Glyphosate excessive use chronically disrupts the shikimate pathway and can affect photosynthesis and yield in citrus trees.

Glyphosate excessive use is reported in Brazilian citrus orchards, whereas there is speculation about its consequences and the published studies are contradictory and inconclusive. This study aimed to describe the possible harmful effects by simulating glyphosate drift directly to the leaves of ∼4-yr-old citrus plants. As major results, glyphosate doses >360 g ae ha-1 increased the shikimate accumulation in leaves (up to 2.3-times above control), which was increased after a second glyphosate application (up to 3.5-times above control), even after a 240-d interval. Interestingly, shikimate accumulation was occasionally related to a dose-response of the herbicide at specific times; however, the doses had their accumulation peak on determined dates. These accumulations were directly correlated to reduced net photosynthesis even months after the glyphosate sprays. Quantum productivity based on electron transport through the photosystem II and apparent electron transport reductions up to 17% were also observed during the entire experiment course. Similarly, quantum productivity based on CO2 assimilation of glyphosate sprayed leaves decreased up to four times compared to the control after the second application. Glyphosate doses >360 g ae ha-1 increased stomatal conductance and transpiration as the carboxylation efficiency decreased, evidencing a carbon drainage in the Calvin-Benson cycle. These metabolic and physiological disturbances suggest possible photooxidative damage and an increase in photorespiration, which may be a mitigation strategy by the citrus plants to glyphosate effects, by the cost of reducing the citrus fruit yield (up to 57%). It is concluded that glyphosate phytotoxicity damages citrus plants over time due to chronic disturbances in the shikimate pathway and photosynthesis, even when there are no symptoms. This study is the first report to demonstrate how glyphosate damages citrus trees beyond the shikimate pathway.

Effects of Deficit Irrigation and Huanglongbing on Sweet Orange Trees.

This study addresses the interactive effects of deficit irrigation and huanglongbing (HLB) infection on the physiological, biochemical, and oxidative stress responses of sweet orange trees. We sought to answer: (i) What are the causes for the reduction in water uptake in HLB infected plants? (ii) Is the water status of plants negatively affected by HLB infection? (iii) What are the key physiological traits impaired in HLB-infected plants? and (iv) What conditions can mitigate both disease severity and physiological/biochemical impairments in HLB-infected plants? Two water management treatments were applied for 11 weeks to 1-year-old-trees that were either healthy (HLB-) or infected with HLB (+) and grown in 12-L pots. Half of the trees were fully irrigated (FI) to saturation, whereas half were deficit-irrigated (DI) using 40% of the water required to saturate the substrate. Our results demonstrated that: reduced water uptake capacity in HLB+ plants was associated with reduced root growth, leaf area, stomatal conductance, and transpiration. Leaf water potential was not negatively affected by HLB infection. HLB increased leaf respiration rates (ca. 41%) and starch synthesis, downregulated starch breakdown, blocked electron transport, improved oxidative stress, and reduced leaf photosynthesis (ca. 57%) and photorespiration (ca.57%). Deficit irrigation reduced both leaf respiration (ca. 45%) and accumulation of starch (ca.53%) by increasing maltose (ca. 20%), sucrose, glucose, and fructose contents in the leaves, decreasing bacterial population (ca. 9%) and triggering a series of protective measures against further impairments in the physiology and biochemistry of HLB-infected plants. Such results provide a more complete physiological and biochemical overview of HLB-infected plants and can guide future studies to screen genetic tolerance to HLB and improve management strategies under field orchard conditions.

Oxidative stress induced by Cu nutritional disorders in Citrus depends on nitrogen and calcium availability.

Nutritional stress caused by copper (Cu) deficiency or toxicity affects fruit production of citrus orchards worldwide, but this could be minimised by fine-tuned fertilisation in the orchards. Two experiments were performed aiming to evaluate the photosynthetic capacity and the antioxidant enzyme activities of Swingle citrumelo seedlings, grown in nutrient solution (NS) with two levels of nitrogen (N) in the first experiment (adequate-N and high-N) and two levels of calcium (Ca) in the second (low-Ca and adequate-Ca). Plants were then exposed to various Cu levels (low, medium and high) for 15 days. Plants under Cu-toxicity exhibited specific effects on reactive oxygen species formation and root-to-shoot plant signalling. Copper absorption was greater with increased Cu concentration in the NS, which reduced plant biomass accumulation, gas exchange measurements, the activity of nitrate reductase and affected Cu partitioning between roots and shoots. Despite these effects, oxidative stress induced by excess-Cu was reduced at the highest N dose when compared to control and, on the contrary, increased with low-Ca supply. Therefore, a rational supply of N or Ca minimises Cu-induced stress damages to roots and leaves of plants, by directly enhancing the antioxidant system and protecting the associated antioxidative enzyme activities, whilst maintaining photosynthesis.

Mechanisms of copper stress alleviation in Citrus trees after metal uptake by leaves or roots.

Nutritional disorders caused by copper (Cu) have affected citrus orchards. Since Cu is foliar sprayed as a pesticide to control citrus diseases, this metal accumulates in the soil. Thereby, we evaluated the effects of Cu leaf absorption after spray of different metal sources, as well as roots absorption on growth, nutritional status, and oxidative stress of young sweet orange trees. Two experiments were carried out under greenhouse conditions. The first experiment was set up with varying Cu levels to the soil (nil Cu, 0.5, 2.0, 4.0 and 8.0 g of Cu per plant as CuSO4.5H2O), whereas the second experiment with Cu application via foliar sprays (0.5 and 2.0 g of Cu per plant) and comparing two metal sources (CuSO4.5H2O or Cu(OH)2). Copper was mainly accumulated in roots with soil supply, but an increase of oxidative stress levels was observed in leaves. On the other hand, Cu concentrations were higher in leaves that received foliar sprays, mainly as Cu(OH)2. However, when sulfate was foliar sprayed, plants exhibited more symptoms of injuries in the canopy with decreased chlorophyll contents and increased hydrogen peroxide and lipid peroxidation levels. Copper toxicity was characterized by sap leakage from the trunk and twigs, which is the first report of this specific Cu excess symptom in woody trees. Despite plants with 8.0 g of Cu soil-applied exhibiting the sap leakage, growth of new plant parts was more vigorous with lower oxidative stress levels and injuries compared to those with 4.0 g of Cu soil-applied (without sap leakage). With the highest level of Cu applied via foliar as sulfate, Cu was eliminated by plant roots, increasing the rhizospheric soil metal levels. Despite citrus likely exhibiting different mechanisms to reduce the damages caused by metal toxicity, such as responsive enzymatic antioxidant system, metal accumulation in the roots, and metal exclusion by roots, excess Cu resulted in damages on plant growth and metabolism when the metal was taken up either by roots or leaves.

Uptake and distribution of soil applied zinc by citrus trees-addressing fertilizer use efficiency with 68Zn labeling.

The zinc (Zn) supply increases the fruit yield of Citrus trees that are grown, especially in the highly weathered soils of the tropics due to the inherently low nutrient availability in the soil solution. Leaf sprays containing micronutrients are commonly applied to orchards, even though the nutrient supply via soil could be of practical value. This study aimed to evaluate the effect of Zn fertilizers that are applied to the soil surface on absorption and partitioning of the nutrient by citrus trees. A greenhouse experiment was conducted with one-year-old sweet orange trees. The plants were grown in soils with different textures (18.1 or 64.4% clay) that received 1.8 g Zn per plant, in the form of either ZnO or ZnSO4 enriched with the stable isotope 68Zn. Zinc fertilization increased the availability of the nutrient in the soil and the content in the orange trees. Greater responses were obtained when ZnSO4 was applied to the sandy loam soil due to its lower specific metal adsorption compared to that of the clay soil. The trunk and branches accumulated the most fertilizer-derived Zn (Zndff) and thus represent the major reserve organ for this nutrient in the plant. The trees recovered up to 4% of the applied Zndff. Despite this relative low recovery, the Zn requirement of the trees was met with the selected treatment based on the total leaf nutrient content and increased Cu/Zn-SOD activity in the leaves. We conclude that the efficiency of Zn fertilizers depends on the fertilizer source and the soil texture, which must be taken into account by guidelines for fruit crop fertilization via soil, in substitution or complementation of traditional foliar sprays.