Molecular markers were used to detect the influence of high concentrations of 2,4 dichlorophenoxyacetic acid (2,4-D) in the callusing media on DNA variations in regenerated rice plants. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) based RFLP analysis were carried out on 12 somaclones of Oryza sativa L. cv. B-370. In vitro culture induced DNA variations were detected in the regenerated plants but the effect of high auxin concentration in the medium could not be revealed. In a second study, fingerprinting of 15 semi-dwarf, high yielding somaclones of B-370 was carried out using RAPD technique. Amplification using 20 random primers produced a total of 167 DNA bands out of which 97 bands were polymorphic. A total of 32 unique DNA bands were detected across all the somaclones and they could be grouped based on their similarity to B-370. RAPD analysis helped to reveal similarity or differences among the somaclones while fingerprinting using additional RAPD markers was not successful.

Proline (Pro) accumulation in leaf discs of the NaCl-treated salt sensitive Lycopersicon esculentum was higher than in the salt tolerant L. pennellii. The magnitude of Pro accumulation differed when leaf discs of both species were floated in the light or under darkness, and in various incubation media: buffer solutions (pH from 3.9 to 7.8), abscisic acid, isobutyric acid, NH4Cl, malate, citrate, and mixtures of NaCl and KCl, NaCl and CaCl2, and NaCl and mannitol. Under darkness, Pro accumulation in L. esculentum was not regulated by salts. Conversely, the light-independent NaCl-induced Pro accumulation observed in L. pennellii became light-dependent when CaCl2 was added. The different expression patterns of Pro accumulation reinforce the proposal of two Pro metabolic pathways in the domestic and the wild tomatoes.

Dry matter production of shoots and roots and the diurnal fluctuation of titratable acidity of single leaves were investigated in the CAM plant Agave attenuata during the first 70 d after germination. The plants were grown either in vermiculite sub-irrigated with a nutrient solution or in in vitro cultures on an inorganic nutrient agar. Two types of culture tube covers were used: either airtight closures or polypropylene caps with membranes permeable to air.
In the earliest ontogenetic phases of development (cotyledon and primary leaf stage), the plants were already able to carry out considerable nocturnal organic acid accumulation. In vitro cultivated plants, from the beginning of their development, were also capable of diurnal acid fluctuation, though of distinctly weaker activity than the pot plants. The mean relative growth rates (RGR) of pot culture plantlets approached a third of perennial herbaceous plants. Plantlets grown in in vitro culture reached only half to the one quarter of the RGR of pot plants. The reduced yield could be attributed to the low CO2 supply in the culture tubes and the less than optimal water and nutrient supply in the agar medium.

Pepper species and cultivars, Capsicum annuum cv. Bell Boy, C. annuum cv. Kulai and C. frutescens cv. Padi, differing in drought tolerance were investigated for their water relations, stomatal responses and abscisic acid (ABA) content during water stress. C. frutescens cv. Padi exhibited a greater osmotic adjustment than C. annuum cultivars. Stomatal conductance of cv. Bell Boy was more sensitive to water stress than that of cvs. Kulai and Padi. In all pepper genotypes, stomatal closure was triggered in the absence of a large decrease in leaf water status. ABA content in xylem sap and leaf was higher in C. annum cultivars compared to C. frutescens cv. Padi.

Total genomic DNA isolated from maize hygromycin B resistant cell line(hygr-G204) was used to transform the maize hygromycin B sensitive cell line(hygs-G204) to the hygr-phenotype using polyethyleneglycol treatment and the transformed calli were selected using hygromycin B. The primary transformant maize plants were regenerated and analysed at the molecular level using DNA hybridization, transgenome rescue and histochemical β-glucuronidase assay. The results indicated that genomic DNA-mediated transformation can lead to transfer, expression and stable integration of a DNA fragment into the host genome.

After treatment with increased quantities of nitrogen and Azotobacter strains, activities of antioxidant enzymes superoxide dismutase, peroxidase and catalase, content of chlorophylls and carotenoids, soluble proteins and dry matter in leaves of sugar beet increased.

The first step during shoot primordia development in Papaver somniferum L. was the differentiation of globular meristemoids, resulting from localized cell division in callus tissue. During meristemoid maturation differences observed were in mitotic activity, cytoplasmic density, starch distribution, and in cell size, cell shape, and nucleus size to cell size ratio between peripheral and central meristemoid cells. Cell specialization within the meristemoids is a prerequisite for shoot primordia development.

Growth rate of fruit of Actinidia deliciosa (A. Chev.) Liang and Fergusoncv. Hayward, measured as increase in fresh mass, followed a simple sigmoidal trend showing two stages in all the seasons studied (1987 - 1989). However, growth profiles were statistically different from year to year in both shape and mean reached. The data suggest an important effect of climatic factors on growth, mainly rainfall, which may explain the differences in growth curves between seasons. Significant correlations between relative growth index and length and/or diameter were found which can be used for a non-destructive determination of Actinidia deliciosa fruit growth.

A spermine-resistant mutant of Arabidopsis thaliana (L.) Heynh. was isolated from M2 population of ethylmethanesulphonate-mutagenized seeds. The mutant was resistant to seed germination inhibition by spermine, but was as sensitive as the wild-type to spermidine and putrescine. In addition, the mutant displayed developmental abnormalities such as frequent cauline leaves, increased number of branches with inflorescence, reduced apical dominance, flowers subtended by bracts, disrupted floral organs with homeotic conversions. Genetic analysis indicated a single recessive nuclear mutation that was allelic to apetala2-1 (AP2-1). The new mutant allele of AP2 locus was accordingly numbered as AP2-10.

Zygotic embryos of Karwinskia parvifolia, isolated from seeds obtained from different regions of Mexico, were cultured on Woody Plant Medium (WPM) supplemented with 0.06 µM indole-3-acetic acid, 0.03 µM gibberellic acid, and 2 µM 6-benzylaminopurine. The growth of embryos and multiplication of shoots from stem segments were achieved. Rooting of excised shoots could be initiated on basal WPM medium with prolonged subculture period to 2 months, or on WPM medium supplemented with 10 µM 1-naphthaleneacetic acid. Multiplication capacity of shoots and rooting of K. parvifolia differed in dependence on the origin of explant material. The shoot multiplication was much lower than that of Karwinskia humboldtiana. The rooting depended on the origin of K. parvifolia seeds. The regenerated plants were successfully transferred to glasshouse.

The physiological responses of alfalfa (Medicago sativa L. cv. Gilboa) to salinity (100 mM NaCl) and some inorganic nutrients (K+, Ca2+ and N as NO3-) were investigated. Salinity caused a substantial reduction in biomass, carbon assimilation rate, stomatal conductance, water use efficiency, leaf area, relative growth rate, NO3- content and nitrate reductase activity, whereas, transpiration rate was slightly affected. Inclusion of K+, Ca2+ and N as NO3- in plant nutrient medium in combination or alone brought about a marked stimulation in control plants and moderated the salinity caused reductions in NaCl treated plants. In addition, plants also exhibited differences in these parameters at two growth stages.

The effects of ferulic acid on L-malate oxidation in mitochondria isolated from soybean (Glycine max L.) seedlings were investigated. Oxygen uptake and the products of L-malate oxidation were measured under two conditions: pH 6.8 and 7.8. At acidic pH, the activity of the NAD+-linked malic enzyme (L-malate:NAD+oxidoreductase [decarboxylating] EC 1.1.1.39) was favoured, whereas at alkaline pH a predominance of the L-malate dehydrogenase activity (L-malate:NAD+oxidoreductase EC 1.1.1.37) was apparent. Ferulic acid inhibited basal and coupled respiration during L-malate oxidation either at acidic or alkaline pH, reducing also the amounts of pyruvate or oxaloacetate produced. The results suggest that the site of ferulic acid action is situated at some step that precedes the respiratory chain. An interference with the L-malate entry into the mitochondria could be an explanation for the effects of ferulic acid, but the possibility of a direct inhibition of both enzymes involved in L-malate oxidation cannot be ruled out.

Effect of two important mycotoxins, aflatoxin B1 and citrinin (concentration 2 g m-3) at various combinations (i.e., 1:1, 1:2, 2:1, 1:3 and 3:1, v/v) on seed germination, seedling growth, chlorophyll, carotenoid, starch, sugar, protein and nucleic acid contents, α-amylase activity, and respiration quotient was studied in maize cv. Suwan composite. The maximum and minimum inhibitions were recorded in most of the above parameters (except starch) at 3:1 and 1:3 combination ratios of these toxins, respectively. However, the inhibition rates varied with the treatments.

We obtained transformed bean shoots by electroporation of intact bean cells with the plasmid pDPG165 containing bar gene conferring herbicide resistance to plants. Transformed shoots were selected from electroporated callus on herbicide containing media. Data of molecular analysis (PCR and Southern blotting) confirmed the insertion of bar gene in the genome of herbicide resistant shoots. Detailed procedures for obtaining regenerative bean callus, optimization of electroporation of intact cells and transgenic shoots are given.

When the proper stimuli are given, somatic plant cells may form adventitious embryos, roots or shoots. The three pathways of regeneration show apparent similarities. They consist of three analogous phases: 1) dedifferentiation (during which the tissue becomes competent to respond to the organogenic/embryogenic stimulus), 2) induction (during which cells become determined to form either a root, a shoot or an embryo), and 3) realization (outgrowth to an organ or an embryo). The first phase may involve a period of callus growth (indirect regeneration), but often cells present in the explant become competent without cell division or without cell division at a large scale (direct regeneration). In an explant, only very few cells show the organogenic/embryogenic response. In direct regeneration, the three regenerative pathways start from cells in different tissues. This is most obvious when the different types of regeneration occur in the same explant. The hormonal trigger for the dedifferentiation phase is a general one, probably auxin. During the induction phase, each pathway requires specific hormonal triggers. During the realization phase, hormones should be absent or at low concentration. The successive steps in the regeneration process coincide with events on the molecular and biochemical levels, but so far no coherent picture has emerged. In particular during the early stages of regeneration, research on these levels is hampered by a technical problem, viz., the very low proportion of cells that participate in the process of regeneration. New methods may overcome this problem.

The localization of peroxidase in cells of horse-radish (Armoracia lapathifolia Gilib.) tumour and teratoma tissues was studied. Both tissue lines were derived from the same primary crown-gall tumour induced on the leaf fragments by a wild type of Agrobacterium tumefaciens B6S3. Enzymatic activity was measured in cell walls, high-density heterogeneous membrane fraction, microsomal and soluble (no particulate) fractions. The subcellular localization of enzymatic activity was distinct for each transformed tissue. Both tumour and teratoma showed similar isoenzyme patterns, but one soluble acidic isoperoxidase could be considered as a marker of cell differentiation.

When seedlings of two rice (Oryza sativa L.) cultivars Ratna and Jaya were raised under 100 and 500 µM cadmium nitrate in the medium, a high proline content was noted in Cd²⁺ stressed seedlings compared to controls. Seedlings grown under 500 µM Cd(NO₃)₂ maintained increased proline level compared to non-stressed seedlings. Kinetic properties of RNase extracted from control grown and Cd²⁺ stressed seedlings showed a marked alteration in Km due to Cd²⁺ treatment. The RNase isoforms were purified from 15-d-old rice seedlings with a total purification of 22.25 fold and 74.75 % yield using conventional biochemical techniques. Three RNase isoforms, namely I, II and III were eluted from DEAE-Sephacel column. The isoform RNase II had Km value of 3.2 mg(RNA) cm^-3. The in vitro osmotic stress created by incorporation of PEG in the enzyme assay medium led to decreased affinity of enzyme towards its substrate with increase in Km. This loss in affinity was partially restored by the addition of 1 M proline in the assay medium, suggesting the possible protective role of proline on RNase under osmotic stress.

In Sedum fabaria, the ovule is anantropus, bitegmic and crassinucellate. The development of the nucellus conforms to the Sedum type. The development of the embryo sac is of the Allium type. The antipodal cells in unfertilized embryo sac occasionally divide and one of them forms four-celled structures resembling embryos and remaining once elongate in the form of haustoria. The entry of the pollen tube is porogamous. After division the primary endosperm nucleus forms two cells: the apical one develops into cellular endosperm according to the Acre type and the basal one acts as the endosperm haustorium of the Sempervivum type. The embryogeny corresponds to the Caryophyllad type.

Secondary somatic embryogenesis and shoot organogenesis from primary somatic embryos of Papaver somniferum L. are described. The embryos became malformed, the root meristem expressed dividing activity without position-dependent cell differentiation, causing abnormal development or arrested growth of primary somatic embryos. The adventitious shoots regenerated from embryo hypocotyl, but secondary somatic embryos had an epidermal origin close to the root meristem. The regeneration occurred without hormonal treatment, indicating endogenous nature of triggering signals. These signals are probably related to the integrity loss of morphogenetic steps during development of primary somatic embryos, which appeared to induce an activation of cells competent to regeneration.