Dihydroflavonol 4-reductase (DFR) is a key enzyme in the anthocyanin biosynthesis. In this study, the localization of 45S rDNA and dfr gene (named as CsDFR-bo) of the blood orange (Citrus sinensis Osbeck cv. Ruby) on chromosomes was investigated by fluorescence in situ hybridization (FISH). A karyotype of C. sinensis was reconstructed based on the length of mitotic metaphase chromosomes. The 45S rDNAs were localized on chromosomes 2p and 7q. The detection ratios of 45S rDNA in the two chromosomes were 69.5 and 77.3 %, respectively. The CsDFR-bo was proved to be a single-copy gene and localized on chromosome 3p. The detection ratio of CsDFR-bo was 8.2 %.

Forty one simple sequence repeats were isolated from two microsatellite enriched libraries of date palm (Phoenix dactylifera L.). After screening, 17 selected microsatellite loci were characterized and evaluated on a set of 31 cultivars and clones from Algerian and Californian germplasm. All primer pairs produced an amplification product of the expected size and detected high polymorphism among the analysed samples. These nuclear simple sequence repeat (SSR) markers are expected to be a very effective tool for evaluating genetic diversity in date palm germplasm. Acrosstaxa amplification showed the usefulness of most SSR markers in 14 other species across the genus Phoenix.

The transformation of potato (Solanum tuberosum L. cv. Désirée) was extended by the Agrobacterium-mediated biolistic method. Using this approach transgenic shoots could be obtained at a similar frequency to that achieved through conventional biolistics. Leaves from shoot cultures were bombarded with gold particles coated in Agrobacterium tumefaciens cells harboring a binary plasmid encoding three genes of interest in the T-DNA. Nine shoots were obtained from 20 shots, with selection of transgenic shoots on a series of media containing progressively increasing concentrations of hygromycin from 5 to 20 mg dm^-3.

A novel protocol for plant regeneration from cotyledon explants of eggplant (Solanum melongena) reducing concentration of sucrose was established. The most efficient bud induction medium consisted of Murashige and Skoog (MS) medium supplemented with 2.0 mg dm^-3 zeatin, 0.1 mg dm^-3 indoleacetic acid and 10 g dm^-3 sucrose. After 15 d, the shoot buds were fragmented and transferred to the shoot elongation MS supplemented with 1.0-2.0 mg dm^-3 gibberellic acid and 4.0-8.0 mg dm^-3 AgNO₃, which promoted shoots elongation. The genetic stability of the regenerated plants was analyzed by flow cytometry, RAPD and SSR molecular markers. The results indicated that almost no somaclonal variation was detected among the regenerants.

Rapid micropropagation was achieved in Chlorophytum borivilianum Santapau and Fernandes using shoot base as explants. Multiple shoots were induced on Murashige and Skoog's (MS) medium supplemented with 3.0 mg dm^-3 6-benzylaminopurine, 0.1 mg dm^-3 1-naphthaleneacetic acid, 150 mg dm^-3 adenine sulphates and 3 % saccharose. Rooting was readily achieved upon transferring the shoots onto half strength MS medium supplemented with 0.1 mg dm^-3 indolebutyric acid and 2 % saccharose. Micropropagated plantlets were hardened in the greenhouse and successfully established in soil. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of the micropropagated plants. Thirty one arbitrary decamers were used to amplify genomic DNA from in vitro and in vivo plant material to assess the genetic stability. All RAPD profile analysis from micropropagated plants was genetically similar to mother plants.

Direct somatic embryogenesis of Frittilaria meleagris L. was induced using leaf base explants excised from in vitro grown shoots. Somatic embryos occurred at the basal part of leaf explants 4 weeks after culture on a Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or kinetin (KIN). The highest number of somatic embryos (SEs) were formed (9.74) from leaf explant on MS medium supplemented with 0.1 mg dm^-3 2,4-D after 4 weeks of culture initiation. An initial exposure to a low concentration of KIN in the medium also enhanced SEs induction. Our observations by light and scanning electron microscopy revealed that SEs originate directly from the epidermal and subepidermal layers of leaf explant. The developmental stages of somatic embryogenesis from the first unequal cell division through the meristematic clusters, multi-cellular globular somatic embryos to the fully formed cotyledonary embryos were determined. After 4 weeks on MS medium without plant growth regulators, SEs developed into bulblets.

Primary leaf explants of aseptically grown seedlings of moth bean [Vigna aconitifolia (Jacq.) Marechal] immersed in water or not were treated in microwave oven (2450 MHz, 800 W cm^-2) for 1, 3, 5 and 7 s before culturing. Callusing and shoot emergence from these explants were enhanced up to microwave exposure lasted 5 s while longer treatment of water-immersed explants delayed callusing. One polypeptide (26.6 kD) was up regulated in the callus derived from microwave treatment in water-immersed explants. RAPD analysis detected alteration in DNA sequences due to microwave treatment in water-immersed explants for 7 s. The frequency of mutation was 1.6 % (4 bands out of 248) over all the cultures analyzed and the same was 13 % (4 bands out of 31), if amplicons generated at 7 s treatment alone were considered.

In this study, photosynthetic performance, pigment content, chlorophyll a fluorescence, and leaf anatomy in peanut (Arachis hypogaea) subjected to zinc (Zn) stress were investigated. Zn stress resulted in reduction of photosynthetic and transpiration rates, pigment contents and root biomass. Zn-induced xerophyte structure in peanut leaves (i.e. thick lamina, upper epidermis, and palisade mesophyll, as well as abundant and small stomata) also contributed to decreased transpiration rate and stomatal conductance. This in turn, partially contributed to the limitation of photosynthesis.

An efficient and reproducible in vitro plant regeneration system from shoot apices was developed in Jatropha curcas. Benzylaminopurine (BAP; 2.5 μM) was most effective in inducing an average of 6.2 shoots per shoot apex. Incorporation of gibberellic acid (GA3; 0.5 μM) to basal medium was found essential for elongation of shoots. The BAP-habituated mother explants continuously produced shoots during successive subculture without any loss of morphogenic potential. The shoots rooted efficiently on half-strength MS medium. The rooted plantlets were acclimatized with more than 98 % success and the plants transferred to soil:compost in nursery showed no sign of variation compared to the seed-grown plants. The whole process of culture initiation to plant establishment was accomplished within 5-6 weeks. A genetic transformation system in J. curcas was established for the first time, using bombardment of particles coated with plasmid pBI426 with a GUS-NPT II fusion protein under the control of a double 35S cauliflower mosaic virus (CaMV) promoter. The β-glucuronidase (GUS) activity in J. curcas shoot apices was significantly affected by the gold particle size, bombardment pressure, target distance, macrocarrier travel distance, number of bombardments, and type and duration of osmotic pre-treatment. The proliferating bombarded shoot apices were screened on medium supplemented with 25 mg dm^-3 kanamycin and surviving shoots were rooted on medium devoid of kanamycin. The integration of the transgene into genomic DNA of transgenic plants was confirmed by PCR and Southern blot hybridization. The transgenic plants showed insertion of single to multiple copies of the transgene.

Short-term treatment of potato (Solanum tuberosum L.) tuber discs with CdCl₂ solution elevated both the StPCS1 transcript level, phytochelatin synthase (PCS) activity and contents of phytochelatin (PC) and abscisic acid (ABA). Similar effects but less marked were noticed after treatment of tuber tissues with ABA solution. Cd-treatment increased also cysteine (CYS) content but did not change glutathione content. If ABA treatment preceded Cd-treatment, the elevation of CYS and PC contents were limited. The data suggest the participation of ABA in the regulation of PCS.

An investigation was undertaken to adjudge the in vitro salt tolerance and biochemical changes due to NaCl on four grape rootstocks (Dogridge, SO4, H-144 and 3309C). The in vitro two-node micro-cuttings from the established cultures were sub-cultured on rooting medium comprising Murashige and Skoog (MS) basal medium supplemented with 0.5 μM indolebutyric acid (IBA), 200 mg dm^-3 activated charcoal and different NaCl concentrations ranging from 0 to 125 mM. The surviving and proliferated cultures were further sub-cultured four times on respective media. Dogridge and H-144 tolerated 125 and 100 mM NaCl, respectively, while SO4 and 3309C survived only up to 75 mM NaCl. Contents of proteins, proline, K⁺ and Na⁺ in tissue increased in all genotypes due to NaCl supplementation, while contents of chlorophyll and total soluble sugars declined. Higher K⁺/Na⁺ ratio was registered in Dogridge and H-144 than in SO4 and 3309C. The relative NaCl tolerance for different grape rootstocks under study could be ranked as Dogridge > H-144 > SO4 and 3309C.

The contents of endogenous free and conjugated polyamines, putrescine (Put) and spermidine (Spd), were determined during 9 week of vernalization (at 5 °C) in winter wheat seedlings cultivated on Murashige and Skoog media without (MS0) and with 2 mg dm^-3 zearalenone (MSZEN). At the 4^th week of chilling treatment, which is sufficient to induce generative development in 30 % of plants, the marked increase in free and conjugated forms of Put and free Spd were observed. The presence of ZEN in medium significantly accelerated the vernalization. About 20 % of plants treated with ZEN flowered already after 2 weeks and 40 % after 3 weeks of chilling. Significantly higher content of free Put was determined in roots grown on MSZEN compared with MS0 during the first 5 weeks of vernalization with maximum at the 4^th week. After germination, a marked decrease in free Spd content was observed both in plants grown on MS0 and MSZEN. Application of ZEN significantly slowed down the Spd decline in leaves and roots during the first and second week of vernalization. The content of Spd and its conjugates decreased in vernalized plants after 1 week of cultivation at 20 °C.

UV-B responses of three rice (Oryza sativa L.) cultivars (Sasanishiki, Norin 1 and Surjamkhi) with different photolyase activity were investigated. Carbon dioxide assimilation data support that Sasanishiki was less sensitive to UV-B than Norin 1 and Surjamkhi. UV-B radiation sharply decreased the content of Rubisco protein in Surjamkhi and has no effect in Sasanishiki. The photochemical activities of photosystem (PS) 1 and PS 2 was slightly affected by UV-B treatment. The content of H₂O₂ and the activities of antioxidant enzymes, catalase (CAT), peroxides (POX) and superoxide dismutase (SOD) were enhanced after UV-B treatment. The activities of CAT and POX isoenzymes in Sasanishiki were more enhanced by UV-B radiation than those in Norin 1 and Surjamkhi.

The aim of this work was to study the short-term effects of salt stress on the antioxidant system and ascorbate peroxidase (APX) expression in two salt sensitive sweet potato cultivars Tainung 57 (TN57) and Tainung 66 (TN66), and one salt-tolerant cultivar Hsusu 18 (HS18). Plants were grown in plastic pots in a greenhouse for 30 d followed by NaCl treatments (0, 150, 300, and 450 mM) for 0, 24 and 48 h in a growth chamber. Young, fully expanded leaves of each treatment and period of time were clipped for enzyme activity measurements. In addition, different tissues (leaves, stems, and roots) were also harvested to analyze the tissue-specific APX gene expression using semiquantitative reverse-transcription polymerase chain reaction (RT-PCR). Three degenerated primers of APX isoforms from cytosol, peroxisomes and chloroplasts were used to amplify the APX complementary DNA of these cultivars. Our results show higher increase in APX activity at 24 and 48 h of salinity (450 mM of NaCl) in salt-stress tolerant genotype than in saltsensitive ones. The expression of APX isoforms in response to salinity was tissue specific and also dependent on stress duration.

The effect of 5-azacytidine (5-azaC) on the alleviation of damaging effects of NaCl treatment was studied in two wheat (Triticum aestivum L.) cultivars differing in salt tolerance (salt-tolerant Dekang-961 and sensitive Lumai-15). The plants were pre-treated or not with 50 μM 5-azaC and then subjected to salt stress induced by 100 or 150 mM NaCl. Salinity caused reduction in biomass accumulation and increase in malondialdehyde content in root tissues in both cultivars, but less in pre-treated seedlings. The activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in the roots of both cultivars increased during salt stress, but the rate of increase was higher in Dekang-961. Plants treated with 5-azaC had higher root SOD, CAT and POD activities under salt stress than untreated plants. Content of 5-methylcytosine (5mC) decreased in both cultivars under salt stress, and the level of demethylation was higher in Dekang-961 than that in Lumai-15. Moreover, the degree of methylation was lower in both cultivars under salt stress after 5-azaC application compared to only salt-treated groups. These findings suggested that 5-azaC could protect plants from salt stress.

Occurrence and genomic organization of dispersed elements containing ZpS1 satellite repeats have been investigated in a wide representation of species of the old plant genus Zamia (Zamiaceae, Cycadales). In Z. paucijuga, the ZpS1 repeat is organized as long satellite DNA arrays and as short arrays inserted into AT-rich dispersed elements. A comparative study by Southern analysis shows that these unusual dispersed elements containing the ZpS1 repeat are present with different organizations in all investigated Zamia species. In some species these elements are present with a low copy number, while in other species secondary amplification events, involving specific sequence clusters, appear to have generated characteristic dispersed elements in a high copy number. Among Zamia species, several groups share similar restriction patterns, as the Zamia loddigesii complex and the Caribbean species suggesting a general correlation between organization and genomic representation of the dispersed repeated sequence and the pattern of phyletic relationships in the genus. However, the finding of different patterns also among closely related species suggests a complex history of amplifications and losses of these dispersed repetitive elements that cannot be always easily traced through the phylogenetic reconstruction of this ancient plant group.

After identifying regions of cDNA conserved between the symbiotic gene DMI1 of the model species Medicago truncatula and the homologous genomic region of Arabidopsis thaliana, universal primers were designed from 8 of 12 exons to allow the routine amplification of plant homologs. As an example, the complete homologous sequence from the pea (Pisum sativum L.) was amplified and sequenced, although the poorly conserved 5'-end and 5'-flanking region of the gene had to be amplified using a modified TAIL-PCR strategy. The identity of this amplified homolog with the SYM8 gene was independently confirmed by the presence of a single nucleotide change in the coding sequence of the mutant line Risnod27 (sym8) that cosegregated with the asymbiotic phenotype. Five insertions in pea introns responsible for increasing the total length of SYM8 by 1443 bp, compared to the M. truncatula homolog DMI1, belong to known transposon and retrotransposon families of pea and legumes in general. In view of the predicted function of SYM8 as an ion channel, the Risnod27 mutation (His309Tyr) appears to be localized in the selectivity filter domain. This finding confirms the essential role of histidine 309 in the symbiotic function of SYM8 and provides a guide to its ionic specificity. In view of the Risnod27 symbiotic phenotype, we hypothesize that SYM8 does not have identical functions in the transduction of rhizobial and mycorrhizal signals. The variability of the N-proximal region of the known legume homologs of DMI1 suggests an interaction with a variable ligand.

Axillary buds sampled from a mature 27-year-old Cornus mas cv. Macrocarpa were grown in vitro on modified woody plant medium (WPM). Adventitious rooting performance of microshoots was assayed on half-strength WPM supplemented with 1-naphthaleneacetic acid (NAA) or indole-3-butyric acid (IBA) under various pH. NAA induced significantly higher rooting frequencies than IBA. The pH of 6.8 inhibited rooting, and differentiated roots were extremely thick and fragile. The highest rooting frequency was recorded on half-strength WPM supplemented with 5.37 µM NAA at the pH value adjusted to 6.2 (73 % of rooted shoots). In the presence of IBA, the formation of adventitious roots was observed only in the basal part of the microshoot dipped into rooting medium. In the case of NAA, however, adventitious roots arose also from the parts of microshoots that were not in contact with medium. The growth of aerial roots was always positively gravitropic. The nuclear microsatellite Cf-G17 gave a monomorphic fingerprinting pattern across the mother shrub and micropropagated plantlets. Acclimatized plants did not show any visually detectable morphological variation and the aerial adventitious root formation was no longer observed.

Two synthetic populations (Syn-5 and Syn-7), four inbred lines and four landraces of alfalfa (Medicago sativa L.) were studied for genetic relatedness. These forms were characterized by different amount of seeds set and green mass yield. Two primer pairs of specific nucleotide sequences of mitochondrial DNA isolated from soybean were used. The mtDNA revealed the existence of significant polymorphism among the investigated forms of alfalfa. The genetic similarity (Dice coefficient) among studied forms of alfalfa ranged from 20.1 to 96.1. The greatest resemblances were noticed between D₅ inbred line and the population of Syn-5. The lowest resemblances were noticed between Syn-7 and E_1/2. The UPGMA dendrogram split investigated forms of alfalfa into two groups: first group include three landraces, the second consist of the rest analyzed forms. There are two landraces distinct with the highest seed set and yield of green mass: one in the first group, another one in the second group.

The present study applies RAPD technique and morphometric analysis to study the diversity of some accessions belonging to section Solanum. A total of 252 products were amplified with 23 12-mer arbitrary primer pairs, among which 210 were found to be polymorphic. Sixteen morphological characters were measured and used to compile a dendrogram. Both the morphological and RAPD marker analysis clearly separated the different accessions into similar groups. The results indicate that the analyzed cultivars with unknown origin could be derived from S. retroflexum. We found morphological differences among the S. scabrum subsp. scabrum accession which were not reflected in the molecular data. Presumably these accessions represent cultivated forms selected for their habit, fruit quantity and/or quality and leaf size, respectively.

Effects of salinity (0, 50, 100 and 250 mM NaCl) on growth, root:shoot dry mass ratio, osmotic potential (ψ_x), electrolyte leakage and contents of Na⁺ and K⁺, polyamines and abscisic acid (ABA) were studied in the grape rootstocks Dogridge, 1613, St. George and Salt Creek. In control rootstocks, the root length was highest in Dogridge and contents of K⁺ and ABA in Salt Creek. Salinity treatments increased root Na⁺ and decreased K⁺ content and St. George exhibited highest Na⁺ content and Na⁺:K⁺ ratio. The root:shoot dry mass ratio in all rootstocks increased upto 100 mM NaCl. With increasing NaCl concentration, putrescine, spermine and spermidine contents showed consistent increase and putrescine increase was highest in St. George and spermidine and spermine in the Dogridge and Salt Creek. Under salinity, the ABA content increased in all the rootstocks but more in Salt Creek and Dogridge than in St. George.

Three genetic similarity coefficients were estimated and compared for their usefulness: simple matching (S _SM), Jaccard's (S _J) and Dice's (S _D), all based on dominant markers data from individuals representing predominantly self-pollinating species. AFLP markers were used to analyze 139 Phaseolus vulgaris L. (common bean) and 67 Lactuca saligna L. (least lettuce) accessions, and RAPD markers were used to analyze 110 Triticum dicoccoides Koern. (wild emmer wheat) accessions. Similar discriminating structure and power based on the three genetic similarity coefficients was found for each of the three species. This discriminating power was high for both P. vulgaris and L. saligna but moderate for T. dicoccoides. With closely related individuals, as in our study, the absence of a band in two individuals should be due to an identical cause inherited from the same ancestor. Accordingly we propose the use of S _SM, which alone out of the three examined coefficients involved shared absence of DNA bands, as contributing to genetic similarity. When RAPDs are employed, inferences about population structure and nucleotide divergence should be made with prudence as the nature of genetic variation uncovered by RAPDs is often unclear.

Seedlings of Sorghum bicolor (L.) Moench were subjected to 180 mM NaCl with or without 0.25 mM spermine (SPM) for 7 d. NaCl treatment resulted in the inhibition of growth and increased the content of free proline, soluble protein and malondialdehyde (MDA). Additionally, it also enhanced the activity of catalase (CAT), peroxidase (POX) in both shoots and roots, while decreased that of glutathione reductase (GR). When exogenous spermine was added to the test solution, the growth of sweet sorghum seedlings was improved, and a smaller increase in the free proline and MDA contents was observed. The addition of spermine also partially increased the activities of POX and GR, but had no effects on soluble protein content or the activity of CAT.

Plantain (Musa ABB CEMSA 3/4) plantlets were micropropagated in temporary immersion bioreactors (TIB) or in gelled medium (GM). After ex vitro transfer ROS accumulation was determined by infiltrating leaves with nitroblue tetrazolium (NBT) and 3,3'-diaminobenzidine (DAB). Stomatal cells were more stained with NBT and DAB in GM plants than in TIB plants, but the difference disappeared at the end of acclimatization. At the end of the in vitro phase, GM plantlets showed higher activities of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR), while activities of catalase (CAT), superoxide dismutase (SOD) and glutathione transferase (GT) were higher in TIB grown plantlets. At the end of acclimatization GT, SOD, CAT and MDHAR stabilized at low values of activity in plantlets derived from both treatments. Concerning the correspondent genes, GM plantlets showed higher expression of all transcripts with the exception of CuZnSOD. The immunobloting of peroxiredoxins (PRXs) showed that chloroplast-located PRXs were expressed at higher levels in TIB plantlets, some showing polymerization. In conclusion, TIB grown plantlets had an improved anti-oxidative response when compared with GM.

This communication presents a simple method for isolation of oxygen evolving photosystem 2 (PS 2) core complex by solubilisation of PS 2 membranes with the nonionic detergent octyl-β-D-thioglucopyranoside (OTG). This complex was free of cytochrome (Cyt) f contamination and also lacked the 22 and 10 kDa proteins that may not be directly required for primary photochemistry of the PS 2 complex and water oxidation. OTG could also remove the Cyt f contamination from the PS 2-core complex isolated using octyl-β-D-glucopyranoside (OGP). The Cyt f contamination in the PS 2 complexes could potentially interfere with spectrophotometric determinations of redox states of Cyt _b559 and its stoichiometry in the PS 2 complex.

The purpose of this work was to evaluate the effect of 2,4-dichlorophenol (2,4-DCP) and pentachlorophenol (PCP) on the activity of antioxidative system and lipid peroxidation in the leaves of reed canary grass (Phalaris arudinacea). The activity of catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), glutathione reductase (GR) and glutathione S-transferase (GST) as well as the content of glutathione, ascorbate and phenolic compounds were determined. An induced-increase in the APX, CAT, GPX and GR activities was stronger for PCP, while a significant increase in the GST activity was noted only for 2,4-DCP. Both compounds increased the content of phenolic compounds, oxidized and reduced glutathione as well as the content of ascorbic acid. PCP induced stronger increase in lipid peroxidation than 2,4-DCP. The observed changes revealed that chlorophenols induce oxidative stress and oxidative damage in the leaves of reed canary grass.

Leucoanthocyanidin reductase (LAR) converts leucoanthocyanidin to (+)-catechin, a precursor of proanthocyanidins abundant in Japanese persimmon (Diospyros kaki Thunb.) fruits. A putative LAR gene (DkLAR) was isolated by rapid amplification of cDNA ends from young fruits. The full-length cDNA of DkLAR gene was 1 356 bp long and encoded an open reading frame of 349 residues. The deduced DkLAR protein was closely related to the homolog in other plant species. The expression of the DkLAR gene in Chinese pollination-constant non-astringent (PCNA) genotype was coincident with the tannin cell development, but was not in Japanese PCNA and Chinese pollination-variant astringent (PCA) genotypes.

The role of acetylsalicylic acid (0, 1 and 10 μM) pre-treatment in amelioration of salt and osmotic stress in a wild species of potato (Solanum stoloniferum) was investigated. We compared the effects of iso-osmotic concentrations of polyethylene glycol 6000 (15 %) and NaCl (80 mM) on the physiological responses of this species in explants grown in the liquid Murashige and Skoog medium. Both salt and drought reduced shoot growth parameters, photosynthetic pigment contents and increased lipid peroxidation, electrolyte leakage, H₂O₂ content and lipoxygenase activity. The effect of NaCl was more severe than that of polyethylene glycol. Salinity also increased Na⁺ content and decreased K⁺ content and K⁺/Na⁺ ratio. Under both stresses, the activities of superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase, catalase and glutathione reductase enzymes were increased. Acetylsalicylic acid pre-treatment alleviated the adverse effects of both stresses on all parameters measured.