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Advancing disease resistance in fruit trees A significant focus of the research collected here is on combating diseases that threaten fruit tree health and productivity.Jia et al. demonstrated a breakthrough in citrus canker resistance by employing the Cas12a/CBE co-editing method to precisely target the susceptibility gene LOB1 in Citrus sinensis cv. Enhancing breeding efficiency and developmental traits Another critical area of exploration is the enhancement of breeding efficiency and manipulation of developmental traits in fruit trees.Guerrero et al. addressed the challenge of the long juvenile phase in olive trees by characterizing olive plants transformed with a flowering locus T (FT) gene from Medicago truncatula. Harnessing genomic tools for precision breeding Advances in genomic tools and molecular techniques are central to the progress in fruit tree improvement addressed in this Research Topic.Deng et al. conducted a comprehensive genome-wide analysis of the GH9 gene family in mulberry, elucidating key genes involved in fruit abscission.

Olive ( Olea europaea L. subsp. europaea ) is one of the most important crops of the Mediterranean Basin and temperate areas worldwide. Obtaining new olive varieties adapted to climatic changing conditions and to modern agricultural practices, as well as other traits such as biotic and abiotic stress resistance and increased oil quality, is currently required; however, the long juvenile phase, as in most woody plants, is the bottleneck in olive breeding programs. Overexpression of genes encoding the ‘florigen’ Flowering Locus T (FT), can cause the loss of the juvenile phase in many perennials including olives. In this investigation, further characterization of three transgenic olive lines containing an FT encoding gene from Medicago truncatula, MtFTa1 , under the 35S CaMV promoter, was carried out. While all three lines flowered under in vitro conditions, one of the lines stopped flowering after acclimatisation. In soil, all three lines exhibited a modified plant architecture; e.g., a continuous branching behaviour and a dwarfing growth habit. Gene expression and hormone content in shoot tips, containing the meristems from which this phenotype emerged, were examined. Higher levels of OeTFL1 , a gene encoding the flowering repressor TERMINAL FLOWER 1, correlated with lack of flowering. The branching phenotype correlated with higher content of salicylic acid, indole-3-acetic acid and isopentenyl adenosine, and lower content of abscisic acid. The results obtained confirm that heterologous expression of MtFTa1 in olive induced continuous flowering independently of environmental factors, but also modified plant architecture. These phenotypical changes could be related to the altered hormonal content in transgenic plants.

Host resistance is the most practical, long-term, and economically efficient disease control measure for Verticillium wilt in olive caused by the xylem-invading fungus Verticillium dahliae ( Vd ), and it is at the core of the integrated disease management. Plant’s microbiome at the site of infection may have an influence on the host reaction to pathogens; however, the role of xylem microbial communities in the olive resistance to Vd has been overlooked and remains unexplored to date. This research was focused on elucidating whether in vitro olive propagation may alter the diversity and composition of the xylem-inhabiting microbiome and if those changes may modify the resistance response that a wild olive clone shows to the highly virulent defoliating (D) pathotype of Vd . Results indicated that although there were differences in microbial communities among the different propagation methodologies, most substantial changes occurred when plants were inoculated with Vd , regardless of whether the infection process took place, with a significant increase in the diversity of bacterial communities when the pathogen was present in the soil. Furthermore, it was noticeable that olive plants multiplied under in vitro conditions developed a susceptible reaction to D Vd , characterized by severe wilting symptoms and 100% vascular infection. Moreover, those in vitro propagated plants showed an altered xylem microbiome with a decrease in total OTU numbers as compared to that of plants multiplied under non-aseptic conditions. Overall, 10 keystone bacterial genera were detected in olive xylem regardless of infection by Vd and the propagation procedure of plants ( in vitro vs nursery), with Cutibacterium (36.85%), Pseudomonas (20.93%), Anoxybacillus (6.28%), Staphylococcus (4.95%), Methylobacterium-Methylorubrum (3.91%), and Bradyrhizobium (3.54%) being the most abundant. Pseudomonas spp. appeared as the most predominant bacterial group in micropropagated plants and Anoxybacillus appeared as a keystone bacterium in Vd- inoculated plants irrespective of their propagation process. Our results are the first to show a breakdown of resistance to Vd in a wild olive that potentially may be related to a modification of its xylem microbiome and will help to expand our knowledge of the role of indigenous xylem microbiome on host resistance, which can be of use to fight against main vascular diseases of olive.

The gene encodes a key component of systemic acquired resistance (SAR) signaling mediated by salicylic acid (SA). Overexpression of confers resistance to biotrophic and hemibiotrophic fungi in several plant species. The gene has also been shown to be involved in the crosstalk between SAR signaling and the jasmonic acid-ethylene (JA/Et) pathway, which is involved in the response to necrotrophic fungi. The aim of this research was to generate transgenic olive plants expressing the gene from to evaluate their differential response to the hemibiotrophic fungus and the necrotroph . Three transgenic lines expressing the gene under the control of the constitutive promoter CaMV35S were obtained using an embryogenic line derived from a seed of cv. Picual. After maturation and germination of the transgenic somatic embryos, the plants were micropropagated and acclimated to conditions. The level of expression in the transgenic materials varied greatly among the different lines and was higher in the -780 line. The expression of did not alter the growth of transgenic plants either or in the greenhouse. Different levels of transgene expression also did not affect basal endochitinase activity in the leaves, which was similar to that of control plants. Response to the hemibiotrophic pathogen varied with pathotype. All plants died by 50 days after inoculation with defoliating (D) pathotype V-138, but the response to non-defoliating (ND) strains differed by race: following inoculation with the V-1242 strain (ND, race 2), symptoms appeared after 44-55 days, with line -780 showing the lowest disease severity index. This line also showed good performance when inoculated with the V-1558 strain (ND, race 1), although the differences from the control were not statistically significant. In response to the necrotroph , all the transgenic lines showed a slight delay in disease development, with mean area under the disease progress curve (AUDPC) values 7-15% lower than that of the control.

Avocado consumption is increasing year by year, and its cultivation has spread to many countries with low water availability, which threatens the sustainability and profitability of avocado orchards. However, to date, there is not much information on the behavior of commercial avocado rootstocks against drought. The aim of this research was to evaluate the physiological and molecular responses of ‘Dusa’ avocado rootstock to different levels of water stress. Plants were deficit irrigated until soil water content reached 50% (mild-WS) and 25% (severe-WS) of field capacity. Leaf water potential (Ψw), net CO2 assimilation rates (AN), transpiration rate (E), stomatal conductance (gs), and plant transpiration rates significantly decreased under both WS treatments, reaching significantly lower values in severe-WS plants. After rewatering, mild- and severe-WS plants showed a fast recovery in most physiological parameters measured. To analyze root response to different levels of drought stress, a cDNA avocado stress microarray was carried out. Plants showed a wide transcriptome response linked to the higher degree of water stress, and functional enrichment of differentially expressed genes (DEGs) revealed abundance of common sequences associated with water stress, as well as specific categories for mild-WS and severe-WS. DEGs previously linked to drought tolerance showed overexpression under both water stress levels, i.e., several transcription factors, genes related to abscisic acid (ABA) response, redox homeostasis, osmoprotection, and cell-wall organization. Taken altogether, physiological and molecular data highlight the good performance of ‘Dusa’ rootstock under low-water-availability conditions, although further water stress experiments must be carried out under field conditions.

Trees have a distinctive and generally long juvenile period during which vegetative growth rate is rapid and floral organs do not differentiate. Among trees, the juvenile period can range from 1 year to 15–20 years, although with some forest tree species, it can be longer. Vegetative propagation of trees is usually much easier during the juvenile phase than with mature phase materials. Therefore, reversal of maturity is often necessary in order to obtain materials in which rooting ability has been restored. Micrografting has been developed for trees to address reinvigoration/rejuvenation of elite selections to facilitate vegetative propagation. Generally, shoots obtained after serial grafting have increased rooting competence and develop juvenile traits; in some cases, graft-derived shoots show enhanced in vitro proliferation. Recent advances in graft signaling have shown that several factors, e.g., plant hormones, proteins, and different types of RNA, could be responsible for changes in the scion. The focus of this review includes (1) a discussion of the differences between the juvenile and mature growth phases in trees, (2) successful restoration of juvenile traits through micrografting, and (3) the nature of the different signals passing through the graft union.

Avocado embryogenic cultures were selected for resistance to the culture filtrate (CF) of Rosellinia necatrix, the causal agent of White Root Rot disease. A resistant callus line was obtained through recurrent selections in progressively increasing concentrations of fungal CF (from 60% to 80%). RNA sequencing (RNA-Seq) technology was used to compare the transcriptomic profiles of the avocado embryogenic-callus-resistant line L3 (capable to survive in the presence of 80% CF) and control line AN-9 (not exposed to CF), after 24 h of growth in a medium containing 40% CF. A total of 25,211 transcripts were obtained, of which 4,918 and 5,716 were differentially expressed in the resistant and control line, respectively. Interestingly, exposure of embryogenic callus lines to 40% of R. necatrix exudates induced genes previously reported to be related to avocado defense against fungal diseases (lignin biosynthesis, Pathogenesis Related (PR) proteins, WRKY (WRKYGQK) Transcription Factor (TF), NAC (NAM, ATAF1/2, and CUC2) TF, proteinase inhibitors and Ethylene Response Transcription Factor (ERF), among others), which were accumulated in greater amounts in the resistant line in comparison to the susceptible one. This research will contribute to the understanding of avocado defense against this pathogen, thereby aiding in the selection of resistant avocado rootstocks.

Regeneration of avocado via somatic embryogenesis is difficult due to poor embryo maturation, resulting in low frequencies of germination. In this study, the influence of semi-permeable cellulose acetate membranes and culture media, containing high levels of sucrose along with coconut water, on maturation and germination of somatic embryos of avocado have been evaluated. The culture of embryogenic calli on top of cellulose acetate membranes significantly increased the number of mature, white-opaque embryos that were recovered after 5 weeks of culture. These embryos showed a much more normal appearance and better quality compared with the control embryos, although the embryo size was significantly reduced. To increase the embryo size and to complete maturation, several two-step maturation treatments were tested. The culture of white-opaque somatic embryos in a modified MS medium with B5 macronutrients gelled with 10 g L−1 agar (B5m10A medium) over a 5-week period, followed by 5 additional weeks in B5m10A with 45 g L−1 sucrose and 20 % coconut water, yielded the best results, reducing the percentage of necrotic embryos and the number of calli formed. The beneficial effects of this maturation treatment were enhanced when using embryos that were pre-matured on cellulose acetate membranes. Following this two-step maturation treatment, the germination rate of the control somatic embryos, which were not cultured on cellulose membranes, was lower than 10 %, but it significantly improved when the embryos had been pre-matured on cellulose acetate membranes for 5 weeks, reaching a germination rate close to 40 %. The water availability was significantly reduced when somatic embryos were cultured on cellulose membranes, and after this pre-maturation treatment, the white-opaque embryos showed lower water potential and ABA content compared with the control embryos. These results suggest that culturing over cellulose membranes causes a controlled embryo desiccation that enhances the recovery of plants.

Olive (Olea europaea L.) is the most characteristic and important oil crop of the Mediterranean region. Traditional olive cultivation is based on few tens cultivars of ancient origin. To improve this crop, novel selections with higher tolerance to biotic and abiotic stress, adaptable to high-density planting systems and resilient to climate change are needed; however, breeding programs are hindered by the long juvenile period of this species and few improved genotypes have been released so far. Genetic transformation could be of great value, in the near future, to develop new varieties or rootstocks in a shorter time; in addition, it has currently become an essential tool for functional genomic studies. The recalcitrance of olive tissues to their in vitro manipulation has been the main bottleneck in the development of genetic transformation procedures in this species; however, some important traits such as fungal resistance, flowering or lipid composition have successfully been manipulated through the genetic transformation of somatic embryos of juvenile or adult origin, providing a proof of the potential role that this technology could have in olive improvement. However, the optimization of these protocols for explants of adult origin is a prerequisite to obtain useful materials for the olive industry. In this review, initially, factors affecting plant regeneration via somatic embryogenesis are discussed. Subsequently, the different transformation approaches explored in olive are reviewed. Finally, transgenic experiments with genes of interest undertaken to manipulate selected traits are discussed.

Avocado globular somatic embryos were transformed with three binary vectors, pK7FNF2, pK7RNR2 and pK7S*NF2, harboring the marker genes gfp, DsRed and a gfp-gus fusion gene, respectively. GFP and DsRed fluorescence was detected in embryogenic lines growing in selection medium 2 months after Agrobacterium inoculation. The fluorescence signal was maintained thereafter in transgenic calli, as well as in mature somatic embryos. Red fluorescence in pK7RNR2 transgenic lines was higher and more easily observable than GFP fluorescence. Furthermore, calli transformed with pK7S*NF2, harboring gfp-gus, showed higher level of fluorescence than those transformed with pK7FNF2, containing two gfp. To improve plant recovery, maturated transgenic embryos that failed to germinate or showed an underdeveloped shoot were cultured for 4 weeks in a medium with 1 mg l−1 TDZ and 1 mg l−1 BA after partial removal of cotyledons. A 50% of embryos developed one or several shoots on the cut surface. These embryos were cultured for 4 additional weeks in a medium with 1 mg l−1 BA for shoot elongation and then, shoots were grafted in vitro onto seedling rootstocks. Culture of micrografts in solid MS medium supplemented with 1 mg l−1 BA allowed a 60–80% success rate. Young leaves from transgenic plants showed GFP or DsRed fluorescence located in the nucleus. The results obtained indicate that fluorescent marker genes, especially DsRed, could be useful for early selection of transgenic material and optimization of the transformation parameters in avocado. Furthermore, the protocol established allowed the successful recovery of transgenic plants, one of the main limiting steps in avocado transformation.