Leaf segments of Prunus persica L. (peach) collected from greenhouse-grown plants and from micropropagated shoots were cultured on a basal medium containing half-strength Murashige and Skoog (MS), Staba vitamins, sucrose (30 g/1) and agar (6.5 g/l); medium adjusted to pH 5.6. The influence of 6 different growth regulators at 3 concentrations (5, 10, 15 μM) were investigated using leaf explants from proliferating shoots of 'Elberta Queen' peach. With thidiazuron (TDZ), compact and multiple green calli were obtained; with benzyladenine and zeatin, lower numbers of small sized calli were obtained; with kinetin, no callus development was observed. Among auxin treatments, both Dicamba and 2,4-D resulted in friable white and yellow calli. Most of the calli produced in all treatments were formed along the cut margins of the explants. In an another experiment, leaf explants of' Bellaire' (greenhouse) and `Elberta Queen' (in vitro shoots) were used to determine the influence of a large scale concentration of TDZ (3 to 23 |iM). Explants from greenhouse and in vitro leaves resulted in higher levels of callus development at TDZ concentrations of 8-13 μM. Higher TDZ levels resulted in necrosis of leaf explants. The-influence of different carbon sources on callogenesis was investigated. We observed more green and compact calli with glucose than with sucrose and fructose at 100 mM. The influence of the glucose at 10 different concentrations (30 to 300 mM) was also investigated.
Veronique Declerck and Schuyler S. Korban
Hak-Tae Lim, Haeng-Soon Lee, and Tage Eriksson
Plant regeneration ability of ginseng (Panax ginseng) via organogenesis was studied morphologically and anatomically. Compact callus was introduced from four different types of explants—leaf, petiole, flower stalk, and root—of in vitro-grown plantlets. Petioles were found to be the best material for callus induction. Calli induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (1.0 mg·L–1) and kinetin (0.1 mg·L–1) were conditioned for 2 weeks on the same medium. These calli differentiated into adventitious shoots when cultured on half-strength MS basal medium plus kinetin at 1.0 mg·L–1 and STS at 2.5 mg·L–1. An addition of GA3 (1.0 mg·L–1) and BA (1.0 mg·L–1) to MS basal medium, however, induced high-frequency in vitro flowering (86.1%) and multiple shoot budding, which affected the normal, complete development of plantlets. Plantlets with well-developed root systems were obtained 6 weeks after regenerated shoots had been transplanted to half-strength MS20 medium containing IBA at 0.25 mg·L–1. Nuclear DNA content was measured to check the stability of their ploidy level. Based on DNA flow cytometric analysis, all of the regenerants were typically diploids as were the mothers plants, indicating that nuclear DNA content remained stable during cell differentiation.
Leighan Howard, Philip Stewart, Amit Dhingra, Craig Chandler, and Kevin Folta
Cultivated strawberry (Fragari×ananassa) is a valuable crop, yet has benefitted little from recent advances in biotechnology and genomics. A high-throughput system for transformation and regeneration would hasten elucidation of gene function for strawberry and possibly the Rosaceae in general. In this report, a protocol for high-frequency octoploid strawberry transformation and regeneration is presented. The protocol uses leaf, petiole, and stolon as explants from a newly selected genotype, `Laboratory Festival #9'. This genotype was selected from progeny of a `Strawberry Festival' self-cross exclusively for its rapid regeneration and robust growth in culture. Direct organogenesis has been achieved from the leaf or from prolific callus with multiple shoots being visible in as few as 14 days. Over 100 viable regenerants may be obtained from a single leaf explant of about 3-cm2 size. This laboratory-friendly genotype allows high-throughput, statistically relevant, studies of gene function in the octoploid strawberry genetic background as well as generation of large transgenic populations.
Nabila S. Karam and Alexander X. Niemiera
The influence of intermittent and continuous irrigation on the growth, substrate nutrient accumulation and leaching from container-grown marigolds was determined. During a three week period. Tagetes erecta L. `Apollo' in a pine bark substrate received 12 irrigations. Each irrigation allotment was applied intermittently (multiple applications) or continuously (single application). Irrigation occurred when bark reached a targeted water content; irrigation water contained a complete nutrient solution. Leachates were cumulatively collected for each container and analyzed for N; plant dry weight. size, and nutrient composition were determined. Compared to continuously irrigated plants, intermittently irrigated plants had 43% greater root dry weight, 0.7% greater N concentration, and 43% more N leached from the substrate. Shoot mass. size. K, and P concentrations, substrate (pour-through extraction) and leachate N concentration were unaffected by irrigation method. Results demonstrated that. compared to conventional irrigation practices, intermittent irrigation was an effective method to reduce fertilizer effluent and increase N absorption for container-grown plants.
S. J. Kays and M. E. Austin
Repression of specific growth parameters of Tetragonia tetragonioides (Pall.) Kuntze (syn. Tetragonia expansa Murray) was studied after application of two types of growth regulators. Succinic acid-2,2-dimethylhydrazide (SADH) decreased stem height and dry weight progressively with increased concentration. Ratio of leaf dry weight to stem dry weight increased with concentration. Maleic hydrazide (MH) caused substantial reduction of fruit weight, with fruit number decreasing only at higher concentrations. Whole plant fresh weight and leaf area decreased only with a multiple 500 ppm application of MH. Leaf to fruit fresh weight ratio increased with concentration and number of applications. A formulation C6 to C12 fatty alcohols (Off-Shoot T) had little effect on fruit development and leaf area.
Tony K. Wolf and Robert M. Pool
Thermal and differential thermal analysis (DTA) are used to detect exotherms that result from the freezing of supercooled tissues (4). They provide a convenient and rapid means of assessing the hardiness of tissues that supercool, such as the floral primordia of Prunus spp. (3) and the compound buds of Vitis spp. (2). The inability to process a large number of tissue samples simultaneously, however, has been a major limitation of DTA. Ashworth et al. (3) described a computer-assisted data-logging system for recording thermal analysis data generated when Prunus flower buds were frozen. Multiple cooper-constantan thermocouples were used to increase the number of buds monitored on a given channel of a multichannel data-logger. Copper-constantan thermocouples, however, were not adequate with our instrumentation to discriminate exotherms generated by the freezing of individual shoot primordia of compound grapevine buds. Furthermore, anatomical barriers to ice propagation may be negated if thermocouples are inserted into buds to increase exotherm detection (5).
Craig K. Chandler, Diane Doud Miller, and David C. Ferree
Experiments were conducted to examine strawberry (Fragaria × anannassa Duchesne) plant renovation practices, singly and in combination, for effects on vegetative growth of greenhouse-grown, potted strawberry plants. The major results of these experiments were as follows: a) most of the plants that were both defoliated and root-pruned after fruit harvest died; b) there was a negative linear relationship between the number of leaves removed and the number of new leaves and runners produced; c) root, leaf, and total plant dry weights were negatively correlated with the severity of root pruning; and d) soil addition after fruit harvest decreased the shoot : root ratio of multiple-crown plants, but had no effect on single-crown plants.
Xiuli Shen, Vladimir Orbović, Manjul Dutt, William S. Castle, and Frederick G. Gmitter Jr.
development at both ends of uncut epicotyls of M. paniculata by 4 weeks culture. ( E ) Multiple shoots formation at one end longitudinally cut epicotyls of M. paniculata . ( F ) Multiple shoots formation at both ends of longitudinally cut epicotyls. ( G
Anita Solar, Jerneja Jakopič, Robert Veberič, and Franci Štampar
= Regalis; C = control; US = upper shoots in the canopy; LS = lower shoots. Means in columns with different letters that are shown under the figure are significantly different according to the Duncan multiple range test ( P ≤ 0.05). Basal
Samir C. Debnath and Danny L. Barney
, data for the control treatment (0 PGR) were excluded from analysis because all explants failed to respond. Statistical F tests were evaluated at P ≤ 0.05. Differences among treatments were further analyzed using Duncan's multiple range test. Shoot