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Outline
N6-methyladenosine (m6A), the most abundant epitranscriptomic modification in eukaryotic RNA, regulates plant growth, development, and stress responses through a dynamic “writer-eraser-reader” regulatory mechanism. Among these regulators, ECT family proteins act as m6A reader by recognizing m6A-modified sites to participate in RNA metabolism regulation. To investigate whether TaECT3 acts as an m6A reader protein in stress responses, this study cloned the coding sequence (CDS) of TaECT3 gene from a wheat cultivar Fielder and conducted comprehensive bioinformatics analyses. Results revealed that the CDS of TaECT3 spans 1 854 bp, encoding a 617-amino acid protein containing the characteristic YTH domain conserved in ECT family members. The TaECT3 protein exhibits a molecular weight of 66.93 kDa and a predicted isoelectric point of 6.48, demonstrating intrinsic hydrophilicity and thermodynamic stability. Secondary structure prediction revealed a predominance of random coils (79.74%) and α-helices (13.45%), while tertiary structure modeling confirmed the presence of a typical m6A-binding pocket. Phylogenetic analysis revealed that TaECT3 exhibits close evolutionary relationship to the ECT proteins of HvECT3, SbECT3, and BdDF3A from barley, sorghum, and brachypodium, respectively. Electrophoretic mobility shift assay (EMSA) confirmed that recombinant TaECT3 specifically binds m6A-modified RNA. Subcellular localization analysis suggested its potential membrane localization. qRT-PCR revealed significant regulation of TaECT3 expression in wheat cultivar Fielder under high-temperature (37 ℃, 6 h) and PEG8000-simulated drought (15%, 6 h) stress conditions, compared to the control. This study demonstrates TaECT3 functions as an m6A reader and participates in response to heat and drought stresses in 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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