Molecular identification and expression patterns of sweet cherry HIPPs and functional analysis of PavHIPP16 in cold stress.

Main conclusion: The HIPP proteins are involved in low-temperature stress, the growth of sweet cherry, and may be potential targets for genetic improvement. PavHIPP16 improved cold resistance in Arabidopsis. In response to abiotic stressors, the heavy metal-associated isoprenylated plant protein (HIPP) proteins play a crucial regulatory role. Although the function of HIPP has been identified in some plants, there have been fewer systematic studies conducted on sweet cherry (Prunus avium L.). Therefore, we performed a comprehensive analysis and expression profiling of PavHIPPs using bioinformatics, RT-PCR, and qRT-PCR techniques. Our findings revealed that 28 PavHIPP genes were unevenly distributed across eight chromosomes. We predicted nine motifs in PavHIPP proteins and observed similar gene structures among highly homologous proteins. The promoter sequences of PavHIPPs contained numerous regulatory elements associated with an adversity of stress. The expression levels of some members showed varying degrees of change under low-temperature treatment. These genes were differentially expressed during flower and fruit development. Arabidopsis overexpressing the PavHIPP16 (OE) gene showed significantly lower relative conductivity and malondialdehyde (MDA) content compared with the wild-type (WT) plants under cold environment. Conversely, peroxidase (POD) activity, superoxide dismutase (SOD) activity, and proline content were significantly higher in OE Arabidopsis than in WT plants. Overall, our results suggest that PavHIPP16 OE Arabidopsis thaliana exhibited enhanced adaptability compared to WT plants under cold conditions. This study provides a foundation for future investigations of the pathways regulating sweet cherry growth and development mediated by the HIPP genes.