Ray Ruby Research Articles

Species assemblage, abundance, and distribution of Phytoseiid mites (Parasitiformes: Phytoseiidae) in Citrus Under Protective Screen.

Citrus Under Protective Screen (CUPS) production system excludes the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, vector of Huanglongbing (HLB), and large predators. However, small pests and beneficial arthropods such as predatory mites may enter the CUPS. Predatory mites from the family Phytoseiidae attack several economically important pests, especially phytophagous mites common in CUPS. The species assemblage, abundance, and distribution of phytoseiid mites in CUPS and open-air (control) were investigated in Ray Ruby grapefruit from 2018 to 2020. Foliar sprays of chemicals for pest suppression were used in both systems and evaluated for effect on predatory mite populations. In the 2 production systems, a significant peak in the phytoseiid mite population was observed in February 2019 and 2020. This increase coincided with the blooming period of grapefruit, which likely provided pollen as an alternative food. A total of 2,234 specimens of 11 species were identified. Amblyseius tamatavensis Blommers and Typhlodromalus peregrinus (Muma) were dominant in both systems. Amblyseius tamatavensis was slightly more abundant in CUPS (50% of total specimens) than T. peregrinus (47%), while T. peregrinus was prevalent (82%) in open-air compared with A. tamatavensis (16%). Shannon index and evenness were significantly higher in CUPS, but species richness was similar. Chlorpyrifos with abamectin and oil, cyantraniliprole with spirodiclofen, copper, and oil as well as pyridaben alone reduced phytoseiid populations between 87% and 96%. Fenbutatin oxide mixed with copper reduced phytoseiids by 48% in open-air only. Our results demonstrate phytoseiid survival, reproduction, and contribution to biological control in CUPS, despite high temperature and humidity.

Grapefruit Root and Rhizosphere Responses to Varying Planting Densities, Fertilizer Concentrations and Application Methods.

Huanglongbing (HLB) disease has caused a severe decline in citrus production globally over the past decade. There is a need for improved nutrient regimens to better manage the productivity of HLB-affected trees, as current guidelines are based on healthy trees. The aim of this study was to evaluate the effects of different fertilizer application methods and rates with different planting densities on HLB-affected citrus root and soil health. Plant material consisted of 'Ray Ruby' (Citrus × paradisi) grapefruit trees grafted on 'Kuharske' citrange (Citrus × sinensis × Citrus trifoliata). The study consisted of 4 foliar fertilizer treatments, which included 0×, 1.5×, 3× and 6× the University of Florida Institute of Food and Agriculture (UF/IFAS) recommended guidelines for B, Mn and Zn. Additionally, 2 ground-applied fertilizer treatments were used, specifically controlled-release fertilizer (CRF1): 12-3-14 + B, Fe, Mn and Zn micronutrients at 1× UF/IFAS recommendation, and (CRF2): 12-3-14 + 2× Mg + 3× B, Fe, Mn and Zn micronutrients, with micronutrients applied as sulfur-coated products. The planting densities implemented were low (300 trees ha-1), medium (440 trees ha-1) and high (975 trees ha-1). The CRF fertilizer resulted in greater soil nutrient concentrations through all of the time sampling points, with significant differences in soil Zn and Mn. Grapefruit treated with ground-applied CRF2 and 3× foliar fertilizers resulted in the greatest bacterial alpha and beta diversity in the rhizosphere. Significantly greater abundances of Rhizobiales and Vicinamibacterales were found in the grapefruit rhizosphere of trees treated with 0× UF/IFAS foliar fertilizer compared to higher doses of foliar fertilizers.

Effect of Huanglongbing on the Volatile Organic Compound Profile of Fruit Juice and Peel Oil in ‘Ray Ruby’ Grapefruit

Along with orange and mandarin, grapefruit production in Florida has declined sharply due to Huanglongbing (HLB), or citrus greening disease, caused by Candidatus Liberibacter asiaticus (CLas). HLB affects the volatile profiles of juice and peel oil in oranges, but there is limited information on grapefruit. In this research, 'Ray Ruby' grapefruit were harvested in 2020 and 2021 from healthy (HLB-) and HLB-affected (HLB+) trees. Peel oil was extracted by hydrodistillation, and the volatiles were analyzed by direct injection of the oil samples into gas chromatography-mass spectrometry (GC-MS). Volatiles in the juice were analyzed by headspace (HS)-solid-phase microextraction (SPME) coupled with GC-MS. HLB significantly altered the volatile profiles of peel oil and juice in 'Ray Ruby' grapefruit. Juice samples of HLB+ fruits had lower decanal, nonanal, and octanal, important citrus juice flavor compounds. HLB+ samples also showed reduced content of nonterpene compounds, other aliphatic and terpene aldehydes, and terpene ketones. Ethanol, acetaldehyde, ethyl acetate, and ethyl butanoate were increased in HLB+ juice samples, indicating an HLB-induced stress response. The most abundant compounds D-limonene and β-caryophyllene, as well as other sesquiterpenes, were increased in HLB+ juice and peel oil samples. On the other hand, the oxidative/dehydrogenated terpenes were increased by HLB in peel oil but decreased in the juice sample. Nootkatone, the key grapefruit volatile was consistently reduced by HLB in both peel oil and juice samples. The impact of HLB on nootkatone deteriorated the quality of both juice and peel oil in grapefruits.

Early Performance of Recently Released Rootstocks with Grapefruit, Navel Orange, and Mandarin Scions under Endemic Huanglongbing Conditions in Florida

Huanglongbing (HLB), which is believed to be caused by the phloem-restricted bacterium Candidatus Liberibacter asiaticus (CLas), has decimated Florida’s citrus production. Grapefruit production has declined 75%, mandarin 78%, and sweet orange 52% due to the high sensitivity of commercial scions and rootstocks to the disease. New combinations of scions and hybrid rootstocks may provide better performance than current commercial selections for Florida’s fresh citrus production, particularly in the Indian River District. The objective of this study was to evaluate and compare University of Florida rootstocks and other recently released rootstocks grafted with grapefruit, navel orange, and mandarin scions by measuring tree growth and HLB tolerance. Three independent large-scale field trials were established in September 2019 in Fort Pierce, FL, USA. Trial 1 (T1) included 36 rootstocks with ‘Ray Ruby’ grapefruit as the scion; Trials 2 and 3 (T2 and T3, respectively) included 30 rootstocks with ‘Glenn’ navel orange F-56-11 and ‘UF-950’ mandarin as the scion, respectively. Tree canopy volume, trunk diameter, CLas titer, HLB severity index, and leaf nutrient concentrations were evaluated during 2020 and 2021. Significant differences among rootstock-scion combinations were found in each trial for most of the assessed traits. In T2, UFR-15 consistently developed the largest ‘Ray Ruby’ grapefruit trees during both years. In T3, ‘Glenn’ navel orange F-56-11 trees were larger on C-22, and US-802. Similarly, US-802 and US-942 generated the largest ‘UF-950’ mandarin trees. Overall, trees had optimum levels of macro- and micronutrients except for calcium. CLas infection and HLB visual index varied among scion-rootstock combinations, especially during the first year of growth when intensive flushing was produced. Generally, trees grew vigorously with WGFT+50-7 and Willits inducing the lowest HLB symptoms in all evaluations. Production and fruit quality need to be evaluated to determine the suitability of potential scion-rootstock combinations that can confer consistent economical and biological advantages under the current HLB scenario in the Indian River District.

Substrate pH Influences the Nutrient Absorption and Rhizosphere Microbiome of Huanglongbing-Affected Grapefruit Plants

The substrate pH directly affects nutrient availability in the rhizosphere and nutrient uptake by plants. Macronutrients such as nitrogen, potassium, calcium, magnesium, and sulfur are highly available at pH 6.0–6.5, while micronutrients become less available at higher, alkaline pH (pH > 7.0). Recent research has indicated that low pHs can enhance nutrient uptake and improve sweet orange (Citrus sinensis) tree health. We designed a study to understand the influence of a wide range of substrate pH values on plant size and biomass, nutrient availability, leaf gas exchange, and rhizosphere microbiome of grapefruit (Citrus paradisi) affected by Huanglongbing (HLB). Two-year-old “Ray Ruby” grapefruit plants grafted on sour orange (Citrus aurantium) rootstock were cultivated indoors in 10-cm wide × 40-cm tall pots with peat:perlite commercial substrate (80:20 v/v). We tested two disease statuses [HLB-free or healthy (negative, HLB–) and HLB-affected (positive, HLB+)] and six substrate pH values (4, 5, 6, 7, 8, 9) in a 2 × 6 factorial arranged on a complete randomized design with four replications. The canopy volume of HLB+ plants was 20% lower than healthy plants, with pHs 7 and 9 resulting in 44% less canopy volume. The root and shoot ratio of dry weight was 25.8% lower in HLB+ than in healthy plants. Poor root growth and a decrease in fibrous roots were found, especially in pH 5 and 6 treatments in HLB+ plants (p < 0.0001). The disease status and the substrate pHs influenced the leaf nutrient concentration (p < 0.05). High substrate pH affects nutrient availability for root uptake, influencing the nutrient balance throughout the plant system. pH values did not affect plant photosynthesis, indicating that pH does not recover HLB+ plants to the photosynthetic levels of healthy plants—even though high pH positively influenced internal CO2. There were collectively over 200 rhizobacterial identified by the 16S rRNA gene sequencing in individual phylogenetic trees. Most rhizobacteria reads were identified in pH 9. Our results indicated no effect of substrate pHs on the plant disease status induced by enhanced nutrient uptake.

‘Ray Ruby’ Grapefruit Affected by Huanglongbing I. Planting Density and Soil Nutrient Management

Huanglongbing (HLB), or citrus greening disease, affects practically all fruit-bearing trees in commercial citrus orchards in Florida with no cure identified yet. High-density plantings and enhanced nutritional programs such as application of controlled-release fertilizer (CRF) with higher micronutrient levels can mitigate disease symptoms and extend the tree life span of sweet oranges (Citrus sinensis). The objective of this study was to evaluate the effects of tree planting density and application of CRF blends differing in N to K ratio and micronutrient content on grapefruit (Citrus paradisi) plant health, canopy volume, fruit yield, and fruit quality in an HLB-affected orchard. A study was conducted in Florida for two growing seasons (2017–18 and 2018–19) to evaluate the response of ‘Ray Ruby’ grapefruit on Kuharske citrange (Citrus sinensis × Poncirus trifoliata) to three planting densities (300, 440, and 975 trees per ha) and two CRF blends [12 nitrogen (N)–1.31 phosphorus (P)–7.47 potassium (K) and 16N–1.31P–16.6K] with different nutrient sources and composition. According to quantitative real-time polymerase chain reaction testing, all sampled trees tested positive for Candidatus Liberibacter asiaticus, the pathogen associated with HLB. Trees planted at 975 trees per ha had 33% lower canopy volume per tree but 160% greater fruit yield per hectare and 190% higher yield of solids compared with 300 trees per ha. Fruit produced in high-density planting (975 trees per ha) was 18% more acidic with higher soluble solid compared with low-density planting (300 trees per ha). The use of a CRF blend with higher amounts of micronutrients along with lower K increased canopy volume in both seasons and resulted in 24% and 29% reduction in fruit yield per hectare and yield of solids, respectively, in 2017–18. Our results indicate that high-density plantings increase fruit yield per area, and regardless of the N to K ratio, the use of CRF blends supplemented with micronutrients may not increase fruit yield in HLB-affected grapefruit.

‘Ray Ruby’ Grapefruit Affected by Huanglongbing II. Planting Density, Soil, and Foliar Nutrient Management

Since the arrival of Huanglongbing (HLB) disease in Florida, several management approaches, including modification of orchard architecture design and nutritional therapy, have been explored. High-density plantings anticipate early economic returns from HLB-affected orchards. With no cure available for HLB, balanced nutrient application through soil and foliar spraying can mitigate the disease. A 2-year study was conducted to investigate the effects of three grapefruit (Citrus paradisi) planting densities [single-row (300 and 440 trees per ha), and double-row high-density (975 trees per ha)], two controlled-release fertilizer (CRF) blends, and foliar-applied micronutrients (FAM) (a blend of B, Mn, and Zn at 0, 1.5, 3, and 6 times the recommended rates) on grapefruit growth and fruit yield, physiological parameters, and foliar nutrient concentrations in an HLB-affected orchard. All the trees tested positive for HLB based on real-time quantitative polymerase chain reaction (qPCR) test. The highest planting density resulted in the lowest trunk diameter and canopy volume. Despite lower yield per tree in 2019–20, 975 trees per ha planting induced the greatest fruit and solid yields per ha. Also, the fruit produced from 975 trees per ha planting tended to be acidic with the deposition of more soluble solids. Use of CRF with higher micronutrients increased canopy volume with the expense of reduced fruit number in 2019–20. FAM did not affect cycle threshold (Ct) value and tree growth parameters. Fruit yield, photosynthesis rate, and stomatal conductance (gS) decreased, and all leaf nutrient concentrations except B increased in 2019–20 with all FAM rates tested. In conclusion, our study showed that high-density planting optimizes yield under HLB-endemic conditions. In addition, supplemental soil and foliar micronutrient application do not enhance yield of HLB-affected trees over a 2-year timeframe, warranting further research for confirmation of results.

Grapefruit Production in Open Hydroponics System

Conventionally managed citrus orchards can be modified to incorporate advanced horticultural practices such as higher plant density and efficient water and fertilizer application known as open hydroponics system (OHS) to increase productivity under Huanglongbing (HLB) endemic conditions. A field study was conducted from 2013 to 2018 to evaluate the effect of an OHS on “Ray Ruby” grapefruit (RR) production under HLB-endemic conditions. We tested a combination of different rootstocks [Sour orange (RR/SO) and US-897 (RR/897)], tree planting densities [standard (STD, 358 trees per ha) and high density staggered (HDS, 953 trees per ha)], fertilization methods (dry granular—dry and fertigation—fert), and irrigation systems (double driplines—DD and microsprinkler—MS), arranged in five treatments: RR/SO_STD_dry_MS, RR/SO_HDS_fert_DD, RR/897_HDS_fert_MS, RR/897_HDS_fert_DD, and RR/SO_HDS_fert_MS. All trees were infected by Candidatus Liberibacter asiaticus five years after planting. Trunk diameter and canopy volume increased over time and were higher under RR/SO_STD_dry_MS compared to other treatments. Total fruit number increased in 2016/17 compared to other seasons; however, 65% of fruit were classified as small (&lt;100 mm). Fruit produced under RR/897_HDS_fert_DD had the highest amount (79%) of adequate size fruit (100–117 mm) compared to other treatments. Fruit yield was similar for both rootstocks planted at HDS using DD and MS fertigation, and 67% higher than the standard treatment (RR/SO_STD_dry_MS). Soluble solid contents (SSC), titratable acidity, and SSC-to-titratable acidity ratio were not affected by the treatments. HDS planting resulted in higher fruit yield, irrespective of rootstock and irrigation system, representing an important advance in grapefruit production. Overall, our results demonstrated that production of grapefruit in high-density using OHS can be used by citrus growers who aim to make the best water and fertilizer management under HLB-endemic conditions.

Comparison of microwave-assisted techniques for the extraction of antioxidants from Citrus paradisi Macf. biowastes.

Microwave-assisted extraction (MAE) and solvent-free microwave extraction and Soxhlet extraction were applied to Ray Ruby grapefruit leaves (Citrus paradisi Macf.) to compare extract efficiency. Face centered composite designs were constructed via response surface methodology. Effects of factors of MAE were investigated on total phenolic content (TPC) and naringin content (NC). The optimized conditions were established as 1.4 kWL-1 for microwave power density, 20.00 gL-1 for solid/solvent ratio, 218.180s for extraction time, while responses were calculated as 14.210mg of gallic acid equivalent per g of the dried leaf (mgGAE g-1DL) and 13.198mg of naringin per g of dried leaf (mgNg-1DL) for TPC and NC, respectively. SFME and classical Soxhlet methods were also conducted for comparison reasons.

Citrus Production Under Screen as a Strategy to Protect Grapefruit Trees From Huanglongbing Disease.

Citrus production under enclosed structures can exclude the Asian citrus psyllid (ACP, Diaphorina citri) and eliminate the negative effects of citrus greening or huanglongbing (HLB) disease caused by Candidatus Liberibacter asiaticus to the grapefruit (Citrus paradisi) fresh fruit industry. Physically impeding the insect vector from accessing trees is a logical method to have disease-free groves. Our objectives were to assess the ability of enclosed screenhouses to exclude the ACP, stop HLB inoculation and dissemination, and improve fruit yield of in-ground and container-grown 6-year-old “Ray Ruby” grapefruit at super-high planting densities relative to open-air trees. We built a large structure to allow commercial-scale trials and tested two production systems (screenhouse and open-air), two planting systems (in-ground and potted), and two rootstocks (“Sour Orange” [Citrus × aurantium] and “US-897” [Citrus reticulata × Poncirus trifoliate]). The experimental design was a randomized complete block design split-split-plot with four replications. Four passively ventilated 1,080-m2 completely enclosed screenhouses were constructed using a 50-mesh monofilament high-density polyethylene screen. The main support for each enclosed, covered structure consisted of pressure-treated, wooden utility poles. Trees were planted in Sept/2013 on a density of 1,957 trees/ha. Irrigation was performed on-demand using two 7.6-LPH drip emitters per tree, and fertigation was applied three times/week using 15N-2.6P-22.4K water-soluble fertilizer at 180 kg N/ha. Psyllids were monitored using sticky cards and detected inside the screenhouses post-Hurricane Irma, which damaged the screen structures in Sept/2017, leaving openings until repairs were completed in Apr/2018. Screen aging and a tropical storm in April/2019 caused another major screen opening fixed in Oct/2019. Despite the weather-related damages to the screens, only trees cultivated in open-air tested positive for Candidatus Liberibacter asiaticus after 6 years. There was fast disease progression for all outside treatments, with 100% infection. Covered, in-ground trees exhibited the highest trunk diameter and canopy volume (P < 0.0001). Trees grown inside screenhouses exhibited higher fruit yield than outside trees, with the highest yield observed for in-ground trees on “US-897” (51,081 kg/ha) (P < 0.0001). Several open-air treatments particularly in containers did not produce any fruit. On the other hand, potted grapefruit trees cultivated inside the enclosures had the highest soluble solids content (P < 0.001). The screenhouses provided disease exclusion, increased fruit yield, and fruit quality, representing an alternative for growers interested in producing high-quality fruit for the fresh market. Production cost and economic viability still need to be evaluated for large-scale implementation.

Fixed-Quat: An Attractive Nonmetal Alternative to Copper Biocides against Plant Pathogens.

In this paper, we report a nonphytotoxic bactericide and fungicide formulation containing a composite of silica and quaternary ammonium compound (quat). The composite material was prepared using an acid-catalyzed sol-gel method. Positively charged quat was associated with a negatively charged silica-gel matrix, producing a stable suspension of fixed-quat gel (FQ-G). The morphology of FQ-G and the interaction of quat with silica were characterized using SEM and FTIR, respectively. Silica gel significantly reduced quat phytotoxicity when tested at 500 and 1000 μg/mL foliar-application rates. The in vitro antimicrobial efficacy of FQ-G was evaluated against Xanthomonas alfalfae, Pseudomonas syringae, and Clavibacter michiganensis, showing comparable efficacies to that of quat itself. In field conditions, its efficacy in controlling the bacterial and fungal diseases citrus canker, scab, and melanose on 'Ray Ruby' red grapefruit was evaluated. Foliar application rates at 100 and 200 μg/mL provided comparable disease control to those of several copper standards, demonstrating the potential for use as an alternative agricultural biocide.

Protected Fresh Grapefruit Cultivation Systems: Antipsyllid Screen Effects on Plant Growth and Leaf Transpiration, Vapor Pressure Deficit, and Nutrition

Completely enclosed screen houses can physically exclude contact between the asian citrus psyllid [ACP (Diaphorina citri)] and young, healthy citrus (Citrus sp.) trees and prevent huanglongbing (HLB) disease development. The current study investigated the use of antipsyllid screen houses on plant growth and physiological parameters of young ‘Ray Ruby’ grapefruit (Citrus ×paradisi) trees. We tested two coverings [enclosed screen house and open-air (control)] and two planting systems (in-ground and container-grown), with four replications arranged in a split-plot experimental design. Trees grown inside screen houses developed larger canopy surface area, canopy surface area water use efficiency (CWUE), leaf area index (LAI) and LAI water use efficiency (LAIWUE) relative to trees grown in open-air plots (P &lt; 0.01). Leaf water transpiration increased and leaf vapor pressure deficit (VPD) decreased in trees grown inside screen houses compared with trees grown in the open-air plots. CWUE was negatively related to leaf VPD (P &lt; 0.01). Monthly leaf nitrogen concentration was consistently greater in container-grown trees in the open-air compared with trees grown in-ground and inside the screen houses. However, trees grown in-ground and inside the screen houses did not experience any severe leaf N deficiencies and were the largest trees, presenting the highest canopy surface area and LAI at the end of the study. The screen houses described here provided a better growing environment for in-ground grapefruit because the protective structures accelerated young tree growth compared with open-air plantings while protecting trees from HLB infection.

Rootstock Effects on Fruit Quality among ‘Ray Ruby’ Grapefruit Trees Grown in the Indian River District of Florida

The objective of this experiment was to compare fruit-quality parameters of ‘Ray Ruby’ grapefruit grown on seven rootstocks. Four recent releases from the United States Department of Agriculture (USDA) rootstock breeding program, ‘US-852’, ‘US-897’, ‘US-942’, and ‘US-812’ (all Citrus reticulata × Poncirus trifoliata hybrids), ‘x639’ (C. reticulata × P. trifoliata), along with industry-standard ‘Sour Orange’ and ‘Swingle’ citrumelo were evaluated in a commercial orchard trial in Indian River County, FL. Fruit-quality data were collected in 2011–12 (eight harvests), 2012–13 (five harvests), and 2014 (single harvest). In each season, rootstock effects on fruit size, total solids, and solids acid ratio were significant. ‘Sour orange’ and ‘Swingle’ produced the largest fruit, whereas ‘US-897’ (a semidwarfing rootstock) produced the smallest fruit. Peel thickness (measured only in the 2011–12 season) was greatest in ‘Sour Orange’ early in the season, but not toward the end of the season. Misshapen (“sheep-nosed”) fruit occurred more frequently on ‘Sour Orange’ than on other rootstocks, although the incidence of sheep-nosing was minor. Analysis of variance (ANOVA) for fruit-quality data collected in January of each of the 3 years confirmed that ‘Sour Orange’ and ‘Swingle’ produced the largest fruit and ‘US-897’ produced the smallest fruit. Total solids were the highest in ‘US-897’ and the lowest in ‘x639’ and ‘US-852’. Taken together, our data indicate that ‘US-942’ and ‘US-897’ rootstocks produced fruit with quality characteristics that equaled or exceeded ‘Sour Orange’ and ‘Swingle’, the two most common rootstocks used in the Indian River district.

Correlation of Diplodia (Lasiodiplodia theobromae) infection, huanglongbing, ethylene production, fruit removal force and pre-harvest fruit drop

Citrus greening or huanglonbing (HLB) is thought to be caused by Candidatus Liberibacter asiaticus (CLas) and is devastating the citrus industry worldwide. One symptom of HLB disease is excessive pre-harvest fruit drop. Recently, higher incidence of Lasiodiplodia theobromae (Diplodia) was found in HLB-symptomatic orange calyx abscission zones (AZ-C) than in non-symptomatic fruit, and the infection was positively correlated with the reduction in fruit detachment force (FDF), suggesting that Diplodia infection may be involved in the HLB-related pre-harvest fruit drop. To verify the hypothesis, we conducted two experiments. Experiment 1 was conducted by shaking HLB-affected ‘Hamlin’ and ‘Valencia’ orange trees during the harvest season (twice for ‘Hamlin’ and once for ‘Valencia’). The fruit that dropped from trees upon shaking were collected (D), and the fruit retained on trees after shaking were harvested (R). Fruit ethylene production was measured, and the levels of Diplodia and CLas in AZ-C of D and R fruit were analyzed. The results revealed significantly higher levels of Diplodia in D compared with R fruit; and ethylene was produced from more than half of the D fruit but none of the R fruit. Ethylene production was positively correlated with Diplodia level in D fruit. In experiment 2, a preliminary trial on the effect of fungicide (Quadris Top) application on incidence of Diplodia infection and fruit drop was investigated. The experiment was conducted in a commercial grove with essentially 100% of the trees being HLB-symptomatic, and included five citrus cultivars (‘Early Gold’ orange, ‘Midsweet’ orange, ‘Murcott’ tangor, ‘Navel’ orange and ‘Ray Ruby’ grapefruit). Diplodia levels were lower and FDF significantly higher in fungicide-treated compared to non-treated ‘Early Gold’, ‘Midsweet’ and ‘Murcott’ fruit, and consequently, the fruit drop was reduced by 45%, 30% and 46% that of non-sprayed controls, respectively. For ‘Navel’ or ‘Ray Ruby’ fruit, there was no significant change between sprayed and non-sprayed controls in the level of Diplodia, FDF or fruit-drop. The results consistently showed a positive correlation between Diplodia infection and fruit drop in HLB-affected fruit, indicating the possible involvement of the fungus in HLB-related excessive fruit drop. This suggests that control of Diplodia fungal infection in the field may reduce HLB-associated pre-harvest fruit drop.

Genetic Transformation of Commercially Important Mature Citrus Scions

ABSTRACTA Mature Citrus Facility (MCF) was established at the University of Florida's Citrus Research and Education Center (CREC) to produce transgenic citrus (Citrus spp.) tolerant to canker and greening bacterial diseases. Plants produced from mature tissue should flower and fruit early. Budsticks of ‘Hamlin’, ‘Valencia’, and ‘Pineapple’ sweet orange [C. sinensis (L.) Osbeck] and ‘Ray Ruby’ grapefruit (C. paradisi Macfad.) were obtained from the Florida Division of Plant Industry (DPI) and introduced into the facility through shoot‐tip grafting (STG). These plants were tested for bacteria, phloem‐limited Huanglongbing (HLB), and indexed for viruses to ensure freedom from disease. Mature buds were grafted onto immature rootstock and the first two flushes of mature scion used in transformation feasibility studies with Agrobacterium strain EHA105 harboring pCAMBIA2301 and strain EHA101 harboring pTLAB21, which carry the β‐glucuronidase (GUS) and green fluorescent protein (GFP) reporters, respectively. β‐glucuronidase assays or GFP fluorescence was used to identify transgenic shoots, which developed through organogenesis from stem explants. Transgenic shoots were micrografted onto immature rootstock, and secondarily micrografted onto 4‐ to 5‐mo‐old rootstock. NPTII enzyme expression in older trees was confirmed using the immunostrip and enzyme‐linked immunosorbent assays (ELISA). Southern blots of a representative group of transgenics showed that most were low copy number with 1 to 3 copies of the T‐DNA integrated into the genome. There were a large number of escapes on kanamycin, and many transgenic shoots were lost in micrografting. Several transgenic trees flowered 18 to 19 mo after cocultivation with Agrobacterium. Transformation of mature citrus with disease resistance genes would be a feasible way to produce citrus tolerant to canker and greening.

Farklı anaçlar üzerine aşılı bazı virüsten ari altıntop çeşitlerinin meyve kalite özellikleri

Sanitation studies against virus and virus like diseases causing large economic losses in citrus plantations in Turkey were begun at Antalya Citrus Research Institute in 1988. As a result of the study, some virus-free grapefruit varieties (Redblush, Rio Red, Ray Ruby, Henderson, Star Ruby and Marsh Seedless) budded on Local sour orange and Troyer citrange rootstocks were planted at breeding parcel in 1995. Effects of rootstocks on fruit quality properties were investigated in these grapefruit varieties. Local sour orange and Troyer citrange affected similarly on the fruit quality of grapefruit cultivars. It could be suggested that these rootstocks could be recommended for grapefruit growing.

Integration of soil applied neonicotinoid insecticides and acibenzolar-S-methyl for systemic acquired resistance (SAR) control of citrus canker on young citrus trees

Soil application of systemic neonicotinoid insecticides for control of psyllid vectors of Huanglongbing disease on young citrus trees also produces season-long SAR control of citrus canker caused by Xanthomonas citri subsp. citri Schaad et al. The neonicotinoids imidacloprid (IMID) and thiamethoxam (THIA) were compared with soil or sprinkler applications of the commercial SAR inducer acibenzolar-S-methyl (ASM) and foliar sprays of copper hydroxide (CH) and/or streptomycin (STREP) to evaluate their effects on the percentage of canker-infected leaves on 2-yr-old ‘Vernia’ orange and 3-yr-old ‘Ray Ruby’ grapefruit trees in Southeast Florida. All treatments significantly reduced the incidence of foliar canker compared to the untreated check. Soil drenches of ASM and season long rotations with IMID and THIA were highly effective for suppressing foliar canker on young grapefruit and orange trees under weather conditions absent of high intensity rains or tropical storms. Sprinkler application of ASM was less effective than soil drench. The level of control for SAR treatments was comparable to eleven 21-day interval sprays of CH and/or STREP. SAR induced by soil-applied insecticides provides substantial benefits for canker disease management on young citrus trees that may be augmented with ASM.

Soil Application of SAR Inducers Imidacloprid, Thiamethoxam, and Acibenzolar-S-Methyl for Citrus Canker Control in Young Grapefruit Trees.

Soil applications of inducers of systemic acquired resistance (SAR) imidacloprid, thiamethoxam, or acibenzolar-S-methyl, at various rates and application frequencies, were evaluated for control of citrus canker caused by Xanthomonas citri subsp. citri in a field trial of 3- and 4-year-old 'Ray Ruby' grapefruit trees in southeastern Florida. Reduction of foliar incidence of canker produced by one, two, or four soil applications of imidacloprid, thiamethoxam, and acibenzolar-S-methyl was compared with 11 foliar sprays of copper hydroxide and streptomycin applied at 21-day intervals. In the 2008 and 2009 crop seasons, canker incidence on each set of vegetative flushes was assessed as the percentage of the total leaves with lesions. By the end of the 2008 season, despite above-average rainfall and a tropical storm event, all treatments significantly reduced foliar incidence of citrus canker on the combined spring-summer-fall flushes. Sprays of copper hydroxide and streptomycin were effective for reducing canker incidence on shoot flushes produced throughout the season compared with the untreated control, whereas soil-applied SAR inducers reduced foliar disease depending on rate, frequency, and timing of application. Except for the treatment of four applications of acibenzolar-S-methyl at 0.2 g a.i. per tree or two applications of imidacloprid, SAR inducers were ineffective for reducing foliar disease on the flushes that were present during the tropical storm. In 2009, all treatments significantly reduced the incidence of foliar canker on the combined spring-summer-fall flushes but not all treatments of spring-summer flushes with SAR inducers were effective compared with the untreated control. Hence, depending on rate, frequency, and timing of application, soil-applied SAR inducers reduced incidence of canker on foliar flushes of young grapefruit trees under epidemic conditions.

Performance of various grapefruit (Citrus paradisi Macf.) and pummelo (C. maxima Merr.) cultivars under the dry tropic conditions of Mexico

Grapefruit growers in the tropics require information about existing and new citrus cultivars with high productivity potential. The objective of this study was to determine the growth, yield, and fruit quality performance of seven pigmented and four white grapefruit cultivars under the dry tropic conditions of Colima, Mexico. The trees were budded on sour orange (Citrus aurantium L.) rootstock and planted at a distance of 8 × 4 m. ‘Oroblanco’ and ‘Marsh Gardner’ white-fleshed grapefruit cultivars and ‘Chandler’, a pink-fleshed pummelo, were the largest trees with the greatest height (5.0–5.6 m), canopy diameter (6.2–6.3 m), trunk diameter (21.9–23.3 cm), and canopy volume (109–123 m3). Lower height (4.3–4.8 m) and canopy volume (73–96 m3), but with similar canopy diameter to the previously mentioned cultivars, were recorded for the remaining pigmented cultivars. ‘Chandler’ pummelo and four pigmented grapefruit cultivars (‘Shambar’, ‘Rio Red’, ‘Ray Ruby’, and ‘Redblush #3’) had yearly productions of 34.8, 34.9, 34.1, 32.7, and 30.6 ton ha−1, respectively. The most productive white grapefruit cultivar was ‘Marsh Gardner’ (30.5 ton ha−1). Grapefruit cultivars having the largest fruit size showed a higher inverse relationship between fruit weight and yield than those with small fruit. Most genotypes had higher values of fruit weight, juice content, and maturity index than those required by the local market. The most promising grapefruit cultivars based on their acceptable growth, yield superior to 30 ton ha−1, and acceptable fruit color were ‘Rio Red’, ‘Shambar’, ‘Ray Ruby’, and ‘Redblush #3’.

Salinity Effects on Florida Grapefruit in the Indian River Region

A study was initiated in the 1997-98 production season to evaluate the effects of salinity on grapefruit yield and fruit quality in the Indian River area of Florida. The experiment was conducted on `Ray Ruby' grapefruit (Citrus paradisi) planted in 1990 on `Carrizo' citrange (C. sinensis × Poncirus trifoliata) and `Swingle' citrumelo (C. paradisi × P. trifoliata) rootstocks. Trees were planted on 15.2-m-wide (50 ft) double-row beds at a spacing of 4.6 m (15 ft) in-row × 7.3 m (24 ft) across-row [286.6 trees/ha (116 trees/acre)]. The control treatment was irrigated via microsprinkler emitters with water from a surficial aquifer well with an electrical conductivity (EC) of 0.7 dS·m-1. Higher irrigation water salinity levels were achieved by injecting a sea water brine mixture into the supply water to achieve ECs of 2.3, 3.9, and 5.5 dS·m-1. A wide range of rainfall and irrigation conditions occurred during the years encompassed by these studies, with rain totaling 1262, 1294, 1462, and 964 mm (49.7, 50.9, 57.6, and 38.0 inches) for 1997, 1998, 1999, and 2000, respectively. Salinity level had little effect on internal juice quality parameters [total soluble solids (TSS), acid, or juice content] at time of harvest. One of the most visible effects of irrigation with high salinity water was the damage to leaves, with leaf chloride (Cl) levels increasing about 0.14% for each 1.0 dS·m-1 increase in EC of the irrigation water for trees on `Carrizo' citrange and 0.02% for trees on `Swingle' citrumelo. For both rootstocks, the number of fruit and the size of the fruit decreased with increasing salinity in the irrigation water. The non-salinized trees had significantly larger fruit compared to the rest of the treatments. In the very dry 2000-01 season, trees on `Carrizo' irrigated with 0.7 dS·m-1 water had about 50% more fruit size 36 [fruit count per 0.028-m3 (4/5 bu) carton] or larger than trees watered with 3.9 or 5.5 dS·m-1 water. For trees on `Swingle' rootstock, trees irrigated with 0.7 dS·m-1 water had 150% to 200% more size 36 and larger fruit than trees watered with 2.3 dS·m-1 water. Over the four seasons, average yields for `Carrizo' were reduced 3200.0 kg·ha-1 (2855 lb/acre) per year for each 1.0 dS·m-1 increase in EC of the irrigation water. For `Swingle' rootstock, the reduction was 2600.3 kg·ha-1 (2320 lb/acre) per year for each 1.0 dS·m-1 increase in EC of the irrigation water. These reductions averaged 7% (`Swingle') and 6% (`Carrizo') for each 1.0 dS·m-1 increase in salinity of the irrigation water.