Late-Season Sweet Orange Selections Under Huanglongbing and Citrus Canker Endemic Conditions in the Brazilian Humid Subtropical Region.

The Brazilian citrus orchards are comprised by few genotypes, which increases the risk of pest and disease outbreaks. The diversification of sweet oranges (Citrus × sinensis) in orchards also generates off-season revenue and extend the fruit processing period. This study aimed to evaluate several horticultural traits of 19 late-season sweet orange selections under citrus canker and huanglongbing (HLB) endemic condition in northwestern Paraná state, Brazil, in a long-term field experiment. Tree size, yield, fruit quality for fresh fruit and industrial markets, estimates of tree density and yield, and citrus canker and huanglongbing (HLB) incidences were assessed. The experimental design was a randomized block with three replicates and five trees per unit. The orchard was drip-irrigated and arranged at tree spacing of 6.5 m × 4.5 m. All scions were graft-compatible with Rangpur lime (C. × limonia). Valencia selections had the tallest trees and largest canopies, particularly Olinda, Frost and #121 with heights and volumes greater than 4.20 m and 43 m3, respectively. Natal África do Sul and Whit's Late Valencia trees were the most productive with cumulative yields above 640 kg per tree. Most of the selections produced fruits of excellent physicochemical quality attending the fresh fruit and industrial market requirements. All selections showed similar horticultural characteristics for the fresh market, while Natal África do Sul and Charmute de Brotas were more suitable for juice processing. Frost Valencia and Valencia Late Fla. had the highest incidence of citrus canker on fruits (>20%), whereas IPR Folha Murcha, Charmute de Brotas and some Valencia selections (Chafeei Late, Campbell 479, Campbell 294, Olinda, Mutação and Whit's Late) exihibed low incidence (3.0-17.7%). At 9 years, Valencia Mutação trees had high HLB incidence (93%). In contrast, Natal IAC and Folha Murcha IAC showed the lowest HLB incidence (13%). Our results revealed that Natal IAC, Folha Murcha IAC, IPR Folha Murcha, Natal Murcha, Campbell 479 Valencia and Valencia Late Fla. had the best horticultural performance in addition to low HLB incidence. Together, these late-season sweet oranges are the most advantageous selections for citrus orchard diversification under citrus canker and HLB endemic conditions in humid subtropical regions.

Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 µg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 µg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 µg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms.

Comparison of Resistance to Asiatic Citrus Canker Among Different Genotypes of Citrus in a Long-Term Canker-Resistance Field Screening Experiment in Brazil.

Asiatic citrus canker (ACC) is caused by Xanthomonas citri subsp. citri. The disease results in yield loss and renders fruit unfit for the fresh market. A 6-year study in Paraná State, Brazil, was conducted to compare the susceptibility of 186 genotypes of citrus representing sweet orange (Citrus sinensis), mandarin (C. reticulata), Mediterranean mandarin (C. deliciosa), Clementine mandarin (C. clementina), Satsuma mandarin (C. unshiu), sour orange (C. aurantium), lemon (C. limon), sweet lime (C. aurantifolia), grapefruit (C. paradisi), and four hybrids (C. reticulata × Citrus sp., C. reticulata × C. paradisi, C. reticulata × C. sinensis, and C. unshiu × C. sinensis). Sweet orange (C. sinensis) was represented by the most genotypes (n = 141). The number of lesions per leaf was assessed 18 times from 2005 to 2010 (up to 4 times per year). The data were analyzed using mixed-model analysis of fixed and random effects, which showed a total of six resistance-susceptibility groupings of species and hybrids. Based on species, the most resistant genotypes, on average, included Satsuma and lemon (mean lesions per leaf = 4.32 and 4.26, respectively), and the most susceptible genotypes were grapefruit and sweet lime, with 14.84 and 10.96 lesions per leaf, respectively. Genotypes of mandarin, sour orange, Mediterranean mandarin, and sweet orange had intermediate severity (5.48 to 9.56 lesions per leaf). The hybrids also showed a range of ACC severity but all were in the more resistant groupings (5.26 to 7.35 lesions per leaf). No genotype was immune to ACC. The most resistant genotype was 'Muscia' (C. reticulata) and the most susceptible was 'Valencia Frost' (C. sinensis) (1.86 and 14.78 lesions per leaf, respectively). Approximately one-sixth of the genotypes showed a negative relationship of mean lesions per leaf with time, suggesting increasing resistance as they aged, due to a reduction in either new flush or plant size and structure. These results of the relative susceptibility of different citrus genotypes can be used in future research and to assist in varietal selection or for breeding purposes both within Brazil and other regions where ACC is an issue.