Citrus Macrophylla Research Articles

A Transmission Assay of 'Candidatus Liberibacter asiaticus' Using Citrus Phloem Sap and Topical Feeding to Its Insect Vector, Diaphorina citri.

'Candidatus Liberibacter asiaticus', the putative causal agent of citrus greening disease, is transmitted by the Asian citrus psyllid, Diaphorina citri, in a propagative, circulative, and persistent manner. Unfortunately, 'Ca. L. asiaticus' is not yet available in pure culture to carry out Koch's postulates and to confirm its etiology. When a pure culture is available, an assay to test its infectivity in both the insect vector and the plant host will be crucial. Herein, we described a transmission assay based on the use of phloem sap extracted from infected citrus plants and topical feeding to D. citri nymphs. Phloem sap was collected by centrifugation, diluted with 0.1 M phosphate buffer pH 7.4 containing 20% (wt/vol) sucrose and 0.1% ascorbic acid (wt/vol) as an antioxidant, and delivered to third through fifth instar nymphs by placing droplets on the mouthparts. Nymphs unfolded the stylets and acquired the phloem sap containing the bacterial pathogen. Nymphs were then placed onto Citrus macrophylla seedlings (10 nymphs per seedling) for an inoculation period of 2 weeks. A transmission rate of up to 80% was recorded at 6 months postinoculation. The method could be a powerful tool to test the transmissibility of the bacterial pathogen after various treatments to reduce the viability of the bacteria or to block its transmission. In addition, it might be a potent assay to achieve Koch's postulates if a pure culture of 'Ca. L. asiaticus' becomes available.

An engineered citrus tristeza virus (T36CA)-based vector induces gene-specific RNA silencing and is graft transmissible to commercial citrus varieties.

A protein-expressing citrus tristeza virus (CTV)-based vector construct, pT36CA-V1.3, obtained from a California isolate of the T36 strain (T36CA), was retooled into a virus induced gene silencing (VIGS) system intended for use with studies of California citrus. VIGS constructs engineered with a truncated Citrus macrophylla (Cm) PHYTOENE DESATURASE (PDS) gene sequence in the sense or anti-sense orientation worked equally well to silence the endogenous CmPDS gene. In a parallel effort to optimize vector performance, two non-synonymous nucleotides in open reading frame 1a of pT36CA-V1.3 were replaced with those conserved in the reference sequences from the T36CA cDNA library. The resulting viruses, T36CA-V1.4 (with one amino acid modification: D760N) and T36CA-V1.5 (with two amino acid modifications: D760N and P1174L), along with T36CA-V1.3 were individually propagated in Nicotiana benthamiana and C. macrophylla plants. Enzyme-linked immunosorbent assay (ELISA) measurements of extracts of the newly emerged leaves suggested that all three viruses accumulated to similar levels in N. benthamiana plants at 5 week-post-inoculation. ELISA values of T36CA-V1.4- and -V1.5-infected C. macrophylla samples were significantly higher than that of T36CA-V1.3-infected samples within an 8 to 12 month-post-inoculation (mpi) window, suggesting a higher accumulation of T36CA-V1.4 and -V1.5 than T36CA-V1.3. However, at 36 mpi, the ELISA values suggested that all three viruses accumulated to similar levels. When C. macrophylla plants infected with each of the three viruses were grafted to commercial citrus varieties, a limited number of receptor plants became infected, demonstrating a weak but nonetheless (the first) successful delivery of T36CA to California-grown commercial citrus.

Effects of Salinity on ‘Fino 95’ Lemon Trees Inoculated with Arbuscular Mycorrhizal Fungi

An experiment was conducted with two-year-old ‘Fino 95’ lemon plants (Citrus limon Burm. f. cv. Fino) grafted on C. macrophylla (Citrus macrophylla Wester) to study the effects of salinity on plants inoculated with arbuscular mycorrhizal fungi (AMF). Half of the inoculated (+AM) and non-inoculated (−AM) plants were irrigated with half-strength Hoagland solution, and the remainder were inoculated with half-strength Hoagland solution + 30 mM NaCl. Ninety-eight days later, results showed that AMF had alleviated the negative effect of salinity on growth. Inoculation with AMF provided some protection against the damage that salinity caused on cellular membranes and improved the plant water status and turgor under saline conditions (Ψx and Π increased by 16% and 48%, respectively). The responses of mineral nutrition to salinity and AMF treatments were complex. P concentrations in the leaves and roots of +AM plants were lower than in those of −AM plants, but inoculation improved Ca2+ (by 20%), Mg2+ (24%), Fe2+ (21%), and Zn2+ (7%) nutrition in roots and also the Mg2+/Na+ ratio in leaves (33%), reducing the antagonistic effect of Na+ on Mg2+ nutrition in salt-treated plants. AMF could protect plants against salt stress through the maintenance of the gas exchange capacity and due to a better antioxidant response. All these positive effects of AMF contributed to mitigating the harmful effects of salinity stress on the plant growth performance of lemon trees grafted on C. macrophylla rootstock under salinity conditions.

Citrus-mediated gene silencing of cytochrome P450 suppresses insecticide resistance and increases mortality in Diaphorina citri.

Asian citrus psyllid, Diaphorina citri, is a hemipteran that vectors the causal pathogen of citrus greening disease, or huanglongbing (HLB). HLB is a tree killing disease that has severely limited citrus production globally. Unfortunately, there is no cure for this disease, and mitigation depends on multiple insecticide applications to reduce vector populations. Silencing of cytochrome P450 expression associated with detoxification enzymes by RNA interference is known to increase susceptibility of D. citri to insecticides. However, dsRNA was previously introduced into psyllids by topical applications. The possible application of this technology for pest management will require effective field delivery of the dsRNA. Therefore, we evaluated a virus vector (Citrus tristeza virus; 'mild strain' T36) to deliver gene silencing directly to this sap-sucking insect via plant phloem. Citrus macrophylla plants inoculated with CTV expressing a truncated consensus sequence of CYP450 (CTV-tCYP450) constantly produced small interfering RNA in the plant phloem that targeted five cytochrome p540 (CYP450) genes in D. citri. Insecticide susceptible D. citri reared on citrus infected with CTV-tCYP450 were subsequently more susceptible to imidacloprid, fenpropathrin, carbaryl, and chlorpyrifos than those reared on citrus infected with wildtype CTV or non-infected negative controls. Additionally, nymph survival and adult lifespan were significantly reduced when psyllids were reared on CTV-tCYP450 citrus plants compared with controls. Interestingly, similar results were obtained after one and two generations of rearing. Finally, field-collected psyllids from areas with known broad-spectrum insecticide resistance were rendered more susceptible to imidacloprid and fenpropathrin after feeding on CTV-tCYP450 citrus trees as compared with those reared on controls. The integration of citrus-mediated RNA inference targeting psyllid detoxification enzymes could function as a resistance management tool and reduce insecticide input in an integrated pest management program for HLB. © 2024 Society of Chemical Industry.

Novel and widely spread citrus rootstocks behavior in response to salt stress

Irrigation water salinity is a major limiting factor in arid and semi-arid environments. The use of rootstock is one of the appropriate agronomic techniques that attempt to support and limit the negative effect of salinity. The objective of this study was to evaluate the behavior of eight one-year-old rootstocks grown in pots under salt stress conditions (30 and 60 mM NaCl). Some genotypes, i.e. C35 citrange, Bitters, Carpenter and Furr citrandarins were recently released or spread and poor information on their behavior in saline conditions is available, while Carrizo citrange, Swingle Citrumelo, Citrus volkameriana and Citrus macrophylla are spread since long ago in the Mediterranean basin. The results clearly demonstrated that the most salt sensitive genotypes were Carrizo and C35 citrange, that reduced morphological and gas exchanges performances. Furr, Bitters and Carpenter citrandarins revealed good physiological and hormonal behavior. An intense antioxidant enzymatic activity was noted in C35 and Carrizo citranges, while Furr showed a decrease in malondialdehyde and antioxidant enzymatic activities at 60 mM NaCl. Its tolerance to saline water was also confirmed by transcriptomic analyses.

The Combination of Citrus Rootstock and Scion Cultivar Influences Trioza erytreae (Hemiptera: Triozidae) Survival, Preference Choice and Oviposition.

Trioza erytreae (Del Guercio, 1918) (Hemiptera: Triozidae) is a citrus pest which produces gall symptoms on leaves and transmits bacteria associated with the citrus disease Huanglongbing, 'Candidatus Liberibacter' spp. In the present work, the biology and behaviour of T. erytreae were studied in different rootstock-cultivar combinations. Six rootstocks were used, Flying dragon (FD), 'Cleopatra' mandarin (CL), Carrizo citrange (CC), Forner-Alcaide no.5 (FA5), Forner-Alcaide no.517 (FA517) and Citrus macrophylla (CM), and six scion cultivars: 'Star Ruby', 'Clemenules', 'Navelina', 'Valencia Late', 'Fino 49' and 'Ortanique'. Survival and oviposition were evaluated in a no-choice trial, and preference in a choice trial, all of them under greenhouse conditions. Trioza erytreae did not show a clear settle preference for any citrus combination. However, it was able to lay more eggs in 'Fino 49' grafted on CC than on FD. In terms of survival, 'Ortanique' grafted onto FA5 was more suitable than when grafted onto FA517, and in the case of 'Valencia Late', when it was grafted onto CM rather than CC. Our results showed that T. erytreae behave differently depending on the citrus combination.

Proline accumulation and antioxidant response are crucial for citrus tolerance to UV-B light-induced stress.

Plants face a wide range of biotic and abiotic stress conditions, which are further intensified by climate change. Among these stressors, increased irradiation in terms of intensity and wavelength range can lead to detrimental effects, such as chlorophyll degradation, destruction of the PSII reaction center, generation of ROS, alterations to plant metabolism, and even plant death. Here, we investigated the responses of two citrus genotypes, Citrus macrophylla (CM), and Troyer citrange (TC) to UV-B light-induced stress, by growing plants of both genotypes under control and UV-B stress conditions for 5 days to evaluate their tolerance mechanisms. TC seedlings had higher sensitivity to UV-B light than CM seedlings, as they showed more damage and increased levels of oxidative harm (indicated by the accumulation of MDA). In contrast, CM seedlings exhibited specific adaptive mechanisms, including accumulation of higher levels of proline under stressful conditions, and enhanced antioxidant capacity, as evidenced by increased ascorbate peroxidase activity and upregulation of the CsAPX2 gene. Phytohormone accumulation patterns were similar in both genotypes, with a decrease in ABA content in response to UV-B light. Furthermore, expression of genes involved in light perception and response was specifically affected in the tolerant CM seedlings, which exhibited higher expression of CsHYH/CsHY5 and CsRUP1-2 genes. These findings underscore the importance of the antioxidant system in citrus plants subjected to UV-B light-induced stress and suggest that CsHYH/CsHY5 and CsRUP1-2 could be considered genes associated with tolerance to such challenging conditions.

Physiological and Growth Responses of W. Murcott Tangor Grafted on Four Rootstocks under Water Restriction

Citrus orchards in semi-arid regions are increasingly exposed to drought conditions due to climate change. This study compared the physiological and growth responses of ‘W. Murcott’ tangor (WM) grafted onto Citrus macrophylla (M), Swingle citrumelo (SC), C-35 citrange (C35), or bitter citrandarin (C22) rootstock subjected to two irrigation treatments: daily irrigation to replace 100% of the water lost daily by evapotranspiration (ET; control treatment) or daily irrigation to replace 75% of the water lost daily by ET (water deficit treatment). For trees in each treatment, leaf gas exchange, relative chlorophyll content, chlorophyll fluorescence, midday stem water potential, trunk cross-sectional area, and shoot length were measured 46 days after treatments were initiated. The results showed that WM on SC or C22 rootstock exhibited isohydric behavior, where decreased stomatal conductance limited transpiration in the water deficit treatment. WM on M rootstock exhibited an anisohydric response in the water deficit treatment, where there was no stomatal control of water loss by transpiration. Among the rootstocks tested for WM, the most tolerant to soil water deficit was SC, whereas trees on M rootstock were the most negatively affected by soil water deficit.

Contribution to the optimization of citrus (Citrus clementina Hort. Ex Tanaka) fruit fertilization using mobile lysimetry in orchards of the Souss-Massa region, Morocco

The growth and development of citrus trees and the quality of their fruits are significantly influenced by the essential role of mineral plant nutrition. This study aimed to improve the productivity of the citrus (Citrus clementina Hort. Ex Tanaka) orchards by optimizing mineral nutrition through well-monitored fertilization and fertigation aspects using lysimetry. The first phase consisted of studying the behavior of the nutrients in the soil solution at the high root concentration level, analysis of the nutritional status of five varieties of clementines (Sidi Aissa, Cadoux, Orogrande, Nules, and Nour), and the variation of leaf composition in major elements (Nitrogen, phosphorus, potassium, magnesium, and calcium) for the five varieties along phenological stages, through the exploitation of the results of analyses carried out for the management of mineral nutrition in 47 plots of citrus fruit in the Souss-Massa region with the use of lysimetry. The results obtained in this first part revealed an important variation of the nutrients in the soil solution (55.24%) in terms of water inputs, fertilizers, and edaphic conditions, as well as a large variation of foliar compositions (62.98%). The second phase, consisting of regular monitoring of the mineral nutrition dynamics targeting the “Nules” variety grafted on “Citrus macrophylla” affirmed the importance of the citrus fertilization approach for determining the availability, distribution, nutrient interactions in soil solution and plant response by regular leaf diagnostics. Thus, mobile lysimetry offered a powerful tool for achieving both productivity and sustainability in citrus fertilization programs.

Conventional vs. Organic: Evaluation of Nutritional, Functional and Sensory Quality of Citrus limon.

Organic farming is growing rapidly worldwide since it is perceived as more respectful of the environment than conventional farming. In this sense, organic agriculture is highly appreciated by consumers since consumers around the world believe that organic food has a higher content of beneficial compounds for health and consider it of higher quality. For that reason, the objective of this research was to evaluate the nutritional, sensorial, and functional quality of the 'Fino 49' lemon grafted on Citrus macrophylla in conventional and organic cultivation. Fatty acids, amino acids, total phenol, and polyphenols were quantified, antioxidant activity was measured, and sensory descriptive analysis was performed. Conventional farming led to an increase in amino acid content (641 mg L-1) and an increase in polyunsaturated fatty acids (254 mg 100 g-1) and monounsaturated fatty acids (37.61 mg 100 g-1). On the other hand, organically produced lemon fruits had better sensory profile (highlighting overall aroma (6.5), lemon odor (6.8), sourness (5.8), floral (0.6), and fresh lemon flavor (9.8)), and lower thrombogenicity index (0.15). The type of cultivation (organic and conventional) had no influence on the antioxidant activity (~1.60, ~3.08, and ~4.16 mmol Trolox L-1 for ABTS+, DPPH•, and FRAP, respectively) and polyphenols content (85.51 and 86.69 conventional and organic, respectively). However, to establish the advantages and disadvantages of different types of cultivation on lemon quality more studies are needed.

Influence of rootstock on yield, morphological, biochemical and sensory characteristics of 'Afourer' variety mandarins

Rootstock's can affect the characteristics and quality of citrus fruit. Knowing which rootstocks produce better quality fruit is very important in late mandarin varieties, due to their high economic value. The objective of this study is to know the effects on yield and the main morphological, biochemical and sensory properties of the late mandarin variety 'Afourer' when cultivated on three different rootstocks [Citrus macrophylla Wester (CM), Mandarino Cleopatra (MC) and Citrange Carrizo (CC)]. The morphological properties (weight and dimensions) and colorimetric characteristics (external and internal fruit color) were analyzed. The main biochemical characteristics, including total antioxidant activity analyzed by ABTS and DPPH methods, organic acids and sugars determined by HPLC, and juice metabolomics by 1H-NMR, were also investigated. In addition, the juice quality was assessed by a sensory panel. The CC rootstock produced up to 77 kg of fruit/tree more than MC. Fruits cultivated on MC and CC had significantly larger dimensions, a greater orange coloration and up to 17 % more juice than fruits cultivated on CM. In terms of biochemical properties, there were no significant differences in most of the properties analyzed among the three rootstocks, highlighting only that the CM rootstock induced 13 % more fructose in the fruits than the rest of the rootstocks. Fruits cultivated on the CC and MC rootstocks had a better taste when evaluated by sensory analysis than those cultivated on the CM rootstock.

Stem-pitting caused by Citrus tristeza virus is associated with increased phloem occlusion

Stem-pitting (SP) disease results from disruption of normal phloem and xylem development. In citrus, a characteristic manifestation of SP caused by Citrus tristeza virus (CTV) is phloem regeneration. We hypothesized that phloem regeneration occurs due to reduced functionality of CTV infected phloem cells. To examine phloem cell occlusions in CTV-SP, we analyzed callose and phloem-protein (PP) accumulation in Citrus macrophylla trees infected with CTV mutants exhibiting different SP phenotypes from very mild (CTVΔp13) to severe (CTVΔp33), in addition to full-length CTV and healthy plants. CTV infection was accompanied by callose and PP accumulation in the phloem. With the increase in the SP symptoms from very mild to severe, there was a constant increase in the levels of callose and PP, accompanied by an increase in PHLOEM-PROTEIN 2 and a decrease in BETA-1,3-GLUCANASE gene expression levels. These results indicate that SP symptom development is associated with increased phloem occlusion.

Gene silencing of cathepsins B and L using CTV-based, plant-mediated RNAi interferes with ovarial development in Asian citrus psyllid (ACP), Diaphorina citri.

Diaphorina citri Kuwayama (Hemiptera: Liviidae) is a vector of the bacteria Candidatus Liberibacter americanus (CLam) and Candidatus Liberibacter asiaticus (CLas), which are phloem-restricted and associated with the most important and destructive worldwide citrus disease, Huanglongbing (HLB). Currently, no cure for HLB has been described. Therefore, measures have focused on reducing D. citri populations. In these insects, cathepsin B (DCcathB) and L (DCcathL) enzymes play an important role in digestion, and are involved in embryogenesis, immune defense, and ecdysis. In this study, we used a CTV-based vector to deliver dsRNA (CTV-dsRNA) into Citrus macrophylla plants targeting DCcathB and DCcathL genes in D. citri that fed on the phloem of these CTV-RNAi infected plants. Subsequently, we evaluated expression of DCcathB and DCcathL genes as well as the Vitellogenin (Vg) gene by RT-qPCR in D. citri fed on CTV-dsRNA occurring in plant phloem. It was found that a defective phenotype in D. citri females as a result of knockdown of DCcathB and DCcathL genes mediated by CTV dsRNA. These results showed that Psyllids fed on plants treated with the CTV-dsRNA exhibited downregulation of the Vg gene, one of the most important genes associated with embryogenic and female development, which was associated with dsRNA-mediated silencing of the two cathepsin genes. Based on our findings, a CTV-based strategy for delivering RNAi via plants that targets DCcathB and DCcathL genes may represent a suitable avenue for development of dsRNA-based tools to manage D. citri that limits the spread of HLB.

Nutritional Performance of Five Citrus Rootstocks under Different Fe Levels.

Iron is an essential micronutrient for citrus, playing an important role in photosynthesis and yield. The aim of this paper was to evaluate the tolerance to Fe deficiency of five citrus rootstocks: sour orange (S), Carrizo citrange (C), Citrus macrophylla (M), Troyer citrange (T), and Volkamer lemon (V). Plants were grown for 5 weeks in nutrient solution that contained the following Fe concentrations (in µM): 0, 5, 10, 15, and 20. At the end of the experiment, biomass (dry weight-DW), leaf area, total leaf chlorophyll (CHL), and the activity of root chelate reductase (FCR) were recorded. Additionally, the mineral composition of roots (R) and shoots (S) was evaluated. Principal component analysis was used to study the relationships between all parameters and, subsequently, the relations between rootstocks. In the first component, N-S, P-S, Ca-S, Cu-S, Zn-S, Mn-S, Zn-R, and Mn-R concentrations were related to leaf CHL and FCR. Increases in leaf CHL, Mg-R, and DW (shoots and roots) were inversely related to Cu-R, which was shown in the second component. The values obtained were consistent for V10, C15, and C20, but in contrast for S0 and S5. In conclusion, micronutrient homeostasis in roots and shoots of all rootstocks were affected by Fe stress conditions. The Fe/Cu ratio was significantly related to CHL, which may be used to assist rootstock performance.

Effects of low nitrogen supply on biochemical and physiological parameters related to nitrate and water, involving nitrate transporters and aquaporins in Citrus macrophylla.

A reduction in chemical N-based fertillizer was investigated in Citrus plants. As N and water uptake are connected, the relationship between the physiological response to reductions in N was studied in relation to N metabolism and water. We examined the response of new and mature leaves and roots of Citrus macrophylla, grown under controlled conditions, and given different concentrations of N: 16, 8 or 4 mM. Differences in growth and development were determined for biochemical (mineral content, photosynthetic pigments, proteins and nitrate and nitrite reductase activity), physiological (photosynthesis and transpiration), and molecular (relative expression of nitrate transporters and aquaporins) parameters. Only plants given 4 mM N showed a reduction in growth. Although there were changes in NR activity, protein synthesis, and chlorophyll content in both 8 and 4 mM N plants that were highly related to aquaporin and nitrate transporter expression. The results revealed new findings on the relationship between aquaporins and nitrate transporters in new leaves of Citrus, suggesting a mechanism for ensuring growth under low N when new tissues are being formed.

Deficit Irrigation Applied to Lemon Trees Grafted on Two Rootstocks and Irrigated with Desalinated Seawater.

The use of desalinated seawater (DSW) for irrigation in semi-arid regions is taking hold. Citrus tolerance to ions that predominate in DSW and water stress depends on the rootstock. Deficit irrigation was applied to DSW-irrigated lemon trees and grafted on rootstocks with different tolerance (Citrus macrophylla (CM) and sour orange (SO)). Plants were irrigated with DSW or Control treatment (distilled water), and, 140 days later, irrigation treatments were started: full irrigation (FI) or DI (50% of the volume applied to FI). After 75 days, differences between CM and SO plants irrigated with DSW and under DI were found. The higher concentrations of Cl- and Na+ in CM and B in SO were the main causes of shoot growth reduction. The osmotic adjustment of CM plants was made possible by the accumulation of Na+, Cl-, and proline, but SO failed to adjust osmotically. In CM and SO plants, photosynthesis reduction was due to lower chlorophyll levels, but also to stomatal factors (CM plants) or alterations of the photochemical machinery (SO plants). Finally, unlike CM, SO had a good antioxidant system. In the future, knowing the different responses of CM and SO under these stressful conditions could be useful in citrus-growing areas.

Influence of Different Rootstocks on Fruit Quality and Primary and Secondary Metabolites Content of Blood Oranges Cultivars.

Blood oranges have high concentrations of bioactive compounds that are beneficial to health. In Europe, the cultivation of blood oranges is increasing due to their excellent nutritional properties. In Citrus crops, rootstocks play an important role in juice and can increase the content of bioactive compounds. The morphological, qualitative and nutritional parameters were analyzed in cultivars 'Tarocco Ippolito', 'Tarocco Lempso', 'Tarocco Tapi' and 'Tarocco Fondaconuovo' grafted onto Citrus macrophylla and Citrus reshni. 'Tarocco Lempso' grafted onto Citrus macrophylla obtained the highest values of weight (275.78 g), caliber (81.37 mm and 76.79 mm) and juice content (162.11 g). 'Tarocco Tapi' grafted onto Citrus reshni obtained the most interesting qualitative parameters (15.40 °Brix; 12.0 MI). 'Tarocco Lempso' grafted onto Citrus reshni obtained the most intense red juice (a* = 9.61). Overall, the highest concentrations of primary metabolites were in proline, aspartate, citric acid, and sucrose. The results showed that 'Tarocco Ippolito' juice grafted onto Citrus reshni had the highest levels of total hydroxycinnamic acids (263.33 mg L-1), total flavones (449.74 mg L-1) and total anthocyanins (650.42 mg L-1). To conclude, 'Tarocco Lempso' grafted onto Citrus macrophylla obtained the best values of agronomic parameters, and the cultivars grafted onto Citrus reshni obtained significantly higher concentrations in primary and secondary metabolites.

Morphological, physiological, and molecular scion traits are determinant for salt-stress tolerance of grafted citrus plants.

Citrus productivity has been decreasing in the last decade in the Mediterranean basin as a consequence of climate change and the high levels of salinity found in the aquifers. Citrus varieties are cultivated grafted onto a rootstock, which has been reported as responsible for plant tolerance to adverse situations. However, other important factors for stress tolerance relying in the scion have been less studied. The aim of this study was to evaluate the effect of the grafted scion on citrus tolerance to salt stress. Four different citrus rootstock/scion combinations were subjected to salt stress for 30 days, using Carrizo citrange (CC) or Citrus macrophylla (CM) as rootstocks, and Navelina orange (NA) or Oronules mandarin (OR) as scions. CM-OR was the most tolerant combination, whereas CC-NA was the most sensitive one. Our results support the idea that the rootstock plays an important role in salt stress tolerance, but scion is also crucial. Thus, photosynthesis and transpiration, processes regulated by abscisic acid and jasmonic acid, are determinant of plant performance. These photosynthetic parameters were not affected in plants of the salt-tolerant combination CM-OR, probably due to the lower intoxication with Cl- ions, allowing a better performance of the photosynthetic machinery under stress conditions. The different stomatal density of the two citrus scions used in this work (higher in the sensitive NA in comparison to the tolerant OR) also contributes to the different tolerance of the grafted plants to this adverse condition. Additionally, CsDTX35.1 and CsDTX35.2, genes codifying for Cl- tonoplast transporters, were exclusively overexpressed in plants of the salt-tolerant combination CM-OR, suggesting that these transporters involved in Cl- compartmentalization could be crucial for salt stress tolerance. It is concluded that to improve citrus tolerance to high salinity, it is important that scions have a versatile photosynthetic system, an adequate stomatal density, and a proper modulation of genes coding for Cl- transporters in the tonoplast.

Insight into chilling stress response of key citrus grafting combinations grown in Egypt

One way to help plants deal with unfavorable temperatures is to graft elite cultivars onto adaptable rootstocks. Citrus macrophylla is a newly introduced rootstock in Egypt. Still, it needs to be evaluated in terms of its response to chilling stress compared to the most widely used rootstock (Citrus volkameriana). Thus, the present work assessed the physiological, biochemical, and anatomical responses of C. volkameriana (Volkamer) and C. macrophylla (Alemow) rootstocks and the cultivars ‘Olinda Valencia orange’ and ‘Murcott Tangerine’ grafted onto them to chilling stress. Plants were subjected to low-night temperatures for 48 h at 20/4 °C ± 1 (day/night), followed by three days of recovery at 30/25 °C ± 1 (day/night). Under low-night temperatures, all plants recorded a significant increment in malondialdehyde (MDA), peroxidase (POX), superoxide dismutase (SOD), proline, and leaf diffusion resistance (LDR). At the same time, a significant reduction was found in the photosynthetic apparatus (stomatal conductance (GS), transpiration (TR), and chlorophyll), polyphenol oxidase (PPO), and catalase (CAT). Moreover, anatomical damage was observed in total leaf thickness, palisade tissue thickness, spongy tissue thickness, midvein vascular bundle thickness, upper epidermis thickness, lower epidermis thickness, and spongy ratio (SR). Our results also indicated that the Volkamer, Olinda Valencia orange/Volkamer, and Murcott Tangerine/Volkamer were slightly damaged and showed a relatively high level of antioxidant activity (SOD and POX), LDR, and a more significant decrease in TR. However, they were less affected anatomically than Alemow rootstock and the scions grafted on it. These results demonstrated that grafting on Volkamer could improve the physiological, biochemical, and anatomical structural responses of the scions grafted on it to low-night temperature stress compared with grafting on Alemow.

Fruit Quality and Primary and Secondary Metabolites Content in Eight Varieties of Blood Oranges

In Europe, the cultivation of blood oranges for fresh consumption is increasing due to their excellent organoleptic and nutraceutical properties, which give this fruit the status of functional food. Blood orange has a higher content of bioactive compounds, which confer additional benefits for human health. The main morphological and qualitative parameters were studied together with the content of primary metabolites by 1H NMR and secondary metabolites by HPLC-ESI-DAD-MSn from eight varieties of blood orange grafted on Citrus macrophylla. Tarocco Dalmuso was the variety with the highest values of weight (350.6 g), caliber (86.4 mm and 88.6 mm) and juice content (214.2 g). Tarocco Gallo obtained the most interesting qualitative parameters (13.95 °Brix; 22.75 MI). The most intense red juice was in Sanguinelli (a* = 9.45) and, in crust, it was in Tarocco Scirè (a* = 40.13). The most abundant primary metabolites were proline, aspartate and asparagine, citric acid and sucrose. The results showed that the juice of the Moro had the highest levels of total flavones and flavanones (90.07 and 592.88 mg L−1, respectively), and Sanguinelli in total anthocyanins (101.06 mg L−1). To conclude, Tarocco Dalmuso obtained the best values of agronomic parameters, and Moro and Sanguinelli in the content of phenolic compounds.