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.
Jeffrey W. Adelberg, Bill B. Rhodes, Halina T. Skorupska and William C. Bridges
Mark H. Brand, Yiqin Ruan and Richard Kiyomoto
To characterize the in vitro behavior of Rhododendron `Montego' with tissue proliferation (TP) to cytokinin and auxin, comparisons were made of normal [TP(–)], dwarf TP [TP(+) dwarf], and long TP [TP(+) long] shoot cultures. On basal medium TP(–) and TP(+), long shoots failed to multiply and had a low relative growth rate (RGR) of 0.1, whereas TP(+) dwarf shoots produced 31.8 shoots per tip, with most shoots being <5 mm long, and RGR was 0.3. Addition of 15 μm 2iP to basal medium induced the production of more than six shoots per TP(–) tip and doubled their RGR; TP(+) long shoots produced 16.8 shoots, most <5 mm long, and had an RGR of 0.3; TP(+) dwarf shoots produced only 16% as many shoots as on basal medium, but still exhibited an increase in RGR. Leaves from TP(–) and TP(+) sources failed to produce shoots on basal medium, but 74% of TP(–) leaves formed shoots when cultured on 1 μm IBA and 30 μm 2iP. TP(+) leaves were able to form shoot meristems on media containing only 5 μm 2iP (26% of explants), but these meristems failed to elongate into shoots. Calli from TP(–) leaves, TP(+) leaves, and TP(+) tumors grown on medium containing 10 μm NAA and 15 μm 2iP had higher RGRs than the same calli on basal medium during the first 8 weeks of culture. Over time, RGR decreased in both TP(–) and TP(+) leaf calli, but increased in TP(+) tumor callus. The increased RGR resulted from differentiation of shoot meristems on 85% of the calli between week 4 and week 8. Our results suggest that TP(+) tissues have altered hormone metabolism or sensitivity that leads to dramatic differences in in vitro behavior and probably contributes to tissue proliferation observed in whole plants. Chemical names used: 6-(γ,γ-dimethylallylamino) purine (2iP); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).
A detached-leaf bioassay was used to evaluate peach [Prunus persica (L.) Batsch] somaclone 122-1 (derived from callus produced on an immature embryo of peach cultivar Redhaven) for resistance to several virulent strains of Xanthomonas campestris pv. pruni [E.F. Sm.) Dows], causal agent of bacterial leaf spot, and to a virulent isolate of Pseudomonas syringae van Hall pv. syringae, causal agent of bacterial canker. The detached-leaf bioassay was also used to evaluate progeny of 122-1 for resistance to X. campestris pv. pruni virulent strain XP1. Somaclone 122-1 was significantly more resistant to most strains of X. campestris pv. pruni than was `Redhaven', and all of its progeny exhibited high levels of resistance to X. campestris pv. pruni strain XP1. Somaclone 122-1 exhibited significantly higher levels of resistance to Pseudomonas syringae pv. syringae than did `Redhaven' and this resistance was retained over time in the greenhouse and following a 2-year cycle of tissue culture propagation.
Jolene Wright, Ann Reilley, Jean Labriola, Stephanie Kut and Thomas Orton
An experiment was conducted to determine the types, extent, and heritability of new phenotypic variants recovered from carrot cell cultures initiated from mature tap-root explants of the male-fertile carrot (Daucus carota L.) `Slendero'. Embryogenic callus was transferred to plant-growth-regulator-free medium 66 days after culture initiation, and regenerated plantlets were harvested and eventually planted in a field. The tap roots of mature regenerated plants were vernalized at 5C for 9 weeks and replanted. Of 31 flowering regenerants, 25 exhibited some form of petaloid male sterility; the remaining six regenerants were male fertile. All plants from the same original explant were either all sterile or all fertile. Three generations of sterile regenerant × petaloid cytoplasmic male sterility (CMS) maintainer (M) progeny tests showed that the new CMS behaved in a similar manner to that previously reported. Comparison of mitochondrial DNA restriction patterns of sterile and fertile regenerants with those of `Slendero', petaloid CMS, petaloid M, and brown anther CMS lines resulted in the following conclusions: 1) the sterile regenerants exhibited patterns identical to the known petaloid CMS and 2) the fertile regenerants were different from the original `Slendero' and the sterile regenerants and nearly identical to a known petaloid CMS M line. The high frequency of CMS among regenerants from `Slendero' carrot cell cultures may provide an efficient method to develop sterile M tandem lines and corresponding new hybrid varieties.
Jodie L. Ramsay, Donald S. Galitz and Chiwon W. Lee
Influences of culture media and sucrose concentrations on plant regeneration from Easter lily (Lilium longiflorum L. cv. Ace) ovary tissues were investigated. Pistils excised from unopened flower buds (3-5 cm long) were sectioned and cultured on either B-5 medium or Murashige and Skoog (MS) medium containing 2%, 5%, or 10% sucrose, with 1 mg·L-1 2,4-D and 2 mg·L-1 BA. Callus formation was most prolific on MS medium containing 5% sucrose. Shoot differentiation was higher on MS medium than on B-5 medium. Rooted plants were transferred into soil medium and grown in a greenhouse. Root tip smears showed that 35% of the regenerated plants showed a variation in chromosome numbers from 10 to 25 per cell, while the rest of the regenerants showed the normal 2n = 2x = 24 chromosomes per cell. The mixoploid condition also existed in different root cells of the same regenerated plant. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); 6-benzylaminopurine (BA).
Dale E. Kester, Kenneth A. Shackel, Warren C. Micke, Mario Viveros and Thomas M. Gradziel
The spatial and temporal pattern of noninfectious bud failure (BF) expression (BFexp) was studied during seven growing seasons in a population of `Carmel' almond trees originating from twelve commercial propagation sources. All progeny trees were grown in a single experimental site with high prevailing summer temperatures. BFexp increased continuously but irregularly in each nursery population as measured as the proportion of trees showing BF and as an average BFexp rating. Populations from the 12 nurseries represented increasing clonal generations from the original seedling tree and showed increasing levels of BF, as well as a decreasing shape value and increasing scale value derived by a failure statistics model. Models for development, distribution and hazard functions were defined for each of the 12 sources studied. Only sources from the original tree and source A demonstrated potential for commercial use. A significant correlation was found between average yearly increase in BFexp and the average daytime temperature for the previous June. The June period coincides with a specific stage in the seasonal growth cycle when vegetative buds mature.
Masood Z. Hadi, Mark P. Bridgen and John P. Sanderson
Procedures were developed to determine if live, adult two-spotted spidermites (Tetranychus urticae Koch) could be surface disinfested before being introduced into in vitro cultures of torenia (Torenia fournieri L.). Three time periods (5, 10, and 15 minutes) and five levels of sodium hypochlorite (0.05% to 0.25%) were evaluated. Surface disinfestation was accomplished by agitating 2 × 3 cm pieces of infested bean leaves in sodium hypochlorite solutions and then drying in a mite drier apparatus. All sodium hypochlorite concentrations disinfested the mites completely, however high concentration levels were lethal to the mites. Exposure periods of 10 and 15 minutes also significantly increased mortality. For optimum disinfestation of two-spotted spidermites with minimum mortality, a concentration of 0.05% sodium hypochlorite and 0.05% Tween-20 for 5 minutes should be used.
Les Frey and Jules Janick
Shoot regeneration in carnation (Dianthus catyophyllus L.) was influenced by genotype, explant source, and plant growth regulator balance. Plants were regenerated from petals, calyxes, nodes, internodes, and leaves, but only petals, calyxes, and nodes were regenerative from all three cultivars examined (`Scania', `Improved White Sire', `Sandra'). Maximum proliferation was achieved with petals on Murashige and Skoog medium supplemented with 0.05 μm TDZ and 0.5 μm NAA. Shoot initiation originated from cells near vascular regions and perhaps from epidermal cells in petals and via organogenic callus from other explants. There was no evidence of chimeral separation from petals or callus, but somaclonal variants (3.3%) were observed involving petal hue and plant dwarfness. Unstable color patterns were observed in tissue-cultured regenerants of `Scania' and `Improved White Sire' similar in type and frequency to propagules derived from cuttings; none were observed for tissue-cultured or cutting-derived plants of `Sandra'. Chemical names used: N-pheny1-N′-l,2,3 -thiadiazol-5-ylurea [thidiazuron (TDZ)]; 1-napthaleneacetic acid (NM).
Maria Luiza De Oliveira, James G. Thomson and Ed Stover
considered the most conservative approach and likely to generate the lowest level of somaclonal variation. However, the rate of propagule increase is much less than with the second method in which axillary budbreak is induced to produce numerous shoots per
Ryutaro Tao and Akira Sugiura
Callus cultures were initiated in the dark from leaf primordia, stem internodes, and young leaves of adult Japanese persimmon (Diospyros kaki L.) to induce adventitious buds. A high frequency of regeneration occurred on Murashige and Skoog medium (MS) with half the normal NH4NO3 and KNO3 concentration (1/2N) and containing 10 μm zeatin or 1 μm 4PU-30 in combination with 0.1 μm IAA, or MS(1/2N) medium containing 0.03 to 0.1 μ m IAA or 0.01 to 0.03 μm NAA combined with 10 μm zeatin. No significant differences in the capacity of regeneration were observed among the calli from different explant sources. Only eight of 16 cultivars formed adventitious buds on MS(1/2N) medium containing 10 μm zeatin and 0.1 μm IAA, with the percentage of explants forming adventitious buds ranging from 2% to 72%. Chemical names used: indole3-acetic acid (IAA); 1-naphthaleneacetic acid (NAA); N-phenyl-N'-(2-chloro-4-pyridyl)urea (4PU-30).