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Susan M. Stieve and Dennis P. Stimart

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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Dennis P. Stimart and John C. Mather

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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James F. Harbage and Dennis P. Stimart

We investigated the role of ethylene on adventitious rooting of `Gala' (easy-to-root) and `Triple Red Delicious' (difficult-to-root) apple (Malus domestica Borkh.) microcuttings. Root count increased significantly as IBA level increased, with highest root counts on `Gala'. Ethylene evolution increased significantly with IBA level without significant differences between cultivars. Basal section removal of microcuttings in the area of root origin reduced root count without changing ethylene evolution. Ethylene treatment of proliferated shoots before microcutting excision failed to enhance rooting. IBA-induced ethylene evolution was eliminated nearly by AVG, but root count remained IBA dependent. ACC reversed IBA plus AVG rooting inhibition, but ACC alone failed to influence root count. Polar auxin transport inhibitors NPA and TIBA stimulated ethylene evolution without increasing root count. Adventitious rooting of apple microcuttings was not associated with ethylene. Chemical names used: 1-H-indole-3-butyric acid (IBA); aminoethoxyvinylglycine (AVG); 1-aminocyclopropane-1-carboxylic acid (ACC); 2,3,5-triiodobenzoic acid (TIBA); N-1-naphthylphthalamic acid (NPA).

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Kenneth R. Schroeder and Dennis P. Stimart

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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Kenneth R. Schroeder and Dennis P. Stimart

Flowering stems from three commercial inbreds and their F1 hybrids of Antirrhinum majus L. were cut when the first eight basal florets opened. Tops of the stems were removed above the eighth floret and florets were removed leaving two, four, six, or eight open florets on a stem. A completely random design with 10 replications was used. Flowering stems were placed in plastic storage containers 35 × 23 × 14 cm (L × W × H) with 2.5 L deionized water for postharvest evaluation. Evaluation took place under continuous cool-white fluorescent light (9 μmol·m–2·s–1) at 24°C Postharvest life was determined as the number of days from cutting to discard when 50% of the open florets on a flowering stem wilted, turned brown, or dried. Results showed postharvest life increased as the number of open florets on a stem decreased. Mean postharvest life increased as much as 4.7 days when only two florets remained on a stem. These results indicate a direct relationship between number of florets on a cut flower stem and postharvest life.

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Kenneth R. Schroeder and Dennis P. Stimart

Three percent hydrogen peroxide (H2O2) was diluted with deionized water (dH2O) to 0.75%, 0.38%, 0.19%, 0.09%, or 0.05% H2O2 plus 1.5% sucrose for use in evaluation of Antirrhinum majus L. (snapdragon) cut flowers. Other vase solutions used as controls included; 300 ppm 8-hydroxyquinoline citrate (8-HQC) plus 1.5% sucrose; dH2O plus 1.5% sucrose; and dH2O. A completely random design with 7 replicationss was used. Flowering stems of three commercial inbreds and one F1 hybrid of snapdragon were cut when the first five basal florets opened. Each stem was placed in an individual glass bottle containing one of the eight different treatments. Flowering stems were discarded when 50% of the open florets wilted, turned brown, or dried. Postharvest life was determined as the number of days from stem cutting to discard. Addition of H2O2 to vase solutions at rates of 0.19 and 0.09% resulted in postharvest life not different from that obtained with 8-HQC plus sucrose. Hydrogen peroxide plus sucrose extended postharvest life of snapdragon cut flowers 6 to 8 days over dH2O and 5 to 7 days over dH2O plus 1.5% sucrose.

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James F. Harbage and Dennis P. Stimart

Involvement of pH and IBA on adventitious root initiation was investigated with Malus domestica Borkh. microcuttings. The pH of unbuffered root initiation medium (RIM) increased from 5.6 to 7 within 2 days. Buffering with 2[N-morpholino] ethanesulfonic acid (MES) adjusted to specific pHs with potassium hydroxide prevented pH changes and resulted in a 2-fold higher root count at pH 5.5 compared to pH 7 or unbuffered medium. As pH decreased, lower concentrations of IBA were required to increase root counts. Colorimetric measurement of IBA in buffered RIM showed greater IBA loss and higher root count were associated with lower pH levels in all cultivars. This suggests that IBA loss from RIM depends on medium pH, which affects root count. Root count differences between easy-to-root through difficult-to-root cultivars were not consistent with amount of IBA loss from RIM. Cultivar differences in root count could not be explained solely by IBA loss from RIM.

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Susan M. Stieve and Dennis P. Stimart

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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Monica E. Figueroa-Cabanas and Dennis P. Stimart

Direct shoot organogenesis (DSO) on Antirrhinum majus L. (snapdragon) was evaluated in vitro to determine the inheritance of genes conditioning this response. One-centimeter-long hypocotyls excised from 2-week-old seedlings started in vitro in the dark on Murashige and Skoog medium served as explants. Optimal conditions for DSO on explants included hypocotyl excision from 10-day-old seedlings, 2.22 μmol BA in the culture medium, and a 21-day culture duration. An adventitious shoot was counted once it developed a stem terminated by at least one leaf appearing to have originated from an apical meristem. Seven populations were evaluated for DSO: parent 1 (P1) with lowest DSO (0.3 shoots); parent 2 (P2) with highest DSO (13.9 shoots); F1 (P1 × P2); F1 (P2 × P1); F2 (self-pollination of F1); P1 × [P1 × P2]; and P2 × [P1 × P2]. P1 and P2 were chosen as parents based on DSO counts being lowest and highest, respectively, of inbreds evaluated. DSO appears to be a trait under nuclear genetic control. High DSO appears to be dominant over low DSO. The trait appears to be simply inherited through one or two genes.

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William J. Martin and Dennis P. Stimart

Cut flowers of Antirrhinum majus L. (snapdragon) P1, P2, F1, F3, and F2 × F2 plants were harvested after the first five flowers were open and were evaluated for postharvest longevity to further evaluate genes conditioning postharvest longevity. F3 progeny evaluated were derived by selfing F2 selections of long keeping, mid-range, and short keeping types. F2 × F2 progeny evaluated were derived from crosses within and between postharvest longevity categories. Populations for evaluation were grown in the greenhouse in winter 1998-1999 in a randomized complete-block design according to standard forcing procedures. Thirty plants of each genotype were held in the laboratory in deionized water under continuous fluorescent lighting at 22 °C for postharvest assessment. The end of postharvest life was defined as 50% of the flowers drying, browning, or wilting. Data will be presented on postharvest longevity and allelic relationships within populations.