Seed germination in the Sicilian subspecies of Dianthus rupicola Biv. (Caryophyllaceae)

Abstract

Five populations belonging to three subspecies of Dianthus rupicola Biv. (D. rupicola subsp. rupicola, D. rupicola subsp. aeolicus, and D. rupicola subsp. lopadusanus) and growing in different geographical areas of Sicily were tested for seed germination at various temperatures. All populations showed high germination rates with an optimum temperature between 15°C and 25°C. Efficient seed germination might contribute significantly to the preservation of these subspecies which are currently exposed to several environmental threats.     

Effect of carbon dioxide enrichment during in vitro cultivation and acclimation to ex vitro conditions

Tobacco and carnation plantlets were grown in vitro on Murashige and Skoog's medium with 2 % saccharose. Carnation plantlets were also grown fully photoautotrophically on a medium without saccharose. The ambient CO2 concentration was increased from 0.6 to 10 or 40 g m-3 during the last 3 weeks of in vitro cultivation or during the first 3 weeks of acclimation to ex vitro condition (plantlets transplanted to pots with sand and nutrient solution) or during both growth phases. CO2 enrichment during in vitro cultivation markedly stimulated growth of tobacco plantlets, and also of carnation plantlets, both with and without saccharose. CO2 enrichment during the acclimation period promoted plant growth more effectively in plantlets grown in vitro at a CO2 concentration of 0.6 g m-3 than in plantlets grown in either growth phase at higher CO2 concentrations. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vitro production of dianthin from crown gall lines of carnation (Dianthus caryophyllus L.)

Tumorous crown gall lines of carnation (Dianthus caryophyllus L.) were obtained by transforming leaf explants by the co-cultivation procedure; transformed lines were checked for their ability to produce in vitro the type 1 ribosome-inactivating proteins dianthins. Crude extracts from cultured callus were able to inhibit protein synthesis in a cell-free system based on rabbit reticulocyte lysate. A protein with an apparent molecular mass comparable to that of dianthin 30 from leaves was identified from its chromatographic behaviour and by Western analysis. Dianthin from callus showed a specific activity comparable to that reported for the leaf isoform. The presence and accumulation in the tumorous line of a 37 kDa protein with no ribosome-inactivating activity and strong antigenic reactivity to dianthins is discussed. Received: 18 July 1997 / Revision received: 10 January 1998 / Accepted: 5 February 1998     

Effect of oxygen concentration on the senescence and energy metabolism of cut carnation flowers

The effect of O₂ concentration on energy metabolism and senescence has been studied in cut flowers of Dianthus caryophyllus L. cv. Scania. As compared to the control (21% O₂), 5% O₂ delays flower senescence as well as decay of nucleotide level and AEC (adenylate energy charge). An atmosphere of 100% O₂ accelerates senescence as well as the decrease of nucleotide level and AEC. While anoxia brings about a faster decrease of ATP and AEC than of total nucleotides, hyperoxia brings about a faster decrease in adenyl nucleotides than in ATP and AEC values. Petal oxygen uptake is over 90% of the maximal value under 4% O₂ and saturates at 10% O₂. The development of senescence is dicussed as a two phase process (first phase-progressive and second phase-catastrophic) triggered by the action of hyperoxia, first on the system for energy utilization and later on the system for energy production, the degradation of which seems to be linked with increase in membrane permeability and withering.     

Cytokinins in cut carnation flowers: I. The complex in ovaries

Tentative identification of the cytokinins present in extracts of Dianthus caryophyllus ovaries using High Performance Liquid Chromatography and Radioimmunoassay techniques, revealed the presence of trans-ribosylzeatin, trans-zeatin, dihydrozeatin and N⁶ (²-isopentenyl)adenine. In addition slow moving compounds (paper chromatography) which could be hydrolysed by -glucosidase were also detected. After hydrolysis the active compounds co-chromatographed with zeatin and ribosylzeatin.     

Regulation of membrane phospholipid catabolism in senescing carnation flowers

Evidence has been obtained for the involvement of μ M levels of Ca²⁺ in phospholipid catabolism during petal senescence by following the breakdown of [U-¹⁴C]-phosphatidylcholine by microsomal membranes from cut carnation ( Dianthus caryophyllus L. cv. White-sim) flowers. Phospholipid degradation was mediated by three membrane-associated lipases, viz. phospholipase D (EC 3.1.4.4), phosphatidic acid phosphatase (EC 3.1.3.4) and lipolytic acyl hydrolase. The activities of phospholipase D and phosphatidic acid phosphatase were stimulated by 30 and 100%, respectively, in the presence of 40 μ M free Ca²⁺, and the Ca²⁺-stimulation of phosphatidic acid phosphatase was calmodulin-dependent. When L-3-phosphatidyl-[2-³H]-inositol and L-3-phosphatidyl-[N-methyl-³H]-choline were used as substrates, inositol and choline accounted for 95 and 99%, respectively, of the water-soluble radiolabelled products. This suggests a predominance of phospholipase D activity over phospholipase C activity in these membranes.
Breakdown of membrane phospholipids in senescing carnations is known to be accelerated by treatment of young flowers with ethylene. To determine whether this involves a specific turnover of phosphatidylinositol as observed in animal systems in response to certain agonists, young flowers pre-labelled with ³²PO^3-₄ were treated with 10 ppm ethylene. All phospholipids incorporated the label, but no enhanced turnover of phosphatidylinositol was observed. Inositol 1,4,5-triphosphate did not release Ca²⁺ from preloaded microsomal vesicles at concentrations known to be effective in animal systems (i.e. < 5 μ M ) although release of Ca²⁺ was observed when a higher (20 μ M ) concentration was used.     

热击对麝香石竹茎段培养物培养效率的影响

本文研究了热击对麝香石竹茎段培养物培养效率的影响。结果表明,43℃2h热击麝香石竹茎段能促进或拟制不定芽的直接分化,提高愈伤组织的诱导率。热击愈伤组织能显著促进愈伤组织在继代培养基上的生长,但抑制愈伤组织在分化培养基上的生长。热击也能提高再生苗和愈伤组织的干重百分率。我们认为,热击所产生的生物学效应是与配合条件、特别是外源激素状况密切相关的。     

Endogenous plant growth regulators in carnation tissue cultures under different conditions of ventilation

Endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinins (zeatin, zeatin riboside, dihydrozeatin, (diH)Z, dihydrozeatin riboside, (diH)[9R]Z, N⁶-isopentenyl adenine and N⁶-isopentenil adenine riboside) levels were evaluated in normal (N) and hyperhydric (H) microplants of Dianthus caryophyllus cultured under different aeration conditions in hormone-free liquid medium. The morphological differences between N and H explants grown under ventilated conditions were correlated with differences in their endogenous hormonal levels: after 15 and 30 days of culture, H explants showed lower IAA and ABA contents than N explants, as well as higher cytokinin levels, mainly of (diH)Z and (diH)[9R]Z. This was associated with less tissue differentiation and with an inability of H microplants to survive under ex vitro conditions. However, these relationships could not be observed between H and N explants grown under non-ventilated conditions probably due to the difficulty in discerning the plant status (N or H) and therefore, an underestimation of H microplants. This assumption is supported by the low ability for acclimatization to ex vitro of N plants grown without ventilation.     

Characterization and heterologous expression of hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase from elicited cell cultures of carnation, Dianthus caryophyllus L.

Benzoyl-CoA:anthranilate N-benzoyltransferase catalyzes the first committed reaction of phytoalexin biosynthesis in carnation (Dianthus caryophyllus L.), and the product N-benzoylanthranilate is the precursor of several sets of dianthramides. The transferase activity is constitutively expressed in suspension-cultured carnation cells and can be rapidly induced by the addition of yeast extract. The enzyme was purified to homogeneity from yeast-induced carnation cells and shown to consist of a single polypeptide chain of 53 kDa. Roughly 20% of the sequence was identified by micro-sequencing of tryptic peptides, and some of these sequences differed in a few amino acid residues only suggesting the presence of isoenzymes. A specific 0.8 kb cDNA probe was generated by RT-PCR, employing degenerated oligonucleotide primers complementary to two of the tryptic peptides and using poly(A)⁺ RNA from elicited carnation cells. Five distinct benzoyltransferase clones were isolated from a cDNA library, and three cDNAs, pchcbt1–3, were sequenced and shown to encode full-size N-benzoyltransferases. The translated peptide sequences revealed more than 95% identity among these three clones. The additional two clones harbored insert sequences mostly homologous with pchcbt1 but differing in the 3-flanking regions due to variable usage of poly(A) addition sites. The identity of the clones was confirmed by matching the translated polypeptides with the tryptic enzyme sequences as well as by the activity of the benzoyltransferase expressed in Escherichia coli. Therefore, carnation encodes a small family of anthranilate N-benzoyltransferase genes. In vitro, the benzoyltransferases exhibited narrow substrate specificity for anthranilate but accepted a variety of aromatic acyl-CoAs. Catalytic rates with cinnamoyl- or 4-coumaroyl-CoA exceeded those observed with benzoyl-CoA, although the corresponding dianthramides did not accumulate in vivo. Thus the cDNAs described represent also the first hydroxycinnamoyltransferases cloned from plants, which classifies the enzymes as hydroxycinnamoyl/benzoyltransferases.