In this report we describe the most suitable protocol for callus formation and plant regeneration for cotton. We screened 15 cotton (Gossypium hirsutum L.) genotypes for metal resistance and two of them, Nazilli M-503 (M503) Nazilli 143 (N-143) selected as Cd, Cu and Ni resistant. The cotyledonary nodes from these genotypes were the best explants for regeneration of shoots (more than 90 %) and roots (50 to 70 %). Shoot apex also gave good shoot regeneration (more than 90 %) but their root regeneration efficiency was low (35 %). These results show that Murashige and Skoog (MS) media containing 0.44 μM naphthaleneacetic acid (NAA) and 0.98 μM indole-3-butyric acid (IBA) was the most suitable recipe for getting high shoot and root regeneration from cotyledonary nodes of N-143 and M503 cotton genotypes.

Solanum nigrum is a model system especially for newcomer to the subject of plant tissue culture. Shoot culture has been easily established from shoot cutting of germinated seeds on Gamborg (B5), or Murashige and Skoog (MS) medium without phytohormones. Direct regeneration was possible using basal media B5, B5C (B5 supplemented with 5 % coconut endosperm milk), Schenk and Hildebrandt (SH), and MS, leaf, stem, shoot tip as explants, cytokinins benzylaminopurine (BAP) or kinetin (KIN) at concentrations from 0.25 to 2 mg dm^-3, and different light treatments (dark, dim and normal light). The best culture condition for shoot formation was the culture of stem internode segments on B5 medium supplemented with 0.5 mg dm^-3 BAP at 16-h photoperiod (irradiance of 100 µmol m^-2 s^-1). Also, root formation was possible under different culture conditions. The best culture condition was the culture of microshoot segments on half strength MS medium supplemented with 1 mg dm^-3 isobutyric acid. Induction of callus formation from young and mature tissues on MS medium supplemented with 0.5 mg dm^-3 BAP, 0.1 mg dm^-3 2,4-dichlorophenoxyacetic acid and 1 mg dm^-3 naphthalene acetic acid, and subsequent plant regeneration on B5C medium supplemented with 0.5 mg dm^-3 BAP was easy. Regeneration of protoplasts isolated from shoot tips and fully expanded leaves was also simple. Finally, the transfer of rooted plantlets to the soil was successful.

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.

A highly reproducible system was developed for efficient rooting of cultivars Boa Casta (BC) and Peneda and a BC seedling-derived clone (BC VII) of almond (Prunus dulcis Mill.). Twenty-four accessions derived from the clone BC VII and subjected to various in vitro culture treatments were screened. The long induction pre-treatment (LIP, 5 d), the brief induction pre-treatment (BIP, 16 h) and the hormonal shock by short dipping in hormone solution (1 min), were tested. BIP was the only that allowed rooting of cultivars. In BC VII, it induced high rooting frequencies (47-100 %) when using a solution of 0.4 mM indole-3-butyric acid solidified with 2 g dm^-3 gellam gum for 16-h. The response to the auxin type was variable depending on the cultivar and the root induction pre-treatment used. Root number was significantly different between the two cultivars and BC VII. Root length was significantly higher when using 0.005 mM IBA in LIP but this concentration induced apical necrosis. The improved acclimatization procedure for up to 4 weeks increased the survival to 45 %. The initiation and development of adventitious roots were proved to be asynchronous.

Barley seedlings were pre-treated with 1 and 5 µM H₂O₂ for 2 d and then supplied with water or 150 mM NaCl for 4 and 7 d. Exogenous H₂O₂ alone had no effect on the proline, malondialdehyde (MDA) and H₂O₂ contents, decreased catalase (CAT) activity and had no effect on peroxidase (POX) activity. Three new superoxide dismutase (SOD) isoenzymes appeared in the leaves as a result of 1 µM H₂O₂ treatment. NaCl enhanced CAT and POX activity. SOD activity and isoenzyme patterns were changed due to H₂O₂ pre-treatment, NaCl stress and leaf ageing. In pre-treated seedlings the rate of ¹⁴CO₂ fixation was higher and MDA, H₂O₂ and proline contents were lower in comparison to the seedlings subjected directly to NaCl stress. Cl^- content in the leaves 4 and 7 d after NaCl supply increased considerably, but less in pre-treated plants. It was suggested that H₂O₂ metabolism is involved as a signal in the processes of barley salt tolerance.

The involvement of plant hormones in the very early response of plants to virus infection was studied in potato plants (Solanum tuberosum L.) infected with potato virus Y^NTN (PVY^NTN). Endogenous plant hormones and compounds mediating a stress response (JA-jasmonic acid, OPDA-12-oxo phytodienoic acid, SA-salicylic acid, IAA-indole-3-acetic acid, ABA-abscisic acid) were simultaneously quantified in susceptible cv. Désirée and resistant cv. Santé, one and three hours after inoculation. Of the hormones analysed, only the contents of endogenous JA and its precursor OPDA changed in a way that could be clearly connected with the early resistant response. In comparison to susceptible cultivar, a much more pronounced increase of JA was detected in virus-inoculated leaves of resistant cultivar at both time points. The same trend of changes was also observed with OPDA. However, there were no significant changes of JA and its precursor in upper intact systemic leaves and roots, at either time point. These findings implicate the jasmonate signalling pathway in a very early local but not systemic resistant defence of potato to PVY^NTN.

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.

The import of ATP into plastids is facilitated by members of the plastidic ATP/ADP transporter (AATP) family. Our results indicate that the cassava (Manihot esculenta Crantz) genome possesses two genes encoding for putative ATP/ADP translocases, which we have designated as MeAATP1 and MeAATP2. Their deduced products are 92 % identical, and phylogenetic reconstructions of plant AATP sequences suggest that MeAATP1 and MeAATP2 are the result of a relatively recent duplication event. Both genes were found to be expressed in a wide range of plant organs via RT-PCR, including young and mature leaves, fibrous and tuberous roots, and green stems. Neither MeAATP1 nor MeAATP2 showed evidence of increased transcription in the presence of exogenous sucrose. Interestingly, the transcriptional activity of MeAATP1 in leaves appeared to be upregulated after wounding, potentially indicating its involvement in the wound response mechanism of cassava.

We isolated and characterized a stress-inducible gene, designated as Slti114, encoding matrix metalloproteinase (MMP) in soybean. The derived amino acid sequences of Slti114 show the 69 % homology with MMP2 from Glycine max (AAL27029). The size of the full-length cDNA of Slti114 is 1194 bp with open reading frame comprised of 394 amino acids. RNA expression of Slti114 was induced by low temperature or wounding. During early stage, Slti114 RNA level was extremely high, but Slti114 RNA was not detectable just after cotyledons became yellowish. Green fluorescent protein fusion expression system confirmed that Slti114-smGFP and H⁺-ATPase-RFP were co-localized to the plasma membrane. Purified glutathione-S-transferase (GST)-Slti114 protein was shown to digest myelin basic protein (MBP) in vitro, but not gelatin. This report provides strong evidence that plasma membrane MMP, Slti114 protein may play a critical role during abiotic stress and senescence in plant.

Rapid shoot multiplication of Nyctanthes arbor-tristis L. was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0-1.5 mg dm^-3 6-benzylaminopurine (BA), 50 mg dm^-3 adenine sulfate (Ads) and 3 % (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA + Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on the MS medium supplemented with 1.5 mg dm^-3 BA, 50 mg dm^-3 Ads and 0.1 mg dm^-3 IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 d on half-strength MS medium supplemented with either indole-3-butyric acid (IBA), IAA or 1-naphthaleneacetic acid (NAA) with 2 % sucrose. Maximum percentage of rooting was obtained on medium having 0.25 mg dm^-3 IBA and 0.1 mg dm^-3 IAA. About 70 % of the rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the field.

A rapid and efficient plant regeneration protocol for a wide range of alfalfa genotypes was developed via direct organogenesis. Through a successive excision of the newly developed apical and axillary shoots, a lot of adventitious buds were directly induced from the cotyledonary nodes when hypocotyl of explants were vertically inserted into modified Murashige and Skoog (MS) medium supplemented with 0.025 mg dm^-3 thidiazuron (TDZ) and 3 mg dm^-3 AgNO₃. When the lower part of shoots excised from explants were immersed into the liquid medium with 1.0 mg dm^-3 α-naphthaleneacetic acid (NAA) for 2 min, and then transferred to hormone free half-strength MS medium, over 83.3 % of the shoots developed roots, and all plantlets could acclimatize and establish in soil. The protocol has been successfully applied to eight genotypes, with regeneration frequencies ranging from 63.8 to 82.5 %.

Mature and immature cotyledon explants of cowpea were cultured on Murashige and Skoog's (MS) medium supplemented with 0.1 - 2.0 mg dm^-3 benzyladenine (BA). Shoot organogenesis was observed from the minimal greenish calli formed at proximal cutting edges of the immature cotyledon explants after 15 - 20 d of culture. Among whole immature zygotic embryo and seven explant types we tested, single whole cotyledon was suitable for shoot organogenesis. Nearly 67.5 % of the explant types produced adventitious shoots on MS medium containing with 1 mg dm^-3 BA, and the shoot number (10.1) per explant was higher than other explant types. From the histological studies, the shoot primordia originated from the procambial strands of immature cotyledon explants. When the shoots were transferred to 1/2 MS basal medium, formed roots within 20 d of culture. The rooted plants were subsequently transferred to the pots and to the field.

A rapid micropropagation protocol through induced multiple shoots from the cotyledonary explant of mulberry (Morus alba L) is described. The highest number of shoots (20.3) was obtained when explants from 14-d-old embryos were cultured on Murashige and Skoog (MS) medium supplemented with 7 μM thidiazuron for 45 d. Of the three cultivars used, cv. S-36 was the best followed by cv. K-2 and S-1. The shoots were transferred to MS medium supplemented with 5 μM 6-benzylaminopurine for elongation. The elongated shoots were rooted on half strength MS medium containing 1 - 7 μM indole 3-butyric acid or 1-naphthalene acetic acid. The rooted plants were transplanted to soil with 90 % success. The emerged shoot primordia probably initiated from the pre-existing meristems since the shoot bud show definite vascular connection to the major vascular tissue.

Agrobacterium-mediated genetic transformation is the most widely used technology to obtain overexpression of recombinant proteins in plants. Molecular events that occur within Agrobacterium during interactions with host plants have been studied extensively, and now we have a reasonable understanding the key factors involved in the regulation of T-DNA nuclear import and genomic integration. By contrast, very little is known about the events that take place in the host cells during genetic transformation by Agrobacterium. Here, we describe the plant-related factors including genotype, genes, proteins, competency of target tissues and phenolic compounds that participate in Agrobacterium-mediated genetic transformation and discuss their possible roles in this process. Because Agrobacterium probably adapts existing cellular processes for its life cycle, identifying the processes in host cells during Agrobacterium infection might contribute to better understanding of basic biological processes as cell communication, intracellular transport and DNA repair and recombination as well as to expanding the host range of Agrobacterium as a genetic engineering tool.

The effects of exogenous abscisic acid (ABA) on lead tolerance in rice (Oryza sativa L.) seedlings were investigated. Pre-treatment with 0.1 g m³ ABA for 2 d restricted amount of Pb translocated from roots to shoots, decreased malondialdehyde and H₂O₂ contents in leaves, and alleviated Pb-induced decrease in plant growth and leaf chlorophyll content. Further, ABA pre-treatment adjusted leaf antioxidative enzyme activities (increased ascorbate peroxidase and catalase activities while decreased superoxide dismutase activity) and so alleviated oxidative stress.

Plant β-glucosidases catalyze the hydrolysis of glycosidic linkages and play a vital role in defense against pathogens and stress. The present work investigated the relationship between leaf development and β-glucosidase protein content in Olea europea L. (cv. Picual) leaves. The total chlorophyll content increased with leaf age in current-season leaves. Immunoblot analysis revealed that the content of 61 kD protein of β-glucosidase also increased with leaf age, and that the enzyme existed in three isoforms (pI 5.8-6.2). Statistical analysis indicated a strong correlation between chlorophyll and β-glucosidase protein contents.

In this communication, we report the binding of abscisic acid responsive elements (ABREs) of rice Osem, namely motif A and motif B, with a cognate trans-acting factor present in the nuclear extract of tobacco leaf. The binding is specific as both the complexes were disrupted with an excess of homologous non-radioactive DNA like motif A or motif B themselves or with cis-elements of rice Rab16A, motif I (ABRE) and motif IIa (non-ACGT ABRE-like sequences). Four tandem repeats of ABRE from wheat Em (4X ABRE) or two tandem repeats of Em ABRE, plus two copies of coupling element (CE1) from barley HVA22 (2X ABRC), also showed specific complexes, that were competed out by an excess of homologous competitors like motif I, motif IIa, motif A, motif B, 4X ABRE and 2X ABRC, but not by the unrelated 4X DRE sequence. Elution of the protein from all the complexes showed a single 26 kDa polypeptide band. Introgression of two of the above synthetic promoters 4X ABRE and 2X ABRC, each fused with minimal promoter of cauliflower mosaic virus 35S (CaMV 35S), could induce the expression of the reporter gene β-glucuronidase (gus) in transgenic tobacco in response to high NaCl concentration, dehydration or abscisic acid, but not at the constitutive level, proving that they can be used as efficient stress-inducible promoters. Our work shows both in vivo and in vitro activity of the promoters from monocot genes in the model dicot plant tobacco.

The toxic aromatic compound 3-hydroxy-4-pyridone (HP) is an intermediate in both synthesis and degradation of mimosine, which is produced by the tree legume Leucaena leucocephala. The L. leucocephala root-nodule symbiont Rhizobium TAL1145 contains a dioxygenase (pydA) and a hydrolase (pydB) gene that produce enzymes necessary for the degradation of HP. In order to coordinately express both genes in plant tissues under a single promoter, three different pydA-pydB fusion constructs (G0, G3, and G7) with varying glycine linkers between the two genes were developed. Prior to transferring the fusion constructs into L. leucocephala, which is highly recalcitrant to genetic transformation, we tested the expression and activity of the hybrid proteins in Nicotiana tabacum, a model plant system that can be easily transformed and analyzed. Seven independent transgenic tobacco lines were generated by Agrobacterium-mediated transformation, and stable integration and expression of pydA-pydB in these transgenic lines were confirmed by polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR) and Western analysis. Only one of the fusion constructs, G3, containing a 9-nucleotide linker between pydA and pydB, provided significant levels of resistance to 3 mM HP, indicating that the hybrid protein produced by this fusion construct could degrade HP.

The leaf water potential, gas exchange and chlorophyll fluorescence were evaluated in five common bean (Phaseolus vulgaris) genotypes A222, A320, BAT477, Carioca and Ouro Negro subjected to moderate water deficit. At the maximum water deficit (10 d of water withholding), the leaf water potential of genotypes A320 and A222 was higher (-0.35 and -0.50 MPa) when compared to the other genotypes (-0.67 to -0.77 MPa). The stomatal conductance and net photosynthetic rate were significantly reduced in all genotypes due to the water deficit. The greater reduction in stomatal conductance of A320 under drought resulted in high intrinsic water use efficiency. Mild water deficit affected the photochemical apparatus in bean genotypes probably by down-regulation since plants did not show photoinhibition. The photochemical apparatus of A222 and A320 genotypes was more sensitive to drought stress, showing reduced apparent electron transport even after the recovery of plant water status. On the other hand, even after 10 d of water withholding, the maximum efficiency of photosystem 2 was not affected, what suggest efficiency of the photoprotection mechanisms.

DNA variations of forty-eight Eucalyptus globulus plants, regenerated by successive culture from seven different explants were assessed by AFLP analysis using 18 primer combinations. At least one variation showed 66.7 % of the analyzed plants, and the numbers of polymorphic bands per plant ranged from 1 to 22. The proportion of polymorphic fragments did not correlate with the numbers of the regenerated plants. However, the more times of successive culture were done the more of polymorphic bands were found within the groups. On average, between 97.39 and 99.88 % of all fragments were shared within the same group. AMOVA analysis showed 39.33 % of the variation was found among the accessions that originated from different calli while 60.67 % was from same calli.

Indian mustard (Brassica juncea L. cv. Vitasso) plants exposed to 10, 30, 50 and 100 µM of Cd for 5 d in hydroponic culture were analysed with reference to the distribution of Cd²⁺, the accumulation of biomass and antioxidants and antioxidative enzymes in leaves. Cd induced a decrease in plant biomass. The maximum accumulation of Cd occurred in roots followed by stems and leaves. Cd induced a decrease in catalase (CAT) and guiacol peroxidase (GPX) activities but an increase in ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR) activities. Enhancement in dehydroascorbate reductase (DHAR) activity was also at 10 µM Cd. Glutathione reductase (GR) activity showed pronounced stimulation after all treatments, but glutathione S-transferase (GST) and glutathione peroxidase (GPOX) activities decreased. The effectiveness of ascorbate-glutathione cycle (AGC) was determined by the ratio of ascorbate to H₂O₂. This ratio decreased in the Cd-treated leaves which indicated that the cycle was disordered.

An efficient, highly reproducible system for plant regeneration via somatic embryogenesis was developed for Cenchrus ciliaris genotypes IG-3108 and IG-74. Explants such as seeds, shoot tip segments and immature inflorescences were cultured on Murashige and Skoog (MS) medium supplemented with 2.0-5.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg dm^-3 N⁶-benzyladenine (BA) for induction of callus. Callus could be successfully induced from all the three explants of both the genotypes. But the high frequency of embryogenic callus could be induced only from immature inflorescence explants. Somatic embryos were formed from nodular, hard and compact embryogenic calli when 2,4-D concentration was gradually reduced and BA concentration increased. Histological studies of somatic embryos indicated the presence of shoot apical meristem with leaf primordia. Ultrastructural details of globular and scutellar somatic embryos further validated successful induction and progression of somatic embryogenesis. Shoots were differentiated upon germination of somatic embryos on MS medium containing 2,4-D (0.25 mg dm^-3) and BA or kinetin (1-5 mg dm^-3). Roots were induced on 1/2 MS medium containing charcoal (0.8 %), and the regenerated plants transferred to pots and established in the soil showed normal growth and fertility.

Plant regeneration through indirect somatic embryogenesis was attempted from leaf, internode, node and shoot-tip derived callus of Leptadenia reticulata. Somatic embryos at the highest frequency was induced on Murashige and Skoog (MS) medium supplemented with 8.87 μM benzyladenine (BA) and 2.46 μM indole-3-butyric acid (IBA). From different explants, only shoot-tip and node explant derived calli induced somatic embryos. Transfer of the embryogenic callus to suspension cultures of the same concentration of growth regulators facilitated the development of embryos. Suspension cultures with reduced concentration of BA (2.22 μM) either alone or in combination with 0.49 μM IBA fostered maturation of embryos. Half-strength MS solid medium with 1.44 μM GA₃ and BA (0.22 or 0.44 μM) facilitated conversion of embryos into plantlets at higher rate compared to that on with BA alone. About 77 plantlets were recovered from 10 mg callus. Plantlets transferred to small cups and subsequently to field survived in 80 %. All the plantlets established in the field exhibited morphological characters similar to that of the mother plant.

To determine the effects of phosphorus nutrition on chilling tolerance of photosynthetic apparatus, tomato (Lycopersicon esculentum Mill. cv. Kenfengxin 2002) plants were raised under different P contents and subjected to 7 d of chilling at 9/7 °C. After chilling (2 h or 7 d) plant growth, P content in tissue, gas exchange and chlorophyll fluorescence were measured. Decreasing P concentration [P] in the nutrient solution markedly reduced plant growth and the chilled plants exhibiting higher optimum [P] than the unchilled plants. Decreasing [P] significantly decreased light saturated net photosynthetic rate (P_Nsat), maximum carboxylation velocity of Rubisco (V_cmax), maximum potential rate of electron transport contributed to Rubisco regeneration (J_max), quantum efficiency of photosystem (PS) 2 (ΠPS2) and O₂ sensitivity of P_Nsat (PS_O2) and this trend was especially apparent in chilled plants.

Multiple shoots of Spilanthes acmella Murr. were induced from nodal buds of in vivo and in vitro seedlings on Murashige and Skoog (MS) medium containing 1.0 mg dm^-3 6-benzyladenine (BA) and 0.1 mg dm^-3 α-naphthalene-acetic acid (NAA). Adventitious shoots were successfully regenerated from the leaf explants derived from the above mentioned multiple shoots. The efficiency of shoot regeneration was tested in the MS medium containing BA, kinetin, or 2-isopentenyl adenine in combination with NAA, indole-3-acetic acid (IAA), or indole-3-butyric acid (IBA) and gibberellic acid. Maximum number of shoots per explant (20 ± 0.47) was recorded with 3.0 mg dm^-3 BA and 1.0 mg dm^-3 IAA. An anatomical study confirmed shoot regeneration via direct organogenesis. About 95 % of the in vitro shoots developed roots after transfer to half strength MS medium containing 1.0 mg dm^-3 IBA. 95 % of the plantlets were successfully acclimatized and established in soil. The transplanted plantlets showed normal flowering without any morphological variation.

Maize (Zea mays L.) is generally considered to be a plant with aerenchyma formation inducible by environmental conditions. In our study, young maize plants, cultivated in various ways in order to minimise the stressing effect of hypoxia, flooding, mechanical impedance or nutrient starvation, were examined for the presence of aerenchyma in their primary roots. The area of aerenchyma in the root cortex was correlated with the root length. Although 12 different maize accessions were used, no plants without aerenchyma were acquired until an ethylene synthesis inhibitor was employed. Using an ACC-synthase inhibitor, it was confirmed that the aerenchyma formation is ethylene-regulated and dependent on irradiance. The presence of TUNEL-positive nuclei and ultrastructural changes in cortical cells suggest a connection between ethylene-dependent aerenchyma formation and programmed cell death. Position of cells with TUNEL-positive nuclei in relation to aerenchyma-channels was described.

This report describes an efficient plant regeneration system for the medicinal plant Lythrum salicaria via direct adventitious shoot development from leaf and stem explants. Leaf explants were much more responsive to regeneration than stem segments. Of the hormonal combinations tested, those involving thidiazuron (TDZ; 0.1, 0.3 or 0.5 mg dm^-3) were more effective than the combinations of other hormones and 0.1 mg dm-3 TDZ combined with either indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was the most productive. Rooting was readily achieved when multiple shoots were singled out and cultured on medium containing different auxins. IAA was the most effective on root development in terms of both the number of roots per shoot and the frequency of rooted shoots. More than 90 % of the regenerants survived after hardening for four weeks at gradually decreased air humidity.

This paper describes a proteome analysis and changes in endogenous abscisic acid (ABA) contents during seed development of Araucaria angustifolia (Bert.) O. Ktze. Megagametophytes and embryonic axis tissues exhibited a similar ABA variation pattern during seed development, reaching maximum values at the pre-cotyledonary stage. The embryonic axis protein content increased until the cotyledonary stage with following stabilization at mature seed. The two-dimensional electrophoresis at the torpedo developmental stage showed approximately 230 polypeptides against 340 in the mature stage. Peptide mass fingerprinting analyses identified three polypeptides, corresponding to an AtSAC4, a late embryogenesis abundant (LEA) and a storage protein, respectively.