DNA methylation and small interference RNAs participate in the regulation of MADS-box genes involved in dormancy in sweet cherry (Prunus avium L.).

Epigenetic modifications can yield information about connections between genotype, phenotype variation and environmental conditions. Bud dormancy release in temperate perennial fruit trees depends on internal and environmental signals such as cold accumulation and photoperiod. Previous investigations have noted the participation of epigenetic mechanisms in the control of this physiological process. We examined whether epigenetic modifications were modulated in MADS-box genes, potential candidates for the regulation of bud dormancy and flowering in sweet cherry (Prunus avium L.). We identified and cloned two MADS-box genes homologous to the already-characterized dormancy regulators DORMANCY-ASSOCIATED MADS-box (DAM3 and DAM5) from Prunus persica (L.) Batsch. Bisulfite sequencing of the identified genes (PavMADS1 and PavMADS2), Methylated DNA Immunoprecipitation and small RNA deep sequencing were performed to analyze the presence of DNA methylations that could be guided by non-coding RNAs in the floral buds exposed to differential chilling hours. The results obtained reveal an increase in the level of DNA methylation and abundance of matching small interference RNAs (siRNAs) in the promoter of PavMADS1 when the chilling requirement is complete. For the first intron and 5' UTR of PavMADS1, de novo DNA methylation could be associated with the increase in the abundance of 24-nt siRNA matching the promoter area. Also, in the second large intron of PavMADS1, maintenance DNA methylation in all cytosine contexts is associated with the presence of homologous siRNAs in that zone. For PavMADS2, only maintenance methylation was present in the CG context, and no matching siRNAs were detected. Silencing of PavMADS1 and PavMADS2 coincided with an increase in Flowering Locus T expression during dormancy. In conclusion, DNA methylations and siRNAs appear to be involved in the silencing of PavMADS1 during cold accumulation and dormancy release in sweet cherry.

A Complex Molecular Interplay of Auxin and Ethylene Signaling Pathways Is Involved in Arabidopsis Growth Promotion by Burkholderia phytofirmans PsJN.

Modulation of phytohormones homeostasis is one of the proposed mechanisms to explain plant growth promotion induced by beneficial rhizobacteria (PGPR). However, there is still limited knowledge about the molecular signals and pathways underlying these beneficial interactions. Even less is known concerning the interplay between phytohormones in plants inoculated with PGPR. Auxin and ethylene are crucial hormones in the control of plant growth and development, and recent studies report an important and complex crosstalk between them in the regulation of different plant developmental processes. The objective of this work was to study the role of both hormones in the growth promotion of Arabidopsis thaliana plants induced by the well-known PGPR Burkholderia phytofirmans PsJN. For this, the spatiotemporal expression patterns of several genes related to auxin biosynthesis, perception and response and ethylene biosynthesis were studied, finding that most of these genes showed specific transcriptional regulations after inoculation in roots and shoots. PsJN-growth promotion was not observed in Arabidopsis mutants with an impaired ethylene (ein2-1) or auxin (axr1-5) signaling. Even, PsJN did not promote growth in an ethylene overproducer (eto2), indicating that a fine regulation of both hormones signaling and homeostasis is necessary to induce growth of the aerial and root tissues. Auxin polar transport is also involved in growth promotion, since PsJN did not promote primary root growth in the pin2 mutant or under chemical inhibition of transport in wild type plants. Finally, a key role for ethylene biosynthesis was found in the PsJN-mediated increase in root hair number. These results not only give new insights of PGPR regulation of plant growth but also are also useful to understand key aspects of Arabidopsis growth control.

Burkholderia phytofirmans PsJN induces long-term metabolic and transcriptional changes involved in Arabidopsis thaliana salt tolerance.

Salinity is one of the major limitations for food production worldwide. Improvement of plant salt-stress tolerance using plant-growth promoting rhizobacteria (PGPR) has arisen as a promising strategy to help overcome this limitation. However, the molecular and biochemical mechanisms controlling PGPR/plant interactions under salt-stress remain unclear. The main objective of this study was to obtain new insights into the mechanisms underlying salt-stress tolerance enhancement in the salt-sensitive Arabidopsis thaliana Col-0 plants, when inoculated with the well-known PGPR strain Burkholderia phytofirmans PsJN. To tackle this, different life history traits, together with the spatiotemporal accumulation patterns for key metabolites and salt-stress related transcripts, were analyzed in inoculated plants under short and long-term salt-stress. Inoculated plants displayed faster recovery and increased tolerance after sustained salt-stress. PsJN treatment accelerated the accumulation of proline and transcription of genes related to abscisic acid signaling (Relative to Dessication, RD29A and RD29B), ROS scavenging (Ascorbate Peroxidase 2), and detoxification (Glyoxalase I 7), and down-regulated the expression of Lipoxygenase 2 (related to jasmonic acid biosynthesis). Among the general transcriptional effects of this bacterium, the expression pattern of important ion-homeostasis related genes was altered after short and long-term stress (Arabidopsis K(+) Transporter 1, High-Affinity K(+) Transporter 1, Sodium Hydrogen Exchanger 2, and Arabidopsis Salt Overly Sensitive 1). In all, the faster and stronger molecular changes induced by the inoculation suggest a PsJN-priming effect, which may explain the observed tolerance after short-term and sustained salt-stress in plants. This study provides novel information about possible mechanisms involved in salt-stress tolerance induced by PGPR in plants, showing that certain changes are maintained over time. This opens up new venues to study these relevant biological associations, as well as new approaches to a better understanding of the spatiotemporal mechanisms involved in stress tolerance in plants.