Screening and Bioinformatic Analysis of Drought-Adaptive-Response Genes in Wild Cerasus humilis

This study aimed to screen genes associated with drought adaptation in Cerasus humilis and analyze their sequence characteristics and evolutionary relationships. The results could provide a theoretical basis for exploring the molecular basis underlying drought adaptation of Cerasus humilis. Two-year-old wild Cerasus humilis plants were cultivated in a greenhouse, and irrigation was completely withheld for 20 consecutive days. Leaves were sampled and a cDNA library of wild Cerasus humilis was constructed. By utilizing high-throughput screening in yeast combined with mannitol-induced simulated drought stress, genes associated with drought adaptation were isolated and identified. Additionally, core candidate genes were cloned and subjected to systematic bioinformatic analysis. The results showed that: (1) the drought adaptation-associated genes ChU40rps27a and ChTp258 were obtained through high-throughput screening of yeast and mannitol-simulated drought resistance test; (2) gene cloning revealed that the open reading frame of ChU40rps27a is 471 bp long, encoding 157 amino acids, and the open reading frame of ChTp258 is 231 bp long, encoding 77 amino acids; (3) multiple sequence alignment of the genes indicated that ChU40rps27a shares 88.5% identity with apple 40S (XM_050288755), and ChTp258 shares 94.4% identity with peach (XM_020567676) in terms of nucleotide sequence; (4) the phylogenetic tree analysis revealed that ChU40rps27a is closely related to Citrus sinensisU40rps27a (XM_006487968); and ChTp258 is closely related to peach (Prunus persica, XM_020567676), apricot (Prunus dulcis, XM_034370534), plum (Prunus mume, XM_008244122), and sweet cherry (Prunus avium, XM_021951344) in the Rosaceae family, clustering in the same clade. This study identified two candidate genes involved in long-term drought adaptation rather than short-term stress responses. These findings clarify their sequence characteristics, sequence conservation and evolutionary status, providing important gene resources and theoretical support for further exploring drought adaptation related molecular processes in Cerasus humilis and developing drought-adapted molecular breeding.