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Petal color of 10 carnation (Dianthus caryophyllus L.) cultivars MINAMI series is divided into two color groups, the acyanic group and the cyanic group. The acyanic group consists of yellow-flowered cultivars 'Poly Minami' and 'Lemon Minami' and off-white-flowered cultivar 'Vanilla Minami'. The cyanic group consists of orange-flowered cultivars 'Orange Minami' and 'Minami', pinkish-flowered cultivars 'Pippi Minami', 'Passion Minami', 'Feminine Minami' and 'Sakura Minami', and reddish-flowered cultivar 'Tommy Minami'. MINAMI series produced from bud mutation has a unique direction of flower color changes. Therefore, the usage of these plant materials might contribute to the elucidation of the mechanism of flower color mutation.

1-Methylcyclopropene (1-MCP) is an antagonist of ethylene for receptor binding sites. Carnation (Dianthus caryophyllus L. 'Amelia' and 'Natila') plants were incubated with 1-MCP (500 nl L -1 and 1000 nl L -1 ) for 24h at 20°C. On the other hand, it was found that the cultivars can be stored at 4°C temperature and 70% humidity in a solution of 1% sucrose. The decreases in quality decay and increases in vase life were obtained from the treatments of 1-MCP doses between 'Amelia' and 'Natila' flowers.

The spray carnation seedlings obtained from the cuttings stored at 4°C during 55 days at cool storage and unstored were transferred to a greenhouse environment and the effects of different doses of Fe-EDTA (50, 100, 200, 300 mg L -1 ) applied via foliar application on the fresh and dry weight of leaf, stem and flowers of plants and the number of flowers per plants were investigated. At the end of the growing season, the effects of storing conditions on the leaf dry weight of the plant and of the Fe-EDTA application on the stem fresh weight and number of flowers per plant were found statistically significant. There were no clear effects observed from Fe-EDTA application on the fresh and dry weight of flowers.

In this study, spray carnation cuttings obtained from storage at 4°C during 55 days in cold storage and unstored cuttings transferred to greenhouse conditions were grown. The effects of different doses of foliar Fe-EDTA application (50, 100, 200, 300 ppm) on stem diameter, fresh and dry weight of stems of carnations were investigated. During the growing period (November, February and May), stem diameter was measured at 3 months interval. In these measurements, stem diameter of unstored carnation cuttings were found to be higher then stored ones. The effect of Fe-EDTA on stem diameter in May was found statistically significant. The effect of Fe-EDTA application doses on fresh weight of carnation was found statistically significant while carnation dry weight was not found statistically significant.

Nano-silver (NS) is nanometer sized (similar to 2-5 nm) silver (Ag+) particles used in other applications as an antimicrobial material. A series of experiments were conducted to investigate effects of pulse and vase solution treatments with NS solutions on the vase lives of commercially important rose (Rosa hybrida), carnation (Dianthus caryophyllus) and gerbera (Gerbera jamesonii) cut-flowers. Both pulse and vase solutions treatments with NS individually and/or in combination extended the vase lives of these three species. Moreover, NS treatments in combination with sugar further prolonged the vase lives of these cut-flowers. In vitro tests and microscope observations indicated that NS treatments inhibited the growth of microorganisms from cut carnation flower vase solution.

Soilless cultures are nowadays an important tool for both (i) large scale synthesis of pharmaceutically active products, and for (ii) ex situ conservation of endemic and/or endangered plants that are also of horticultural importance. (i) Epilobium hirsutum explants were grown on both solid- and shaken liquid-medium, for multiplication and for greater biomass production, respectively. Secondary metabolites extracted from the biomass thus obtained were mainly polyphenols (galloylglucose and monomeric and dimeric ellagitannins), which are pharmaceutically important in prostate adenoma treatment, among others. Seedlings' dry biomass production increased from 1.5 g DW L -1 to 22 g DW L -1 after 15 days when nodes/inocula were grown on shaken liquid-medium, in light, and using different phytohormonal conditions. Secondary metabolites yield (polyphenols) of seedlings was equal to or higher (under some conditions) than for intact plants grown normally, in soil, under normal conditions. (ii) In vitro conservation on solid media resulted in a collection of 12 species that are endemic and/or endangered for Europe, and 7 species for Romania, respectively. Included in this collection are: Dianthus spiculifolius, D. nardiformis, D. henteri, D. glacialis subsp. gelidus, D. haematocalyx subsp. pindicola, D. seguieri, D. sternbergii, Achillea pyrenaica, Paradisea liliastrum. Dianthus petraeus subsp. simonkaianus was the species with the best multiplication rate (110 shoots/inoculum).

As in many ornamentals, also in carnation the number of varieties in common knowledge is large and identification throughout the chain from breeder to consumer using plant material from different stages and organs may be needed. Results in this study on the use of microsatellite markers from Dianthus caryophyllus L. for the characterization of carnation varieties as well as the construction and evaluation of a molecular database show that these markers show potential for identification purposes

Experiments were conducted to determine the effects of Thidiazuron (TDZ) applied preharvest under glasshouse conditions on Dianthus caryophyllus 'Lunetta'. Thidiazuron at 0, 1, 10, 100, and 1000 mu M was applied as a foliar spray in two separate experiments. 15 plants were used for each concentration in both experiments. Experiments were arranged in completely randomized designs. Time to flowering was recorded, and relative stem length, total nitrogen, and tissue water content were measured at harvest. Postharvest vase life, relative fresh weight changes, and solution uptake changes were also measured. TDZ treatments sometimes decreased relative stem length compared to the control (0 mu M). TDZ treatment tended to decrease total nitrogen and water content of tissues slightly, but not significantly (P > 0.05). In the first experiment, TDZ at 100 mu M significantly increased the vase life of cut carnation flowers compared to the control. In the second experiment, TDZ applied preharvest at both 10 and 100 mu M significantly increased their vase life. TDZ treated flowers tended to maintain higher relative fresh weight, with positive differences for the 100 mu M TDZ treatment being apparent at days 5, 7 and 9 of vase life. Solution uptake was sometimes higher in TDZ treated flowers.

Regenerative callus was established and was successfully applied to Agrobacterium-mediated genetic transformation. The callus was isolated from the base of adventitious shoots on cotyledon explants of Dianthus hybrida cv. 'Telstar Scarlet' cultured on MS media supplemented with 1 mg L -1 TDZ and 0.1 mg L -1 NAA. This callus culture was successfully applied to Agrobacterium-mediated genetic transformation. Transformed shoots were obtained from inoculated calli selected on MS medium containing 1.0 mg L -1 TDZ, 0.1 mg L -1 NAA, 20 g L -1 sucrose, 2 g L -1 gellan gum and 20-80 mg L -1 G-418 within 20 weeks from inoculation. All transformants grew, flowered normally and produced seeds when crossed with other Dianthus species.

Cryptic plant viruses are seed-borne dsRNA-viruses which co-exist with their host plants throughout their lives without inducing any symptoms. Since changing the growth conditions or altering the host-virus combination (cultivar, strain, isolate, etc.) may have a strong effect on virus multiplication we wanted to find out whether and if so what effect long term tissue culture has on the survival of carnation cryptic virus (CarCV). In vitro cultures of 18 different carnation species of a Hungarian germplasm collection were initiated from sterilised seeds 16 years ago and the plantlets have been grown aseptically since then. Genomic dsRNAs of CarCV were detected in Dianthus caryophyllus, D. superbus, D. carthusianorum and D. gratianopolitanus. In addition, in several cases 1-3 dsRNA-species ranging from 1100 to 3600 bp were seen. The origin of the latter dsRNA is unknown. In addition to CarCV we have also been able to show that the beet cryptic viruses BCV1, -2 and -3 survive under in vitro conditions. After 5-7 years of cultivation in some but not all sugar beet plantlets quite high concentrations of BCV dsRNAs were detected. Our results indicate that cryptic viruses are so well adapted to their hosts that despite the dramatic change of environment they may survive for more than a decade of tissue culturing.

The studies conducted on this topic, upon Dianthus genus, are justified both by its exceptional horticultural importance and by the fact that this genus has a great phytogeographic importance, because of the high rate of endemic species comprised in it. Also, many of the taxa included in this genus can be found on the national and European Red Lists. The in vitro biotechnology can successfully accomplish the needs for conservation of these species. 9 endemic or rare taxa from the Romanian and European Flora were studied. In vitro cultures were induced starting from seeds or caulinar, nodal and apical fragments. By successive transfers of apical and nodal explants on culture media having a varied hormonal balance (diverse combinations between NAA, IBA, IAA and BAP, K, 2iP, TDZ) a micropropagation of these species was achieved, with a maximum multiplication rate of 110 neoplantlets/explant (Dianthus petraeus ssp. simonkaianus). Ex vitro acclimatization was facilitated by an in vitro photoautotrophic phase, prior to the transfer in non-sterile conditions. At present, the studied species are acclimatized or they are in progress of being acclimatized in the Alexandru Borza Botanical Garden of Cluj-Napoca.

Within the framework of the European project LIFEOONAT/IT/7216 for conservation of environments of endangered bats, in vitro propagation of Dianthus balbisii Ser. ssp. liburnicus (Bartl.) Pign. (Caryophyllaceae) from wild plants in the Onferno Natural Reserve has been planned to restore foraging habitats. A satisfactory percentage of not contaminated explants has been achieved after double sterilisation of nodal segments. Shoot proliferation occurred on a MS based medium enriched with 0.5 mg L -1 BAP and 0.7% commercial agar (B&V, Parma, Italy, S1000). After a short in vitro phase on a medium enriched with 1 mg L -1 IBA for root induction, acclimatisation in the greenhouse was successful. Plants have already been transplanted to outdoor flowerbeds in the Reserve. Different characteristics shown during proliferation and after acclimatisation by clones, each derived from single initial shoot tip, are still under observation.

The effects of silver thiosulfate (STS; AVB) and 1-methylcyclopropene (1-MCP; Ethylbloc) were determined on 14 commonly-grown cut flower species, represented by one to three cultivars per species. Stems were unpacked, sorted, and placed in either deionized water (DI) and subjected to 1-MCP (740 nL L') or ambient air for 4 h or DI plus STS at either 0.1 mM (Alstroemeria) or 0.2 mM (all other species) for 4 h. After treatment, stems were removed, placed in polyethylene sleeves and stored either wet in DI water or dry in plastic-lined floral boxes at 5°C in the dark for 4 days. After storage bunches were placed in DI water under 12 h (76 to 100 μmol m -2 s -1 ) light per day. Flowers were monitored daily to determine the end of wholesale vase life, which was designated as the first day a change was noticed in the flower or inflorescence that would typically prevent it from being sold by a wholesaler or retailer. The consumer vase life was also recorded for each stem and was designated as the day a typical consumer would dispose of it. The 19 cut flower taxa could be organized into four groups based on effectiveness of STS and 1-MCP: (1) Both STS and 1-MCP increased vase life but STS was more effective: Dianthus caryophyllus (all three cultivars), Bouvardia, Lilium (Asiatic), and Lathyrus odorata. (2) Both STS and 1-MCP prevented the negative effects of dry storage: Freesia (both cultivars) and Chamelaucium (one cultivar). (3) STS increased vase life while 1-MCP did not: Alstroemeria, Delphinium, Matthiola, and Gypsophila. (4) STS and 1-MCP either had no effect or a negative effect: Consolida, Eustoma, Ranunculus, Antirrhinum, and Chamelaucium (one cultivar).

We studied application of 1-methylcyclopropene (1-MCP) in various ornamentals, with the aim of finding optimum application temperature, dose, and duration. We also studied its competition with ethylene, and its synergistic effect with silver thiosulfate (STS). Ethylene (5 µl l -1 ) was simultaneously applied with 1MCP (15 or 150 nl l -1 ) at 4, 12 or 20oC to carnation (Dianthus caryophyllus cv. ‘Yellow Candy’). At these application temperatures ethylene negatively affected flower quality. 1-MCP applied at 15 nl l -1 neutralized the adverse ethylene effects when applied at 4oC, but not at 12 or 20oC. However, when applied at 150 nl, l -1 1MCP successfully eliminated ethylene effects at all temperatures. It seems, therefore, that 1-MCP is a better competitor with ethylene at low temperatures. 1-MCP pre-treatment (0.2-1 µl l -1 /2-20 h), followed by a subsequent exposure to ethylene (1-5 µl l -1 /24 h/20oC or 0.5 µl l -1 /12 days/12oC), significantly improved the quality of cut flowers (carnations cv. ‘Yellow Goldy’, Cymbidium orchids) and potted plants (Ficus ‘Green island’), respectively. In Limonium hybrid ‘Beltlaard’ cut flowers the combined use of 1-MCP and STS resulted in highest quality. This combined treatment also enabled storage under sea transport conditions (8 days at 2oC). When applied alone, 1-MCP (0.4 µ µ µ µl l -1 /4 h) was also very effective in improving flower opening and the length of vase life in several rose cultivars. The results suggest that 1-MCP may be very useful in preserving quality of various ornamentals and that it protects against ethylene effects.

In senescing carnation (Dianthus caryophyllus L.) flowers, ethylene is produced from the gynoecium, and acts as a diffusible signal received by petals to induce the expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase (DC-ACS1) and ACC oxidase (DC-ACO1) genes in the petals. This results in autocatalytic ethylene production in the petals. We investigated ethylene production in cut flowers of cv. White Candle, which produce ethylene only in trace amounts and have a long vase life. The low ethylene production in these flowers was due to low ethylene production in the gynoecium, accompanied by low accumulation of ACC synthase transcripts. These findings further support the importance of ethylene production from the gynoecium in the senescence of carnation flowers. Cv. White Candle flowers had water relations that were different from those in cv. Light Pink Barbara, a cultivar that showed the normal climacteric rise of ethylene production during senescence. We discuss a factor that possibly induces ethylene production in the carnation gynoecium.

Microcuttings (microshoots) from transformed carnation (Dianthus caryophyllus L. cv. Tanga) plantlets, originated in vitro from leaf explants and carrying the bar gene with resistance to the herbicide BASTA®, achieved 47% rooting in vitro in MS medium supplemented with 3 μM IBA plus 100 μg l -1 kanamycin as a selection agent. Microcuttings from non-transformed plantlets did not root at all using the same selection agent. For the acclimatization stage, the rooted microcuttings were planted to a soilless mixture in boxes and maintained in a growth chamber with controlled environmental conditions for 2 months. Young plants produced from these transformed microcuttings were moved to greenhouse and then cuttings were taken from them for rooting. Rooted cuttings, which were transplanted to pots in a soil mixture, grew successfully to plants and produced red flowers typical of the cv. Tanga.

Resistance of carnation to Fusarium wilt (Fusarium oxysporum f. sp. dianthi) is of major importance for breeding new cultivars. New germplasm and combination between ornamental traits and high resistance to Fusaruim wilt is a main achievement in the selection of this crop. The investigation of the response of parental forms and new selected hybrid material is a main step in breeding programs. Interspecific hybridization of carnation allows increasing the diversity in germplasm. Five spray carnation cultivars and two breeding lines of D. caryophyllus f. spray Hort. (Krassina, Regina, Fea, Naslada, Barbara, L-169 and L-230) used as a mother plant and seven Dianthus species used as a father plant (D. carthusianorum, D. gratianopolitanus, D. sequieri, D. knappii, D. silvestris, D. plumarius and D. chinensis) were evaluated for their resistance to Fusarium wilt after artificial inoculation with the causal pathogen of the disease. The resistance of wild Dianthus species is much lower compared to this of spray carnation cultivars. Hybridisation was performed after castration and compulsory, forced pollination with pollen collected from different species under isolation. All tested breeding lines showed a very high level of resistance to F. oxysporum f. sp. dianthi. The level of resistance in F1 and BC1 is variable. Clones of individual plants in some crosses showed a very high level of resistance, reaching complete resistance. Interspecific hybridization gives an opportunity to select plant forms combining variable ornamental traits with high level of resistance to Fusarium wilt.

In vitro shoots without roots from different media compositions were selected and transferred into test tubes supplemented with MS medium without hormone for root induction. After 4 weeks, full grown plantlets were transferred to sterile soil for acclimatization process. The plantlets were maintained in the culture room at 25±1°C with 16 hours light and 8 hours dark. Plantlets were then planted in pots and covered with polystyrene plastic bags. They grew well and after 4 weeks in the culture room, well grown and healthy plantlets were transferred to the greenhouse. Survival rate of the acclimatized plantlets was 70% and from observation it showed that most of the plantlets successfully survived after transferred to the greenhouse.

The influences of the medium composition (gelling agent, BA concentration and NO 3 -/NH 4 + ratio) at different recultures on vitrification of carnation in vitro shoots proliferation have been investigated. It was found that the Gelrite-containing medium produced a higher multiplication rate and higher percentage of vitrification shoots than the Difco agar-containing medium. The percentage of vitrified shoots decreased with increasing concentrations of Difco agar in the medium while the multiplication rate decreased. Shoot production increased with increasing benzyladenine (BA) concentration (from 0 to 4 mg/l), however it also resulted in a proportional increase in shoot vitrification. The effect was more pronounced at high BA concentrations. Higher ratio of NO 3 -/NH 4 + in the medium prevented the vitrification of carnation plantlets. Moderate ratios of 1/1 to 2/1 resulted in a higher number of adventitious shoots at the fourth reculture only and no significant difference was found at the first reculture. Nevertheless, shoots of the fourth reculture showed a higher percentage of vitrification than those of the first reculture for all the factors tested in this study. In conclusion, the medium containing 1/2 MS (Murashige & Skoog, 1962) basic salts supplemented with 3% sucrose, 0.9% agar, and 0.5 mg/l BA (pH 5.7) was ideal for both mass propagation and for reducing vitrification of carnation. Shoot tips that havefewer recultures were more beneficial for preventing shoot vitrification during mass propagation of carnation in vitro than shoots that have been recultured many times.