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PLANT PHYSIOLOGY , Vol 102, Issue 2 473-480, Copyright © 1993 by American Society of Plant Biologists
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METABOLISM AND ENZYMOLOGY |
Polyuronides in Avocado (Persea americana) and Tomato (Lycopersicon esculentum) Fruits Exhibit Markedly Different Patterns of Molecular Weight Downshifts during Ripening
D. J. Huber and E. M. O'Donoghue
Horticultural Sciences Department, University of Florida, Gainesville, Florida 23611-0690
Avocado (Persea americana) fruit experience a rapid and extensive loss of
firmness during ripening. In this study, we examined whether the chelator
solubility and molecular weight of avocado polyuronides paralleled the
accumulation of polygalacturonase (PG) activity and loss in fruit firmness.
Polyuronides were derived from ethanolic precipitates of avocado mesocarp
prepared using a procedure to rapidly inactivate endogenous enzymes. During
ripening, chelator (cyclohexane-trans-1,2-diamine tetraacetic acid
[CDTA])-soluble polyuronides increased from approximately 30 to 40 [mu]g of
galacturonic acid equivalents (mg alcohol-insoluble solids)-1 in preripe
fruit to 150 to 170 [mu]g mg-1 in postclimacteric fruit. In preripe fruit,
chelator-extractable polyuronides were of high molecular weight and were
partially excluded from Sepharose CL- 2B-300 gel filtration media. Avocado
polyuronides exhibited marked downshifts in molecular weight during
ripening. At the postclimacteric stage, nearly all chelator-extractable
polyuronides, which constituted from 75 to 90% of total cell wall uronic
acid content, eluted near the total volume of the filtration media.
Rechromatography of low molecular weight polyuronides on Bio-Gel P-4
disclosed that oligomeric uronic acids are produced in vivo during avocado
ripening. The gel filtration behavior and pattern of depolymerization of
avocado polyuronides were not influenced by the polyuronide extraction
protocol (imidazole versus CDTA) or by chromatographic conditions designed
to minimize interpolymeric aggregation. Polyuronides from ripening tomato
(Lycopersicon esculentum) fruit extracted and chromatographed under
conditions identical with those used for avocado polyuronides exhibited
markedly less rapid and less extensive downshifts in molecular weight
during the transition from mature-green to fully ripe. Even during a 9-d
period beyond the fully ripe stage, tomato fruit polyuronides exhibited
limited additional depolymerization and did not include oligomeric species.
A comparison of the data for the avocado and tomato fruit indicates that
downshifts in polyuronide molecular weight are a prominent feature of
avocado ripening and may also explain why molecular down-regulation of PG
(EC 3.2.1.15) in tomato fruit has resulted in minimal effects on fruit
performance until the terminal stages of ripening.
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