Selected factors affecting somatic embryogenesis efficiency have been studied, namely genotype, explant type and its orientation in the medium, different basal media, different auxins for somatic embryo induction, and two ways of donor plant cultivation. The key role is played by genotype and auxin used, the minimum effect was observed due to basal media. In the series of subsequent experiments we have found the best combination of individual factors as follows: cv. Altona, 10 uM 2,4-D, L2 basal medium, central part of immature cotyledon as initial expiant oriented by adaxial side down on the agar medium, and field grown donor plants. This combination exhibited 100 % embryogenic explants with 5.43 ± 0.65 somatic embryos per expiant,i.e. somatic embryogenesis efficiency 5.43.

Multiple shoots (16-20 shoots per expiant) were induced from cotyledonary node region ofAnogeissus acuminata (Roxb. ex DC.) Guill. & Perr. on Murashige and Skoog's (MS) medium containing IAA 0.1 mg 1^-1 + BAP 1.5 mg 1^-1 and ascorbic acid 50 mg 1^-1, citric acid 25.0 mg 1^-1, arginine 25 mg 1^-1 and adenine sulphate 25 mg 1^-1. From the first node of seedling only 4-6 shoots per expiant were proliferated. Segments ofin vitro produced shoots were used as expiants for further multiplication of shoots upto 16 successive cultures at an interval of 4 week on MS medium with IAA 0.1 mg 1^-1 + BAP 1.0 mg 1^-1 and additives. The original cotyledonary expiant was repeatedly subcultured upto 4 times after harvesting crop of shoots, each time.In vitro produced shoots were rooted on half strength MS medium containing 0.5 mg 1^-1 IBA. Plantlets were transferred to pots. Other expiants (cotyledons, hypocotyl, and leaf) produced callus on medium containing auxins and cytokinins. The calluses differentiated into embryo like structures or roots on MS medium.

Floral induction in the long day plant spinach (Spinacia oleracea) has been shown to be accompanied by a thickening of plasmalemma. This change was observed at early evocation, in both shoot apices and leaves, as well as after inducing GA₃ treatment. To get further information on this thickening, plasma membranes from spinach leaves were isolated, in the present study, using aqueous two phase partitioning and the effect of variousin vitro treatments on their thickness was investigated. The average plasmalemma thickness was unaffected by Na⁺ and K⁺ ions. It was increased upon the effect of either Ca²⁺ or gibberellic acid. A thickening of plasmalemma was also observed when plasma membranes from vegetative plants were incubated with a cytosolic preparation from photoinduced plants. The results were discussed in relation with the plasmalemma modifications previously reported in spinach.

The fungal colonization of the angiosperm root parasiteCynomorium coccineum and the halophytic hostsLimonium delicatulum andArthrocnemum glaucum were investigated in a Mediterranean salt marsh in March 1992. The main fungal inhabitants on the leaves or shoot surface of the test plants wereAspergillus niger, Penicillium chrysogenum andCladosporium herbarum. The qualitative analysis of the fungal species associating the parasite, the hosts and the non-infected plants showed similar pattern. However, the total population exhibited quantitative differences coupled with the amount and the chemical composition of the exudates on plant surface and the quantity of transpired water. The fungal catch from the aerial shoot (inflorescence) of the parasite was higher than that collected from either the leaves or aerial shoots of non-infected or host plants. The fungal density on the leaves ofL. delicatulum was higher than those isolated from the aerial shoots ofA. glaucum. Infection byC. coccineum caused a marked drop in the total fungal population on leaves or shoot surfaces of the hosts as compared to the corresponding non-infected individuals. The stimulative effect of washings on spore germination of some isolated fungal species was matched with the density of fungi on the target plants.

The effect of salinity on vacuolar pH was studied in carrot (Daucus carota L.) cells grown in liquid suspension culture either in the absence or presence of 150 mM NaCl. Both vacuolar and cytoplasmic pH were determined by several independent techniques. These techniques were NMR spectrometry, distribution of radioactive probes and spectrophotometric measurement of the absorbance changes of a naturally occurring vacuolar pH indicator. There was no difference in the cytoplasmic pH between cells grown in the presence or the absence of NaCl, but the vacuolar pH of cells grown in the presence of NaCl was higher by 0.38 to 1.05 pH units (depending on the technique that was used) than the vacuolar pH of cells grown in the absence of NaCl.

Ten major host-encoded pathogenesis-associated proteins have been found in extracts of intercellular sap of cucumber cotyledons infected with tobacco necrosis virus. By native and two-dimensional SDS-polyacrylamide gel electrophoresis, five major acidic virus-elicited proteins have been identified and classified on the basis of their molecular mass in three groups: group 1 included one protein of 14 kD; group 2, one protein of 28 kD and group 3, three proteins of 37-40 kD. Another five viruselicited proteins were basis, and were again classified in three groups: group 1, included two proteins of 15-16 kD; group 2, two proteins of 22-23 kD; group 3, one protein of 40 kD.

Several factors influencing reliability of the quantitative fluorimetric β-glucuronidase (GUS) assay in transgenic plant tissue have been investigated. We obtained linear dependence of fluorescence on both the duration of hydrolysis and the extract concentration. The stability of the enzyme in the homogenate was fairly high, the same as the stability of the substrate solution and of the final reaction product. The modification of the extraction/incubation buffer was proposed, resulting in several times higher activity in comparison with original procedure.

Transport of ions in young maize plants was affected by the temperature gradient (gradT). This influence was dependent on the age of plants and the solute concentrations in compartments at the two sides of plants. When gradT was increased, current, volume and heat flows rose during 20 min and then declined. On the other hand, the diffusive flow only declined. The character of the flow changes was given by the changes in permeability and reflection coefficients. In young plants under low solute concentration on both sides of plants, the permeability of the plant tissues shifted from positive to negative values. Under higher solute concentrations on both sides of the plants such phenomenon was not observed. At high gradT the reflection coefficients of the plant tissues were low, but they rose during levelling the temperature in the two compartments.

Vanadium bioaccumulation calculated as the ratio of its content in plant biomass to that in the substrate (Vbi-index) was studied in pea. Vbi was on average 11.275, 11.770 and 13.153 in the roots, and 0.809, 0.467 and 0.749 in the shoots of the cultivars Opal, Laser and Ramir, respectively. This indicates cultivar differences in vanadium uptake, and low translocation rates from roots to shoots. Vanadium (3 to 30 mg 1^-1) decreased shoot and root fresh and dry masses of the three cultivars. Seedlings of the cv. Opal were the most susceptible to higher concentrations of vanadium (20 to 30 mg 1^-1), whereas seedlings of cv. Laser were the most resistant.

NaCl salinity affected growth, the contents of chlorophyll, carotenoids, saccharides and some minerals (Na, K, Ca, Mg, P) in wheat (Triticum vulgaris L.) plants. Irrigation of soil with CaCl₂ or KCl greatly ameliorated the adverse effects of NaCl salinity. This counteraction was associated with an increase in contents of saccharides, proteins and Ca, Mg and P which might explain their role in osmotic adjustment.

Thidiazuron incorporated into MS medium stimulated rosettes formation only in some treatments. This effect was more pronounced in cultures ofMorus alba thanPrunus sp. Mulberry cultures responded to the optimal concentration of thidiazuron (0.2 mg I^-1) not only with shoot formation but also, with growth of large leaves and poor development of callus tissue. In cultures of both investigated genera the shoot elongation was inhibited. Shoots of mulberry cultures growing on proliferation medium supplemented with thidiazuron formed roots, in many cases.

Investigation was made on a use of microcrystal cellulose as a new and inexpensive gelling agent instead of agar. Microcrystal cellulose in concentration 20 % forms a suitable structure of nutrient medium for in vitro cultivation. The higher humidity in the culture container with microcrystal cellulose causes partial vitrification of Nicotiana tabacum L. plants, cv. Zlatna arda. It is proved by reduced chlorophyll content, changes in protein synthesis and strongly reduced isoenzyme spectrum of peroxidase.

l-alkyl-l-ethylpiperidinium bromides inhibited light induced Mg²⁺-adenosin-triphosphatase activity of isolated thylakoids in both Phaseolus vulgaris L. and Zea mays L. The short chain detergents (C₅ to C₇) were less effective in maize than in bean.

The effects of decreasing Ca²⁺ concentrations (Na/Ca ratios were 24, 49, 99 and 199) of the saline growth medium (NaCl concentration 200 mM) on three grass speciesCenchrus pennisetiformis Hochst. & Steud,Leptochloa fusca L. Kunth. andPcmicum turgidum Forssk. were assessed after 7 weeks growth in sand culture.L. fusca produced the highest dry biomass of all the species at varying Na/Ca ratios. Number of tillers per plant and number of green leaves per tiller were reduced significantly only inC. pennisetiformis. Leaf water potential ofC. pennisetiformis decreased at all external Na/Ca ratios, whereas inL. fusca it decreased only at an Na/Ca ratio of 99. Leaf osmotic potential ofL. fusca consistently decreased at all Na/Ca ratios, whereas that of the other two species remained unchanged. The shoot and root total sugars of all species remained unaffected at all decreasing Ca²⁺ concentrations. InP. turgidum chlorophyllb and total chlorophyll decreased consistently at all Na/Ca ratios, but inL. fusca they decreased only at the highest Na/Ca ratios. The leaf soluble proteins of all the species remained unaffected at all Na/Ca ratios. The leaf free amino acids decreased significantly inL. fusca with the increase in Na/Ca ratios. The leaf proline content was only decreased inL. fusca at the highest Na/Ca ratio. The significant correlations between the growth of the three grass species and other variables determined in this study were not found.

The full length cDNA clone of leghemoglobin I gene fromLupinus luteus was placed under dual promoter into the plant expression vector pCB1399 and the resulting vector (pCB1415) was transfered into theAgrobacterium strain LBA4404 (pAL4404). The binary system LBA4404 (pAL4404, pCB1415) was then used for transformation ofNicotiana plumbaginifolia andLotus corniculatus. In both species kanamycin-resistant plants have been selected and regenerated. The synthesis of LbI protein in transformed plants has not been shown.

The effects of different nitrogen salts on nitrate reductase activity and protein contents were investigated in three Yugoslav cultivars of wheat. The nitrate salts appeared to be a better form of nitrogen than ammonium in respect of the increase of the nitrate reductase activity and root total protein contents, whereas the treatment with ammonium salt resulted in a comparably higher shoot total protein contents. KNO₃ was the best in respect of the level of nitrate reductase activity. Different concentrations of nitrate and ammonium ions in nutrient solution, showed very similar effects on investigated parameters. NS Rana 2 cultivar had the highest values of nitrate reductase activity and protein contents.

The efficiency ofAgrobacterium-based transformation technique in oilseed rape and cauliflower was influenced by cultivar specificity, donor plant age and explant type. Marked differences in demands for plant hormone contents in the regeneration medium were recorded already among different types of nontransformed explants. The highest regeneration capacity was recorded with stem and leaf segments isolated from one-month-old aseptically grown plants. The regeneration was markedly species-dependent. Regeneration of transformed plants from stem segments and thin layers isolated from field-grown oilseed rape plants (at the most 2% of regenerating explants) and from oilseed rape hypocotyls (0.8% of regenerating explants) and cauliflower (1.2% of explant regenerated transformed shoots) was achieved after disarmedAgrobacterium treatment. Hypersensitive reaction of explants could be prevented by using prolongedin vitro precultivation and delayed application of the selective agent.

Relative competition among various plant parts for N during water stress,i.e. nitrogen distribution index (NDI) was determined in relation to specific nitrogenase activity (SNA) and nodule and soil nitrogen in both indeterminate (H-77-216) and determinate (ICPL-151) types of pigeonpea (Cajanus cajan L.) under greenhouse conditions. Two levels of water stress,i.e. moderate (soil Ψ_w) -0.77 MPa) and severe (soilΨ_w -1.34 MPa) were created by witholding the irrigation at vegetative (40 DAS) and flowering (70 DAS) stages. At vegetative stage under moderate stress the highest NDI was in nodules of cv. H-77-216 and in leaf of cv. ICPL-151. Under severe stress both the cultivars showed negative values of NDI, with maximum loss of N from root and nodules. Cultivar ICPL-151 behaved differently at flowering and vegetative stages. Very high loss of N from different plant parts was seen at flowering under severe stress. All the plant parts showed gain in N during rehydration. Loss and gain in N at both the stages under stress and rehydration respectively, correlated with available N in soil. Specific nitrogenase activity (SNA) and nodule N were maximum at moderate stress and related with NDI values of leaf and nodules.

Changes in the levels of free indol-3-ylacetic acid (IAA) and free cytokinins were determined in the course of dormancy and sprouting period in potato tubers(Solanum tuberosum L., cv. Nevskii) stored at 4 °C. The same analyses were performed in potato tubers after Ethrel application, which prolongs dormancy. No significant changes were found in free IAA level during dormancy followed by a rapid decrease during sprouting. After Ethrel application a significant lower IAA level was found 3 weeks after application, but further on no changes in free IAA level between treated and non-treated tubers were detected. Cytokinin level was relatively low and constant till sprouting and increases then by about 55 %, mainly due to an increase in the level of zeatin riboside and isopentenyladenosine. Ethrel application decreased cytokinin level during dormancy slightly, but postpones the increase coupled with sprouting by about 1 month. Thus, IAA does not seem to have a significant effect on tuber dormancy, while cytokinins are probably necessary for sprouting initiation.

The angiosperm root parasiteCynomorium coccineum and the halophytic hostsLimonium delicatulum andArthrocnemum glaucum were invesrigated under natural conditions in a Mediterranean salt marsh in March 1992. The diurnal transpiration rate of the parasite was low and parallel to the climatic factors. The non-infected plants showed higher transpiration rates than the parasite and its hosts. Compared to the non-infected plants, it is apparent that the perennial herbL. delicatulum is more sensitive to infection since its reduction in the fresh and dry mass was higher than that found inA. glaucum.

Using a membrane potential probe, Oxonol VI, it was possible to demonstrate generation of ATP- and NADH-dependent membrane potential across the plasmalemma, with membrane vesicles derived from parenchyma cells of Jerusalem artichoke tubers(Helianthus tuberosus L.). It was shown that ATP- and NADH-dependent membrane potential generation was higher in dormant material than in non-dormant tissue and that the effects of ATP and NADH on membrane potential generation were additive. ATP-dependent potential generation was sensitive to vanadate, an inhibitor of plasmalemma ATPase activity. The results are discussed in relation to the properties of the different enzymes bound to the plasma membrane, the morphogenetic potentialities of tuber buds and the hypothesis that tuber dormancy could be an extreme case of nutrient deficiency induced by short-distance intercellular relationships.

RNA synthesis as measured by the incorporation of tritiated uridine into trichloroacetic acid insoluble material was studied in the leaf protoplasts of cv. Kennebec and its parthenogenically derived dihaploid. Protoplasts of cv. Kennebec incorporated tritiated uridine at a greater rate and accumulated more than twice the amount of radioactive materials than did the dihaploid over a 6-h incubation period. Poly(A)⁺RNA, isolated from the total RNA of the tetraploid and of the dihaploid, by oligo(dT)-cellulose column chromatography, was present in amounts of 11.3 and 5.2 % respectively. The tetraploid synthesized 4.8 times the amount of poly(A)⁺RNA that was synthesized by the dihaploid.