Fig. 3: Identification and verification of the balance modules and the key candidate gene, AsHSFA2c.

a A schematic illustration that outlines the status of gene families identified in the ‘balance’ module, Cluster 12. Gene families in Cluster 12 were classified into three groups: those that play significant roles in plant growth and development (growth and development module), those crucial for drought resistance (drought resistance module), and those important for both growth and drought resistance (balance module). Each node represents a gene family, with the node size indicating the number of genes within that family. The connections between nodes represent the average correlation of co-expression between genes within the families. The green half of (a) represents the growth and development module, and the blue half of (a) represents the drought resistance module. The green nodes represent the previously reported gene families related to plant growth and development. The blue nodes represent the gene families potentially related to drought resistance. The pink nodes represent gene families potentially important for balancing drought resistance and growth/development. b Most of the genes in the balance module responded to both drought and IAA treatments. qRT-PCR analysis of gene expression levels before and after drought and IAA treatments. c AsHSFA2c responded to both drought and IAA treatments. d qRT-PCR analysis of target gene AsHSFA2c expression level in AsHSFA2c knock-down mutant lines. e The malfunction of AsHSFA2c promoted plant growth and negatively regulated plant drought tolerance. “Watered” represents well-watered conditions. “After drought” represents a 2-day period of recovery with full irrigation post-drought treatment. Scale bars, 5 cm. f The up part: schematic representation of the pUBI::AsHSFA2c-GFP vector with the Ubi promoter and Nos terminator; the down part: Detection of GFP fluorescence signal in callus induced from mature embryos infected with the pUBI::AsHSFA2c-GFP vector (leaf). Scale bar, 2 cm. Regeneration phenotypes of mature embryos infected with the pUBI::AsHSFA2c-GFP vector (right). Scale bar, 2 cm. g Detection of the expression of AsHSFA2c in the pUBI::AsHSFA2c-GFP genetic plants. h The over-expression of AsHSFA2c increased plant tolerance to drought. “Watered” represents well-watered conditions. “After drought” represents a 2-day period of recovery with full irrigation post-drought treatment. Scale bars, 3 cm. Student’s t-test was performed to determine statistical significance. Error bars represent the SD of three biological replicates. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, p ≥ 0.05.