Mitogen-activated protein kinase kinase kinase kinases (MAP4Ks) are a class of highly conserved serine/threonine-protein kinases in eukaryotes. They participate in the typical MAPK cascade system and various signal transduction pathways regulating biological processes in plants, during stressful conditions. To date, genome-wide identification of MAP4Ks in cotton has not been reported. In this study, 77 MAP4K genes were identified in four Gossypium species. Protein characteristics, gene structures, conserved motifs and gene expression analysis were carried out. Genome-wide or fragment duplication has played an important role in the expansion of the GhMAP4K. Promoter cis-acting elements and expression patterns indicated that GhMAP4Ks are related to plant hormones (ABA, MeJA, GA, IAA, SA) and various stresses (drought, hypothermia and wound). Overexpressing GhMAP4K13 in Arabidopsis showed higher stem length in response to drought and salt stress. The wilting degree in virus-induced GhMAP4K13 gene silenced plants was substantially greater than wild type plants under drought and salt stress. Transcriptomic analysis showed that most differentially expressed genes were involved in the MAPK signaling pathway, carbon metabolism and porphyrin metabolism. Additionally, transgenic Arabidopsis and VIGS cotton showed that GhMAP4K13 was positively responsive to drought and salt stresses. This study will play an important role in understanding the function of the MAP4K gene family in response to abiotic stress in cotton.
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