Changes in ascorbate and glutathione contents and the activities and isoenzyme patterns of enzymes of the ascorbate-glutathione cycle were investigated in embryo axes and cotyledons of germinating lupine (Lupinus luteus L.) seeds. Ascorbate content was not significantly affected over the initial 12 h of imbibition in embryo axes, but afterwards increased, with the most rapid accumulation coinciding with radicle emergence. A somewhat similar trend was observed for glutathione with significant increase in embryo axes shortly before radicle protrusion followed by decline in the next hours. In cotyledons the ascorbate pool rose gradually during germination but the amount of glutathione showed fluctuations during a whole germination period. The activity of ascorbate peroxidase (APX) rose progressively in embryo axes, while activities of dehydroascorbate reductase (DHAR) and glutathione reductase (GR) showed transient increase during germination. New isoforms of APX and GR were synthesized, suggesting that they play a relevant role during germination. All analyzed enzymes were already present in dry seeds which allowed them to be active immediately after imbibition.

Agrobacterium tumefaciens mediated vacuum infiltration transformation in planta has been established in pakchoi, a kind of Chinese cabbage, but the transformation frequency in harvested seeds has varied in the range of 0.5 to 3.0 × 10^-4 over several years and is much lower than the transformation frequency in Arabidopsis thaliana. To understand that, the distribution and vitality changes of A. tumefaciens in plant tissues were examined. Results revealed that there was a majority of A. tumefaciens in the flower compared with that in the stem and in the leaf at all times after infiltration. As fact of transformants in the upper part of the treated plant (T₀) stalk and fact of the survival of A. tumefaciens in the plant were proved, possibilities of optimizing the transformation conditions to increase the transformation frequency in pakchoi was discussed.

In vitro regeneration from leaf, cotyledon and hypocotyl explants of six cultivars belonging to three species of Capsicum was achieved by direct organogenesis. The cultivar Umorok showed the best response while Meiteimorok, Haomorok, Mashingkha and Uchithi showed intermediate response and the cultivar Chiengpi was the least responsive. Leaf and cotyledon explants regenerated more shoots than hypocotyl explants and the maximum number of shoots were produced on Murashige and Skoog (1962) medium containing 8.8 µM 6-benzylaminopurine (BAP) with 11.4 µM indole-3-acetic acid (IAA). Elongation of shoot buds derived from different explants was achieved on medium containing 2.8 µM IAA and the elongated shoots were rooted on medium containing 2.8 or 5.7 µM IAA and 2.4 or 4.9 µM indole-3-butyric acid (IBA). Four-week old rooted plantlets were hardened and transplanted to the soil. The plantlets showed 90 % survival during transplantation.

Effects of various stress treatments such as NaCl, hydrogen peroxide, hydroxyl free radical, and high irradiance (HI, 1 000 µmol m^-2 s^-1) on the photosystem (PS) 2 mediated electron transport rate and the degradation of D1 protein in the thylakoid membranes of barley were studied. The applied stresses caused significant reduction in the PS 2-mediated electron transport and a degradation of D1 protein that was highest during the HI-treatment. Presence of 2,6-dichlorophenol indophenol (DCPIP), which is an artificial electron acceptor from water, significantly minimizes the HI-induced deleterious effect on the PS 2-mediated electron transport rate, disarrangement of PS machinery, and degradation of the D1 protein. HI in the absence of an acceptor resulted in production of reactive oxygen species due to electron transfer to oxygen.

Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides Pall. was here developed. The protoplasts were isolated directly from the leaves of the hairy root-induced plants. The highest yield of protoplasts was obtained from fully expanded leaves of young plants. Their viability was up to 72 ± 2.3 %. The highest division frequency (32.4 ± 0.13 %) and sustained divisions were obtained in Durand, Potrykus and Donn (DPD) medium supplemented with 2.0 mg dm^-3 2,4-dichlorophenoxyacetic acid, 0.2 mg dm^-3 6-benzylaminopurine, 0.3 M mannitol, 2 % sucrose and 500 mg dm^-3 casein hydrolysate at the plating density of 3.0 × 10⁵ cm^-3. The frequency of shoot differentiation from protocalli reached to 91.75 ± 3.1 %. Opine synthesis and polymerase chain reaction analysis confirmed that T-DNA still existed in the protoplast regenerated plants.

Suppressive subtraction hybridization (SSH) was used to identify differentially expressed genes caused by a chlorophyll-reduced mutation in B. napus. The cDNA fragments, derived from SSH positive subtractive library (tester: normal wild type, driver: mutant) were cloned into pMD18-T vector. Two hundred SSH cDNA clones were screened by dot blot array, and 151 clones were identified as differentially expressed cDNA fragments in Cr3529 line. Thirty-six positive clones which showed marked expression differences were selected and sequenced. After redundant cDNAs were removed, 33 differentially expressed unique cDNA section clones were obtained. Among the 33 clones, two clones possess different parts of the cDNA sequence of the same gene coding geranylgeranyl reductase, four clones belong to unknown proteins, and the rest share homology to genes of diverse class. Sequence analysis showed that at least 12 genes were discovered to be related to the photosynthesis, seven of them coded the proteins which belong to the subunit of photosystem 2. RNA gel blot analysis showed that compared with 3529, the gene expression of the chlorophyll a/b-binding protein Lhcb2 in photosystem 2 declined markedly in the cotyledons and seedling leaves of Cr3529, indicating that the reduced light-harvesting complex 2 accumulation in thylakoid membrane of Cr3529 was due to the decrease of the related gene mRNA level for translation.

An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm^-3 BA and 0.2 mg dm^-3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings with age of about 18-27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal MS medium supplemented with 0.5 mg dm^-3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where 100 % survived after 1 month of acclimatization.

In our present study assessment of genetic diversity and identification of pigeonpea cultivars has been done by employing 76 random amplified polymorphic DNA (RAPD) primers. Out of 796 amplified products, 587 showed polymorphism (73.7 %) and an average of 10.47 bands were amplified per primer. Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped all the cultivars into three clusters. The cluster I consists of 7 cultivars, cluster II of 11 cultivars in 4 sub-clusters and cluster III 4 cultivars. Two cultivars were not included in any cluster. The clustering was strongly supported by high bootstrap values. Furthermore, high values of the average heterozygosity (H_av) and marker index (MI) also indicated the efficiency of RAPD as a marker system.

Gametophores of mosses Mnium undulatum and Polytrichum commune were submerged in distilled water or in calcium chloride solution (0.9 mM Ca²⁺) to induce hypoxia. The net photosynthetic (P_N) and dark respiration rate (R_D) were measured in the air containing 300-400 µmol(CO₂).mol^-1(air) and 0.21 mol(O₂).mol^-1(air). P_N of M. undulatum gametophores decreased to 58 % of the control after 1-h submersion in water, whereas to 80 % of the control in P. commune gametophores. A smaller decrease in P_N was observed when the gametophores were immersed in CaCl₂ solution. In hypoxia, R_D in the tested mosses species was a little higher than in the control.

Effects of high night temperature on the lipid and protein compositions in the tonoplasts isolated from the leaves of two Crassulacean acid metabolism (CAM) plants, Ananas comosus (pineapple) and Kalanchoë pinnata were studied. The results showed that the phospholipids/protein ratios in the tonoplasts isolated from pineapple and K. pinnata leaves decreased from 1.82 to 1.21 and 2.63 to 1.50, respectively, as the night temperature increased from 20 to 37 °C. Under high night temperature, relative amount of total unsaturated fatty acids in K. pinnata was increased by 6 %, which was mainly caused by increased C18:2 and C18:3, whereas unsaturated fatty acids, C18:2 and C18:3 in pineapple did not show significant change. The distribution patterns of tonoplast proteins in the two CAM species were different between normal and high night temperature and in K. pinnata, especially those with molecular mass ranging from 66.2 to 97.4 KDa. Compared with normal night temperature, more proteins were found in pineapple, but no difference was found in K. pinnata. Thus, above result indicated that the pineapple tonoplasts could keep higher rigidity under high night temperatures compared to the K. pinnata.

The effects of amino acids (arginine, aspargine, cysteine, glutamine, glycine and proline) and polyamines (putrescine and spermidine) on embryogenesis and plant regeneration from cultured anthers of Guizotia abyssinica (L. f.) Cass. cv. Ootacamund was studied. Supplementation of amino acids (0.5-5.0 mM) to the induction medium individually and in combination, improved embryo yield. B5 medium supplemented with 2 mM proline, 10 µM 2,4-dichlorophenoxyacetic acid, 2 µM kinetin and 0.2 M sucrose induced highest number of embryos (63 per 60 anthers cultured). Addition of polyamines (5-200 µM) to the same medium also enhanced the rate of embryogenesis.

An experiment was taken up to find out possibilities of manipulating the in vitro flowering in moth bean. Abscisic acid (ABA) and proline both alone and in combination influenced days to flower induction, number of flowers per plant, number of pods per plant and seeds per pod. Frequency of flowering plants approached 100 % at 1 and 3 µM ABA and 800 µM proline. The range of flowering period (3 to 23.6 d) has also been influenced by various treatments.

A quantitative trait loci (QTL) approach was applied to dissect the genetic control of the common wheat seedling response to osmotic stress. A set of 114 recombinant inbred lines was subjected to osmotic stress from the onset of germination to the 8^th day of seedling development, induced by the presence of 12 % polyethylene glycol. Root, coleoptile and shoot length, and root/shoot length ratio were compared under stress and control conditions. In all, 35 QTL mapping to ten chromosomes, were identified. Sixteen QTL were detected in controls, 17 under stressed conditions, and two tolerance index QTL were determined. The majority of the QTL were not stress-specific. In regions on five chromosome arms (1AS, 1BL, 2DS, 5BL and 6BL) the QTL identified under stress co-mapped with QTL affecting the same trait in controls, and these were classified as seedling vigour QTL, in addition to those expressed in controls. Tolerance-related QTL were detected on four chromosome arms. A broad region on chromosome 1AL, including five QTL, with a major impact of the gene Glu-A1 (LOD 3.93) and marker locus Xksuh9d (LOD 2.91), positively affected root length under stress and tolerance index for root length, respectively. A major QTL (LOD 3.60), associated with marker locus Xcdo456a (distal part of chromosome arm 2BS) determined a tolerance index for shoot length. Three minor QTL (LOD < 3.0) for root length and root/shoot length ratio under osmotic stress were identified in the distal parts of chromosome arms 6DL (marker locus Xksud27a) and 7DL (marker locus Xksue3b). Selecting for the favourable alleles at marker loci associated with the detected QTL for growth traits may represent an efficient approach to enhance the plants' ability to maintain the growth of roots, coleoptile and shoots in drought-prone soils at the critical early developmental stages.

6-Methoxy-2-benzoxazolinone (MBOA) inhibited germination of rice (Oryza sativa L.), wheat (Triticum aethiopicum Jakubz), rye (Secale cereale L.), onion (Allium cepa L.), wild oat (Avena fatua L.), barnyard grass [Echinochloa crus-galli (L.) Beauv.], ryegrass (Lolium rigidum Gaudin), cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), tomato (Lycopersicum esculentum Mill.), carrot (Daucus carota L.) and amaranth (Amaranthus retroflexus L) and the inhibition increased with increasing MBOA concentrations. MBOA also inhibited the induction of α-amylase in these plant seeds and the inhibition increased with increasing MBOA concentrations. There were variations in sensitivity of these plant species to MBOA, and species of family Poaceae (barnyard grass, wild oat, rice, rye, ryegrass, and wheat) were less sensitive to MBOA than the other plant species.

A bacterial artificial chromosome (BAC) library of banana (Musa acuminata) was used to select BAC clones that carry low amounts of repetitive DNA sequences and could be suitable as probes for fluorescence in situ hybridization (FISH) on mitotic metaphase chromosomes. Out of eighty randomly selected BAC clones, only one clone gave a single-locus signal on chromosomes of M. acuminata cv. Calcutta 4. The clone localized on a chromosome pair that carries a cluster of 5S rRNA genes. The remaining BAC clones gave dispersed FISH signals throughout the genome and/or failed to produce any signal. In order to avoid the excessive hybridization of repetitive DNA sequences, we subcloned nineteen BAC clones and selected their 'low-copy' subclones. Out of them, one subclone gave specific signal in secondary constriction on one chromosome pair; three subclones were localized into centromeric and peri-centromeric regions of all chromosomes. Other subclones were either localized throughout the banana genome or their use did not result in visible FISH signals. The nucleotide sequence analysis revealed that subclones, which localized on different regions of all chromosomes, contained short fragments of various repetitive DNA sequences. The chromosome-specific BAC clone identified in this work increases the number of useful cytogenetic markers for Musa.

An efficient micropropagation protocol was developed for elite male and female genotypes of Simmondsia chinensis using nodal segments. Bud initiation was found to be best on Murashige and Skoog's (MS) medium supplemented with 4.44 µM 6-benzylaminopurine (BAP) and 88.8 µM adenine. Upon sub-culture, 10-15 shoots per explant were obtained when 4.44 µM BAP and 74.0 µM adenine were incorporated in the medium. Increase in KNO₃ concentration in the medium improved shoot multiplication rate and in vitro flowering in 20 % of male cultures. Elongated shoots were harvested, pulse treated for 48 h on liquid medium supplemented with 49.0 µM indole-3-butyric acid, 5.40 µM α-naphthaleneacetic acid and 5.71 µM indole-3-acetic acid for root induction and rooting (92 %) was achieved on hormonal free half-strength MS medium supplemented with 1.37 µM chlorogenic acid, 1 % activated charcoal and 2 % sucrose. After successful hardening, plantlets were transferred to greenhouse with 99 % establishment.

A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in cotton (Gossypium hirsutum L.) cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming frequency (82.86 %) was obtained in protoplast cultures from suspension culture cells in KM₈P medium with 0.45 µM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 µM kinetin (KIN), 1.5 % glucose and 1.5 % maltose. Protocolonies formed in two months with plating efficiency of 14 %. However, the callus-forming efficiencies from other two explants were low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7 % of normal plantlets were obtained on medium contained 3 % maltose or 1 % of each sucrose + maltose + glucose, 2.46 µM indole-3-butyric acid (IBA) and 0.93 µM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were transferred to the soil and the highest survival rate (95 %) was observed in transplanting via a new method.

A rapid and efficient method for the regeneration of Eucommia ulmoides Oliver has been developed. The ability of hypocotyl segments to produce adventitious buds varied depending upon their position, seedling age and culture medium. The most adventitious buds were induced from the hypocotyl segments near to the root of 2-week-old seedlings on the Murashige and Skoog (MS) basal medium supplemented with 10 µM benzylaminopurine (BAP). However, the MS medium with 6 µM BAP and 6 µM gibberellic acid (GA₃) was most suitable for shoot multiplication and further growth. Shoots longer than 2 cm cultured on the half-strength MS medium supplemented with 0.5 µM naphthaleneacetic acid (NAA) produced a maximum number of roots per shoot. Regenerated plantlets could be successfully acclimatized.

It was observed that the unpollinated flowers of Cymbidium pendulum (Roxb.) Sw. and C. aloifolium (L.) Sw. stayed fresh for 20 and 18 d, respectively, but attained senescence in 8 and 7 d, respectively, after pollination. The higher content of total soluble sugars, reducing sugars and free amino acids was observed in all the floral organs of pollinated flowers than in unpollinated ones. Pollination also up-regulated the activity of hydrolytic (α-amylase, β-amylase, invertase) and proteolytic enzymes (proteases) in floral organs. Amongst floral organs, the lip and perianth possessed highest contents of metabolites. Application of auxin inhibitor (0.25 µM triiodobenzoic acid) and ethylene inhibitor (0.25 µM AgNO₃) to the pollinated flowers partially prevented the process of senescence.

Carnation plantlets (Dianthus caryophyllus L.) cultured in vitro often develop morphological and physiological anomalies, a phenomenon called hyperhydricity, which impairs their survival ex vitro. When the agar concentration of the growth medium was increased (from 0 to 12 g dm^-3), thereby reducing water availability, the hyperhydricity of those adventitious shoots regenerated from carnation petals decreased. This was accompanied by a progressive fall in the water content of shoots (94.9 to 91.4 %), fresh mass (from 57.2 to 1.8 mg), number of leaf parenchyma cell layers (from 9.3 to 7.7), and the size of these cells (from 968 to 254 µm²). However, the number of regenerated shoots also decreased (17.7 in 2 g dm^-3 agar to 4.3 in 12 g dm^-3). Similarly, in ventilated tubes, which exhibit a lower relative humidity than tightly closed tubes, shoot organogenesis diminished up to 28 %, in tandem with shoot water content. Thus, relative humidity and water availability in culture vessels do not only influence shoot hyperhydricity in carnations, but also greatly affect adventitious shoot organogenesis.

Inheritance of 10 morphological and quantitative traits related to plant and fruit development and resistance to the pathogen Phytophthora capsici was studied in an intraspecific cross between a non-pungent, susceptible Capsicum annuum parent (cv. Americano) and a wild, pungent and resistant line (Serrano Criollo de Morelos-334). Data were obtained from the segregation of 166 F₂ plants and 50 F₃ plants in four years. Three of the traits analyzed (necrosis length, leaf width and leaf length) exhibited a transgressive segregation. A multiple linear regression analysis was applied in order to establish a relationship between necrosis length and some of the morphological traits measured such as length and width of leaf, length, diameter and mass of fruit, capsaicin content in fruits, and presence of hair on leaves and stems. The results identified a linear dependence between necrosis length (as an inverse measurement of resistance) and leaf width, fruit diameter and hair presence in the stem. Pungency was not related with resistance.

To dissect differences in both lipid accumulation and composition and the role of these modifications during drought stress, four grapevine cultivars exhibiting differential tolerance to drought were subjected to water shortage. Tolerant cultivars, Kahli Kerkennah and Cardinal, exhibited higher leaf water potential (Ψ_w), and lower lipid peroxidation compared to the sensitive cultivars Guelb Sardouk and Superior Seedless during stress. Total lipid amounts increased during stress only in the leaves of the tolerant cultivars. Drought induced increases in the ratios digalactosyldiacylglycerol/monogalactosyldiacylglycerol and phosphatidylcholine/phoshatidylethanolamine of almost all the drought stressed cultivars. Moreover, the overall analysis of the composition of fatty acids revealed that a linolenic acid was prevalent in grapevine and the unsaturation level of lipids increased under water stress in all the cultivars. Specific adjustments in the lipid composition during stress could compromise stress tolerance.

Phylogenetic analysis was conducted based on sequences of the internal transcribed spacer region (ITS) of nuclear ribosomal DNA in 17 species of Kengyilia, together with those of 18 species from Pseudoroegneria, Agropyron, Roegneria and Douglasdeweya by the maximum parsimony, maximum likelihood and neighbor-joining distance methods. The results indicate that species of Kengyilia had close affinities to species of Douglasdeweya and Agropyron. The species in Kengyilia was identified as two subgroups with regard to geographic distribution, indicating that species from the same distribution had a closer phylogenetic relationship. The genus Kengyilia was found as a ligament-group between Roegneria and Agropyron. The ITS sequence is a useful tool for studying the phylogeny of closely related species.

Different lines of cell suspension cultures of Taxus × media Rehd. and Taxus floridana Nutt. were cryopreserved with a two-step freezing method using a simple and inexpensive freezing container instead of a programmable freezer. Four to seven days old suspension cell cultures were precultured in growth medium supplemented with 0.5 M mannitol for 2 d. The medium was then replaced with cryoprotectant solution (1 M sucrose, 0.5 M glycerol and 0.5 M dimethylsulfoxide) and the cells incubated on ice for 1 h. Before being plunged into liquid nitrogen, cells were frozen with a cooling rate of approximately -1 °C per min to -80 °C. The highest post-thaw cell viability was 90 %. The recovery was line dependent. The cryopreservation procedure did not alter the nuclear DNA content of the cell lines. The results indicate that cryopreservation of Taxus cell suspension cultures using inexpensive freezing container is possible.

Waterlogging is a serious problem, which affects crop growth and yield in low lying rainfed areas. The main cause of damage under waterlogging is oxygen deprivation, which affect nutrient and water uptake, so the plants show wilting even when surrounded by excess of water. Lack of oxygen shift the energy metabolism from aerobic mode to anaerobic mode. Plants adapted to waterlogged conditions, have mechanisms to cope with this stress such as aerenchyma formation, increased availability of soluble sugars, greater activity of glycolytic pathway and fermentation enzymes and involvement of antioxidant defence mechanism to cope with the post hypoxia/anoxia oxidative stress. Gaseous plant hormone ethylene plays an important role in modifying plant response to oxygen deficiency. It has been reported to induce genes of enzymes associated with aerenchyma formation, glycolysis and fermentation pathway. Besides, nonsymbiotic-haemoglobins and nitric oxide have also been suggested as an alternative to fermentation for maintaining lower redox potential (low NADH/NAD ratio), and thereby playing an important role in anaerobic stress tolerance and signaling.

An efficient micropropagation protocol was established for Capsicum chinense Jacq. cv. Umorok, a pungent chilli cultivar. Shoot-tip explants were cultured on Murashige and Skoog (MS) medium containing cytokinins (22.2-88.8 µM 6-benzylaminopurine, BAP, 23.2-93.0 µM kinetin, Kin, or 22.8-91.2 µM zeatin, Z) alone or in combination with 5.7 µM indole-3-acetic acid (IAA). Maximum number of shoots were induced on medium containing 91.2 µM Z or 31.1 µM BAP with 4.7 µM Kin. The separated shoots rooted and elongated on medium containing 2.5 or 4.9 µM indole-3-butyric acid (IBA). Axillary shoots were induced from in vitro raised plantlets by decapitating them. The axillary shoot-tip explants were used for further multiple shoot buds induction. A maximum of about 150 plantlets were obtained from a single seedling. Hardened and acclimatized plantlets were successfully established in the soil.

Biological control of wilt of egg plant (Solanum melongena L.) caused by Fusarium solani was made with the application of five Trichoderma species, T. harzianum, T. viride, T. lignorum, T. hamatum and T. reesei. The effect of volatile and non-volatile antibiotics of Trichoderma origin on growth inhibition of the wilt pathogen was studied. T. harzianum showed maximum growth inhibition (86.44 %) of the pathogen through mycoparasitism. The non-volatiles produced by the Trichoderma species exhibited 100 % growth inhibition of the pathogen under in vitro condition. Production of siderophores and fungal cell wall degrading enzymes, chitinase and β-1,3-glucanase were found. Treatments with two most efficient Trichoderma species, T. harzianum and T. viride resulted in the decreasing population of Fusarium solani in soil thereby deterring disease incidence in field condition.

The effects of 28-homobrassinolide (HBL) on nickel uptake, protein content and activities of antioxidative enzymes were determined in the seedlings of Brassica juncea L. The seeds were treated with different concentrations (0, 0.01, 1 and 100 nM) of HBL for 8 h and then sown in the Petri plates containing various concentrations (0, 25, 50 and 100 mg dm^-3) of nickel. After 7 d, observations were made on shoot and root length, Ni uptake, protein content and activities of antioxidative enzymes (guaiacol peroxidase, catalase, glutathione reductase, ascorbate peroxidase and superoxide dismutase). The growth of seedlings was inhibited by Ni, however, less after HBL pre-treatment. The protein content and antioxidative enzyme activities were also increased by HBL treatment.