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A missense mutation in the VHYNP motif of a DELLA protein causes a semi-dwarf mutant phenotype in Brassica napus

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Abstract

Although dwarf genes have been widely used to improve lodging resistance and enhance harvest index in cereal crops, lodging is still a serious problem in rapeseed (Brassica napus) production. A semi-dwarf B. napus mutant, ds-1, was identified through EMS mutagenesis of a microspore-cultured DH line. The mutant had a significant reduction in height due to a lower first branch position and shorter internodes when compared with wild-type cultivars. This dwarfism was inherited as a single semi-dominant gene, ds-1. DS-1 locus was mapped to chromosome A6, and co-segregated with a microsatellite marker BnEMS1125 derived from the gene BnRGA. BnRGA encodes a DELLA protein that functions as a GA signaling repressor. The expression of a mutant BnRGA allele from ds-1, Bnrga-ds, caused dwarf phenotypes in Arabidopsis. Comparative sequencing of RGA open-reading frames (ORFs) of ds-1 and wild-type cultivars revealed a single proline (P)-to-leucine (L) substitution that may lead to a gain-of-function mutation in GA signaling. The expression of the Arabidopsis homolog, Atrga-ds, bearing this site-directed mutation also rendered dwarf phenotypes in Arabidopsis, which demonstrated that the P-to-L mutation in the VHYNP motif of Bnrga-ds is responsible for the dwarfism. A yeast two-hybrid assay confirmed that this mutation inhibited the interaction between Bnrga-ds/Atrga-ds and the GA receptor, AtGID1A, in the presence of GA3, suggesting that the conserved proline residue in the VHYNP motif of DELLA protein directly participates in DELLA-GID1 interaction. Identification and characterization of the dwarf gene ds-1 will facilitate its utilization in improving lodging resistance in Brassica breeding.

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Acknowledgments

This research was supported by the National High-Technology Research and Development Program (the “863” Program) of China (2006AA10A112) and the National Science Foundation of China (30623012). We thank M. Mazourek for critical review of this manuscript.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, 430070, China

    Chao Liu, Jilin Wang, Tiandai Huang, Fang Wang, Fang Yuan, Xiaomao Cheng, Yan Zhang, Shuwen Shi, Jiangsheng Wu & Kede Liu

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  1. Chao Liu
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  2. Jilin Wang
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  7. Yan Zhang
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Correspondence to Jiangsheng Wu or Kede Liu.

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Communicated by H. Becker.

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Liu, C., Wang, J., Huang, T. et al. A missense mutation in the VHYNP motif of a DELLA protein causes a semi-dwarf mutant phenotype in Brassica napus . Theor Appl Genet 121, 249–258 (2010). https://doi.org/10.1007/s00122-010-1306-9

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  • DOI: https://doi.org/10.1007/s00122-010-1306-9

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