Physiological characterization of the wild almond Prunus arabica stem photosynthetic capability

Leaves are the major plant tissue for transpiration and carbon fixation in deciduous trees. In harsh habitats, atmospheric CO2 assimilation via stem photosynthesis is common, providing extra carbon gain to cope with the detrimental conditions. We studied two almond species, the commercial Prunus dulcis cultivar “Um-el-Fahem” and the rare wild Prunus arabica. Our study revealed two distinctive strategies for carbon gain in these almond species. While, in P. dulcis, leaves possess the major photosynthetic surface area, in P. arabica, green stems perform this function, in particular during the winter after leaf drop. These two species' anatomical and physiological comparisons show that P. arabica carries unique features that support stem gas exchange and high-gross photosynthetic rates via stem photosynthetic capabilities (SPC). On the other hand, P. dulcis stems contribute low gross photosynthesis levels, as they are designed solely for reassimilation of CO2 from respiration, which is termed stem recycling photosynthesis (SRP). Results show that (a) P. arabica stems are covered with a high density of sunken stomata, in contrast to the stomata on P. dulcis stems, which disappear under a thick peridermal (bark) layer by their second year of development. (b) P. arabica stems contain significantly higher levels of chlorophyll compartmentalized to a mesophyll-like, chloroplast-rich, parenchyma layer, in contrast to rounded-shape cells of P. dulcis's stem parenchyma. (c) Pulse amplitude-modulated (PAM) fluorometry of P. arabica and P. dulcis stems revealed differences in the chlorophyll fluorescence and quenching parameters between the two species. (d) Gas exchange analysis showed that guard cells of P. arabica stems tightly regulate water loss under elevated temperatures while maintaining constant and high assimilation rates throughout the stem. Our data show that P. arabica uses a distinctive strategy for tree carbon gain via stem photosynthetic capability, which is regulated efficiently under harsh environmental conditions, such as elevated temperatures. These findings are highly important and can be used to develop new almond cultivars with agriculturally essential traits

TRANSCRIPTOME DYNAMICS IN MANGO FRUIT PEEL REVEALS MECHANISMS OF CHILLING STRESS

Cold storage is considered the most effective method for prolonging fresh produce storage. However, subtropical fruit is sensitive to cold. Symptoms of chilling injury in mango include red and black spots that start from discolored lenticels and develop into pitting. The response of ‘Keitt’ mango fruit to chilling stress was monitored by transcriptomic, physiological and microscopic analyses. Transcriptomic changes in the mango fruit peel were evaluated during optimal (12°C) and suboptimal (5°C) cold storage. Two days of chilling stress upregulated genes involved in the plant stress response, including those encoding transmembrane receptors, calcium-mediated signal transduction, NADPH oxidase, MAP kinases and WRKYs, which can lead to cell death. Indeed, cell death was observed around the discolored lenticels after 19 days of cold storage at 5°C. Localized cell death and cuticular opening in the lumen of discolored lenticels were correlated with increased general decay during shelf-life storage, possibly due to fungal penetration. We also observed increased phenolics accumulation around the discolored lenticels, which was correlated with the biosynthesis of phenylpropanoids that were probably transported from the resin ducts. Increased lipid peroxidation was observed during chilling injury by both the biochemical malondialdehyde method and a new non-destructive luminescent technology, correlated to upregulation of the α-linolenic acid oxidation pathway. Genes involved in sugar metabolism were also induced, possibly to maintain osmotic balance. This analysis provides an in-depth characterization of mango fruit response to chilling stress and could lead to the development of new tools, treatments and strategies to prolong cold storage of subtropical fruit

ToBRFV Infects the Reproductive Tissues of Tomato Plants but Is Not Transmitted to the Progenies by Pollination

Tomato brown rugose fruit virus (ToBRFV), a newly identified Tobamovirus, has recently emerged as a significant pathogen of tomato plants (Solanum lycopersicum). The virus can evade or overcome the known tobamovirus resistance in tomatoes, i.e., Tm-1, Tm-2, and its allele Tm-22. ToBRFV was identified for the first time only a few years ago, and its interactions with the tomato host are still not clear. We investigated ToBRFV’s presence in the reproductive tissues of tomato using fluorescent in situ hybridization (FISH) and RT-PCR. In infected plants, the virus was detected in the leaves, petals, ovary, stamen, style, stigma, and pollen grains but not inside the ovules. Fruits and seeds harvested from infected plants were contaminated with the virus. To test whether the virus is pollen transmitted, clean mother plants were hand pollinated with pollen from ToBRFV-infected plants and grown to fruit. None of the fruits and seeds harvested from the pollinated clean mother plants contained ToBRFV. Pollen germination assays revealed the germination arrest of ToBRFV-infected pollen. We concluded that ToBRFV might infect reproductive organs and pollen grains of tomato but that it is not pollen transmitted

Sucrose Synthase and Fructokinase Are Required for Proper Meristematic and Vascular Development

Sucrose synthase (SuSy) and fructokinase (FRK) work together to control carbohydrate flux in sink tissues. SuSy cleaves sucrose into fructose and UDP-glucose; whereas FRK phosphorylates fructose. Previous results have shown that suppression of the SUS1,3&4 genes by SUS-RNAi alters auxin transport in the shoot apical meristems of tomato plants and affects cotyledons and leaf structure; whereas antisense suppression of FRK2 affects vascular development. To explore the joint developmental roles of SuSy and FRK, we crossed SUS-RNAi plants with FRK2-antisense plants to create double-mutant plants. The double-mutant plants exhibited novel phenotypes that were absent from the parent lines. About a third of the plants showed arrested shoot apical meristem around the transition to flowering and developed ectopic meristems. Use of the auxin reporter DR5::VENUS revealed a significantly reduced auxin response in the shoot apical meristems of the double-mutant, indicating that auxin levels were low. Altered inflorescence phyllotaxis and significant disorientation of vascular tissues were also observed. In addition, the fruits and the seeds of the double-mutant plants were very small and the seeds had very low germination rates. These results show that SUS1,3&4 and FRK2 enzymes are jointly essential for proper meristematic and vascular development, and for fruit and seed development

Proximal and Distal Parts of Sweetpotato Adventitious Roots Display Differences in Root Architecture, Lignin, and Starch Metabolism and Their Developmental Fates

Sweetpotato is an important food crop globally, serving as a rich source of carbohydrates, vitamins, fiber, and micronutrients. Sweetpotato yield depends on the modification of adventitious roots into storage roots. The underlying mechanism of this developmental switch is not fully understood. Interestingly, storage-root formation is manifested by formation of starch-accumulating parenchyma cells and bulking of the distal part of the root, while the proximal part does not show bulking. This system, where two parts of the same adventitious root display different developmental fates, was used by us in order to better characterize the anatomical, physiological, and molecular mechanisms involved in sweetpotato storage-root formation. We show that, as early as 1 and 2 weeks after planting, the proximal part of the root exhibited enhanced xylem development together with increased/massive lignin deposition, while, at the same time, the distal root part exhibited significantly elevated starch accumulation. In accordance with these developmental differences, the proximal root part exhibited up-regulated transcript levels of sweetpotato orthologs of Arabidopsis vascular-development regulators and key genes of lignin biosynthesis, while the distal part showed up-regulation of genes encoding enzymes of starch biosynthesis. All these recorded differences between proximal and distal root parts were further enhanced at 5 weeks after planting, when storage roots were formed at the distal part. Our results point to down-regulation of fiber formation and lignification, together with up-regulation of starch biosynthesis, as the main events underlying storage-root formation, marking/highlighting several genes as potential regulators, providing a valuable database of genes for further research

Alternate bearing in citrus: changes in the expression of flowering control genes and in global gene expression in ON- versus OFF-crop trees.

Alternate bearing (AB) is the process in fruit trees by which cycles of heavy yield (ON crop) one year are followed by a light yield (OFF crop) the next. Heavy yield usually reduces flowering intensity the following year. Despite its agricultural importance, how the developing crop influences the following year's return bloom and yield is not fully understood. It might be assumed that an 'AB signal' is generated in the fruit, or in another organ that senses fruit presence, and moves into the bud to determine its fate-flowering or vegetative growth. The bud then responds to fruit presence by altering regulatory and metabolic pathways. Determining these pathways, and when they are altered, might indicate the nature of this putative AB signal. We studied bud morphology, the expression of flowering control genes, and global gene expression in ON- and OFF-crop buds. In May, shortly after flowering and fruit set, OFF-crop buds were already significantly longer than ON-crop buds. The number of differentially expressed genes was higher in May than at the other tested time points. Processes differentially expressed between ON- and OFF-crop trees included key metabolic and regulatory pathways, such as photosynthesis and secondary metabolism. The expression of genes of trehalose metabolism and flavonoid metabolism was validated by nCounter technology, and the latter was confirmed by metabolomic analysis. Among genes induced in OFF-crop trees was one homologous to SQUAMOSA PROMOTER BINDING-LIKE (SPL), which controls juvenile-to-adult and annual phase transitions, regulated by miR156. The expression pattern of SPL-like, miR156 and other flowering control genes suggested that fruit load affects bud fate, and therefore development and metabolism, a relatively long time before the flowering induction period. Results shed light on some of the metabolic and regulatory processes that are altered in ON and OFF buds

Shoot Regeneration Is Not a Single Cell Event

Shoot regeneration is a key tool of modern plant biotechnology. While many researchers use this process empirically, very little is known about the early molecular genetic factors and signaling events that lead to shoot regeneration. Using tobacco as a model system, we found that the inductive events required for shoot regeneration occur in the first 4–5 days following incubation on regeneration medium. Leaf segments placed on regeneration medium did not produce shoots if removed from the medium before four days indicating this time frame is crucial for the induction of shoot regeneration. Leaf segments placed on regeneration medium for longer than five days maintain the capacity to produce shoots when removed from the regeneration medium. Analysis of gene expression during the early days of incubation on regeneration medium revealed many changes occurring with no single expression pattern evident among major gene families previously implicated in developmental processes. For example, expression of Knotted gene family members increased during the induction period, whereas transcription factors from the Wuschel gene family were unaltered during shoot induction. Expression levels of genes involved in cell cycle regulation increased steadily on regeneration medium while expression of NAC genes varied. No obvious possible candidate genes or developmental processes could be identified as a target for the early events (first few days) in the induction of shoot regeneration. On the other hand, observations during the early stages of regeneration pointed out that regeneration does not occur from a single cell but a group of cells. We observed that while cell division starts just as leaf segments are placed on regeneration medium, only a group of cells could become shoot primordia. Still, these primordia are not identifiable during the first days

Characterization of Two Ethephon-Induced IDA-Like Genes from Mango, and Elucidation of Their Involvement in Regulating Organ Abscission

In mango (Mangifera indica L.), fruitlet abscission limits productivity. The INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) peptide acts as a key component controlling abscission events in Arabidopsis. IDA-like peptides may assume similar roles in fruit trees. In this study, we isolated two mango IDA-like encoding-genes, MiIDA1 and MiIDA2. We used mango fruitlet-bearing explants and fruitlet-bearing trees, in which fruitlets abscission was induced using ethephon. We monitored the expression profiles of the two MiIDA-like genes in control and treated fruitlet abscission zones (AZs). In both systems, qRT-PCR showed that, within 24 h, both MiIDA-like genes were induced by ethephon, and that changes in their expression profiles were associated with upregulation of different ethylene signaling-related and cell-wall modifying genes. Furthermore, ectopic expression of both genes in Arabidopsis promoted floral-organ abscission, and was accompanied by an early increase in the cytosolic pH of floral AZ cells—a phenomenon known to be linked with abscission, and by activation of cell separation in vestigial AZs. Finally, overexpression of both genes in an Atida mutant restored its abscission ability. Our results suggest roles for MiIDA1 and MiIDA2 in affecting mango fruitlet abscission. Based on our results, we propose new possible modes of action for IDA-like proteins in regulating organ abscission

Gibberellin Promotes Sweetpotato Root Vascular Lignification and Reduces Storage-Root Formation

Sweet potato yield depends on a change in the developmental fate of adventitious roots into storage-roots. The mechanisms underlying this developmental switch are still unclear. We examined the hypothesis claiming that regulation of root lignification determines storage root formation. We show that application of the plant hormone gibberellin increased stem elongation and root gibberellin levels, while having inhibitory effects on root system parameters, decreasing lateral root number and length, and significantly reducing storage root number and diameter. Furthermore, gibberellin enhanced root xylem development,caused increased lignin deposition, and, at the same time, decreased root starch accumulation. In accordance with these developmental effects, gibberellin application upregulated expression levels of sweet potato orthologues of Arabidopsis vascular development regulators (IbNA075, IbVND7, and IbSND2) and of lignin biosynthesis genes(IbPAL, IbC4H, Ib4CL, IbCCoAOMT, and IbCAD), while down regulating starch biosynthesis genes (IbAGPase and IbGBSS) in the roots. Interestingly, gibberellin down regulated root expression levels of orthologues of the Arabidopsis BREVIPEDICELLUS transcription factor (IbKN2 and IbKN3), regulator of meristem maintenance. The results substantiate our hypothesis and mark gibberellin as an important player in regulation of sweet potato root development, suggesting that increased fiber formation and lignification inhibit storage root formation and yield. Taken together, our findings provide insight into the mechanisms underlying sweet potato storage-root formation and provide a valuable database of genes for further research

High temperature environment reduces olive oil yield and quality.

Global warming is predicted to have a negative effect on plant growth due to the damaging effect of high temperatures. In order to address the effect of high temperature environments on olive oil yield and quality, we compared its effect on the fruit development of five olive cultivars placed in a region noted for its high summer temperatures, with trees of the same cultivars placed in a region of relatively mild summers. We found that the effects of a high temperature environment are genotype dependent and in general, high temperatures during fruit development affected three important traits: fruit weight, oil concentration and oil quality. None of the tested cultivars exhibited complete heat stress tolerance. Final dry fruit weight at harvest of the 'Barnea' cultivar was not affected by the high temperature environment, whereas the 'Koroneiki', 'Coratina', 'Souri' and 'Picholine' cultivars exhibited decreased dry fruit weight at harvest in response to higher temperatures by 0.2, 1, 0.4 and 0.2 g respectively. The pattern of final oil concentration was also cultivar dependent, 'Barnea', 'Coratina' and 'Picholine' not being affected by the high temperature environment, whereas the 'Koroneiki' and 'Souri' cultivars showed a decreased dry fruit oil concentration at harvest under the same conditions by 15 and 8% respectively. Regarding the quality of oil produced, the 'Souri' cultivar proved more tolerant to a high temperature environment than any other of the cultivars analyzed in this study. These results suggest that different olive cultivars have developed a variety of mechanisms in dealing with high temperatures. Elucidation of the mechanism of each of these responses may open the way to development of a variety of olives broadly adapted to conditions of high temperatures