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Molecular cloning and expression analysis of a RGA-like gene responsive to plant hormones in Brassica napus

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Abstract

DELLA proteins are negative regulators of GA-induced growth. DELLA protein family is characterized by a DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. A full-length cDNA encoding a putative DELLA protein with high sequence homology to Arabidopsis thaliana RGA (AtRGA), designated as BnRGA, was isolated from Brassica napus. The full-length cDNA of BnRGA contained a 1,740 bp open reading frame (ORF) encoding a precursor protein of 579 amino acid residues. Comparative and bioinformatics analyses revealed that BnRGA showed a high degree of homology with DELLA proteins and contained the DELLA domain, TVHYNP domain, VHIID domain and RVER domain. Using real-time PCR, the expression patterns of BnRGA and two our previously isolated genes, BnGID1a and BnSLY1 in B. napus, were analyzed by adding exogenous gibberellins acid-3 (GA3), GA biosynthetic inhibitor paclobutrazol (PAC) and abscisic acid (ABA). The results showed that the expression of BnGID1a and BnSLY1 was down-regulated after treated by GA3 and induced by PAC and ABA. These results suggest that the expression of BnGID1a and BnSLY1 may be negatively regulated by the level of endogenous GA in B. napus. Moreover, BnRGA was not significantly regulated by GA3, PAC and ABA in the low concentrations. These suggest that GA-GID1-SCF-DELLA complex may have a mechanism of self-regulation, thereby preserving the stability of the expression level of BnRGA in B. napus.

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Fig. 1

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Abbreviations

GA:

Gibberellins

ABA:

Abscisic acid

PAC:

Paclobutrazol

GID1:

Gibberellin insensitive dwarf 1

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Acknowledgments

This work was supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the National ‘863’ Program (SN: 2001AA241104) and the 10th ‘five-year’ key task project in crop breeding of Sichuan Province (SN: 200107001).

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Authors and Affiliations

  1. College of Bioscience and Biotechnology, Key Laboratories of Crop Genetics and Physiology of the Jiangsu Province and Plant Functional Genomics of the Ministry of Education, Yangzhou University, Yangzhou, 225009, Jiangsu, People’s Republic of China

    Yong Gao & Jianmin Chen

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

    Yun Zhao, Tingting Li & Maolin Wang

Authors
  1. Yong Gao
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  2. Jianmin Chen
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  3. Yun Zhao
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  4. Tingting Li
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  5. Maolin Wang
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Corresponding author

Correspondence to Maolin Wang.

Electronic supplementary material

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11033_2011_943_MOESM1_ESM.jpg

Fig. S1 Comparison of the putative amino acid sequences of BnRGA with some representative DELLA proteins. The identical amino acids were showed in white with black background and the conserved amino acids were showed in white with gray background. The specific sites for DELLA domain, TVHYNP domain, VHIID domain and RVER donain were presented. The aligned DELLA proteins sequences were from A. thaliana (RGA1, GenBank accession no. NP_178266; GAI, GenBank accession no. NP_172945; RGL1, GenBank accession no. NP_176809; RGL2, GenBank accession no. NP_186995; RGL3, GenBank accession no. NP_197251). Supplementary material 1 (JPEG 1849 kb)

11033_2011_943_MOESM2_ESM.jpg

Fig. S2 Phylogenetic tree analysis of BnRGA from plants by MEGA version 3.1. The neighbor-joining method was used to construct the tree. Numbers on nodes indicated the bootstrap values after 1,000 replicates. The genes used in phylogenetic tree analysis were from plants including A. thaliana RGA1 GenBank accession no. NM_126218; GAI GenBank accession no. NM_101361; RGL1 GenBank accession no. NM_105306; RGL2 GenBank accession no. NM_111216; RGL3 GenBank accession no. NM_121755. (JPEG 197 kb)

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Gao, Y., Chen, J., Zhao, Y. et al. Molecular cloning and expression analysis of a RGA-like gene responsive to plant hormones in Brassica napus . Mol Biol Rep 39, 1957–1962 (2012). https://doi.org/10.1007/s11033-011-0943-7

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  • DOI: https://doi.org/10.1007/s11033-011-0943-7

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