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  • Author or Editor: Dennis P. Stimart x
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Leaf explants of Nicotiana alata Link and Otto. were surface disinfested and cultured on Murashige and Skoog (MS) medium containing 2.66 μm N6-benzyladenine (BA) to promote shoot proliferation. After 5 weeks, proliferated shoots were removed and remaining callus saved. Callus was inoculated with Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase which catalyzes cytokinin synthesis. Following inoculation, the callus was cocultivated for 6 days on BA medium. Selection for transgenics was done on BA medium plus 100 mg Kanamycin and 400 mg Ticarcillin (antibiotics) per liter. Proliferating shoots were rooted on MS medium containing antibiotics. Rooted cuttings were transplanted to soil, acclimated and flowered in the greenhouse. Transgenics were outcrossed to a commercial N. alata hybrid. Seed was germinated in vitro on half-strength MS medium plus antibiotics. Segregation of transgenics to nontransgenics was 1:1. Evaluation of leaf senescence on 5-month-old plants showed 2 to 14 times fewer senesced leaves on the transgenic than the nontransgenic plants.

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Cut flowers of a short (S)-lived (3-day) inbred, a long (L)-lived (15-day) inbred and their hybrid (F1, 7.3 days) of Antirrhinum majus L. were evaluated for fresh weight and ethylene evolution change postharvest when held in deionized water. Fresh weight change of all accessions increased 1 day postharvest then declined over the remainder of postharvest life. The loss of fresh weight was most rapid for S and less rapid for F1 and least rapid for L. Ethylene release postharvest for S and F1 started on day 1, but for L ethylene release started on day 9. Once ethylene evolution began it continued through postharvest life. On the last day of postharvest life, ethylene release from S and F1 were similar, but L was twice the level as S and F1. It appears that a slower decline in fresh weight, a delay in outset of ethylene release and higher final amount of ethylene release at senescence are heritable and associated with longer keeping time of A. majus.

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In an attempt to analyze genetically the interaction of endogenous auxin concentration and adventitious root formation, an EMS mutagenized M2 population of Arabidopsis thaliana was screened for mutants with altered abilities to form adventitious roots. A selected recessive nuclear mutant, rooty (rty), is characterized by extreme proliferation of roots, inhibition of shoot development and other morphological alterations suggestive of auxin or ethylene effects. The rty phenotype occurs in wild type seedlings grown on auxin containing medium and relatively normal growth is stimulated in rty seedlings growing on cytokinin containing medium. Analysis by GC-MS found that endogenous IAA concentrations in rty are 2 to 17 times higher than in wild type depending on tissue type and IAA form. Dose response experiments with IAA and NAA indicated that rty does not express increased sensitivity to auxin. These data suggest that the rty phenotype is due to elevated endogenous auxin. A genetic map location for rty and possible roles for the wild type RTY gene product in regulating auxin concentration will be presented.

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Eighteen commercially used white Antirrhinum majus (snapdragon) inbreds, a hybrid of Inbred 1 × Inbred 18 (Hybrid 1) and an F2 population (F2) of Hybrid 1 were evaluated for stomatal size and density and transpiration rate to determine their affect on postharvest longevity. Stems of each genotype were cut to 40 cm, placed in distilled water and discarded when 50% of florets wilted or browned. Postharvest longevity of inbreds ranged from 3.7 to 12.9 days; Hybrid 1 and the F2 averaged 3.0 and 9.1 days postharvest, respectively. Leaf impressions showed less than 3% of stomata were found on the adaxial leaf surface. Inbred abaxial stomatal densities ranged from 128.2 to 300.7 stomata mm-2; Hybrid 1 and the F2 averaged 155 and 197 stomata mm-2, respectively. Transpiration measurments on leaves of stems 24 hr after cutting were made with a LI-COR 1600 Steady State Porometer. Statistical analysis showed inbreds were significantly different based on postharvest longevity, stomatal size and density and transpiration of cut stems.

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Many physiological responses in plants are influenced by pH. The present chemiosmotic hypothesis suggests that auxin uptake into plant cells is governed by pH. Since auxin is used widely to enhance rooting, the influence of pH on 1H-indole-3-butyric acid (IBA) induced adventitious root formation was examined. Roots were initiated aseptically in 5 node apical shoot cuttings of micropropagated Malus domestica 'Gala'. Initiation was induced using a four day pulse in IBA and 15 g/L sucrose at pH 5.6 and 30C in the dark. Observations showed pH rose to 7.0 or greater within 1 to 2 days from microcutting placement in unbuffered initiation medium. Root numbers from shoots in media containing 1.5 μM IBA buffered with 10 mM 2[N-morpholino] ethanesulfonic acid (MES) to pH 5.5, 6.0, 6.5 or 7.0 with KOH resulted in average root numbers of 14.2, 10.9, 8.7, and 7.1, respectively, while unbuffered medium yielded 7,6 roots per shoot. Comparison of MES buffered medium at pH 5.5, 6.25 or 7.0 in factorial combination with IBA at 0, 0.15, 1.5, 15.0, and 150.0 μM resulted in a significant pH by IBA interaction for root number. At 0, 0.15 and 1.5 μM IBA root numbers were greatest at pH 5.5. At 15.0 μM IBA, pH 6.25 was optimal and at 150.0 μM IBA all three pH levels produced equivalent root numbers. A calorimetric assay to measure IBA removal from the initiation medium by microcuttings of `Gala' and `Triple Red Delicious' showed more IBA removal at pH 5.5 than at pH 7.0. Possible reasons for the effect of pH on adventitious root formation will be discussed.

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Selecting for increased postharvest longevity through use of natural variation is being investigated in Antirrhinum majus (snapdragon) in order to decrease postharvest chemical treatments for cut flowers. The postharvest longevity of eighteen white commercial inbreds was evaluated. Twelve stems of each inbred were cut to 40 cm and placed in distilled water. Stems were discarded when 50% of spike florets wilted or browned. Postharvest longevity ranged from 3.0 (Inbred 1) to 16.3 (Inbred 18) days. Crossing Inbred 18 × Inbred 1 yields commercially used Hybrid 1 (6.6 days postharvest). The F2 population averaged 9.1 days postharvest (range 1 to 21 days). F3 plants indicate short life postharvest may be conferred by a recessive gene in this germplasm. Populations for generation means analysis as well as hybrids between short, medium and long-lived inbreds were generated and evaluated for postharvest longevity.

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Actively growing shoots from Pulmonaria L. `Roy Davidson' were cultured in vitro on Murashige and Skoog medium containing benzyladenine (BA) to establish proliferating cultures. BA at 0, 0.4, 0.8, 4.4, 8.8, and 44.4 μm was compared for shoot proliferation and rooting response. Shoot count was highest on 8.8 μm BA with root count highest on 0 or 0.4 μm BA. Subculture 4 weeks later of shoots to the same treatments resulted in highest shoot counts on 44.4 μm BA. Optimum level for micropropagation was 8.8 or 44.4 μm BA. Greatest rooting was at 0 or 0.4 μm BA.

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Cotyledons from developing embryos 6 to 8 weeks old of Liatris spicata (blazing star) were cultured on Murashige-Skoog (MS) medium containing 0, 0.4, 4.4, and 44.4 μ M benzyladenine (BA) or 0, 0.2, 2.2, and 22.2 μ M thidiazuron (TDZ) to induce adventitious shoot formation. The highest percent of cotyledons forming shoots with highest shoot counts was on medium containing 2.2 μ M TDZ. Vitreous shoots formed on medium with 22.2 μ M TDZ. Callus derived from cotyledons and cultured on medium containing 4.44 μ M BA or 2.2 μ M TDZ formed adventitious shoots with highest shoot counts on 4.44 μ M BA. Adventitious shoots derived from cotyledons and callus were rooted on MS medium with 5.0 μ Mindole-3-butyric acid, acclimatized and grown ex vitro. All micropropagated plants appeared similar to each other.

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Abstract

Zinnia elegans Jacq. ‘Carved Ivory’, ‘Cherry Ruffles’, and ‘Rosy Future’ were grown under short days (SD) or long days (LD) to determine the effect of photoperiod on flower initiation and morphological development. Plants grown under LD (14, 16, 18, or 24 hours) were greater in height, leaf size, flower diameter, and node number and flowered later compared to plants grown under SD (8, 10, or 12 hours). The greatest increases in plant responses occurred between 12- and 14-hour treatments. Supplemental irradiation (day continuation, night break, and predawn lighting) resulted in LD responses but did not equally affect days to flowering, stem diameter, or node number. Height and ray petal number were greatest for all cultivars under day continuation and flower diameter was largest under day continuation and predawn lighting. More LD before SD resulted in increases in height, stem and flower diameter, node number, ray petal number, and days to flowering, indicating a cumulative effect of LD on morphological development. Results demonstrate that Z. elegans is a facultative SD plant for floral initiation and development and that photoperiodic responses vary between genotypes.

Open Access

Abstract

Bulblets of Lilium longiflorum generated in vitro in the dark at 30°C produced leaves more rapidly and more completely after transplanting to vermiculite than those generated in vitro at 25°. The product of temperature × time (days) was equalized by varying the number of days at the 2 temperatures to make treatments comparable. Therefore, the effect of temperature in vitro was not due to a linear relationship between bulblet development and the product of temperature × time. Bulblets formed at 30°C for half of the temperature × time product of the tissue culture period produced leaves after transplanting at rates similar to those of bulblets generated at a constant 30°C. Exposure of bulblets generated in vitro at 25°C to 4° for 2 weeks before planting in vermiculite resulted in leaf production not significantly different from that of chilled or non-chilled bulblets generated in vitro at 30°C. However, chilling significantly increased fresh weight of bulblets produced in vitro at either temperature. The data suggest that temperature during in vitro development affects Easter lily bulblet dormancy as measured by leaf production.

Open Access