Browsing by Author "Cutting, Jonathan Garth Melville."

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Optimization, validation and application of radioimmunoassays for plant growth substances in avocado (Persea americana Mill.) fruits.
(1984) Cutting, Jonathan Garth Melville.; Wolstenholme, B. Nigel.; Lishman, Arthur William.
The objective was to develop, optimize and validate radioimmunoassays (RIA) for several plant growth substances (PGS) and then apply the RIA's to determine PGS trends in 'Fuerte' avocado fruits from fruit set to fruit ripening. Antibodies to the cytokinin isopentenyl adenosine (IPA) were obtained from rabbits inoculated with a periodate-derived IPA-BSA conjugate. The antiserum cross-reacted (25%) with only 2iP (isopentenyl adenine). The RIA measuring range was from 0,1 to 100 ng. Anti-IPA serum was used to develop a RIA for 2iP, with a measuring range from 0,5 to 100 ng. Using Dowex 50W-X8 and cellulose TLC purified avocado fruit extract, 20 samples per day could be processed. The RIA for abscisic acid (ABA) was developed from rabbit antibodies from an inoculated carbodiimide-derived, (+ -) ABA-BSA conjugate. The free active ABA component was isolated prior to quantitation by RIA by solvent partitioning and silica gel TLC. The indole-acetic-acid (IAA) RIA was established from sheep-produced antibodies to a formaldehye-derived IAA-BSA conjugate, after repeated inoculations. For both the ABA and IAA RIA's, contaminants in the avocado tissue were removed by solvent partitioning. Developing avocado fruits, and especially young fruits were rich sources of IAA and 2iP in particular with seed concentrations exceeding those of the fruit flesh. The concentration of ABA rose throughout fruit development, reaching 100 ng g(-1) in the flesh at fruit maturity. Just prior to seed and fruit maturity, relatively high levels of IAA, 2iP and IPA were associated with the thick, fleshy testa, rapidly declining to zero as the testa dried. The avocado fruit is physiologically dependent on the seed right up to this stage, and testa maturity correlates well with minimum "legal" maturity of 80% moisture content of flesh. In ripening avocado fruit, the concentration of free ABA rose as softening progressed. Total ABA concentrations fell initially, and then rose after a firmometer reading between 50 and 60 (100 coincides with eating ripeness). Later harvested fruit had double the ABA concentration of early harvested fruit. The ripening stimulus appeared to be related to moisture stress in the fruit. This study has confirmed the prime advantages of sensitivity, specificity and rapidity of RIA, as well as its usefulness in multi-PGS studies and batch-type analysis.
Some aspects of cold storage of 'Fuerte' avocados (Persea americana Mill.) grown in the Natal midlands.
(1995) Donkin, Derek John.; Wolstenholme, B. Nigel.; Cutting, Jonathan Garth Melville.
The South African avocado industry is largely export orientated and export by sea to European markets necessitates cold storage for up to 4 weeks at temperatures around 5.5°C. Avocado fruit is subject to chilling injury which is manifested as mesocarp discolouration, and pitting and blackening of the rind. Of the South African cultivars exported, 'Fuerte' is the most susceptible to chilling injury, and accounts for > 50 % of avocado exports. A number of temperature regimes where temperature was reduced in a step-wise fashion from 8.5 or 7.5°C to 4.5 or 5.5°C during 3 to 5 weeks of storage were tested weekly throughout the 1993 and 1994 'Fuerte' harvesting seasons in the Natal Midlands (a cool mesic subtropical area), in attempt to find cold storage temperature regimes which would minimise chilling injury. No definite trends with regard to certain temperature regimes resulting in fruit with less chilling injury were evident. Overall, stepped down temperature regimes produced fruit of quality no better than storage for 5.5°C for 4 weeks. There was no significant difference in concentration of total phenolics in 'Fuerte' fruit mesocarp throughout the 1994 harvesting season (P < 0.05). Levels of ethylene evolution during 4 weeks of storage at 7.5 and 5.5°C ranged from 0 to 5 µl.kg(-l).h(-l), and peaked at 109 and 75 µl.kg(-l).h(-1) in fruit stored at 7.5 and 5.5°C respectively at room temperature on removal from cold storage. Rapid moisture removal from 'Fuerte' fruit after harvest and before cold storage by placing the fruit in glass jars to which a suction of -75 kPa was applied, resulted in increased susceptibility to external chilling injury, the severity of which was proportional to the amount of moisture removed from the fruit. Pre-storage heat treatments with a view to decreasing sensitivity of fruit to cold storage were carried out on 'Fuerte' fruit. Dry heat and warm water baths at temperatures of 36 to 40°C caused rind blackening of varying severity, depending on temperature and duration. Vapour heat treatments at temperatures of 36 to 48°C for 10 min to 48 h also caused rind blackening, with the exception of 10 min at 48°C and 1.5 and 3 h at 40°C which produced fruit of higher overall quality after 4 weeks of cold storage at 3.5°C than fruit not heat treated. These treatments however, could not be repeated in 1994 to confirm the results obtained as the harvesting season was over by the time the trial was completed.