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To investigate the effects of two alternative splicing variants of wheat receptor-like kinase gene TaNAK1 on agronomic and yield-related traits, this study used the winter wheat cultivar Xiaoyan 6, the wild-type spring wheat cultivar Fielder, and its transgenic lines overexpressing TaNAK1.1 (OE1-1, OE1-9, OE1-11） and TaNAK1.2 (OE2-1, OE2-2, OE2-4). The expression patterns of TaNAK1.1 and TaNAK1.2 in wheat were examined using semi-quantitative RT-PCR (sqRT-PCR) and quantitative real-time RT-PCR (qRT-PCR), respectively. Agronomic and yield-related traits of the transgenic lines under field conditions, along with seedling drought tolerance, were evaluated. The results showed that TaNAK1.1 and TaNAK1.2 exhibited distinct spatiotemporal expression profiles across wheat organs and tissues under normal conditions; TaNAK1.1 displayed relatively high expression levels in grains at 10 and 15 days post-anthesis, whereas TaNAK1.2 showed relatively high expression levels in seedling leaves and flag leaves. Under drought and low-temperature stress conditions, TaNAK1.2 was significantly up-regulated, while the expression of TaNAK1.1 was not induced by these abiotic stresses. The overexpression of TaNAK1.1 and TaNAK1.2 differentially affected agronomic and yield-related traits under field conditions. Compared to wild-type wheat Fielder, TaNAK1.1 transgenic lines showed no significant differences in heading and flowering time or flag leaf area. In contrast, heading and flowering time in TaNAK1.2 transgenic lines were delayed by approximately 5 d, and flag leaf area was reduced by 12.50% to 14.45%. Both TaNAK1.1 transgenic and TaNAK1.2 lines exhibited significant reductions in plant height, tiller number, spike length, number of spikelets per spike, grain size, thousand-grain weight, and grain weight per plant; however, the extent of these reductions varied between the two types of overexpressing lines, with TaNAK1.1 transgenic lines showing greater reductions in grain weight per plant and biomass per plant. The effects of TaNAK1.1 and TaNAK1.2 overexpression on wheat seedling drought tolerance were similar, with both significantly enhancing drought tolerance. Specifically, under drought stress, compared to the wild-type wheat Fielder, the leaves of TaNAK1.1 transgenic and TaNAK1.2 transgenic lines showed significantly increased contents of osmotic adjustment solutes (soluble sugars and proline) and relative water content, along with significantly decreased levels of H2O2 and the membrane lipid peroxidation product MDA. Furthermore, their survival rate was also significantly enhanced following extreme drought stress treatment. These results not only reveal the important roles of TaNAK1 alternative splicing in regulating growth, development, and adaptation to abiotic stress in wheat, but also provide new genetic resources for molecular breeding of stress-tolerant wheat.
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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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