

Coding sequence

PCR:

Polymerase chain reactions

RT–PCR:

Reverse transcription polymerase chain reactions

LRT:

Likelihood ratio test

TR:

Two ration model

HWSB:

Hot water-soluble boron

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (30771283, 30971861) and the National 863 High Technology Program of China (2007AA10Z117).

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China

    Jinhua Sun, Lei Shi, Chunyu Zhang & Fangsen Xu

  2. Microelement Research Centre, Huazhong Agricultural University, Wuhan, 430070, China

    Jinhua Sun, Lei Shi & Fangsen Xu

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  1. Jinhua Sun
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  2. Lei Shi
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Correspondence to Fangsen Xu.

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11033_2011_930_MOESM1_ESM.png

Supplementary figure Fig. S1 Multiple alignment of BnBOR1 s with AtBOR1 (NP_850469) Dots indicate identical residues with the BnBOR1;1a; grey box and star indicate possible positive selective sites detected by M8 model in PAml analysis; line indicates the conserved HCO3_cotransporer domain (PNG 160 kb)

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Sun, J., Shi, L., Zhang, C. et al. Cloning and characterization of boron transporters in Brassica napus . Mol Biol Rep 39, 1963–1973 (2012). https://doi.org/10.1007/s11033-011-0930-z

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