John K. Fellman

The BioCassava Plus Program: Biofortification of Cassava for Sub-Saharan Africa

More than 250 million Africans rely on the starchy root crop cassava (Manihot esculenta) as their staple source of calories. A typical cassava-based diet, however, provides less than 30% of the minimum daily requirement for protein and only 10%-20% of that for iron, zinc, and vitamin A. The BioCassava Plus (BC+) program has employed modern biotechnologies intended to improve the health of Africans through the development and delivery of genetically engineered cassava with increased nutrient (zinc, iron, protein, and vitamin A) levels. Additional traits addressed by BioCassava Plus include increased shelf life, reductions in toxic cyanogenic glycosides to safe levels, and resistance to viral disease. The program also provides incentives for the adoption of biofortified cassava. Proof of concept was achieved for each of the target traits. Results from field trials in Puerto Rico, the first confined field trials in Nigeria to use genetically engineered organisms, and ex ante impact analyses support the efficacy of using transgenic strategies for the biofortification of cassava.

A role for jasmonates in climacteric fruit ripening

Abstract. Jasmonates are a class of oxylipins that induce a wide variety of higher-plant responses. To determine if jasmonates play a role in the regulation of climacteric fruit ripening, the effects of exogenous jasmonates on ethylene biosynthesis and color, as well as the endogenous concentrations of jasmonates were determined during the onset of ripening of apple (Malus domestica Borkh. cv. Golden Delicious) and tomato (Lycopersicon esculentum Mill. cv. Cobra) fruit. Transient (12 h) treatment of pre-climacteric fruit discs with exogenous jasmonates at low concentration (1 or 10 μM) promoted ethylene biosynthesis and color change in a concentration-dependent fashion. Activities of both 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase and ACC synthase were stimulated by jasmonate treatments in this concentration range. The endogenous concentration of jasmonates increased transiently prior to the climacteric increase in ethylene biosynthesis during the onset of ripening of both apple and tomato fruit. The onset of tomato fruit ripening was also preceded by an increase in the percentage of the cis-isomer of jasmonic acid. Inhibition of ethylene action by diazocyclopentadiene negated the jasmonate-induced stimulation of ethylene biosynthesis, indicating jasmonates act at least in part via ethylene action. These results suggest jasmonates may play a role together with ethylene in regulating the early steps of climacteric fruit ripening.

Preharvest factors influencing flavor of fresh fruit and vegetables

Abstract Perception of fruit and vegetable flavor is a composite of sensory responses in the nose and mouth to aroma and taste. A diverse array of fruit and vegetable constituents including acids, sugars, volatiles and many other compounds individually elicit sensory responses that are recognized in total as flavor. Accumulation of these compounds during development as well as dynamic changes during ripening and/or senescence are determined in large part by the genetics of each species as well as developmental stage at harvest. However, other factors that influence development prior to harvest subsequently impact flavor. For horticultural crops, environment, cultural practices, agrichemicals and nutrition are some of the factors impacting flavor through effects on plant development.

Relationship of harvest maturity to flavor regeneration after CA storage of ‘Delicious’ apples

Red Delicious producers in the USA face increased pressure to produce fruit with optimum ‘on-shelf’ dessert quality following storage. In addition to firmness and soluble solids measurement other ripening-related events affect flavor perception. Ester production is always closely linked to the onset of climacteric ripening, while prolonged low-oxygen storage is usually detrimental to volatile production. Experiments focused on timing of the optimum harvest for maintaining sweetness, sourness and aroma generating capacity during CA storage. In multiple-harvest experiments with ‘Redchief Delicious’ apples, we investigated the relationship between the internal ethylene maturity indicator and flavor retention and regeneration after storage for different lengths of time. Using a cohort of untrained panelists, it was found that overall flavor perception and perceived fruit ripeness begins to increase at the onset of the climacteric. CA conditions reduce post-storage volatile production when compared with those stored in refrigerated air although not to a level below those displayed in ‘over-mature’ fruit at harvest after 3 months in storage. As harvest maturity advanced, the time required to regenerate aroma volatiles to an ‘optimum’ level after removal from CA storage decreased markedly. A linear relationship between attainment of optimal eating quality and time out of storage was established. Firmness, soluble solids, and titratable acidity of fruit from all harvest dates remained at acceptable levels throughout the post-storage ripening period.

Ester biosynthesis in “Rome” apples subjected to low-oxygen atmospheres

Abstract The relationship between acetate ester-forming activity of acetyl CoA alcohol transferase (ACAT), non-ethylene volatile emission, and flesh volatile content of “Rome” apples ( Malus domestica Borkh.) after removal from 9 months storage in low-oxygen environments was investigated. Apple samples held at 0–1°C were removed from refrigerated air (RA) or controlled-atmosphere (CA) storage at 1.0 or 0.5% v/v O 2 /1% CO 2 and placed in ambient laboratory conditions. Every three days fruit flesh and headspace were analyzed for volatile compounds using capillary gas chromatography. Acetate ester forming activity was assayed spectrophotometrically on partially-purified extracts of cortical tissue. Patterns of acetate ester formation depended upon storage environment and alcohol moiety precursor. Ethyl acetate content was always higher in the headspace and flesh of RA fruit, regardless of time after storage. Propyl acetate concentrations in CA stored apple flesh were higher until post-storage day 12 when they decreased. Headspace propyl acetate levels were higher in RA controls until days 9–15 when more emanated from CA fruit. Butyl acetate concentrations were lower in flesh and headspace of CA fruit until day 15, when RA fruit headspace levels decreased. Headspace and flesh concentrations of 2-methyl-1-butyl acetate were higher in CA apples. Acetate ester-forming activity was detectable at day 0 in 1.0% oxygen-stored fruits, but not in 0.5% oxygen-stored fruits. By post-storage day 9, ACAT activity in CA apples reached maximal levels, only to decrease by day 15. RA-stored apples had more ACAT activity at day 0, but did not substantially increase in activity like the 0.5% oxygen-stored apples removed from CA storage. It is unknown whether the ACAT protein is reactivated by exposure to ambient oxygen or synthesized de novo.

Non-contact bruise detection in apples by thermal imaging

Abstract Thermal imaging is a non-destructive and non-contact infrared sensing technique. Such imaging creates a bit-map called a thermogram by detecting infrared radiation emitted from an object. Up to 100% of apple bruises were detected using thermal imaging during warming of the fruits by discriminating surface temperature between bruised and sound tissues. Apples were bruised by dropping them from 0.46 m onto a smooth concrete floor and then were held at 26 °C and 50% RH for 48 h. They were then thermally imaged using a ThermaCam™ PM390 (FLIR Systems, Inc., Portland, OR) during heating and cooling treatments. Thermal images of bruised tissue showed at least 1–2 °C difference from sound tissue within 30–180 s. The temperature differences between bruised and sound tissues were possibly due to the differences in thermal diffusivity. Under steady-state temperature, thermal imaging did not detect bruises, indicating that the temperature differences were not due to emissivity differences. The technique could provide a basis for automatic bruise sorting, and possibly a better understanding of bruised tissue.

Inhibition of Salmonella Typhimurium and Listeria monocytogenes in mung bean sprouts by chemical treatment.

This study was undertaken to compare the efficacies of chlorous acid (268 ppm), sodium hypochlorite (200 ppm), and lactic acid (2%) in eliminating total mesophilic microorganisms, Salmonella Typhimurium, and Listeria monocytogenes on commercial mung bean sprouts immediately after treatment and during posttreatment refrigerated storage. Treatment with sodium hypochlorite for 10 min did not reduce the total aerobic count. However, treatment with lactic acid and chlorous acid for 10 min initially reduced the total aerobic count by 0.6 and 0.8 log CFU/g, respectively, and maintained the same level or a lower level of the total aerobic count during the storage time. Treatment with chlorous acid reduced Salmonella Typhimurium from 5.0 log to undetectable levels (<0.48 log CFU/g), and the pathogen remained undetectable over a 9-day storage period. Treatment with lactic acid resulted in an initial 3-log reduction and further reduced the number of Salmonella Typhimurium cells to undetectable levels after 3 days. For L. monocytogenes, treatment with chlorous acid resulted in an initial 5-log reduction, and treatment with lactic acid resulted in a 2-log reduction at the beginning and undetectable levels after 9 days. When chemically injured cells were investigated by the selective overlay method, no statistical difference was observed (P < 0.05) between the number of injured cells recovered following treatment with chlorous acid and the number of bacteria counted on selective media, whereas sodium hypochlorite generated more injured cells than the other treatments did. These data suggest that treatment with chlorous acid may be useful in reducing total mesophilic microorganisms, Salmonella Typhimurium, and L. monocytogenes in commercial mung bean sprouts.

The influence of nitrogen and biotin interactions on the performance of Saccharomyces in alcoholic fermentations.

Aim:  To study the impact of assimilable nitrogen, biotin and their interaction on growth, fermentation rate and volatile formation by Saccharomyces.

Inhibition of apple fruit 1-aminocyclopropane-1-carboxylic acid oxidase activity and respiration by acetylsalicylic acid

Summary Acetylsalicylic acid (aspirin) inhibited ethylene production of apple (Malus domestica Borkh. cv. Granny Smith) fruit discs in a concentration and time-dependent fashion. Activity of 1-aminocyclopropane-1-carboxylic acid (ACQ oxidase was inhibited both in vivo and in vitro by acetylsalicylic acid, both Vmax and Km decreased as acetylsalicylic acid concentration increased. The apparent Ki was 0.12 mmol·L−1. We conclude acetylsalicylic acid inhibits ethylene production by inhibiting ACC oxidase activity. For unknown reasons, acetylsalicylic acid also inhibited fruit respiration.

Alteration of foliar flavonoid chemistry induced by enhanced UV-B radiation in field-grown pinus ponderosa, Quercus rubra and Pseudotsuga menziesii

Chromatographic analyses of foliage from several tree species illustrate the species-specific effects of UV-B radiation on both quantity and composition of foliar flavonoids. Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii were field-grown under modulated ambient (1x) and enhanced (2x) biologically effective UV-B radiation. Foliage was harvested seasonally over a 3-year period, extracted, purified and the flavonoid fraction applied to a mu Bondapak/C(18) column HPLC system sampling at 254 nm. Total flavonoid concentrations in Quercus rubra foliage were more than twice (leaf area basis) that of the other species; Pseudotsuga menziesii foliage had intermediate levels and P. ponderosa had the lowest concentrations of total flavonoids. No statistically significant UV-B radiation-induced effects were found in total foliar flavonoid concentrations for any species; however, concentrations of specific compounds within each species exhibited significant treatment effects. Higher (but statistically insignificant) levels of flavonoids were induced by UV-B irradiation in 1- and 2-year-old P. ponderosa foliage. Total flavonoid concentrations in 2-year-old needles increased by 50% (1x ambient UV-B radiation) or 70% (2x ambient UV-B radiation) from that of 1-year-old tissue. Foliar flavonoids of Q. rubra under enhanced UV-B radiation tended to shift from early-eluting compounds to less polar flavonoids eluting later. There were no clear patterns of UV-B radiation effects on 1-year-old P. menziesii foliage. However, 2-year-old tissue had slightly higher foliar flavonoids under the 2x UV-B radiation treatment compared to ambient levels. Results suggest that enhanced UV-B radiation will alter foliar flavonoid composition and concentrations in forest tree species, which could impact tissue protection, and ultimately, competition, herbivory or litter decomposition.