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A GA-insensitive dwarf mutant of Brassica napus L. correlated with mutation in pyrimidine box in the promoter of GID1

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Abstract

A dwarf mutant from Brassica napus, namely NDF-1, which was derived from a high doubled haploid (DH) line ‘3529’(Brassica napus L.) of which seeds were jointly treated with chemical inducers and fast neutron bombardment, was revealed that dwarfism is under the control of a major gene(designated as ndf1) with a mainly additive effect and non-significant dominance effect. The germination and hypocotyls elongation response of dwarf mutants after exogenous GA and uniconazol application showed NDF-1 was a gibberellin insensitive dwarf. We cloned the Brassica napus GID1 gene, named BnGID1, and found it was the ortholog of AtGID1a. The sequence blasting of the BnGID1 genes from NDF-1 and wild type showed there was no mutant in the gene. But the quantitative RT-PCR analysis of GID1 EST pointed out the mutation was caused by the low-level expression of BnGID1 gene. After sequenced the BnGID1 gene’s upstream, we found three bases mutated in the pyrimidine box (P-box) of the BnGID1 promoter, which is linkage with the dwarf mutant.

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Abbreviations

GA:

Gibberellins

ORF:

Open reading frame

GID1 :

Gibberellin insensitive dwarf 1

EST:

Expressed sequence tags

BrUBQ:

Brassica rape ubiquitin

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Acknowledgements

This work was supported by the National ‘863’ Programme (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. Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610064, Sichuan, People’s Republic of China

    Huapeng Li, Yun Wang, Xiaocheng Li, Yong Gao, Zhijun Wang, Yun Zhao & Maolin Wang

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  1. Huapeng Li
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  2. Yun Wang
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  3. Xiaocheng Li
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  4. Yong Gao
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  5. Zhijun Wang
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  7. Maolin Wang
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Correspondence to Maolin Wang.

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Supplemental Fig. S1

The forward primer according to the upstream sequences of AtGID1a (about −3000, −2000, −1000, −500) and the reverse primer 5 test according to the BnGID1 sequence(129 bp after the initiation codon) (JPG 8 kb)

Supplemental Fig. S2

Sequential alignment of BnGID1 with AtGID1s and OsGID1. The sequence of BnGID1 was compared with that of AtGID1a (AGI code At3g05120), AtGID1b (At3g63010), AtGID1c (At5g27320) and OsGID1 (DDBJ/EMBL/GenBank accession no. AB211399). The two framed amino acid residues were Gly-196 and Arg-251, which were essential for maintaining the GA-binding activity of GID1s. (JPG 152 kb)

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Li, H., Wang, Y., Li, X. et al. A GA-insensitive dwarf mutant of Brassica napus L. correlated with mutation in pyrimidine box in the promoter of GID1 . Mol Biol Rep 38, 191–197 (2011). https://doi.org/10.1007/s11033-010-0094-2

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  • DOI: https://doi.org/10.1007/s11033-010-0094-2

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