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  • Author or Editor: Gary F. Polking x
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Our recent research has focused on the control of genes and enzymes involved with the synthesis of chlorophyll, especially as it relates to tomato fruit development and ripening. Glutamate-1-semialdehyde-2,1-aminomutase (GSAAM) is one of the first committed enzymes in the chlorophyll biosynthetic pathway, and it is one of three enzymes that catalyze the conversion of glutamate into 5-aminolevulinic acid. We have isolated a full-length cDNA clone of GSAAM from a tomato fruit library. The tomato primary sequence shows extensive homology to GSAAM sequences found in other plant species. The primary structure also predicts a 46.7-kDa, 437-amino acid, mature protein and a transit peptide of 44 amino acids. Southern analysis indicated that GSAAM was present as a single copy. Northern blot analysis showed that GSAAM was expressed differentially in various tomato organs and that GSAAM transcripts decreased with increased fruit age. Immunoblot analysis also indicated that GSAAM protein decreased dramatically with increased fruit age. These results show that there is developmental regulation of the expression of GSAAM in tomato fruits.

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Germinated Impatiens wallerana Hook. f. `Super Elfin Salmon Blush' seeds were exposed to subatmospheric O2 concentrations for 12, 24, or 48 hours at 25C. Suppression of radicle growth during a subsequent 24-hour simulated shipping period was monitored, as was plant growth during a subsequent growth cycle. One percent to 2% O2 for 12 or 24 hours limited radicle elongation to <1.0 mm during the simulated shipping period (darkness, ambient O2) and caused no permanent damage to seedlings. Suppression of radicle elongation with low O2 was greater with a 24-hour than a 12-hour exposure. Oxygen at 0% for 24 hours or at 0% to 1.5% O2 for 48 hours damaged seedlings irreversibly. These results show that specific subatmospheric O2 treatments can restrict radicle elongation of germinated seeds during subsequent shipment to a grower and that the low O2 treatment does not decrease subsequent plant growth.

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Generation of pregerminated seeds at a central facility and subsequent mechanical sowing at a remote facility require that radicle elongation during shipment be minimized. Low-oxygen treatments were applied to pregerminated `Super Elfin Salmon Blush' impatiens seeds to suppress radicle growth during a subsequent one-day simulated shipping period in air. In the first experiment, O2 concentrations of 0, 3, 7, 10, 13, and 20% were applied for 24 and 48hr. The 0% O2/24-hr and the 0% O2/48-hr treatments held the radicle length close to the desired length of 1.0mm, but both of these treatments decreasad 7-day hypocotyl length and percentage normal seedlings, when they were compared with the control treatment (untreated pregerminated seeds) In the second experiment, O2 concentrations of 0, 1, 2, 3, 7, and 20% were applied for 24 and 48hr. Five treatments (0, 1, and 2% O2/24-hr and 0 and 1% O2/48-hr) held the radicle to a length <1.0mm. Of these five treatments, only the 2% O2/24-hr treatment resulted in recovery parameters (7-day hypocotyl length and percentage normal seedlings) that compared with those of the control treatment.

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Activity of 5-aminolevulinic acid (ALA) dehydratase [ALAD, (EC 4.2.1.24)] and soluble protein content were determined in `Rutgers' tomato (Lycopersicon esculentum Mill.) fruit pericarp extracts during development and ripening. ALAD activity in several organs of tomato plant also was determined. Fruit tissue was analyzed at 5-day intervals between days 10 and 60 postanthesis. ALAD activity in fruit tissue declined over time, with the most pronounced decrease occurring between days 10 and 25. At the mature green stage (day 40), and before the breaker stage (day 45), activity of ALAD had declined to a steady-state minimum, and it remained detectable at residual levels throughout ripening (days 40 to 60). Soluble protein content declined less rapidly than did ALAD activity. Immunoblot analysis showed that ALAD protein existed as a doublet of isozymes. One isozyme decreased in abundance, whereas the other isozyme remained constant during development and ripening. ALAD activity was greatest in extracts of chlorophyllous organs (stems, leaves, immature fruit) but only marginally detectable in extracts of nonchlorophyllous organs (roots, overripe fruit). The pH optimum and Km for tomato fruit ALAD were similar to those of ALAD isolated from other sources. Abbreviations: ALA, 5-aminolevulinic acid; ALAD, ALA dehydratase; PBG, porphobilinogen; SDS-PAGE, sodium dodecyl sulfate-polyacry - lamide gel electrophoresis.

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