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  • Author or Editor: Richard J. Gladon 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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Tomato fruit ripening is characterized by a decrease in chlorophyll content and an increase in lycopene synthesis. 5-Aminolevulinic acid (ALA) dehydratase (ALAD) is the fruit committed enzyme in the chlorophyll and heme biosynthetic pathways, and it catalyzes the dimerization of two ALA molecules into porphobilinogen We have focused our attention on the potential pivotal role of ALAD in the developmental regulation of chlorophyll biosynthesis during tomato fruit growth, development, and ripening. We have standardized an assay procedure for measuring the enzymatic activity of ALAD in tomato fruit tissues. The activity of ALAD was assayed from ten days past anthesis to day 60, when fruits where void of chlorophyll. We observed a several-fold decline in ALAD activity to residual levels during fruit ontogeny. Our data also show greater ALAD activity in chlorophyllous organs (leaves, stems, immature fruits) than in nonchlorophyllous organs (roots, ripe fruits), where heme production is predominant.

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Tomato fruit ripening is characterized by a decrease in chlorophyll content and an increase in lycopene synthesis. We are interested in the role of chlorophyll metabolism as it relates to tomato fruit ripening. 5-Aminolevulinic acid dehydratase (ALAD) is the first committed enzyme in the chlorophyll biosynthetic pathway, and it catalyzes the conversion of two 5-aminolevulinic acid molecules into porphobilinogen. We have isolated a full-length tomato ALAD cDNA clone from a tomato fruit library. Sequence analysis showed that this tomato ALAD was highly homologous to ALAD found in spinach and pea, and the analysis predicted a protein of 46.8 kDa. Southern analysis indicated that 1 to 3 copies of the ALAD gene are present in the tomato genome. Northern analysis suggested that the gene is expressed constitutively throughout tomato fruit development. Currently, we are subcloning the fragment into an E. coli expression vector in order to obtain protein for antibody production for Western analysis.

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Energy conservation in controlled-environment agriculture is a major concern for both commercial and research facilities as well as extraterrestrial facilities for food production. Supplying optimal irradiance by using electrical lighting for the greatest edible biomass production potentially is the greatest draw on energy during earth-based or extraterrestrial food production in controlled environments. Our objective was to determine the optimal irradiance for greatest edible biomass production of three cultivars of basil (Basilicum ocimum L.) in a controlled-environment production system. Seedlings of the three cultivars were transplanted into soilless medium, one plant per pot, and grew for 17 days in reach-in growth chambers maintained at 25 ± 4 °C with a 16-h photoperiod. Canopy-level irradiances of 300, 400, 500, and 600 μmol·m−2·s−1 were provided by cool-white fluorescent and incandescent lamps. Shoot growth was measured as height, diameter, and number of leaves 0.5 cm long or greater; and edible biomass was measured as leaf fresh weight, shoot fresh weight, and shoot dry weight. There was no irradiance × cultivar interaction, but main effects of irradiance and cultivar were observed. Plant growth and edible biomass production were least at 300 μmol·m−2·s−1 and greatest at 500 or 600 μmol·m−2·s−1. In several cases, 400 μmol·m−2·s−1 yielded intermediate growth or edible biomass. Within the main effect of cultivar, Italian Large Leaf produced greater edible biomass than ‘Genovese’, and ‘Nufar’ yielded an intermediate amount of shoot fresh weight and dry weight. Under our environmental conditions that included ambient CO2 concentration and ambient relative humidity, the rate of growth peaked at 500 μmol·m−2·s−1, and no additional accumulation of edible biomass occurred at 600 μmol·m−2·s−1. Based on our results, canopy-level irradiance of 500 μmol·m−2·s−1 provides maximum edible biomass production of basil in a controlled-environment production system.

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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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Impatiens (Impatiens wallerana Hook. f.) is the most important annual bedding plant in the US, based on wholesale dollar volume. Production of high-quality plants requires optimization of the nutrition regimen during growth, especially the total nitrogen (N) concentration and the ratio of N sources. Our objective was to determine the N concentration and ratio of N sources that optimize bedding-plant impatiens growth and development. We used four N concentrations (3.5, 7, 10.5, and 14 mmol·L-1 of N) in factorial combination with four ratios of nitrate-N (NO3 --N) to ammonium-N (NH4 +-N) (4:0, 3:1, 1:1, and 1:3). Application of treatments began at day 30, and every-other-day applications were conducted until day 60. From day 60 to day 70 only deionized water was applied. N concentration and source displayed interation for most growth parameters. When N was supplied at a concentration ≤7 mmol·L-1, the NO3 --N to NH4 +-N ratio did not affect growth. When N was supplied at a concentration ≥10.5 mmol·L-1, a 1:3 NO3 --N to NH4 +-N ratio yielded the greatest shoot dry weight, shoot fresh weight, plant diameter, and number of flower buds per plant. With a NO3 --N to NH4 +-N ratio of 4:0, these growth parameters decreased. To produce high-quality, bedding-plant impatiens, N should be applied at NO3 --N to NH4 +-N ratios between 1:1 and 1:3 in combination with an N concentration of 10.5 mmol·L<-1 at each fertigation from day 30 to day 60 of the production cycle.

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'Honeoye' (June-bearing) and 'Tristar' (day-neutral) strawberries were harvested, graded, and then stored for 7 days et 2C or 21C in air (control) or each of these 8 modified atmospheres: 1.5% O2, 3.5% O2, 15% CO2, 25% CO2, 1.5% O2 + 15% CO2, 1.5% O2 + 25% CO2, 3.5% O2 + 15% CO2, and 3.5% O2 + 25% CO2; all balance N2. When compared with storage at 21C, storage at 2C reduced weight loss and gray mold growth in all corresponding sets of storage atmosphere treatments. The combination of increased CO2 and decreased O2 controlled weight loss and gray mold growth more effectively than treatment with reduced O2 alone. Storage at 2C (versus 21C) reduced respiration of both cultivars. Respiration decreased as the O2 concentration decreased. 'Tristar' did not produce C2H4 at either temperature, whereas `Honeoye' produced more C2H4 at 21C than it did at 2C. Increased CO2 and/or decreased O2 concentrations in the storage atmosphere are not satisfactory substitutes for proper low-temperature storage of strawberries.

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Chlorophyll a and b contents were determined in developing tomato fruit (Lycopersicon esculentum Mill. `Heinz 1350') at 5-day increments from 10 or 15 days past anthesis to fulIy ripe (55 to 60 days). When presented on a whole-fruit basis, chlorophyll a and b contents increased from 15 days past anthesis to 35 days and then decreased to zero at 55 days. Porphobilinogen (EC 4.2.1.24; PBG) deaminase activity was measured in extracts from the fruit, and changes in PBG deaminase activity correlated with changes in chlorophyll and protein contents with respect to fruit age. Partial characterization of tomato PBG deaminase enzyme showed similarities to PBG deaminase enzymes isolated from other sources.

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Water uptake by impatiens (Impatiens wallerana Hook. f. cv. Super Elfin Coral) seeds was measured as an increase in fresh weight every 24 hours during 144 hours of germination. Seeds absorbed most of the water required for germination within 3 hours of imbibition and germinated at 60% to 67% moisture on a dry-weight basis. Germination started at 48 hours and was complete by 96 hours at 25C. Water stress of -0.1, -0.2, -0.4, and -0.6 MPa, induced by polyethylene glycol 8000, reduced germination by 13%, 49%, 91%, and 100%, respectively, at 96 hours. Under the same water-stress conditions, increases in fresh weight were inhibited by 53%, 89%, 107%, and 106%, respectively. Three distinct groups of storage proteins were present in dry seed; their estimated molecular weights were 1) 35, 33, and 31 kDa; 2) 26, 23, and 21 kDa; and 3) two bands <14 kDa. Major depletion of storage proteins coincided with the completion of germination. Water potentials that inhibited germination also inhibited degradation of storage proteins. During germination under optimum conditions, the soluble protein fraction increased, coinciding with a decrease in the insoluble fraction.

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