Gary A. Luzio

Effect of Liberibacter Infection (Huanglongbing Disease) of Citrus on Orange Fruit Physiology and Fruit/Fruit Juice Quality: Chemical and Physical Analyses

More than 90% of oranges in Florida are processed, and since Huanglongbing (HLB) disease has been rumored to affect fruit flavor, chemical and physical analyses were conducted on fruit and juice from healthy (Las -) and diseased (Las +) trees on three juice processing varieties over two seasons, and in some cases several harvests. Fruit, both asymptomatic and symptomatic for the disease, were used, and fresh squeezed and processed/pasteurized juices were evaluated. Fruit and juice characteristics measured included color, size, solids, acids, sugars, aroma volatiles, ascorbic acid, secondary metabolites, pectin, pectin-demethylating enzymes, and juice cloud. Results showed that asymptomatic fruit from symptomatic trees were similar to healthy fruit for many of the quality factors measured, but that juice from asymptomatic and especially symptomatic fruits were often higher in the bitter compounds limonin and nomilin. However, values were generally below reported taste threshold levels, and only symptomatic fruit seemed likely to cause flavor problems. There was variation due to harvest date, which was often greater than that due to disease. It is likely that the detrimental flavor attributes of symptomatic fruit (which often drop off the tree) will be largely diluted in commercial juice blends that include juice from fruit of several varieties, locations, and seasons.

Enzymatic modification of a model homogalacturonan with the thermally tolerant pectin methylesterase from Citrus: 1. Nanostructural characterization, enzyme mode of action, and effect of pH.

Methyl ester distribution in pectin homogalacturonan has a major influence on functionality. Enzymatic engineering of the pectin nanostructure for tailoring functionality can expand the role of pectin as a food-formulating agent and the use of in situ modification in prepared foods. We report on the mode of action of a unique citrus thermally tolerant pectin methylesterase (TT-PME) and the nanostructural modifications that it produces. The enzyme was used to produce a controlled demethylesterification series from a model homogalacturonan. Oligogalacturonides released from the resulting demethylesterified blocks introduced by TT-PME using a limited endopolygalacturonase digestion were separated and quantified by high-pressure anion-exchange chromatography (HPAEC) coupled to an evaporative light-scattering detector (ELSD). The results were consistent with the predictions of a numerical simulation, which assumed a multiple-attack mechanism and a degree of processivity ∼10, at both pH 4.5 and 7.5. The average demethylesterified block size (0.6-2.8 nm) and number of average-sized blocks per molecule (0.8-1.9) differed, depending upon pH of the enzyme treatment. The mode of action of this enzyme and consequent nanostructural modifications of pectin differ from a previously characterized citrus salt-independent pectin methylesterase (SI-PME).

Formation of Stylet Sheaths in āere (in air) from Eight Species of Phytophagous Hemipterans from Six Families (Suborders: Auchenorrhyncha and Sternorrhyncha)

Stylet sheath formation is a common feature among phytophagous hemipterans. These sheaths are considered essential to promote a successful feeding event. Stylet sheath compositions are largely unknown and their mode of solidification remains to be elucidated. This report demonstrates the formation and solidification of in āere (in air) produced stylet sheaths by six hemipteran families: Diaphorina citri (Psyllidae, Asian citrus psyllid), Aphis nerii (Aphididae, oleander/milkweed aphid), Toxoptera citricida (Aphididae, brown citrus aphid), Aphis gossypii (Aphididae, cotton melon aphid), Bemisia tabaci biotype B (Aleyrodidae, whitefly), Homalodisca vitripennis (Cicadellidae, glassy-winged sharpshooter), Ferrisia virgata (Pseudococcidae, striped mealybug), and Protopulvinaria pyriformis (Coccidae, pyriform scale). Examination of in āere produced stylet sheaths by confocal and scanning electron microscopy shows a common morphology of an initial flange laid down on the surface of the membrane followed by continuous hollow core structures with sequentially stacked hardened bulbous droplets. Single and multi-branched sheaths were common, whereas mealybug and scale insects typically produced multi-branched sheaths. Micrographs of the in āere formed flanges indicate flange sealing upon stylet bundle extraction in D. citri and the aphids, while the B. tabaci whitefly and H. vitripennis glassy-winged sharpshooter flanges remain unsealed. Structural similarity of in āere sheaths are apparent in stylet sheaths formed in planta, in artificial diets, or in water. The use of ‘Solvy’, a dissolvable membrane, for intact stylet sheath isolation is reported. These observations illustrate for the first time this mode of stylet sheath synthesis adding to the understanding of stylet sheath formation in phytophagous hemipterans and providing tools for future use in structural and compositional analysis.

Changes in Volatile and Non-Volatile Flavor Chemicals of “Valencia” Orange Juice over the Harvest Seasons

Florida “Valencia” oranges have a wide harvest window, covering four months after first reaching the commercial maturity. However, the influence of harvest time on juice flavor chemicals is not well documented, with the exception of sugars and acids. Therefore, we investigated the major flavor chemicals, volatile (aroma), non-volatile (taste) and mouth feel attributes, in the two harvest seasons (March to June in 2007 and February to May in 2012). Bitter limonoid compounds, limonin and nomilin, decreased gradually. Out of a total of 94 volatiles, 32 increased, 47 peaked mid to late season, and 15 decreased. Juice insoluble solids and pectin content increased over the season; however, pectin methylesterase activity remained unchanged. Fruit harvested in the earlier months had lower flavor quality. Juice from later harvests had a higher sugar/acid ratio with less bitterness, while, many important aroma compounds occurred at the highest concentrations in the middle to late season, but occurred at lower concentrations at the end of the season. The results provide information to the orange juice processing industry for selection of optimal harvest time and for setting of precise blending strategy.

Use of an Evaporative Light Scattering Detector Coupled to MALLS for Determination of Polysaccharide Molecular Weights

Abstract A method to calibrate and use an evaporative light scattering detector (ELS) as a mass detector for molecular weight determination of polysaccharides using a multi‐angle laser light scattering detector (MALLS) is described. The calibration of the ELS is performed under isocratic conditions using concentration values obtained from an interferometric refractive index detector (RI). The observed response fit a power trendline (y= a x b ) for the concentration response of the RI versus the voltage response of the ELS. The combined average log a coefficient was (5.472, s.d.=0.020) and the average b coefficient was (1.372, s.d.=0.004) for all the runs. Band broadening, which could occur between the detectors, was not observed and did not affect the calibration values. Without adjustment the ELS was used as a mass detector for MALLS to accurately determine molecular weights (MW) at elevated buffer concentrations. Mention of a trademark or proprietary product is for identification only and does not imply a guarantee or warranty of the product by the US Department of Agriculture. The U.S. Department of Agriculture prohibits discrimination in all its programs and activities on the basis of race, color, national origin, gender, religion, age, disability, political beliefs, sexual orientation and marital or family status.

Determination of degree of methylation of food pectins by chromatography.

BACKGROUND In Florida, 90% of citrus is processed into juice, leaving large amounts of peel waste that could be processed into food ingredients such as pectins for other applications. Pectins of low degree of methylation (DM < 50) have important functional properties for use in foods and pharmaceuticals. Thus determination of DM is important, but DM analyses are difficult to perform and assays can have interference due to the presence of salts or neutral sugars. RESULTS A chromatography method has been developed for determination of DM of food pectin using alkaline hydrolysis followed by pectin digestion with a commercial pectinase. Analysis was performed by ion exclusion chromatography (IEC) to obtain methanol (MeOH) concentrations and by anion exchange chromatography (AEC) to obtain galacturonic acid (GalA) concentrations. Data were compared with values obtained using other DM analysis procedures. For example, a 55-75 DM pectin assayed as 55.5, 59.6, 60.1 and 57.0 DM by IEC, AEC, colorimetric and conductivity detection methods respectively. CONCLUSION The chromatography procedure described in this paper provides an alternative method for determination of DM of food pectins for accurate and efficient analysis when confronted with limited quantities or large numbers of samples.