Monochromic Radiations Provided by Light Emitted Diode (LED) Modulate Infection and Defense Response to Fire Blight in Pear Trees.

Pathogenesis-related (PR) proteins are part of the systemic signaling network that perceives pathogens and activates defenses in the plant. Eukaryotic and bacterial species have a 24-h 'body clock' known as the circadian rhythm. This rhythm regulates an organism's life, modulating the activity of the phytochromes (phys) and cryptochromes (crys) and the accumulation of the corresponding mRNAs, which results in the synchronization of the internal clock and works as zeitgeber molecules. Salicylic acid accumulation is also under light control and upregulates the PR genes expression, increasing plants' resistance to pathogens. Erwinia amylovora causes fire blight disease in pear trees. In this work, four bacterial transcripts (erw1-4), expressed in asymptomatic E. amylovora-infected pear plantlets, were isolated. The research aimed to understand how the circadian clock, light quality, and related photoreceptors regulate PR and erw genes expression using transgenic pear lines overexpressing PHYB and CRY1 as a model system. Plantlets were exposed to different circadian conditions, and continuous monochromic radiations (Blue, Red, and Far-Red) were provided by light-emitting diodes (LED). Results showed a circadian oscillation of PR10 gene expression, while PR1 was expressed without clear evidence of circadian regulation. Bacterial growth was regulated by monochromatic light: the growth of bacteria exposed to Far-Red did not differ from that detected in darkness; instead, it was mildly stimulated under Red, while it was significantly inhibited under Blue. In this regulatory framework, the active form of phytochrome enhances the expression of PR1 five to 15 fold. An ultradian rhythm was observed fitting the zeitgeber role played by CRY1. These results also highlight a regulating role of photoreceptors on the expression of PRs genes in non-infected and infected plantlets, which influenced the expression of erw genes. Data are discussed concerning the regulatory role of photoreceptors during photoperiod and pathogen attacks.

Day-Length Is Involved in Flooding Tolerance Response in Wild Type and Variant Genotypes of Rootstock Prunus cerasifera L.

Current and predicted climate changes scenarios require crops with an improved adaptability to mutable environmental features, such as, hypoxia for the root system. In order to overcome the reduction of oxygen, plants activate coping mechanisms and strategies. Prunus spp. are hypoxia-sensitive woody species and although many information has been gathered over the last decades, many physiological mechanisms remain unclear. To verify whether anoxic plant responses are also regulated by photoperiod, plants of Mr.S.2/5-WT plum, and its variant genotypes S.4 tolerant (plus) and S.1 sensitive (minus) to flooding, were grown in a greenhouse and were submitted to natural photoperiod (NP) and to constant photoperiod (CP) from mid-July until the first 10 days of October. From mid-September plants from each genotype, grown under the two photoperiods, were divided into two groups, and one of them underwent long-term flooding. Gas exchange parameters, energetic and biochemical activities, leaf chlorophyll contents, and stress symptoms were measured at different times, whereas soluble sugars were quantified in leaves and roots 14 days after flooding, when stress symptoms in WT and S.1 became prominent. Seasonal changes in the photoperiod played a role in the adaptability to anoxia, although flooding stress response differed among the three genotypes. Anoxia affected leaf gas exchange and S.4 flooded-leaves retained higher ACO2 under conditions of NP and CP. Leaf soluble sugar concentration differed among genotypes. Regardless the photoperiod, S.4 anoxic-leaf sugar concentration was the lowest, except for sorbitol. S.4 anoxic-roots under CP accumulated the highest levels of sucrose and sorbitol. Influences of the photoperiod were observed in WT and S.1 anoxic-leaves, whereas S.1 anoxic roots accumulated the lowest concentration of sugars, regardless of photoperiod. Leaf and root respiratory activity in flooded-plants was highest in S.4, and ADH activity increased in all flooded plants under CP but the highest activity was observed only in S.1 under NP during flooding. Results are consistent with the hypothesis that the S.4 genotype has a plastic adaptability to flooding stress, escaping from the photoperiod regulatory cross-talk system, and can better cope with the new scenarios generated by climate changes.

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging.

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.

Study of rhizosphere and phyllosphere bacterial community and resistance to bacterial canker in genetically engineered phytochrome A cherry plants.

The cherry rootstock 'Colt' line was transformed with a phytochrome A rice gene with the aim of altering light perception. Three transgenic events were chosen because of a modified developmental behavior. When red enriched light was supplied horizontally to stems, the PD3 transgenic line showed an increased rate of phytomer formation associated to a superior rate of plant growth compared to wild type (WT). Under the same light conditions, the PO1 and PA lines were less altered in morphology and development. When far-red enriched light was supplied, all transgenic lines had a reduced rate of growth, with the PD3 line being the most similar to the WT. The influence of the alien gene on root and leaf-associated bacteria was studied for a duration of 1 year. Significantly more culturable bacteria were recovered from PA lines than from PO1, PD3 and WT lines. On average, significantly more fluorescent pseudomonads were recovered from the rhizosphere of PA and PO1 lines than from PD3 and WT. No significant differences were detected in the number of bacteria recovered from the phyllosphere of transgenic and WT plant lines. A total of 143 Pseudomonas fluorescens strains isolated from rhizosphere of transgenic and WT lines were tested for their antagonistic activity against Phytophthora nicotianae and differences between bacteria derived from transgenic and WT were not detected. Fluorescent pseudomonads strains isolated from phyllosphere of PA and PO1 lines showed antagonistic activity against P. syringae pv. syringae, whereas no difference among the transgenic and WT lines was detected when fluorescent Pseudomonas strains were tested against P. syringae pv. mors-prunorum. Pathogenicity tests were conducted on rooted and micropropagated plants with P. s. pv. syringae and P. s. pv. mors-prunorum: in all assays, the PO1 lines were the most tolerant to P. s. pv. Syringae, and the PO1 and PD3 were tolerant to P. s. pv. mors-prunorum.