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Transgenic tomatoes (Lycopersicon esculentum Mill. `Ohio 8245') expressing an antisense catalase gene (ASTOMCAT1) were used to test the hypothesis that modification of the reactive oxygen species scavenging mechanism in plants can lead to changes in oxidative stress tolerance. A 2- to 8-fold reduction in total catalase activity was detected in the leaf extracts of transformants. A 2-fold increase in levels of H₂O₂ was observed in the transgenic plants with reduced catalase activity. Electrophoretic characterization of multiple catalase isoforms revealed the specific suppression of CAT1 in transgenic plants. Homozygous plants carrying the antisense catalase transgene were used to study the effect of alteration in the expression of catalase on stress tolerance. Transgenic plants treated with 3% H₂O₂ showed visible damage within 24 hours and subsequently died. In contrast, wild-type and azygous control plants recovered from the treatment. Transgenic plants did not survive 4 °C chilling stress compared to control wild-type and azygous lines. Physiological analysis of these plants indicated that suppression of catalase activity in transgenic tomato led to enhanced sensitivity to oxidative stress. Our data support a role for catalase in oxidative stress defense system in tomato.

Following a compatible pollination in carnation (Dianthus caryophyllus L. `White Sim'), a signal that coordinates postpollination events is translocated from the style to the ovary and petals. In this paper the roles of ethylene and its direct precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), in this signaling were investigated. Following pollination, ethylene and ACC increased sequentially in styles, ovaries, and petals. Ethylene and ACC were highest initially in the stigmatic region of the style but by 24 hours after pollination were highest in the base. Activity of ACC synthase correlated well with ethylene production in styles and petals. In ovaries, ACC synthase activity decreased after pollination despite elevated ethylene production. Lack of ACC synthase activity in pollinated ovaries, coupled with high ACC content, suggests that ACC is translocated within the gynoecium. Further, detection of propylene from petals following application to the ovary provided evidence for movement of ethylene within the flower. Experiments that removed styles and petals at various times after pollination suggest there is a transmissible pollination signal in carnations that has reached the ovary by 12 hours and the petals by 14 to 16 hours.

Treatment of cut carnation (Dianthus caryophyllus L. `White Sim') flowers with the synthetic cytokinin benzyladenine (BA) at concentrations >1.0 μm induced premature petal senescence. Flowers treated with 100 μm BA exhibited elevated ethylene production in styles and petals before untreated flowers. The gynoecia of BA-treated flowers accumulated 1-aminocyclopropane-l-carboxyllc acid (ACC) and enlarged before untreated flowers. Removal of the gynoecium (ovary and styles) or styles prevented BA-induced petal senescence and resulted in a substantial delay in petal senescence. In contrast, removal of the gynoecium had no effect on timing of petal senescence in flowers held in water. These results indicate BA stimulates petal senescence by inducing premature ACC accumulation and ethylene production in the gynoecium.

Abstract

‘Forever Yours’ roses (Rosa Hybrid Tea) were grown in recirculating nutrient solutions at 1.0 and 10.0 meq/liter K in combination with 10.0 meq/liter NO₃-N or NH₄-N. Low K limited the growth and flower production, regardless of N form. Ammonium-N fertilized plants showed NH₄-N toxicity symptoms as interveinal chlorosis of the lower leaflets. An increased K supply reduced NH₄-N toxicity symptoms. Concentrations of Ca and Mg were lower, while P was higher, in the tissue of NH₄-N fertilized plants, as compared to NO₃-N fertilized plants. Total N, alcohol insoluble N, soluble organic N, and NH₄-N were higher in the tissue of plants which received NH₄-N, as compared to NO₃-N, regardless of K level. An increased K supply from 1.0 to 10.0 meq/liter resulted in higher NO₃-N in NO₃-N fertilized plants and lower NH₄-N in NH₄-N fertilized plants.

Diurnal variation in the chilling sensitivity of `Rutgers' tomato (Lycopersicon esculentum Mill.) seedlings was examined. Chilling sensitivity was highest in seedlings chilled at the end of the dark period, and these seedlings became more resistant to chilling injury on exposure to the light. The development of chilling tolerance in tomato seedlings was a response to light and not under the control of a circadian rhythm. The recovery of leaf gas exchange following chilling was faster in seedlings chilled at the end of the light period. Diurnal variation in chilling sensitivity was associated with changes in catalase and superoxide dismutase activities. An increase in catalase and superoxide dismutase activities was observed at the end of the light period. Catalase activity was significantly higher in all stages of chilling following the light period compared to those chilled after the end of the dark period. Forty-eight hours of 14 °C acclimation or pretreatment with hydrogen peroxide conferred increased chilling tolerance to tomato seedlings. Hydrogen peroxide-treated seedlings showed little evidence of a diurnal variation in chilling sensitivity. These results support a role for light and oxidative stress in conferring increased chilling tolerance to tomato seedlings.

Abstract

Rooted cuttings of Chrysanthemum × morifolium Ramat. ‘Gt. #4 Indianapolis White’ were grown in a greenhouse in a sand culture and supplied with either 3.75 or 15.0 mm ${\text{NO}}_3^{-}$ . Changes in dry matter, reduced N and ${\text{NO}}_3^{-}$ of the leaves, stems (plus petioles), roots, and inflorescence and in vivo ${\text{NO}}_3^{-}$ reductase activity (NRA) of leaves were determined at various stages of development. A decrease in the ${\text{NO}}_3^{-}$ supply caused a decrease in the accumulation of plant dry matter, reduced N and ${\text{NO}}_3^{-}$ . Plants receiving 3.75 mm ${\text{NO}}_3^{-}$ remobilized a significant amount of reduced N from vegetative tissues during inflorescence development, suggesting that newly absorbed N was inadequate to supply the flower. At both ${\text{NO}}_3^{-}$ fertilization levels, the ${\text{NO}}_3^{-}$ content of the leaves and stems declined during inflorescence development, suggesting an increased dependence on previously accumulated ${\text{NO}}_3^{-}$ for reduction. The highest NRA of the leaves (3.4 μmoles NO₂ gFW⁻¹ hr⁻¹) was associated with early vegetative growth. NRA, however, was detectable throughout plant development. Nitrate reductase activity was greater at 15 mM ${\text{NO}}_3^{-}$ than at 3.75 mM ${\text{NO}}_3^{-}$ during vegetative growth and visible bud stages, but not at later stages of growth.

Mature pollen from Petunia hybrida contains significant levels of 1-aminocyclopropane-1-carboxylic acid (ACC), and this ACC is thought to play a role in pollination-induced ethylene by the pistil. We investigated the developmental accumulation of ACC in anthers and pollen. The level of ACC in anthers was very low until the day before anthesis, at which time it increased 100-fold. A 1.1-kb partial ACC synthase cDNA clone (pPHACS2) was amplified from total RNA isolated from mature anthers by reverse transcriptase, followed by polymerase chain reaction using oligonucleotide primers synthesized to conserved amino acid sequences in ACC synthases. The expression of pPHACS2 mRNA during anther development was correlated with the accumulation of ACC and was localized to the pollen grain. The pPHACS2 cDNA was used to identify the PH-ACS2 gene from a library of genomic DNA fragments from Petunia hybrida. PH-ACS2 encoded an ACC synthase transcript of four exons interrupted by three introns. The ACC synthase protein encoded by the PH-ACS2 gene shared >80% homology with ACC synthases from tomato (LE-ACS3) and potato (ST-ACS1a). A chimeric PH-ACS2 promoter-β-glucuronidase (GUS) gene was used to transform petunia and transgenic plants were analyzed for GUS activity. GUS staining was localized to mature pollen grains and was not detected in other tissues. Despite similarities to LE-ACS3, we did not detect GUS activity under conditions of anaerobic stress or in response to auxin. A series of 5-prime-flanking DNA deletions revealed that sequences within the PH-ACS2 promoter were responsible for pollen-specific expression.