Effect of fluoride (10 and 50 mM) on the activities of sucrose metabolizing enzymes, alkaline inorganic pyrophosphatase, and transaminases in relation to the accumulation of free sugars, starch, and soluble protein was studied in detached ears of wheat cultured in a liquid medium. Culturing for 5 d in the presence of fluoride reduced the amount of grain starch whereas contents of total free sugars, particularly sucrose, and soluble protein increased. Fluoride inhibited the activities of soluble acid and neutral invertases, as well as sucrose synthase acting in the cleavage direction. Uptake of uniformly labelled ¹⁴C-sucrose or fructose was also drastically reduced by fluoride. Glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) activities also increased with fluoride addition in correspondence with an increase in soluble protein. Apparently, the wheat grain responds to fluoride-mediated disruption of carbon metabolism by a compensatory effect on nitrogen metabolism.

A novel approach for simultaneous localization of two DNA sequences on plant chromosomes is described. The approach is based on a combined use of primed in situ DNA labelling (PRINS) with fluorescent in situ hybridization (FISH). Traditionally, this has been done using FISH with two probes labelled by two different marker molecules. Compared to this method, the combined PRINS-FISH procedure is faster. Furthermore, because one of the DNA sequences is localized by PRINS with specific primers, only one labelled probe is needed.

Potato (Solanum tuberosum L. cv. Bintje) was transformed with a cDNA clone encoding an osmotin-like protein. Transgenic and non-transgenic in vitro plants were subjected to NaCl for 3 weeks. The shoot and root development was slightly affected by salinity indicating that the salt condition used was a mild stress. The endogenous proline content of the osmotin-like transformed clone only raised slightly as compared to the non-transformed genotype, where a marked increase in proline content could be observed as a result to salt stress. These data provide evidence for the involvement of osmotin-like proteins in the mechanisms of salt tolerance in potato plants.

Tobacco cv. White Burley was transformed with disarmed expression vector pCB1314 containing dimeric cDNA of potato spindle tuber viroid (PSTV, severe strain) in plus orientation regulated from the mannopine promoter. Amount of PSTV specific (-) and (+) sequences and PSTV circular forms was measured in transformed tobacco stock and compared with PSTV content in untransformed tomato and tobacco grafts. It follows from the results that lower rate of accumulation of PSTV in tobacco as compared with tomato is due to less intensive viroid transportation through the cytoplasm and/or cell to cell moverment, whereas both, viroid replication and processing showed comparable characteristics in tomato and tobacco with respect to accumulation of minus and plus strands and circular forms in infected tissues. Despite of accumulation of viroid in comparable amount in both transformed tobacco and infected tomato, no expression of any morphological symptom of disease was observed in transgenic tobacco.

Differential display of mRNA has been recently developed as a tool to detect and characterize changes in gene expression. We applied this technique to fruit trees plantlets induced to root in vitro, in order to isolate and study genes involved in root induction. A reproducible pattern of polymerase chain reaction (PCR) products was obtained, both in almond and apple, in vertical polyacrylamide gels stained with ethidium bromide. Differences in PCR fingerprinting were detected in mRNAs of basal part of either auxin induced or non induced microcuttings. Thus, we suggest that this technique can be used in woody species to detect changes among mRNA populations during root induction.

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.

Primed in situ labelling (PRINS) of nucleic acids was developed as an alternative to traditionally used fluorescence in situ hybridization (FISH). Compared to FISH, PRINS is faster and does not require preparation of labelled probes. Nevertheless, the number of applications for physical mapping of DNA sequences on plant chromosomes remains low. This is due to the fact that there are a number of factors which influence the specificity and sensitivity of the reaction. The purpose of this work was to analyse the effect of some of them, including the age of slides, type of Taq DNA polymerase, number and concentration of primers, the presence and concentration of bovine serum albumine and MgCl₂ in the reaction mixture. Furthermore, the effect of various pre-treatments on signal intensity and non-specific fluorescence was studied. A consensus Arabidopsis-type telomeric sequence and Vicia faba mitotic chromosomes were used as a model system. We have found that the age of slides was critical and that under optimal conditions it was possible to achieve relatively high signal to noise ratio.

The immature zygotic embryos of reciprocal maize hybrids (CHI-31 x GF1 and CHI-31 × GE2) were used as the initial material for induction of somatic embryogenesis in vitro. Histological analysis of somatic embryogenesis revealed high developmental variability. The arising formations were classified into 5 groups: A) somatic embryos phenotypically similar to zygotic embryos, B) polyembryos, C) formations with radicle but without meristematic plumule, D) formations with radicle without differentiated plumule, and E) formations with plumule without radicle. The formatioms A and B regenerated directly into plants. Plant regeneration from formations E required preculture on the rooting medium. Formations C and D failed to develope into plants possibly because of early loss of meristematic cell character during the embryo axis differentiation. The reverse sequence of radicle and plumule differentiation in somatic embryos in comparison with zygotic ones was noted. The epigenetic character of the scutellum, coleoptile, coleorhiza and leaves primordia development was discussed.

Aqueous salicylate solutions stimulated ethylene formation only when injurious, or potentially injurious, concentrations were exogenously supplied to soybean cuttings. Stimulation occurred via the biochemical sequence involving ACC as an intermediate, and was attributable to stimulation of ACC synthesis but not of EFE activity. Similar results were obtained by testing wound-induced ethylene, whereas the production of virus-induced ethylene was not affected by salicylate. Prolonged salicylate treatments which did not produce evident injurious effects inhibited soybean growth and rooting, probably through the moderate antiauxinic property attributable to salicylates. These findings are discussed in relation to other results obtained from similar or different plant materials.

21-day old plants ofChenopodium rubrum L. ecotype 374 were used. Organ relationships in the shoots were investigated by³²P distribution, which indicated different organ correlations in plants grown in continuous light and in plants treated with flower-inducing and non-inducing dark periods. Dark periods were associated with a low³²P distribution in young leaves and a high one in axillary buds. In the following light period the high³²P distribution in axillary buds continued whereas the³²P distribution in the leaves on the main axis increased and was similar to that in plants grown in continuous light. The high³²P distribution in axillary buds was brought about by both, flower-inducing and non-inducing dark treatments. Decapitation resulted in a high³²P distribution in buds, in continuous light an increased³²P distribution was also found in leaves. These effects were not fully cancelled by IAA application.
The results are discussed with respect to an assumption that decrease of apical dominance represents a step in a sequence of events leading to flowering.

Plasticity of the tobacco genome was studied by testing the DNAs of protoplast-derived regenerants with three different repetitive DNA sequences by the method of quantitative DNA/DNA hybridizations. A large population of 91 regenerants belonging to 35 different protoclones was analysed and a high degree of heterogeneity in the contents of the different DNA repeats was detected. The contents of middle repetitive sequences of two types were more stable or changed in the same direction, while the highly repetitive sequence varied independently and displayed a significant reduction in comparison with the two other sequences. Comparing the variation within the subpopulations of plants of the same clonal origin and the variation among the protoclones led to a conclusion that the pre-existing DNA variability in the starting plant material and/or thein vitro stress during the very early stages of protoclone regeneration played a decisive role in the formation of modified genomes in regenerants.

Potato root cultures transformed by_pRiA4 T-DNA do not regenerate plants in vitro spontaneously, but regeneration can be induced by a sequence of callogenesis and regeneration media. Both T_l and T_r-DNA were found by Southern blotting. Regenerated transformants showed characteristic morphological deviations in the in_vitro cultures. Plants in the soil showed differences in the inflorescence and tuber development.

Ultrastructural changes following increasing periods of water stress induced by means of polyethylene glycol 4000 (from 5 min to 18 h) were investigated in young epidermal cells of the primary roots ofZea mays. The sensitivity of the individual cell components to water stress was considered according to the time sequence in which their alterations appeared.
Endoplasmic reticulum (ER) and mitochondria proved to be the most sensitive cell components, their structure changing after 5 min of water stress. By 8 h of stress, the condensation of nuclear chromatin in some cells was apparent, preceding polyribosome degradation but not all the other changes of the stressed cells. Similar types of structural alterations appeared slightly earlier in the more differentiated epidermal cells. The rapid changes in the structure of ER and mitochondria coincided with the rapid decrease of water potential of the root tips and the immediate cessation of the root growth.

A binary vector system inA. tumefaciens for the introduction of foreign genes into the plant genome was developed. The first component is the Ri plasmid and the second component is a small vector plasmid, replicating inAgrobacterium, which carries the 25 bp terminal sequence of the Ti plasmid, theNos gene as the selectable marker and neighbouring sequences of the Ti plasmid. Functions necessary for integration are providedin trans by the virulence region of the Ri plasmid. Transformed cells are selected on the basis of hairy root tumor proliferation and agropine synthesis. They also showNos activity coded by the gene on the small part of Ti T-DNA.

Treatment of etiolated and excisedCucurbita cotyledons with exogenous cytokinin (benzyladenine) in darkness or light results in a marked stimulation of Rubisco activity, content of enzyme protein, and incorporation of labelled precursors into it, indicating cytokinin-stimulatedde novo synthesis of the enzyme. Cell-free translations of RNA in the wheat germ andE. coli systems show an increase in both large and small subunit mRNA amounts relative to the increase of total RNA under the influence of the phytohormone and light. This increase in the level of translatable RNA is confirmed by RNA hybridization with the Rubisco large subunit gene of spinach. In addition, our results demonstrating additive effects of benzyladenine and light in cotyledon and chloroplast development suggest that the two factors co-act independently in the causal sequence of Rubisco gene expression. The data are discussed in a general view of cytokinin action in gene expression steps.

The changes in cell division rate were studied in different components of the shoot apex ofChenopodium rubrum during short-day photoperiodic induction and after the inductive treatments. Induced and vegetative apices were compared. Accumulation of metaphases by colchicine treatment was used to compare the mean cell cycle duration in different components of the apex. A direct method of evaluating the increase in cell number obtained by anticlinal or periclinal divisions was applied if the corresponding components of induced and non-induced apices had to be compared. The short-day treatment prolonged the cell cycle more in the peripheral zone than in the central zone and still more in the leaf primordia. The importance of changing growth relations for floral transition was shown particularly if the induced plants were compared with the vegetative control with interrupted dark periods. Induced plants transferred to continuous light showed further changes in the rates of cell division. The cell cycle was shortened more in the central zone than in the peripheral zone,i.e. there was a further shift in growth relations within the apical dome. The cell cycle in the leaf and bud primordia was also shortened if compared with the vegetative control, the acceleration being stronger in the bud primordia. There was a subsequent retardation in cell division in the leaf primordia formed during and after the inductive treatment if the plants were fully induced. An inhibition of the oldest bud primordia was observed in fully induced apices, as well.

Influence of high intensive flashes on the yield of free radicals in intact seeds and excised embryonic axis, endosperm, and seed coat, and its resulting effect on seedling growth, total biomass production and phosphorus metabolism in wheat (Triticum aeativum), vetch (Vicia sativa L.) and onion (Allium cepa L.) was studied. Free radicals (f.r.) were formed mainly in seed coat and not in the endosperm. Vetch seeds after irradiation had 20.76 X 10¹³ f.r. g^-1 dry intact seed and 17.30 X 10¹³ f.r. g^-1 dry seed coat. Excised seed coats exposed to irradiation also yielded 17.28 × 10¹³ f.r. g^-1 dry matter. High irradiance "white light" flashes induced more f.r. than a monochromatic one of the same photon content. Red (650 nm), farred (750 nm) and even infra-red (1100 nm) radiation did not initiated f.r. formation but resulted in their decay in samples irradiated earlier by "white", blue and green parts of the spectrum. Blue irradiation of seeds led to the decrease in the length of shoots and roots in comparison to "white", green and red irradiation but their biomass increased faster than in the seedlings obtained from non-irradiated or irradiated with "white" and green radiation. The quantity of total acid soluble phosphorus followed a sequence with respect to wavelength of radiation: 436 nm > 650 nm> > 540 nm > non-irradiated > 300-800 nm. Quantity of inorganic phosphorus remained unaffected by different spectral character of radiation. The quantity of organic acid soluble nucleic phosphorus and acid insoluble polyphosphates was higher in samples irradiated with red beams (650 ± 6 nm).

A study was made on the mitotic cycle times in meristematic cells of Vicia faba root tips and on the relationship between their duration and the position of a certain cell in the column of proliferating cells. For the demonstration of the sequence and duration of mitotic oycles a single-column model was used. The results of experiments show that the great variability in the duration of mitotic cycles (from 12 to l20 h)is the result of a different program of the apical meristem cells. The long duration of the cycle of initials corresponds to the sum of cycle times of their descendants.

Populations ofScenedesmus quadricauda grown in a continuous chemostatic regime and synchronized by light and dark intervals were exposed to continuous illumination. The effect of light on synthetic and reproduction processes during the time of the omitted dark period and in the subsequent cell cycle was studied. In general, the sequence of cellular processes and their mutual coupling remain the same as in the darkened population. Synthetic processes and photosynthetic activity are depressed during the period of protoplast fission also in the light. The synchronizing effect of the dark period in chlorococcal algae consists in reducing the developmental variability in the population. The developmental state of daughter cells at the end of the dark interval varies in the span of two (as a rule) or four (at maximum) genomic cycles, while at the end of the light period this variability comprises up to eight genomic states. The more advanced autospores start the next cycle with a greater lag.

Taking into consideration simultaneous infection of tomato leaves withCladosporium fulvumCooke and TMV, the fungus infection inhibits the TMV reproduction. This inhibition occurs in the plant irrespective of the sequence of pathogen inoculation and reaches even 70 per cent. The fungus spores applied to the TMV purificate inhibit the TMV infection, as well.

The effect of acute gamma rays (⁶⁰Co; 1 to 10 kR) on the vascular differentiation in stems of seedlings of Capsicum annuum L. long fruited cultivar at 8-loaf stage of development. Prooambium, phloem and xylem of irradiated seedlings showed an early initiation and maturation in terms of distance from the tip. Magnitude of these differences increased with the increasing exposures and time following irradiation. In irradiated seedlings there is a ohange in the developmental sequence of metaphloem and metaxylem when they first appear (in terms of number of leaf primordia). There is also a general increase in the number of procambial cells, sieve elements and vessel elements after irradiation.
The early initiation and maturation of the vascular tissue is correlated with the growth inhibition resulting from cell death and suppression of cell division activity in the shoot apex.

The purpose of this paper is to determine the sequence of the formation of the lignified cell wall and to confirm the hypothesis expressed previously on the alternate deposition of cellulose and lignin in a 24-hour period. In this connection, the inner surface of the cell wall of the fibre cells from the youngest wood ofPopulus nigra L. 'Italica' taken during a typical summer day in two-hours' intervals was investigated electron microscopically.
The fine structure of the inner surface of the growing secondary cell walls was found to change regularly within 24 hours and to be determined by the structure of the components just being deposited. Cellulose and lignin were identified unambiguously, hemicelluloses could not be identified by their structure. The cellulose was characterised by the parallel microfibrillar structure; lignin was detected as an amorphous deposition.
The identified structural components are being deposited separately in certain intervals of the day cycle: cellulose in the afternoon hours (12 a. m. - 6 p. m.) and lignin after midnight (0 - 4 a.m.); in the morning hours (6-10 a.m.), neither lignin nor cellulose are deposited, however, the structure of the cellulose formed previously rises as a microrelief under the amorphous layer.
The alternate deposition of both structural components corresponds to variations of the biophysical and biochemical condition of the cells observed in the cambial zone during 24 hours. This periodicity appears to be autonomous and affected only partly by environmental factors. The results confirm the presumption that each lamella of the secondary wall consists of a cellulosic and a non-cellulose (lignin) part. Lignin permeates through the cellulosic part formed previously, impregnates it and cements it to the preceding lamella.

The paper presents results obtained from studying the linkages of the Rf₁ gene causing the restoration of fertility of pollen in the T type of cytoplasmic sterility of maize with signal genes of the 3rd chromosome in the F₂ and B₁ hybrid generations. Crossing of the fertility-restorer lines obtained on the basis of T cytoplasm with signal lines carrying recessive genes of the 3rd linkage group was carried out during the coupling phase.
The data obtained on the linkage of genes indicates that the Rf₁ gene is located at the proximal end of the long arm of the 3rd chromosome between loci d₁ and ba₁. The linear sequence of genes and the map of their distances were determined on the basis of experimental recombined values as follows: d₁-26-Rf₁-16(?)-ba₁-42-sh₂.

The sequence of intensities with which pollen tubes utilize tested sugars from their 0.3 M solutions, is the following: sucrose >glucose >invert sugar > >fructose. The same sequence is maintained in sugar-agar media with the exception of the first two hours of incubation when sucrose, glucose and fructose are all utilized at a practically equal rate. During this period no pollen tubes were formed in the presence of fructose, in common with the sugar-free control, while in the presence of sucrose they reached a length of up to 450 µ. If sucrose was used as carrier for radioactive sugars, fructose-¹⁴C was utilized up to 12 times and glucose-¹⁴C as much as 6 times more actively than sucrose-¹⁴C. When sucrose + glucose or sucrose + fructose (in molar ratio 1 : 1) were used the pollen tubes utilized sucrose-¹⁴C at a slower rate than the corresponding monosaccharide and also at a slower rate than observed with sucrose alone. When glucose or fructose was used as carrier for sucrose-¹⁴C . the carbon dioxide-¹⁴C production by the pollen tubes was (during several time intervals of the experiment) several tens per cent higher than when sucrose served as carrier. Fructose is utilized preferentially from a medium with invert sugar.
It thus appears that the capacity of the pollen enzymic systems for incorporating the sugars tested into their respiratory pathways is as follows: fructose >glucose > sucrose, this sequence being the opposite to that found for the intensity of their growth effect and to that established for the rate of their utilization, unless present in combination. The specific growth effect of sucrose cannot thus be due primarily to the rate of its absorption or to the intensty of its utilization. The rapid rate of utilization of sucrose alone is due to the more intense growth in the presence of sucrose. The results obtained further suggest that sucrose is utilized above all via its inversion when the fructose component is respired preferentially.

1. The aim of the present paper was an investigation of pollen grain size of different species of the Triticum L. natural polyploid series.
2. We determined the pollen grain size of 15 specimens belonging to 8 species. The determination was carried out on permanent acetocarmine preparations.
3. The pollen grain size of differentTriticum species tends to grow with the increasing number of chromosomes. The following mean volumes were computed for the different groups: 62.10³ μ ³ for the diploid group, 96.10³ μ ³ for the tetraploid group (of which 73.10³ μ ³ forT. timopheevi Zhuk.), and 118.10³ μ ³ for the hexaploid group.
4. As can be seen, the volume of the pollen grain does not represent a corresponding multiple of the chromosome number. The reason for this disagreement with the theoretically expected linear relation between the chromosome number and the pollen grain size lies apparently in the allopolyploidy of the polyploid series of the genusTriticum. It seems that the genetically different genoms A, B, D, and G also exert a different effect on the pollen grain size.
5. Our calculations show genom B to have 62%, genom D 29%, and genom G 18% of the effect of genom A (100%). These genoms form a sequence A>B>D>G with respect to their effect on pollen grain volume.