Chile is an important producers of sweet cherry (Prunus avium L.), with a total of 356,385 t exported in the 2021 to 2022 season. The production area includes most of the country's regions. Bacteri...

Salinity is a major constraint limiting the yield of tomatoes. However, grafting strategies may help to overcome the salt toxicity of this important horticultural species if appropriate rootstocks are identified. The present study aimed to test a new rootstock, JUPAFORT1, obtained by crossing the glycophyte Solanum lycopersicum (cv. Poncho Negro) with the halophyte wild-related species Solanum chilense to improve the salinity tolerance of the Chilean tomato landrace Old Limachino Tomato (OLT). Intact OLT plants were exposed to 0, 80, or 160 mM of NaCl for 21 days at the vegetative stage and compared with self-grafted (L/L) and Limachino plants grafted on JUPAFORT1 rootstock (L/R) under a completely randomized design. JUPAFORT1 increased OLT scion vigor in the absence of salt but did not significantly increase fresh weight under stress conditions. However, JUPAFORT1 confers to the scion an anisohydric behavior contrasting with the isohydric behavior of L and L/L plants as indicated by measurements of stomatal conductance; L/R plants were able to maintain their metabolic status despite a slight decrease in the leaf’s relative water content. JUPAFORT1 rootstock also enabled the maintenance of photosynthetic pigment concentrations in the scion in contrast to L and L/L plants, which exhibited a decrease in photosynthetic pigments under stress conditions. L/R plants encountered oxidative stress at the highest stress intensity (160 mM of NaCl) only, while L and L/L plants suffered from oxidative damage at a lower dose (80 mM of NaCl). L/R plants behaved as includer plants and did not sequester Na⁺ in the root system, in contrast to L and L/L, which behaved as excluder plants retaining Na⁺ in the root system to avoid its translocation to the shoots. The expression of genes coding for ion transporters (HKT1.1, HKT1.2, LKT1, SKOR, SOS2, and SOS3) in the root system was not modified by salinity in L/R. In contrast, their expression varied in response to salinity in L and L/L. Overall, L/R plants exhibited higher physiological stability than L/L or L plants in response to an increasing NaCl dose and did not require additional energy investment to trigger an adaptative response to salinity. This suggests that the constitutive salinity tolerance of the halophyte S. chilense was maintained in the interspecific rootstock. JUPAFORT1 issued from S. lycopersicum x S. chilense may thus improve salt-stress resilience in OLT tomatoes. Additional studies are required to identify the molecular components involved in the root-to-shoot signaling pathway in this promising material.

Pseudomonas syringae pv. syringae (Pss) is an economically significant bacterial pathogen that causes canker in sweet cherry trees. In Chile, sweet cherry (Prunus avium L.) is a key crop whose exponential production growth has increased phytosanitary pressure. However, the genetic diversity and adaptive mechanisms of local Pss populations have remained poorly understood. This study characterized 41 Pss isolates from major Chilean production regions. Their genomes were sequenced and compared with 152 public genomes from the PG2 phylogenetic group. The analysis revealed a predominance of the PG2d subgroup among the Chilean isolates, with a population structure defined by at least 18 genomic clusters, some of which are exclusive to Chile. A characteristic feature of this entire PG2d subgroup is the presence of indole-3-acetic acid (IAA) synthesis genes (iaaM and iaaH). Furthermore, this subgroup displayed a marked increase in ancestral gene gain and loss events, indicating extensive remodeling of the shell genome and supporting a model of lineage-specific adaptive evolution. We also identified lineage-specific orthogroups, structural variants of the T-PAI pathogenicity island, and a differential distribution of Hop-type effector proteins. Furthermore, an extended copper resistance operon (cop and cus systems) was detected in a subset of strains, and a dominant lineage was found to have a dual i1-type of T6SS system. These findings highlight the local diversification of Pss in Chile, likely driven by agro-environmental pressures. This study provides crucial insights into the evolution, adaptation, and pathogenic potential of this important pathogen in a crop of high strategic value.

Abscisic acid (ABA) plays a key role in the ripening process of non-climacteric fruits, triggering pigment production, fruit softening, and sugar accumulation. Transcriptomic studies show that ABA modifies the expression of several ripening-related genes, but to date, the epigenetic approach has not been utilized to characterize the role of ABA during this process. Therefore, this work aimed to perform transcriptomic and DNA methylation analyses of fruit samples treated with ABA during the fruit ripening process in the non-climacteric sweet cherry model. RNA-seq analyses revealed an overrepresentation of transcripts annotated in functional categories related to ABA response, secondary metabolism, and sugar synthesis. In contrast, Whole Genome Bisulfite Sequencing (WGBS) revealed DNA hypomethylation in the 5’UTR region of genes related to carotene catabolism. Genes encoding xyloglucan enzymes were regulated transcriptionally and epigenetically during ripening. ABA treatment enhanced color development and ripening. GO analysis of DEGs in the RNA-seq of the ABA treatment revealed expression variations in genes encoding members of the Aux/IAA and ARF families. In the WGBS, genes encoding enzymes of the cytokinin biosynthesis had differential DNA methylation after the ABA treatment. Our work shows the genetic factors modulated by ABA at the genetic and epigenetic levels during non-climacteric ripening.

Prunus rootstock belonging to subgenera Amygdalus (peach), Prunus (plum) and Cerasus (cherry) are either from the same species as the scion or another one. The number of inter-species (including inter-subgenera) hybrids have increased as a result of efforts to broaden the genetic basis for biotic and abiotic resistance/tolerance. Identifying genes associated with important traits and responses requires expression analysis. Relative quantification is the simplest and most popular alternative, which requires reference genes (housekeeping) to normalize RT-qPCR data. However, there is a scarcity of validated housekeeping genes for hybrid Prunus rootstock species. This research aims to increase the number of housekeeping genes suitable for Prunus rootstock expression analysis. Twenty-one candidate housekeeping genes were pre-selected from previous RNAseq data that compared the response of root transcriptomes of two rootstocks subgenera to hypoxia treatment, ‘Mariana 2624’ ( P. cerasifera Ehrh.× P. munsoniana W. Wight & Hedrick), and ‘Mazzard F12/1’ ( P. avium L.). Representing groups of low, intermediate or high levels of expression, the genes were assayed by RT-qPCR at 72 hours of hypoxia treatment and analyzed with NormFinder software. A sub-set of seven housekeeping genes that presented the highest level of stability were selected, two with low levels of expression ( Unknown 3, Unknown 7 ) and five with medium levels ( GTB 1, TUA 3, ATPase P, PRT 6, RP II ). The stability of these genes was evaluated under different stress conditions, cold and heat with the hybrid ‘Mariana 2624’ and N nutrition with the hybrids ‘Colt’ ( P. avium × P. pseudocerasus Lindl.) and ‘Garnem’ [ P. dulcis Mill.× ( P. persica L.× P. davidiana Carr.)]. The algorithms of geNorm and BestKeeper software also were used to analyze the performance of these genes as housekeepers. Stability rankings varied according to treatments, genotypes and the software for evaluation, but the gene GBT 1 often had the highest ranking. However, most of the genes are suitable depending on the stressor and/or genotype to be evaluated. No optimal number of reference genes could be determined with geNorm software when all conditions and genotypes were considered. These results strongly suggest that relative RT-qPCR should be analyzed separately with their respective best housekeeper according to the treatment and/or genotypes in Prunus spp rootstocks.