Transgenic Sweet Orange Expressing the Sarcotoxin IA Gene Produces High-Quality Fruit and Shows Tolerance to 'Candidatus Liberibacter asiaticus'.

Huanglongbing (otherwise known as HLB or greening) is currently the most devastating citrus disease worldwide. HLB is primarily associated with the phloem-inhabiting bacterium 'Candidatus Liberibacter asiaticus' (CLas). Currently, there are no citrus species resistant to CLas. Genetic transformation is one of the most effective approaches used to induce resistance against plant diseases. Antimicrobial peptides (AMPs) have shown potential breakthroughs to improve resistance to bacterial diseases in plants. In this paper, we confirm the Agrobacterium-mediated transformation of Pera sweet orange expressing the AMP sarcotoxin IA (stx IA) gene isolated from the flesh fly Sarcophaga peregrina and its reaction to CLas, involving plant performance and fruit quality assessments. Four independent transgenic lines, STX-5, STX-11, STX-12, and STX-13, and a non-transgenic control, were graft-inoculated with CLas. Based on our findings, none of the transgenic plants were immune to CLas. However, the STX-5 and STX-11 lines showed reduced susceptibility to HLB with mild disease symptoms and low incidence of plants with the presence of CLas. Fruit and juice quality were not affected by the genetic transformation. Further, no residues of the sarcotoxin IA protein were found in the juice of the STX-11 and STX-12 fruits, though detected in the juice of the STX-5 and STX-13 lines, as revealed by the immunoblotting test. However, juices from all transgenic lines showed low traces of sarcotoxin IA peptide in its composition. The accumulation of this peptide did not cause any deleterious effects on plants or in fruit/juice. Our findings reinforce the challenges of identifying novel approaches to managing HLB.

Interplay between Abscisic Acid and Gibberellins, as Related to Ethylene and Sugars, in Regulating Maturation of Non-Climacteric Fruit.

In this review, we address the interaction between abscisic acid (ABA) and gibberellins (GAs) in regulating non-climacteric fruit development and maturation at the molecular level. We review the interplay of both plant growth regulators in regulating these processes in several fruit of economic importance such as grape berries, strawberry, and citrus, and show how understanding this interaction has resulted in useful agronomic management techniques. We then relate the interplay of both hormones with ethylene and other endogenous factors, such as sugar signaling. We finally review the growing knowledge related to abscisic acid, gibberellins, and the genus Citrus. We illustrate why this woody genus can be considered as an emerging model plant for understanding hormonal circuits in regulating different processes, as most of the finest work on this matter in recent years has been performed by using different Citrus species.

Rootstock selection for 'Swatow' Mandarin trees grown at different locations throughout the Brazilian subtropics.

Evaluating citrus rootstocks is of paramount importance in determining their suitability for a certain region and promoting resilience in orchards by increasing the genetic pool, thereby potentially contributing to a more strategic establishment of new plantings. This long-term field study (2000-2013) aimed to evaluate different rootstocks for 'Swatow' mandarin grown at two locations (Paranavaí and Londrina) in the Brazilian subtropics. Nine rootstocks were evaluated, including 'Rangpur' lime, 'Swingle' citrumelo, 'Volkamer' lemon, 'Caipira DAC' sweet orange, 'Cleopatra' and 'Sunki' mandarins, 'Trifoliate' orange, 'Carrizo', and 'Fepagro C-13' citranges. Trees were assessed for vegetative growth, yield, fruit quality, density, and yield estimates. The experimental design was a randomized block arranged in a 9 × 2 setting (rootstock × location) with 6 replicates and 4 trees per plot. 'Swatow' trees grew more vigorously in Londrina than Paranavaí, particularly for 'Cleopatra' and 'Sunki' pairings. Tree vigor was reduced with 'Trifoliate', resulting in higher tree density estimates and yield efficiency. This rootstock, along with 'Rangpur', 'Swingle', and 'Carrizo' provided superior yield to the scion. All tested rootstocks conferred good fruit quality. Fruits were larger and heavier in 'Sunki' pairings, showing higher soluble solids (SS) content, along with 'Caipira DAC', 'Trifoliate', 'Swingle', and 'Carrizo' at both locations. Our findings confirm the suitability of 'Trifoliate' orange, 'Carrizo' citrange, or 'Caipira DAC' orange rootstocks as promising candidates for 'Swatow' mandarin cultivation in humid subtropical and analogous regions. Further investigations are invoked to improve the horticultural performance of 'Swatow' mandarin trees grafted onto these rootstocks.

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.

Determining Seed Viability During Fruit Maturation to Improve Seed Production and Availability of New Citrus Rootstocks.

In recent years, the pressure for replanting and resetting huanglongbing (HLB or citrus greening) affected citrus groves has led to an inadequate seed supply for the most popular rootstock cultivars in the State of Florida, United States. Early fruit harvesting of citrus rootstock source trees might reduce fruit losses and enhance seed availability, especially in HLB-endemic and hurricane susceptible areas, if the physiological quality of the seeds is adequate. The effects of fruit maturity on seed quality and seedling performance of US-802, US-897, and US-942 citrus rootstocks were investigated for two consecutive growing seasons. The study included the evaluation of seed germination and nursery performance of the citrus rootstock seedlings. The germination test was performed in vitro, where seeds were hand-peeled, surface-sterilized and placed in culture tubes containing basal Murashige and Skoog medium. For the emergence test, seeds were sown in seedling trays containing sterilized growing substrate in a greenhouse with controlled-environment conditions. Rootstock fruits from all three varieties harvested in August and September had seeds with higher germination potential, as more than 90% of the seeds generated seedlings. US-942 had more % of emergence than US-802 and US-897, resulting in faster seed germination; in contrast, US-802 had the faster shoot growth rate. Assays on fruit abscission response showed that by August, fruit from all three varieties were responsive to ethylene and abscised, although response varied and was higher in US-942, suggesting the seeds were mature enough. Taken together, our findings indicate that fruits these three rootstocks can be harvested as early as August in contrast to the current procedures without losing germination potential. This will result in an increase in available seeds for nurseries in Florida.

Five Rootstocks for "Emperor" Mandarin Under Subtropical Climate in Southern Brazil.

Rootstocks modulate several characteristics of citrus trees, including vegetative growth, fruit yield and quality, and resistance or tolerance to pests, diseases, soil drought, and salinity, among other factors. There is a shortage of scion and rootstock cultivars among the combinations planted in Brazil. "Ponkan" mandarin and "Murcott" tangor grafted on "Rangpur" lime comprise the majority of the commercial mandarin orchards in Brazil. This low genetic diversity of citrus orchards can favor pest and disease outbreaks. This study aimed to evaluate the agronomic performance, Huanglongbing (HLB) tolerance, and fruit quality of "Emperor" mandarin on five different rootstocks for nine cropping seasons under the subtropical soil-climate conditions of the North region of the state of Paraná, Brazil. The experimental design was a randomized block, with six replications, two trees per block, and five rootstocks, including "Rangpur" lime, "Cleopatra," and "Sunki" mandarins, "Swingle" citrumelo, and "Fepagro C-13" citrange. The evaluations included tree growth, yield performance, fruit quality, and HLB disease incidence. "Emperor" mandarin trees grafted on "Rangpur" lime and "Swingle" citrumelo had early fruiting and high yield efficiency. "Rangpur" lime also induced the lowest tree growth, but low fruit quality. Trees on "Swingle" citrumelo and "Fepagro C-13" citrange showed low scion and rootstock affinity and produced fruits with high total soluble solids (TSS), with a lower number of seeds for those from trees on "Fepagro C-13" citrange. "Cleopatra" and "Sunki" mandarins induced higher juice content, while fruits from trees on "Cleopatra" also had higher TSS/titratable acidity (TA) ratio. "Emperor" mandarin trees were susceptible to HLB regardless of the rootstocks. Overall, "Cleopatra" and "Sunki" mandarins, "Swingle" citrumelo, and "Fepagro C-13" are more suitable rootstocks for "Emperor" mandarin under Brazilian subtropical conditions than "Rangpur" lime.

Analysis of the phenolic composition and yield of 'BRS Vitoria' seedless table grape under different bunch densities using HPLC-DAD-ESI-MS/MS.

Grapes are considered to be a major source of phenolic compounds as compared with other fruits. To improve the quality of table grapes, some techniques like thinning can be used. In addition, grape cultivars with distinct characteristics are directly linked to its phenolic profile. This study aimed to identify and quantify the phenolic compound profile and yield of the hybrid 'BRS Vitoria' seedless table grape under different bunch densities, using a combination of solid-phase extraction (SPE) methodologies and analytical high-performance liquid chromatography-diode array detector with tandem mass spectrometry (HPLC-DAD-ESI-MS/MS). A trial was carried out in 2016, in a commercial vineyard at Marialva, state of Parana (South Brazil). Three weeks after anthesis, the following bunch densities were evaluated: 4.0, 4.5, 5.0, 5.5, and 6 bunches per m2 (corresponding to an estimation of 16, 18, 20, 22, and 24 tons ha-1). The randomized block design was used as a statistical model with each treatment was replicated four times, with one vine per plot. Different characteristics were evaluated at harvest, e.g., soluble solids content (SS), total acidity (TA), maturation index (MI = SS/TA), bunch and berry masses, yield, as well hydroxycinnamic acid derivative (HCAD), anthocyanin, flavonol, and flavan-3-ol contents by HPLC-DAD-ESI-MS/MS analysis. The evaluated bunch densities did not interfere with the physicochemical characteristics of the berries, such as SS and MI. Under the density of 6.0 bunches per m2, the highest yield of 25 tons ha-1 was reached. Under all bunch densities, the phenolic profile presented the same compounds, but at different concentrations. Under a density of 5.0 bunches per m2, the compounds belonging to the anthocyanin and flavonol families were present in high concentrations. In contrast, at the densities of 4.0 and 4.5 bunches per m2, there was a reduction in the flavan-3-ol content. With respect to stilbenes, only the trans-piceid and its cis- isomer were detected. However, their concentrations had no significant influence on the evaluated bunch densities.