Genome-wide identification and expression profiling of WRKY gene family in grain Amaranth (Amaranthus hypochondriacus L.) under salinity and drought stresses.

Background: The WRKY gene family plays a significant role in plant growth, development, and responses to biotic and abiotic stresses. However, the role of the WRKY gene family has not been reported in Amaranthus hypochondriacus. This study presents a comprehensive genome-wide analysis of the WRKY gene family in grain amaranth (A. hypochondriacus L.), a resilient crop known for its high nutritional value and adaptability to challenging environments.

Results: In this study, 55 WRKY genes (AhyWRKY1-55) were identified in A. hypochondriacus and distributed unevenly across 16 scaffolds. Of these, 50 contained conserved WRKY domains and were classified into three main groups. Group II was further divided into five subgroups (IIa-IIe) based on phylogenetic analysis, with each clade being well supported by conserved motifs. Additionally, the gene structure analysis revealed variations in exon-intron organization. In contrast, motif analysis showed the presence of conserved domains that were similar within the group but differed between groups, suggesting their functional diversity. Cis-acting elements related to plant growth and development and light, hormones, and stress responses were identified. Synteny analysis revealed that 34 (61.8%) of the genes originated from tandem duplication, indicating the role of tandem duplication in the expansion of the A. hypochondriacus WRKY gene family. Protein-protein interaction analysis suggested that AhyWRKY3, AhyWRKY27, AhyWRKY28, AhyWRKY36, and AhyWRKY52 were hub genes involved in the complex protein interaction network. Using in silico and real-time quantitative PCR, expression analysis revealed tissue- and condition-specific expression patterns of AhyWRKY genes. Notably, under drought stress, AhyWRKY39, AhyWRKY40, AhyWRKY54, and AhyWRKY01 showed increased expression, while under salt stress, AhyWRKY40, AhyWRKY54, AhyWRKY39, AhyWRKY49, and AhyWRKY8 were upregulated at 30 days, suggesting that these genes may play key role in response to salinity stress.

Conclusions: The present study provides valuable insights into the organization and evolutionary patterns of the WRKY gene family in amaranth. It also identifies putative candidate WRKY genes that may play a role in conferring drought and salt tolerance. Overall, this study lays a foundation for further functional validation of these WRKY candidate genes, facilitating their exploitation in the amaranth genetic improvement programs to develop stress-resilient varieties.