Key message
Here we report the enhancement of tolerance to salt stress in Brassica rapa (Chinese cabbage) through the RNAi-mediated reduction of GIGANTEA ( GI ) expression.
Abstract
Circadian clocks integrate environmental signals with internal cues to coordinate diverse physiological outputs. The GIGANTEA (GI) gene was first discovered due to its important contribution to photoperiodic flowering and has since been shown to be a critical component of the plant circadian clock and to contribute to multiple environmental stress responses. We show that the GI gene in Brassica rapa (BrGI) is similar to Arabidopsis GI in terms of both expression pattern and function. BrGI functionally rescued the late-flowering phenotype of the Arabidopsis gi-201 loss-of-function mutant. RNAi-mediated suppression of GI expression in Arabidopsis Col-0 and in the Chinese cabbage, B. rapa DH03, increased tolerance to salt stress. Our results demonstrate that the molecular functions of GI described in Arabidopsis are conserved in B. rapa and suggest that manipulation of gene expression through RNAi and transgenic overexpression could enhance tolerance to abiotic stresses and thus improve agricultural crop production.
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Acknowledgments
This work was supported by grants from the Research Program for Agricultural Science & Technology Development, National Academy of Agricultural Science, (Project No. PJ10025) to JAK and from Rural Development Administration, Republic of Korea, BioGreen 21 Program (Project No. PJ01106901, PJ01106902, and PJ009615) to YWK, JAK, and CRM.
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Supplementary Figure 1. BrGI structure and phylogenetic comparison with other plant GI proteins. (A) Alignment of the GI protein sequences of Brassica rapa (BrGI) and Arabidopsis thaliana (GI) using ClustalW. B. Exon–intron structure of the BrGI and GI genes. Black boxes indicate exons; lines represent introns. C. Phylogenetic comparison of the GI proteins from several dicot and monocot species. The GI protein sequences used were A. thaliana (GI; NM_102124.2), Populus trichocarpa (PtGI; XM_002307480.1), Triticum aestivum (TaGI1; AF543844.1), Hordeum vulgare (HVGI; AY740524.1), Ricinus communis (Rc; XM_002524295.1), Ipomoea nil (InGI; AB265781.1), Allium cepa (AcGIa; GQ232756.1 and AcGIb; GQ232757.1), and Oryza sativa (OsGI; NM_001048755.1). Trees were constructed using the neighbor-joining method in the MEGA software, version 4.0 (Kumar et al. 2004). Bar represents 0.05 residue substitutions per site
Supplementary Figure 2. Germination and growth of the indicated B. rapa genotypes under the indicated salt conditions. Seeds were sown on ½ MS salts, 2% sucrose, and 1% agar supplied with the indicated NaCl concentrations and grown for 2 weeks
Supplementary Figure 3. GIGANTEA protein abundance in 2-week-old transgenic B. rapa measured by Western blot analysis using an antibody directed against Arabidopsis GI.Arabidopsis wild-type (Col-0) harvested at ZT1 and ZT13 and 35S::GI-HA (constitutive GI overexpression line) plants were used as GI protein controls. B. rapa transgenic plants were harvested at ZT12.
Supplementary Figure 4. Seeds of the indicated genotypes were sown in sponge cubes and grown in nutrient solution for 2 weeks. After two to three true leaves had developed, the nutrient solution was replaced with that including either 0 or 150 mM NaCl every 2 days for 1 week.
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Kim, J.A., Jung, He., Hong, J.K. et al. Reduction of GIGANTEA expression in transgenic Brassica rapa enhances salt tolerance. Plant Cell Rep 35, 1943–1954 (2016). https://doi.org/10.1007/s00299-016-2008-9
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DOI: https://doi.org/10.1007/s00299-016-2008-9