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Shailaja Sinha and Madhuri Sharon

Somatic embryogenesis from apical meristem, cell elongation zone, and cell differentiation zone of roots of Punica granatum L. var. Ganesh was obtained. The basal medium used was Gamborg's B5. 2, 4-D induced white globular callus in root tip explants, which on further subculture to medium containing 0.5 mg/L BAP, produced somatic embryos from the outer surface of the callus. Direct somatic embryogenesis occurred from all the three zones of the root in presence of 2 mg/L kinetin. BAP induced embryogenic callus in elongation and cell differentiation zone segments of the roots, which, on further subculture onto the same medium, produced somatic embryos. NAA caused rhizogenesis in all the three root segments. Differentiation of somatic embryos into plantlets took place on B5 medium supplemented with 0.01 mg/L NAA + 0.5 mg/l BAP + 2 mg/L kinetin.

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Chamchuree Sotthikul and Pimchai Apavatjrut

Propagation of Curcuma roscoeana in vitro was done successfully by culturing 0.5 × 1.0 mm shoot tips from young buds onto modified MS (1962) + 0.25 mg·L–1 kinetin. The bud-derived new plantlets could be multiplied on a new medium. Stem explants 10 mm in size, measured from base of the plantlets longitudinally cut in half, were the most suitable culture explants providing 2.8 new healthy plantlets/cultured explant. Explants from 4, 6, and 8 weeks old plantlets were more suitable than those of 2 weeks old when grown on agar or in liquid medium. From a histological study, it was found that new buds developed from preexisting meristems. The buds, like root initiation, could also occur directly from initial culture explants, not through callus. The plantlets obtained could successfully be transferred into growing pots, having a 95% survival rate.

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A. James Downer, Matthew Shaw, and Dennis Pittenser

Coast Live Oak (Quercus agrifolia) and White oak (Q. lobata) are landscape trees which are prone to sudden branch drop. The purpose of this work was to determine the effect of various pruning techniques on growth reduction of one member of a pair of codominant stems. Forty trees of each species were selected with codominant stems. One stem of each pair was pruned by: 1. removing all apical meristems; 2. thinning 50% of branches; 3. heading back to a 10cm stub or, 4. unpruned. Calipers of thinned or headed branches were most retarded while tipped and unpruned branches grew most. Results suggest that pruning by thinning may be an alternative to removal of codominant branches when training young trees.

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Audrey I. Gerber, Karen I. Theron, and Gerard Jacobs

Protea L. sp. can be assigned to groups according to similar times of flower initiation and harvest. The stages occurring during flower initiation and their synchrony relative to shoot growth were investigated for three cultivars when flower initiation occurred on the spring growth flush. For all three cultivars, the spring flush was preformed and enclosed in the apical bud before spring budbreak. During elongation of the spring flush, the apical meristem produced floral primordia which differentiated into involucral bracts. After completion of the spring flush, meristematic activity continued and produced floral bracts with florets in their axils. The different cultivars were characterized by differences or similarities in the time of budbreak, and the rates of shoot growth, appendage formation, and flower development. Insight into the time of flower initiation relative to vegetative growth could be useful in making management decisions, as well as forming a basis for manipulation of the flowering process.

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Jeffrey W. Adelberg, Bill B. Rhodes, Halina T. Skorupska, and William C. Bridges

Adventitious and axillary shoots of melon (Cucumis melo L.) were cultured from explants on a modified Murashige and Skoog medium containing 10 μm BA. Explants were diversified with regard to genetic source (breeding lines Miniloup, L-14, and B-line), seed parts (apical and cotyledon tissue), seed maturity (10-40 days after pollination; DAP), and cotyledon sections with respect to apical-radicle axis (distal and proximal). Plants were screened for ploidy level by pollen morphometry. Immature cotyledons produced more tetraploid regenerants than mature cotyledons from seed of breeding line Miniloup; the highest frequency of tetraploid regenerant plants was from cotyledons of embryos harvested 18 and 22 DAP. Explants from the apical meristem of the same seeds produced fewer or no tetraploid plants. Proximal sections from immature cotyledons of three genotypes (Miniloup, L-14, B-line) produced higher frequencies of tetraploids than whole mature cotyledons or whole immature cotyledons.

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Ryoichi Nakajima, Shungo Otagaki, Katsuhiro Shiratake, and Shogo Matsumoto

Previously, we showed that a reduction of FaTFL2 (homolog of Arabidopsis thaliana TFL1) expression is a key signal for flowering in domesticated octaploid strawberries (Fragaria ×ananassa). Since FaTFL2 expression seemed to be regulated by temperature rather than by photoperiod, we investigated whether down regulation of FaTFL2 and floral meristem generation occurred at different temperature conditions. In addition to the conditions for a normal super-forcing cultivation system of an 8-hour photoperiod and day/night temperatures of 31.2 or 30/15 °C, flowering also was generated under the same photoperiod and day/night temperatures of day/night-half/night-half temperatures of 30/15/25 °C conditions. We demonstrate that the new super-forcing cultivation system is energy saving based on the reduction of FaTFL2 expression.

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Terri Woods Starman

Manually pinched plants of 18 cultivars of Impatiens hybrids (Keintzler New Guinea impatiens) were compared to control plants to determine the effect of apical meristem removal on flowering, growth and branching. Pinching delayed days to anthesis (first flower) of all cultivars, however, further delay in days to marketability (5 flowers open) was dependent upon cultivar. Plant area and fresh and dry weight were not affected by pinching plants of any cultivar. Cultivar influenced response to pinching treatments for plant height and plant width. Secondary branch number was increased by approximately 3 branches for all cultivars when plants were pinched. There were interactions between cultivar and treatment for primary, tertiary, and total branch number. Measured improvements in plant form determined two cultivars, Sylvine and Thecla, should be pinched. Chemically pinching these two cultivars with dikegulac at 780 mg·liter-1 was comparable to manually pinching plants.

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Michael Parker and Eric Young

Controlling vegetative growth resulting from a long growing season in the southeast is difficult while trying to promote early fruiting. This study was initiated in 1990 to evaluate higher density leader training techniques, cultivar interactions, and the benefits of pre-plant fumigation on apple replant sites. Another objective was to evaluate these management parameters in four regions with very different climates and elevations. The training techniques evaluated were, weak leader renewal, bending of the leader during the growing season, and partial terminal leaf removal every 10 inches of leader growth (without injuring the apical meristem). The cultivars used were Jonagored, Spur Gala, and Red Fuji, all on Mark rootstock. First and second year branching was not significantly different between the various training techniques. Yields during the third year did not appear to differ between the leader training techniques. Pre-plant fumigation appeared to be beneficial in increasing tree growth in only two of the four sites.

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E.B. Peffley, M. Hart, Z. Xiang, Z. Hiang, A. El-Sharif, N. Dong, and G.C. Phillips

Particle bombardment using the Bio-Rad PDS-1000/HE system is being investigated as a means to develop a stable transformation system for the bulb onion. Donor materials for bombardments included sterile meristems and radicals from newly germinated seeds, callus of Allium cepa `TG1015', `Sunlite', and `Buffalo', A. fistulosum `Heshiko', and suspension-cultured cell lines with confirmed regeneration capability, as well as regenerated plants of an F1 interspecific hybrid. Transient expression assays using the B-glucuronidase (GUS) reporter gene system were used to optimize the conditions for transformation. Various promoters combined with the GUS coding sequence were tested. Results indicate genotype specificity for promoter expression. Some tissue continued to exhibit GUS activity after several months in culture, indicating potential for achieving stable transformation of onion.

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Ellen B. Peffley and Melanie A. Hart

Particle bombardment was investigated as a potential transformation method for onion. Seeds of Allium cepa `TG 1015' were planted onto BDS medium and placed in a dark incubator at 25C for germination. Two to 3 weeks after the seeds were germinated, meristems (1 to 2 mm) were excised and placed onto BDS medium containing 2 mg 2,4-D/liter for callus initiation. Callus was transferred monthly onto fresh BDS medium containing 2,4-D until bombardment. The reporter gene for B-glucuronidase (GUS) expression was used to assess efficiency of gene delivery in all particle bombardments. Characteristics examined were target distance and helium pressure (particle velocity). Tissues were subjected to the 5-bromo-4-chloro-3-indoyl-B-D-glucuronide (xgluc) test for detection of GUS activity. Measurements were taken on particle dispersion as affected by target distance and helium pressure. GUS expression was detected in putatively transformed tissues.