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June Liu, Zhimin Yang, Weiling Li, Jingjin Yu and Bingru Huang

). Additionally, Bertin and Bouharmont (1997) reported enhanced chilling tolerance in rice ( Oryza sativa ) using somaclonal variations and in vitro selection. Cold tolerance of warm-season turfgrass could be improved through in vitro selection for somaclonal

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Brian W. Trader*, Hope A. Gruszewski, Norman I. Barclift, Richard E. Veilleux and Holly L. Scoggins

Coreopsis grandiflora (tickseed) regenerates readily from leaf segments allowing the possibility to exploit somaclonal variation as a means to develop novel phenotypes. We used true leaves from in vitro seedlings of Coreopsis grandiflora `Domino' grown on MS basal medium as explants in a series of experiments to evaluate the effect of media, leaf explant orientation, and genotype on shoot regeneration. Genotype accounted for most of the variation with two particular seedlings regenerating freely and eight others generally recalcitrant. From these two seedlings, designated E2 and H2, shoots were regenerated and acclimated to the greenhouse over a period of weeks. Once the plants had established (≈6 weeks after acclimatization) they were vernalized by moving them to a lighted bench (12-h photoperiod) in a walk-in cooler at 4 °C .On transfer back into the greenhouse, the plants flowered within a few weeks; 15 of 175 somaclones were selected based on distinct differences in flower orientation and appearance. The selected somaclones were propagated by division and transplanted to the field in May 2002 in a randomized complete-block design with three-plant plots and three replications, to determine if the novel characteristics persisted through an additional propagation cycle. In the field, plant height, leaf dimension, flowering, and flower dimensions were scored in June-July 2003. Significant differences were found between somaclones and the original E2 and H2 similarly propagated seedlings for desirable (more petals per flower, greater flowering, shorter plants), undesirable (less flowering, smaller flowers), and neutral (narrower leaves, taller plants) traits.

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Mark Bridgen

The potential value of somaclonal variation for economically important plants is well-documented. The process of somaclonal variation can arise from a controlled or a random source of variation. Variability can be obtained by applying cellular pressures and selection. Valuable resistance to diseases and nematodes has already been accomplished with somaclonal variation; now, plant tolerance to pests has been realized. Tetranychus urticae, the two-spotted spidermite, and Trialeurodes vaporariorum, the greenhouse whitefly, were disinfected and introduced to aseptic shoot cultures of Torenia fournieri. These pests were allowed to feed until such time that their populations decreased due to the absence of food. The plant cells that remained after feeding were induced to form adventitious shoots and plantlets. These regenerated plantlets were acclimated to greenhouse conditions and evaluated for tolerance to the pest to which they were subjected in vitro. Highly significant differences were found in somaclones for both the two-spotted spidermite and greenhouse whitefly when compared to control plants. A wide range of variability was observed among the somaclonal population. There were significantly fewer mite eggs laid on plants regenerated from in vitro cultures screened with two-spotted spidermites than on seed-sown controls. Regenerants from cultures screened with whiteflies in vitro had fewer eggs, immatures and live adults than controls. The potential for somaclonal variation to be used as a method to develop insect resistant plants will be discussed.

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Brian W. Trader, Hope A. Gruszewski, Holly L. Scoggins and Richard E. Veilleux

Coreopsis species (tickseed) can be regenerated from leaf segments allowing the possibility to exploit somaclonal variation as a means to develop novel phenotypes. We used true leaf explants from in vitro seedlings of perennial C. grandiflora (A. Gray) Sherff `Domino' and `Sunray' grown on Murashige and Skoog (MS) basal medium. Two of ten seedlings of `Domino' regenerated freely and others were generally recalcitrant. From these two seedlings, designated E2 and H2, shoots were regenerated and acclimatized to the greenhouse. About 175 plants were established and vernalized from which somaclones were selected based on distinct differences in flower orientation and appearance. The selected somaclones were propagated by division and transplanted to the field in August 2001 in a randomized complete block design with three-plant plots and three replications to determine whether novel characteristics persisted through an additional propagation cycle. In the field, plant height, leaf dimension, flowering, and flower dimensions were scored in June and July 2003. Differences were found between somaclones and similarly propagated E2 and H2 for desirable (more petals per flower, greater flowering, shorter plants), undesirable (less flowering, smaller flowers), and neutral (narrower leaves, taller plants) traits. Open-pollinated (OP) seed was collected and germinated and the seedlings from somaclones that differed significantly from E2 and H2 were evaluated. These maternally selected seedlings were overwintered then planted in the field in May 2004. Most traits that differentiated somaclones from E2 and H2 did not persist in the OP seedling population; however variation that was likely introduced through outcrossing resulted in desirable phenotypes with potential for new cultivar development.

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Jude W. Grosser*, J.L. Chandler and R.M. Goodrich

Sweet orange (Citrus sinensis L. Osbeck) is the most horticulturally important and widely grown Citrus species in Florida and worldwide, and `Valencia' is the most important cultivar for processing. Frozen concentrate orange juice has been the primary product of the Florida and Brazilian industries, but recently there has been a strong shift to not from concentrate (NFC) product in Florida. The higher quality NFC has a greater consumer appeal, and brings a higher market price. The development of higher quality oranges with expanded maturity dates will facilitate this change and should increase the competitive ability of the Florida industry. No true sweet orange cultivars have been developed by conventional breeding due to biological impediments, and alternative methods to obtain genetic variation are being investigated, including studies of somaclonal variation. We have produced nearly 1000 somaclones of `Valencia' sweet orange using organogenesis, somatic embryogenesis, and protoplasts. Following several years of fruit evaluation, early and late maturing high quality somaclones have been identified based on juice analytical data (brix, acid, ratio, juice percentage, juice color, and lbs. solids). These clones have also performed exceptionally in taste panel evaluations comparing them with the traditional mid- and late-season cultivars. Second generation trees of the most promising clones have been propagated for further evaluation, and superior processing clones will be released to the Florida industry in the near future. An overview of this program including pilot plant juice quality data and taste panel results will be presented.

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Jason J. Goldman, Wayne W. Hanna and Peggy Ozias-Akins

`TifEagle' (2n = 3x = 27) hybrid bermudagrass [Cynodon dactylon (L.) Pers. (2n = 4x = 36) × Cynodon transvaalensis Burtt-Davy (2n = 2x = 18)] is an ultradwarf cultivar for greens, and `TifSport' (2n = 3x = 27) is a more versatile hybrid used on fairways, athletic fields, and lawns. To develope a transformation system and determine if somaclonal variation was present in regenerated plants, both cultivars were tested for their ability to produce embryogenic callus from which plants could be regenerated. Sliced nodes of both cultivars and immature inflorescences from `TifSport' were used as the explant sources. Cultures were initiated on Murashige and Skoog medium supplemented with 6.79 μm 2,4-D and 0.044 μm BA (`TifSport' and `TifEagle') or 6.79 μm 2,4-D plus 200 mg.L-1 casein hydrolysate (`TifSport'). In total, 51 plants were regenerated from callus of a single node of `TifEagle'. Nodes from `TifSport' did not produce embryogenic callus. In total, 29 plants were regenerated from callus of `TifSport' produced from immature inflorescences. These plants were grown in the field for at least one season, and 5-cm-diameter plugs were harvested, repotted in a greenhouse, and allowed to reestablish. Data on canopy height, leaf width, leaf length, and number of stolons were collected. Seven `TifEagle'-derived entries (14%) were not significantly different (α = 0.05) from `TifEagle' harvested from the breeder plot in Tifton, Ga., for all measured traits, and 41%, 24%, and 22% differed by one, two, or three measurements, respectively. Flow cytometry indicated that 33% (13 plants) of the `TifEagle' regenerants were hexaploid (2n = 6x = 54) and the rest remained triploid. One `TifSport' regenerant was significantly different (α = 0.05) for plant height.

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Jericó J. Bello-Bello, Lourdes G. Iglesias-Andreu, Susana A. Avilés-Viñas, Eunice Gómez-Uc, Adriana Canto-Flick and Nancy Santana-Buzzy

genetic variability relatively quickly without sophisticated technology ( Larkin and Scowcroft, 1981 ; Shu and Lagoda, 2007 ). The term “somaclonal variation” describes the tissue culture-induced stable genetic, epigenetic, or phenotypic variation in

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W.A. Mackay, T.J Ng and F.A. Hammerschlag

Cucumis melo L. (muskmelon) is susceptible to Myrothecium roridum at all stages of growth with no known source of resistance. Screening of regenerants from long-term cotyledonary-derived callus cultures of muskmelon cv. Hales Best (tolerant), Iroquois (susceptible), and Perlita (intermediate) was carried out to determine if novel plants with increased levels of resistance could be obtained. A detached-leaf bioassay was used to screen greenhouse-grown regenerants and seedlings of the three cultivars. Resistance was determined by measuring necrotic lesion diameter and chlorotic plus necrotic lesion diameter. No change in the level of resistance to M. roridum has thus far been observed. Thus, screening for somaclonal variation may not be aviable approach to recover resistance in muskmelon to M. roridum.

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Samira Samarfard, Mihdzar A. Kadir, Saleh B. Kadzimin, Halimi M. Saud, Seyed Ali Ravanfar and Mahmoud Danaee

multiplication of PLBs. This study was therefore designed to evaluate the influence of chitosan and TDZ combination on PLB proliferation of Phalaenopsis gigantea . In the case of Phalaenopsis gigantea , somaclonal variation may result in changes in the

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Susan P. Martin, Carol A. Bobisud and Terry T. Sekioka

The objective of this project is to propagate disease-free clones of fresh market pineapples for growers. To expedite the distribution process, sterile cultures were obtained from the USDA/ARS Germplasm Repository, Hilo, Hawaii of pineapple clones 119, 155 and 159. Plantlets were placed on modified Murashige and Skoog (MS) medium with one half of the salts and 2 uM BAP and 1 uM IAA. The cultures were transferred to modified MS medium with one half of the salts, no growth regulators, and solidified with Phytagel. Plantlets were transplanted at one-inch size into peatlite mix. At 4-6 inch size, the plants were transplanted in growers fields. Early observations indicate difference between clones in kinds and frequency of “off-types.” Compared with surrounding plants, “off-types” were categorized as being dwarfed, variegated, thorny leaved, and pigmentation modified.