Physiological and Molecular Mechanisms of ABA and CaCl2 Regulating Chilling Tolerance of Cucumber Seedlings

Cold stress is a limiting factor to the growth and development of cucumber in the temperate regions; hence, improving the crop's tolerance to low temperature is highly pertinent. The regulation of low-temperature tolerance with exogenous ABA and CaCl2 was investigated in the cucumber variety Zhongnong 26. Under low-temperature conditions (day/night 12/12 h at 5 degrees C), seedlings were sprayed with a single application of ABA, CaCl2, or a combination of both. Our analysis included a calculated chilling injury index, malondialdehyde (MDA) content, relative electrical conductivity, antioxidant enzyme activities (SOD, CAT, and APX), leaf tissue structure, and expression of cold-related genes by transcriptome sequencing. Compared with the water control treatment, the combined ABA + CaCl2 treatment significantly improved the superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) of the seedlings by 34.47%, 59.66%, and 118.80%, respectively (p < 0.05), and significantly reduced the chilling injury index, relative electrical conductivity, and MDA content, by 89.47%, 62.17%, and 44.55%, respectively (p < 0.05). Transcriptome analysis showed that compared with the water control treatment, 3442 genes were differentially expressed for the combined treatment, 3921 for the ABA treatment, and 1333 for the CaCl2 treatment. KEGG enrichment analysis for both the ABA and combined ABA + CaCl2 treatments (as compared to the water control) showed that it mainly involves genes of the photosynthesis pathway and metabolic pathways. Differentially expressed genes following the CaCl2 treatment were mainly involved in plant hormone signal transduction, plant-pathogen interaction, MAPK signaling pathway-plant, phenylpropanoid biosynthesis, and circadian rhythm-plant. qRT-PCR analysis and RNA-seq results showed a consistent trend in variation of differential gene expression. Overall, this study demonstrated that although all three treatments provided some protection, the combined treatment of ABA (35 mg/L) with CaCl2 (500 mg/L) afforded the best results. A combined ABA + CaCl2 treatment can effectively alleviate cold-stress damage to cucumber seedlings by inducing physiological changes in photosynthesis and metabolism, and provides a theoretical basis and technical support for the application of exogenous ABA and CaCl2 for low-temperature protection of cucumber seedlings.