A phenol-sulfuric acid assay was used to quantify non-specific neutral carbohydrates in Antirrhinum majus L. flowering stems of three inbreds and their hybrids. Flowering stems 40 cm long were harvested with five to six florets open and flower, leaf, and stem tissue separated, freeze-dried, and finely ground. Carbohydrates were extracted from the tissue with 95% ethanol in a 70 °C water bath and combined with a 5% w/v phenol solution and concentrated sulfuric acid. Glucose equivalents were determined with a spectrophotometer at absorbance of 490 nm. Averaged over tissue type, results were genotype dependent, ranging from 213 to 291 μg glucose equivalent per mg dry tissue with a LSD0.05 = 13. Flowers had the highest concentration of 340 μg/mg dry tissue, followed by stems, then leaves with 36% and 38% lower concentrations, respectively. Carbohydrate concentrations in two inbreds were compared when grown under cool (16 °C) and warm (29 °C) conditions. A genotype x environment interaction exists with inbred 3 exhibiting no reduction, 6% increase, and a 45% reduction in carbohydrate concentration when grown in warm conditions, while inbred 2 exhibited 15%, 23%, and 37 % reductions for flowers, leaves, and stems, respectively. Overall, there were 10% and 21% reductions in carbohydrate concentration for inbreds 2 and 3, respectively, when plants were grown under warm conditions.
Kenneth R. Schroeder and Dennis P. Stimart
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.
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.
Kenneth R. Schroeder and Dennis P. Stimart
Hypocotyls from Antirrhinum majus L. were excised at 2 weeks of age from seedlings grown under a 16-hour photoperiod or continuous darkness. Explants were cultured on modified Murashige-Skoog (MS) medium containing 0, 0.44, 2.22, 4.44, 8.88, or 44.4 μm BA to investigate adventitious shoot formation. Excised hypocotyls from eight commercial cultivars, three inbred lines, and an F1 hybrid between two of the inbreds were cultured on MS medium containing 2.22 μm BA to assess genotypic effects on adventitious shoot formation. The influence of seedling age was assessed by excising hypocotyls from seedlings at 6, 10, 14, 18, 22, 26, or 30 days. Optimal conditions for adventitious shoot formation on excised hypocotyls included: seedling growth in a lighted environment, use of hypocotyls from 10-day-old seedlings, and culture on medium containing 2.22 μm BA for 3 weeks. Under these conditions, up to a 5-fold improvement in number of shoots per hypocotyl over previous studies was achieved. Adventitious shoot formation was genotype-dependent and appeared to be a dominant trait. Chemical name used: N 6-benzyladenine (BA).
William J. Martin and Dennis P. Stimart
Stomatal density is being investigated as a highly correlated trait to postharvest longevity (PHL) and subsequently may be used for selection in early generations of breeding germplasm. To this end, leaf imprints were created from Antirrhinum majus L. (snapdragon) P1, P2, F1, BC1 (F1×P1), BC2 (F1×P2), F2, and F3 plants and evaluated for stomatal densities. Cut flowers of P1, P2, F1, BC1 (F1×P1), BC2 (F1×P2), and F3 were harvested after the first five flowers opened and evaluated for PHL. Additionally, cut flowers from these lines were evaluated for leaf surface area. Populations for evaluation were grown in the greenhouse in winter and spring 1999-2000 in a randomized complete-block design according to standard forcing procedures. Twenty-five cut flowering stems of each genotype were held in the laboratory in deionized water under continuous fluorescent lighting at 22 °C for PHL assessment. The end of PHL was defined as 50% of the flowers drying, browning, or wilting. Data will be presented on the correlation between stomatal density and PHL.
Wendy S. Higgins and Dennis P. Stimart
Lilium longiflorum Thunb. `Ace' bulblets generated in vitro at 25 or 30C were stored at 4C for O, 1, 2, 4, or 6 weeks after removal from culture and before planting to ascertain the effects of in vitro generation temperature and post-in vitro cold storage duration on bulblet growth responses during 36 weeks of greenhouse growth. Increasing post-in vitro storage duration decreased the number of days to first leaf emergence and percentage of plants producing shoots within 36 weeks, but increased the number of days to shoot emergence and anthesis, leaf number, and flower bud number. The length of time required for bulblet development from planting to shoot emergence was affected by storage duration more than periods from shoot emergence to visible bud and anthesis. It is feasible to produce high-quality L. longiflorum pot plants from in vitro-produced bulblets.
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.
Jaime A. Weber, William J. Martin, and Dennis P. Stimart
Progeny of 158 F5 × F5 crosses of Antirrhinum majus (snapdragon) selected within and among cut flower postharvest longevity (PHL) categories (long = 12.6-16.8 days, middle = 9.3-12.1 days, and short = 4.8-8.9 days) were evaluated for PHL and quality traits. Results were compared with previous studies involving F2 × F2 progeny, and F3, F4, and F5 inbred lines. Heritability of PHL in F5 × F5 progeny (0.77 ± 0.11) agrees with that of inbred lines (0.79 to 0.81) but is higher than in F2 × F2 progeny (0.41). Therefore, selection for increased PHL should progress more rapidly and predictably through application of inbred lines rather than F2 individuals. Significant differences between F5 × F5 progeny PHL categories confirm PHL is heritable with a significant additive component. Heritabilities of quality traits in A. majus are high, suggesting selection for quality traits should progress without difficulty. Phenotypic and genotypic correlations of PHL with quality traits are not consistently significant across PHL studies in A. majus. Discrepancies between studies suggest most traits may not be correlated to PHL or are subject to strong environmental influence.
Joseph J. King, Lloyd A. Peterson, and Dennis P. Stimart
Ammonium and NO3 uptake from hydroponic solutions containing 1 mm each of (NH4)2SO4 and Ca(NO3)2 were measured during development of Dendranthema ×grandiflorum (Ramat.) Kitamura `Iridon', `Sequoia', and `Sequest'. Nitrogen depletion from solutions approximated a 1 NH4: 1 NO3 ratio throughout a 90-day growth cycle (r = 0.96). Although harvest date cultivar interactions were significant for both forms of N, overall patterns of N uptake were similar among cultivars. Nitrogen removal from hydroponic solutions (milligrams per plant) was greatest from days 40 to 60; however, N removal (milligrams per gram of tissue dry weight) was greatest in the first month of development and decreased steadily until day 90. From day 40 to 60, new leaf development ceased while inflorescence buds developed to ≈1.0 cm in diameter. After this time, N uptake decreased rapidly as inflorescences expanded. Correlations between morphological changes and N demand could maximize the efficiency of applied N by matching form and application timing with plant needs.
Kenneth R. Schroeder, Dennis P. Stimart, and Erik V. Nordheim
Nicotiana alata Link and Otto (Jasmine tobacco) was transformed with an autoregulated senescence-inhibition gene construct PSAG12-IPT encoding isopentenyl transferase via Agrobacterium-mediated transformation. Transformation was confirmed by polymerase chain reaction. Transgenic plants exhibited up to 2- to 4-fold fewer senesced leaves, 29% longer in situ flower life, 26% more shoot dry weight, and a 32% to 50% reduction in flowers per branch. Additionally, transgenics were 28% shorter and had up to 174% more branches, indicative of cytokinin overproduction and a lack of tight autoregulation of PSAG12-IPT. Variation among independent transgenics suggests selection for enhanced PSAG12-IPT is feasible. Our observations of increased branching and in situ flower longevity, as well as reduced plant height and flowers per branch provide new information on PSAG12-IPT and its potential value for biological study and horticultural application.