The amount of free glucose and sucrose within the leaves of Stevia rebaudiana has been evaluated under different photoperiods and related to steviol glycoside (SVgly) accumulation. The diurnal dynamics of glucose and sucrose in leaves, internodes, apices, and roots revealed considerable fluctuations. Within leaves, glucose and sucrose content decreased up to three-fold during the night whereas SVgly amounts did not show any significant diurnal fluctuations. Ontogenetic variations in glucose content was markedly different under long-day (LD) and short-day (SD) regimes. Under LD, glucose content increased two-fold after the onset of flower bud formation, whereas under SD, a stagnation or small decrease was observed. This increase under LD was already noticed in the upper sink leaves at the end of the vegetative stage, possibly acting as a metabolic trigger for the phase transition to reproductive development. Under SD, glucose content appeared more controlled resulting in a better linear relationship with dry matter and SVgly content. The increased sink demands of the transitional apex led to a rapid decline in leaf sucrose which showed little or no correlation with SVgly amounts. From our results, it becomes clear that the correlation between glucose or sucrose as substrates and SVglys as products is very dynamic and is significantly influenced by day-length and ontogeny.

When exposed to an acute ozone stress, cell death occurred in leaves of the O₃ sensitive Populus deltoides × maximowiczii clone Eridano. After treatment (5 h fumigation and 24 h recovery), the damaged areas covered more than 50 % of the leaf surface. At cellular level, these lesions were preceded by some apoptotic hallmarks and by biochemical and physiological alterations evoked by the apoplastic O₃ dissociation. The cell death pattern was highly localized and involved an increase of membrane permeability, externalization of phosphatidylserine, DNA fragmentation, callose accumulation, polyphenol production, and a biphasic oxidative burst accompanied by NO overproduction. These results indicate a process of programmed cell death that could have the biological significance of limiting the spreading the oxidative burst triggered by ozone dissociation in apoplastic environment. Moreover, materials derived from cell dismantling could be remobilized toward developing structures which can conclude their ontogenetic program after the stress.

Lingonberry (Vaccinium vitis-idaea L. ssp. vitis-idaea Britton) cultivars Regal, Splendor, and Erntedank were obtained by conventional softwood cuttings (taken as a control), by in vitro shoot proliferation of node explants, and by adventitious shoot regeneration from excised leaves of micropropagated shoots. In the plants propagated in vitro, the total ascorbate content increased and its pool was more oxidized, the total glutathione content also increased but its pool became more reduced. The leaves of plants obtained from the in vitro culture showed significantly higher antioxidant enzyme activities except for dehydroascorbate reductase which was at a similar level in all plants. Total soluble phenolics, tannins, and flavonoids were enhanced in fruits of in vitro-propagated plants whereas in leaves, the levels of these metabolites (except flavonoids) were higher in ex vitro derived plants. The total radical scavenging capacity was enhanced in berries of the in vitro propagated plants. It is suggested that the active morphogenetic process, characterized by intensive formation and scavenging reactive oxygen species is reflected in the activities of antioxidant enzymes and metabolites. The reduction potential of glutathione is the most important parameter which determines patterns of growth and differentiation in the investigated plants.

Homogeneous low phosphorus availability was reported to regulate root architecture in Arabidopsis via auxin, but the roles of auxin in root architecture plasticity to heterogeneous P availability remain unclear. In this study, we employed auxin biosynthesis-, transport- and signalling-related mutants. Firstly, we found that in contrast to low P (LP) content in the whole medium, primary root (PR) growth of Arabidopsis was partially rescued in the medium divided into two parts: upper with LP and lower with high P (HP) content or in the reverse arrangement. The down part LP was more effective to arrest PR growth as well as to decrease density of lateral roots (DLR) than the upper LP, and effects were dependent on polar auxin transport. Secondly, we verified that auxin receptor TIR1 was involved in the responses of PR growth and lateral root (LR) development to P supply and loss of function of TIR1 inhibited LR development. Thirdly, effects of heterogeneous P on LRD in the upper part of PR was dependent on PIN2 and PIN4, and in the down part on PIN3 and PIN4, whereas density of total LRs was dependent on auxin transporters PIN2 and PIN7. Finally, heterogeneous P availability altered the accumulation of auxin in PR tip and the expression of auxin biosynthesisrelated genes TAA1, YUC1, YUC2, and YUC4. Taken together, we provided evidences for the involvement of auxin in root architecture plasticity in response to heterogeneous phosphorus availability in Arabidopsis.

In order to set up large-scale acclimatization protocols of micropropagated plants, an in-depth knowledge of their physiological responses during in vitro to ex vitro transfer is required. This work describes the photosynthetic performance of Ulmus minor micropropagated plants during acclimatization at high irradiance (HI; 200 ± 20 μmol m^-2 s^-1 or low irradiance (LI; 100 ± 20 μmol m^-2 s^-1). During this experiment, leaf pigment content, chlorophyll a fluorescence, gas exchange, stomata morphology, the activity of the Calvin cycle enzymes and saccharides were measured in persistent and new leaves. The results indicated that HI induces a higher photosynthetic performance compared to LI. Therefore, plants acclimatized under HI are likely to survive better after field transfer.

Changes in membrane lipid composition is a fundamental strategy for plants to resist low-temperature stress. We compared members of 11 membrane glycerolipid classes in Thellungiella salsuginea and its close relative Arabidopsis thaliana at normal growth temperature, and during cold acclimation (CA), freezing (FR), and post-freezing recovery (PFR). The results showed several properties of T. salsuginea distinct from that in A. thaliana, which included: 1) low relative content of phosphatidic acid (PA) and a rapid increase and decrease of PA during FR and PFR respectively; 2) insensitivity of lyso-phospholipids to freezing; and 3) high ratio of phosphatidylcholine to phosphatidylethanolamine. All these properties were in favour of maintaining membrane integrity and stability and therefore enable T. salsuginea to be more tolerant to freezing than A. thaliana.

In transgenic plants, for many applications it is important that the inserted genes are expressed in a tissue-specific manner. This in turn could help better understanding their roles in plant development. Germin-like proteins (GLPs) play diverse roles in plant development and defense responses. In order to understand the functions and regulation of the GLP13 gene, its promoter (762 bp) was cloned and fused with a β-glucuronidase (GUS) reporter gene for transient expression in Arabidopsis thaliana and tobacco (Nicotiana tabacum cv. K326). Histochemical analysis of the transgenic plants showed that GUS was specifically expressed in vascular bundles predominantly in phloem tissue of all organs in Arabidopsis. Further analyses in transgenic tobacco also identified similar GUS expression in the vascular bundles.

The alleviative effects of exogenous calcium on copper phytotoxicity were investigated in Elodea canadensis plants. There was a significant accumulation of Cu in the plants after their exposure to 0.01 mM Cu accompanied by many symptoms of toxicity. Increased uptake of Cu severely reduced content of photosynthetic pigments, soluble proteins, and free proline. The total antioxidant capacity (T-AOC), reduced glutathione (GSH), and non-protein thiol (NP-SH) were severely suppressed in Cu-stressed plants resulting in a rapid increase in content of superoxide anion (O₂ ^.-), hydrogen peroxide, lipid peroxidation, and cell death. Simultaneous application of Ca markedly increased the content of photosynthetic pigments, soluble proteins, free proline, T-AOC, GSH, and NP-SH, and reduced oxidative damage as indicated by lowered content of MDA, O₂ ^.-, and H₂O₂; and decreased cell death. Furthermore, application of Ca reduced Cu uptake and effectively reversed the Cu-induced nutrient imbalance.

Zinc is the most widespread deficient micronutrient in the tea growing soils of India which affects growth of the plants. In order to investigate the structural, physiological, and biochemical changes under Zn stress (i.e. both deficient and excess supply) of tea [Camellia sinensis (L.) O. Kuntze cv. T-78] plants, we treated young plants with ZnSO₄ at 0 (deficiency), 0.3, 3 (optimum), and 30 μM (toxic) concentrations for 8 weeks. Zn deficiency and excess resulted in considerable decrease in shoot and root fresh and dry masses, and transmission electron microscopy (TEM) revealed disorganization of some cellular organelles. Further, Zn-stress decreased net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), and content of chlorophylls a and b. On the other hand, content of superoxide anion, malondialdehyde, hydrogen peroxide, and phenols, and electrolyte leakage were elevated in stressed plants. The activities of ascorbate peroxidase, catalase, superoxide dismutase, and peroxidase as well as expression of respective genes were up-regulated under Zn-stress. Nevertheless, antioxidant system as a whole did not afford sufficient protection against oxidative damage.

The localization, biotransformation, and chemical speciation of copper in root tips of cucumber (Cucumis sativus) were investigated using synchrotron-based micro X-ray fluorescence (μ-XRF) and micro X-ray absorption near edge structure (μ-XANES). The highest content of Cu was found in root cap and meristematic zone whereas low Cu content in elongation and maturation zone. There was a dramatic increase of Cu content in root cap and meristematic zone after treatment with 100 μM CuSO₄ for 72 h. The μ-XANES analysis revealed that most of Cu in root tip was bound with alginate, citrate, and cysteine-like ligands whereas rarely deposited in form of CuO. From root cap to maturation zone, the proportion of Cu bound with alginate-like ligands increased whereas that bound with citrate-like ligands decreased. The proportion of Cu bound with cysteine-like ligands increased from root cap to elongation zone but sharply declined in maturation zone. The results suggested that Cu was chelated by S ligands in the cell walls which protect protoplasm against possible damage caused by Cu excess.

Plant C2 domain proteins play important roles in diverse cellular processes including growth, development, and membrane targeting, as well as in abiotic and biotic stress adaptations by sensing intracellular Ca²⁺ signals. In this study, we isolated a novel C2 domain protein gene, TaERG3, from wheat infected by Puccinia striiformis f. sp. tritici. TaERG3 was predicted to encode a 144 amino acid protein with molecular mass of 15.68 kD and isoelectric point of 3.93. Analysis of the deduced amino acid sequence of TaERG3 using InterProScan revealed the presence of an N-terminal calciumdependent phospholipid-binding module (C2 domain, 5 to 103). Transient expression analysis showed that the TaERG3 protein was predominately and uniformly localized in the plasmalemma and nucleus of onion epidermal cells. Quantitative real-time PCR analyses indicated that TaERG3 transcript was differentially induced in both incompatible and compatible interactions, as well as by applied abscisic acid (ABA) and CaCl₂. However, the significant transcript changes induced by methyl jasmonate, ethylene, and salicylic acid treatments were not as dramatic as those induced by ABA. TaERG3 was also up-regulated by environmental stimuli including low temperature and high salinity. These results imply that TaERG3 might be involved in wheat defence responses against stripe rust and abiotic stresses in an ABA-dependent signalling pathway.

Cucumber (Cucumis sativus L.) is considered a model plant for the investigation of Fe deficiency responses, since it strongly exhibits typical strategy I, i.e. increased activities of Fe(III)-chelate reductase, H⁺-ATPase and iron regulated transporters. In this study, cDNA amplified fragment length polymorphism analysis was employed to identify genes differentially expressed in the root apex following Fe deficiency. The expression patterns of the most interesting transcript derived fragments were validated by semiquantitative reverse trascriptase - polymerase chain reaction. A set of new genes overexpressed under Fe deficiency, such as those coding for calmodulin, SNAP, TIM23 and V-PPase were identified. Furthermore, we also observed that calmodulin protein accumulated in Fe-deficient root apexes.

Treatment of potato (Solanum tuberosum) plants with cadmium or abscisic acid (ABA) enhanced the content of StPCS 1 transcript and activity of phytochelatin synthase (PCS) in roots.transcript and activity of phytochelatin synthase (PCS) in roots. These treatments enhanced the contents of ABA and expression of genes coding 9-cis-epoxycarotenoid dioxygenase 1 (NCED1) and basic leucine zipper (b-ZIP).expression of genes coding 9-cis-epoxycarotenoid dioxygenase 1 (NCED1) and basic leucine zipper (b-ZIP). Simultaneous treatment of potato plants with Cd and fluridone (Flu), an inhibitor of ABA biosynthesis, completely halted the Cd-induced transcription of StPCS1, NCED1 and StbZIP genes and limited the increases in PCS activity and ABA content. The data suggest that ABA participates in transduction of the Cd signal to the cells of potato roots.

Information on plant responses to combined ozone and cadmium stresses are scarce and limited to herbaceous species. In this research, two poplar clones (I-214 and Eridano), differently sensitive to O₃, were grown for 5 weeks in pots supplied with 0, 53.5, and 160.5 mg(Cd) kg^-1 (soil d.m.) and then exposed to 15-d O₃ fumigation (0.06 mm³ dm^-3, 5 h a day). The effects of the two stressors, alone or in combination, on Cd, Ca, Fe, and Zn accumulation in above-nad below-ground organs, photosynthesis, leaf pigments, and accumulation of H₂O₂ and NO were investigated. Cadmium induced a reduction in stomatal conductance and a significant accumulation of H₂O₂ and NO in both clones nad negatively affected the carotenoid content in I-214. Ozone, on the other hand, counteracted Cd accumulation in the above-ground organs and significantly increased the xanthophyll de-epoxidation state indicating photoinhibition in O₃-treated plants. Surprisingly, O₃ alone or in combination with Cd decreased H₂O₂ accumulation in I-214. The NO production was generally stimulated by Cd, whereas it decreased following O₃ exposure in I-214. The overall data indicate that Cd and O₃ induced clone specific responses. Moreover, when they were applied in combination, antagonistic rather than synergistic effects were observed.

The pollen tube grows rapidly, exclusively at its tip, to deliver its sperm for fertilization. The polarized tip growth of pollen tubes is dependent on the highly dynamic actin cytoskeleton. Plant LIM proteins (named after initials of containing proteins Lin11, Isl-1, and Mec-3) have been shown to regulate actin bundling in different cells, however, their roles in pollen tube growth have remained obscure. Here, we report the function of Arabidopsis LIM proteins PLIM2a and PLIM2b in pollen tube growth. The PLIM2a mutation resulted in short and swollen Arabidopsis pollen tube with defective actin bundles. The expression of the construct green fluorescent protein (GFP)-PLIM2b led to fluorescence of the actin bundles in germinating pollen and also the long actin bundles along the growing pollen tubes in Arabidopsis, but not of the short and sparse actin bundles that characterize the tip regions of the pollen tubes. There is a partially redundant function between PLIM2a and PLIM2b in the shank actin bundle organization during Arabidopsis pollen tube growth, as PLIM2b could rescue for the defective shank actin bundles in PLIM2a mutation pollen tubes. This report suggests critical roles of PLIM2a/PLIM2b in actin configuration during Arabidopsis pollen germination and tube growth.

Sunflower broomrape (Orobanche cumana Wallr.) is considered a self-fertilizing species, but there is no indication as to whether it is strictly self-fertilized or that it presents some extent of cross-fertilization. The objective of this research was to measure the rate of cross-fertilization in O. cumana using an unpigmented recessive mutant as a visual marker. A pot and a field experiment in which single unpigmented plants were surrounded by a large number of pigmented plants were conducted. Occurrence of F₁ hybrids, readily distinguishable from unpigmented plants in the progenies of unpigmented plants provided a direct measurement of the cross-fertilization rate. Progenies of unpigmented plants contained 21.5 % of F₁ hybrids in the pot experiment and 28.8 % in the field experiment. The results revealed that O. cumana is a partially allogamous species, which has great relevance for understanding the genetic structure and dynamics of populations and, ultimately, race evolution in this parasitic plant.

Annexins play a crucial role in plant development and response to environmental stimuli. In this study, a total of 23 annexin genes (GmANN1 - GmANN23) were identified from the soybean genome database and for two of them (GmANN11 and GmANN14), complete cDNAs were cloned. GmANN1 - GmANN23 encoded a set of predicted proteins which showed high similarity to other known annexins. Most GmANN genes contained four putative annexin repeats. Generally, a type II Ca²⁺-binding site is found to exist in the first and fourth repeats. GmANN1, 10, 11, 12, and 14 showed different organ-specific expression patterns. Furthermore, expression of these five GmANNs was significantly induced by drought and abscisic acid. Expression of four annexins (GmANN1, 11, 12, and 14) was induced by cold and expression of three annexins (GmANN1, 11, and 12) responded to high salinity.

The changes in the activities of antioxidant enzymes and amounts of proteins, phenols, and flavonoids in regenerating and non-regenerating calli during organogenesis of Sterculia urens were monitored. Maximum growth of calli and the most efficient regeneration of shoots occurred on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm^-3 benzylaminopurine (BAP) + 2 or 4 mg dm^-3 naphtalene acetic acid (NAA). Peroxidase (POD), catalase, and superoxide dismutase activities increased in the regenerating calli but decreased in the non-regenerating calli. Six POD isoenzymes were detected. Protein content decreased in the non-regenerating calli and increased significantly during regeneration of shoots from callus. Total phenols and flavonoids increased in the non regenerating calli. SDS-PAGE analysis revealed a role of many proteins in organogenesis.

A suppression subtraction hybridization (SSH) cDNA library of alfalfa (Medicago sativa L.) cv. Zhongmu NO.1 had been constructed to identify differentially expressed genes under stress. Based on the sequence of a 460 bp expressed sequence tags (ESTs), a cDNA of 1652 bp was cloned by rapid amplification of cDNA ends (RACE) method. This gene (MsPBL) was predicted to encode a 434-amino-acid protein, which contained a Phox and Bem1 (PB1) domain. PB1 domain is a functional domain comprising about 80 amino acid residues, which exists in many signal transduction proteins and mediates dimerization in the proteins. PB1 domain is mostly involved in two cell signal transduction pathways: MAPK and NF-KB. When fused to the green fluorescent protein, we found MsPBL localization in the nucleus of onion (Allium cepa L.) epidermal cells. The transcripts of MsPBL rose significantly when alfalfa was treated with 300 mM NaCl, 0.1 mM ABA, and 20 % polyethylene glycol (PEG-6000). These results indicated that MsPBL may be functional within the nucleus as a signal transduction protein to allow alfalfa to rapidly respond to the environmental stress signals.

Seedlings of mangrove plant Bruguiera gymnorrhiza cultured in sand with Hoagland's nutrient solution were treated with 1 to 30 mM Cd(NO₃)₂ or Pb(NO₃)₂ for 2 months. In all Cd/Pb treatments, the malondialdehyde content increased while the chlorophyll content declined. Peroxidase (POD) and superoxide dismutase (SOD) activities in roots increased at moderate Cd/Pb concentrations (1-10 mM), whereas decreased at higher concentrations (20-30 mM). Catalase (CAT) activity in roots was inhibited by 1-10 mM Cd but enhanced by 1-10 mM Pb. The activities of POD, SOD and CAT in leaves were less affected by Cd and Pb than in roots. A new SOD and three CAT isoenzymes were induced by Pb. In contrast, no additional SOD and CAT isoenzymes were induced by Cd.

The manipulation of cytokinin contents via Agrobacterium-mediated transformation is an efficient tool for delaying leaf senescence and improving the resistance to environmental stresses. In the present study, cotton transformants harbouring the Agrobacterium tumefaciens isopentenyl transferase (IPT) gene under the control of the promoter of Gossypium hirsutum cysteine proteinase (Ghcysp) were generated. PCR and Southern blot analysis indicated that the foreign DNA fragment was successfully integrated into the cotton genome. The chlorophyll and cytokinin contents, and ROS-scavenging enzymatic activities were significantly increased in transgenic cotton lines, which resulted in a significant delay in leaf senescence. The growth characteristics of transgenic cotton lines resembled the non-transgenic lines except delaying premature senescence and the lint yield and fiber quality of transgenic lines were improved. In addition, the transgenic lines had higher biomasses, IPT transcripts, and endogenous cytokinin contents compared with those of non-transgenic lines under 200 mM NaCl stress.

Changes in antioxidative enzymes activities during Tacitus bellus direct shoot organogenesis from leaf explants were examined. During the early stages of shoot organogenesis there was a decrease in superoxide dismutase (SOD) and an increase in catalase (CAT) activity, and later during organogenesis there was an increase in peroxidase (POD) and polyphenol oxidase (PPO) activity. Two highly regulated turning points may be distinguished regarding activities and isoforms of antioxidative enzymes: the initiation of shoot organogenesis and the shoot bud formation. Our data suggest the role of specific CAT, POD, SOD and PPO isoforms in separate processes during T. bellus direct shoot organogenesis.

In order to provide additional information on the coloration of chrysanthemum flowers, the flavonoid composition and the expression of six structural genes involved in anthocyanin pathway in the ray florets of a pink flowering (cv. H5) and two white flowering (cvs. Keikai and Jinba) Chrysanthemum grandiflorum cultivars were examined. HPLCDAD/ESI-MSⁿ analysis showed that cyanidin 3-O-(6"-O-malonylglucoside) and cyanidin 3-O-(3",6"-O-dimalonylglucoside) were the two major flavonoids presented in H5, while white flowering cultivars contained flavones instead of anthocyanins. Nine flavone derivatives were detected in the three cultivars, the amount of each flavone varied upon cultivars, and seven of these were identified as luteolin 7-O-arabinosylglucuronide, apigenin 7-O-glucoside, luteolin 7-O-malonylglucoside, apigenin 7-O-malonylglucoside, chrysoeriol 7-O-malonylglucoside, acacetin 7-O-rutinoside and acacetin 7-O-malonylglucoside. The two white flowering cultivars showed similar total flavonoid content, which was about two fold higher than that in H5. A high expression of the genes encoding dihydroflavonol 4-reductase and 3-O-glucosyltransferase was detected only in H5 but not in Keikai or Jinba. Chalcone synthase, chalcone isomerase, flavanone 3-hydroxylase, and flavonoid 3'-hydroxylase were expressed in all flowers, suggesting that the lack of anthocyanin in white flowering cultivars cannot be due to any blockage of their expression.

The aim of this study was to test the hypothesis that salinity can affect indirectly the nitrate acquisition by a negative modulation triggered by glutamine accumulation. Cowpea plants were exposed to a mild NaCl concentration (50 mM) in order to restrict growth and N-demand. After 21 d, pretreated plants and control plants were supplied with 0, 5 and 10 mM of Ca(NO₃)₂ for 3 d in absence of NaCl. Salt pretreated plants showed a great limitation in acquisition of NO₃ ^-, indicated by decline in the nitrate uptake rate, NO₃ ^- accumulation, nitrate reductase activity and protein content. The restriction of NO₃ ^- utilization was positively associated with increased glutamine synthetase activity and glutamine accumulation, especially in roots.

Histone H3 lysine 4 methylations catalyzed by histone lysine methyltransferases (HKMTs), like the Arabidopsis thaliana ATX1 and ATX2, are important epigenetic modifications related to chromatin decondensation and gene activation. In order to study this epigenetic mechanism in monocot cereal plants, we performed homology searches of ATX1 and ATX2 against the Brachypodium distachyon L. Beauv and rice (Oryza sativa L. spp. japonica) genomes, discovering single homologues for each cereal crop representing both Arabidopsis sequences. Using this information, we employed the rolling circle amplification - rapid amplification of cDNA ends (RCA-RACE) method to isolate, clone and characterize HvTX1 from RNA extracted from barley (Hordeum vulgare L.) tissues and studied its expression during seed development and under drought stress. The cloned cDNA sequence contained a 3 093 bp ORF homologous to ATX1 and ATX2. Characterization of the translated HvTX1 transcript sequence revealed the multi-domain nature of the putative protein, including all conserved regions characteristic for ATX1 and ATX2. By comparative genomic analysis and homology searches in EST databases we located, with high probability, the gene coding for HvTX1 on the barley chromosome 5H. Constant elevation of HvTX1 expression was observed during seed development. Expression of HvTX1 after drought stress was analyzed by quantitative real-time polymerase chain reaction (qPCR) in two different barley cultivars with varying drought stress tolerance, revealing HvTX1 drought-induction in a tolerance-specific manner.

Cucumber seedlings were pretreated with 3 μM 5-aminolevulinic acid (ALA) followed by cultivation at normal (25/18 °C) or high (42/38 °C) day/night temperature to investigate the protective effects of ALA on heat stress in plants. Heat elevated the contents of malondiadehyde (MDA), superoxide radical (O₂ ^.-) and hydrogen peroxide (H₂O₂) in leaves of all plants but less in ALA-pretreated plants. Heat treatment resulted in higher antioxidant enzyme activities and proline and soluble sugar contents and weaker growth inhibition in ALA-pretreated plants than in those treated with heat alone. These results indicate that ALA pretreatment increased the tolerance of seedlings to heat stress.

An efficient in vitro regeneration protocol was developed for medicinally important aromatic plant Anethum graveolens. Nodal segments were cultured onto Murashige and Skoog (MS) basal medium supplemented with different auxins and cytokinins singly as well as in combinations. The optimum callus induction (93.33 %) was obtained on medium fortified with 2.2 μM N⁶-benzyladenine (BA) and 0.21 μM α-naphthaleneacetic acid. The best shoot regeneration (85.7 %) with 12.86 shoots per explant was achieved in two weeks when callus was subcultured on MS medium amended with 2.2 μM BA and 1.85 μM kinetin. The average length of regenerated shoots varied from 3.15 to 4.8 cm. The rooting of regenerated shoots was nearly 100 % on 1/4 MS augmented with 4.9 μM indolebutyric acid with a maximum root length of 5.1 cm. Plantlets were successfully acclimatized with 60 % survival rate. During organogenesis, catalase and ascorbate peroxidase activity increased while superoxid dismutase activity decreased. Clonal fidelity of in vitro raised plants has been checked by random amplified polymorphic DNA using 10 selected decamer primers. It has been found that regenerated plants are true to type plants.

Although most plastid transformation studies have focused on chloroplast expression, plastid transformation can also be used to express genes in plastids of a wide variety of plant tissues by using appropriate plastid promoters. Based on the sequence of the Gossypium hirsutum chloroplast genome, we developed primers and amplified segments of 20 different plastid genes. The PCR products were labeled and used in filter dot blot hybridization studies to characterize their expression levels and patterns in total RNA isolated from light- and dark-grown cotton tissues at different developmental stages. A subset of 6 genes among these was further characterized by real time PCR. Highest expression levels were observed for rrn16 and psbA. Four genes were expressed in all samples at relatively constant levels: accD, atpA, matK and rrn16. Expression in root tissue was generally low. The results of our study can be used to predict which operons and promoters are most likely to be preferentially expressed in the plastids of tissues of interest at levels that would result in the desired phenotype, facilitating the development of plastid transformation vectors.

Fruit crops have a growing economic importance worldwide and molecular genetics might be useful in solving many problems that arise during commercial production. One of the fields that have attracted intense attention is the molecular basis of self-incompatibility that may result in low fruit set. In tree fruits of the Rosaceae family, the incompatibility reactions take place between the pistil S-ribonuclease (S-RNase) and the pollen-expressed S-haplotype specific F-box (SFB) proteins. In most cases, the loss of self-incompatibility was associated with mutations in the S-RNase or SFB genes. A total of 27 non-functional S-haplotypes have been identified and characterized, most (24) of which emerged as a consequence of natural mutations. In the Prunoideae, most haplotypes are pollen-part mutants (50 %), while 8 are stylar-part mutants (36 %), one haplotype shows both pollen- and stylar-part mutations, and molecular changes for two haplotypes still have not been clarified. In contrast, non-functional natural haplotypes in the Maloideae are all stylar-part mutants. The analysis of such mutants may shed light on underlying molecular mechanisms as was the case with the establishment of the general inhibitor model that describes interactions between pollen and pistil S-proteins. However, several other molecules were supposed to contribute to the molecular interactions, at least in Solanaceae, a family with a similar self-incompatibility system. This review also endeavours to delineate the evolutionary implications of the S-locus mutations and collect limited data on non-S-locus molecular interactions and signaling events after self- and cross-pollination of fruit tree species.