Drought is one of the main factors affecting sweet cherry yields, and cherry rootstocks can provide a range of tree vigor levels to better match sweet cherries with the characteristics of the soil. To investigate the molecular events of the cherry to water deficiency, we performed transcriptomic and metabolomic analyses of Prunus mahaleb CDR-1 (drought-tolerant cherry rootstock (DT)) and P. cerasus × P. canescens Gisela 5 (drought-susceptible cherry rootstock (DS)), respectively. The results revealed 253 common drought-responsive genes in leaves and roots in DT and 17 in DS; 59 upregulated metabolites were explored in leaves in DT and 19 were explored in DS. Differentially expressed metabolites related to the cyanoamino acid metabolism pathway and phenylpropanoid biosynthesis pathway may be key factors in the difference in drought resistance in the two rootstocks. Moreover, six central metabolites—3-cyanoalanine, phenylalanine, quinic acid, asparagine, p-benzoquinone, and phytosphingosine—were identified as potential biological markers of drought response in cherries and may be key factors in the difference in drought resistance, along with caffeic acid and chlorogenic acid. We also selected 17 differentially expressed genes as core candidate genes and the mechanism of DT in response to drought is summarized.

Alternaria alternata is a necrotrophic fungal pathogen with a broad host range that causes widespread and devastating disease in sweet cherry (Prunus avium). We selected a resistant cultivar (RC) and a susceptible cultivar (SC) of cherry and used a combined physiological, transcriptomic, and metabolomic approach to investigate the molecular mechanisms underlying the plant’s resistance to A. alternata, of which little is known. We found that A. alternata infection stimulated the outbreak of reactive oxygen species (ROS) in cherry. The responses of the antioxidant enzymes and chitinase to disease were observed earlier in the RC than in the SC. Moreover, cell wall defense ability was stronger in the RC. Differential genes and metabolites involved in defense responses and secondary metabolism were primarily enriched in the biosynthesis of phenylpropanoids, tropane, piperidine and pyridine alkaloids, flavonoids, amino acids, and α-linolenic acid. Reprogramming the phenylpropanoid pathway and the α-linolenic acid metabolic pathway led to lignin accumulation and early induction of jasmonic acid signaling, respectively, in the RC, which consequently enhanced antifungal and ROS scavenging activity. The RC contained a high level of coumarin, and in vitro tests showed that coumarin significantly inhibited A. alternata growth and development and had antifungal effect on cherry leaves. In addition, differentially expressed genes encoding transcription factors from the MYB, NAC, WRKY, ERF, and bHLH families were highly expressed, they could be the key responsive factor in the response of cherry to infection by A. alternata. Overall, this study provides molecular clues and a multifaceted understanding of the specific response of cherry to A. alternata.

Background: Crown gall disease, caused by the pathogenic bacteriumAgrobacterium tumefaciens, is responsible for extensive economic losses in orchards. Cherry rootstock ‘CDR-1’ (Prunus mahaleb) shows high resistance but the mechanism remains unclear. Here, we examined the morphology of pathogen-infected root neck surface, determined the activity of 10 defense-related enzymes and the content of salicylic acid (SA) and jasmonic acid (JA), and also applied transcriptome analysis, transient expression and transgenic verification to explore the crown gall resistance genes in ‘CDR-1’ plants.Results: In our study, peroxidase increased in the first 10 days, while phenylalanine ammonialyase and lipoxygenase increased in the first 15 days post-infection. Four key enzymes in the AsA-GSH cycle also responded, to a certain extent; although JA content increased significantly after the treatment, the SA content did not. In a follow-up transcriptome analysis, the differentially expressed genesPm4CL2,PmCYP450,PmHCT1,PmHCT2, andPmCADwere up-regulated. Based on the above results, we focused on the lignin biosynthetic pathway, and further measured lignin content, and found it increased significantly. ThePm4CL2gene was used to conduct transient expression and transgenic experiments to verify its function in crown gall disease resistance. It showed the relative expression of the treatment group was almost 14-fold that of the control group at 12 h post-treatment. After the infection treatment, clear signs of resistance were found in the transgenic lines; this indicated that under the higher expression level and earlier activation ofPm4CL2, plant resistance was enhanced.Conclusions: The crown gall resistance of ‘CDR-1’ is likely related to the lignin biosynthetic pathway, in whichPm4CL2functions crucially during the plant defense response to the pathogenA. tumefaciens. The results thus offer novel insights into the defense responses and resistance mechanism of cherry rootstock ‘CDR-1’ against crown gall disease.

Bacterial contamination is a major and constant threat to the establishment and subculture of in vitro plant culture. In this study, we used a slightly modified qualitative disk diffusion method to screen optimal antibiotics to control the growth of bacterial contaminants isolated from explants of cherry rootstock ‘Gisela 6’. Bacterial susceptibility to eight different antibiotics was tested. The results showed that tetracycline was the most effective antibiotic for controlling bacterial growth; cefotaxime, carbenicillin, kanamycin, and streptomycin were less effective, whereas ampicillin, penicillin, and cefazolin did not inhibit growth of the isolated bacteria. Using the quantitative E test, the minimal inhibitory concentration (MIC) of tetracycline was determined to be 1.0 µg mL⁻¹. We also measured the F_v/F_m values, chlorophyll content, and enzymatic activity of superoxide dismutase and peroxidase to explore the effect of different tetracycline concentrations, 0, 0.064, 0.5, 1.0, 16, and 256 µg mL⁻¹, on the growth of bacteria and explants over 30 days. Results indicated that 1.0 µg mL⁻¹ tetracycline was effective in restricting bacterial growth, with non-significant negative effects on explants at low concentrations, but were enhanced negative effects at high concentrations. The application of the disk diffusion method and E test enabled the identification of an antibiotic and its MIC value effective for eliminating bacterial contaminants while causing minimal damage to explants, indicating a high potential of these methods to control bacterial contaminants in in vitro plant culture.