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Cytokinin response regulators (RR), as important components of the cytokinin signaling pathway, perform critical functions in cytokinin signaling output and regulation of downstream gene signaling responses. In order to investigate the role of wheat RR genes in response to low-temperature stress, we cloned the A-type cytokinin responsive regulator gene TaRRA6-D from the wheat cultivar Dongnongdongmai 1 (Dn1), performed bioinformatics analysis, subcellular localization analysis, and promoter activity analysis under low-temperature. We also genetically transformed Arabidopsis wild type to create overexpression lines for phenotypic observation of root length and cold resistance evaluation. Bioinformatics analysis showed that the TaRRA6-D gene (TraesCS5D02G140200) was 1 088 bp in total length, with a coding sequence of 696 bp, which was localized to wheat chromosome 5D, and the encoded protein was an unstable hydrophobic protein. The phylogenetic tree analysis showed that the wheat TaRRA6-D protein was most closely related to the RR proteins of Aegilops tauschii and Hordeum vulgare. Transient expression analysis of transformed tobacco showed that TaRRA6-D protein was localized in the nucleus. Analysis of cis-acting elements in the promoter region of the gene showed that the TaRRA6-D promoter region contained cis-acting elements responsive to low-temperature, drought, light, and a variety of hormones. Expression pattern analysis of the transformed Arabidopsis thaliana lines showed that TaRRA6-D promoter activity was induced to express by low-temperature. Phenotypic observations showed that root length of the overexpression lines increased significantly compared with the wild type, with or without the addition of 6-BA. Seedling freezing resistance assay showed that seedlings of overexpression lines were significantly resistant to low-temperature compared with wild type, and 6-BA treatment significantly improved the freezing resistance of seedlings.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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