László Heszky

Novel male-specific molecular markers (MADC5, MADC6) in hemp

Decamer RAPD primers were tested on dioecious and monoecious hemp cultivars to identify sex-specific molecular markers. Two primers (OPD05 and UBC354) generated specific bands in male plants. These two DNA fragments were isolated, cloned and sequenced. Both markers proved to be unique, since no sequence with significant homology to OPD05961 and UBC354151 markers were found in databases. These markers were named MADC3 (OPD05961) and MADC4 (UBC354151) (Male-Associated DNA from Cannabis sativa). The markers were converted into sequence-characterized amplified region (SCAR) markers. The SCAR markers correlated with the sex of the segregating F2 population and proved the tight linkage to the male phenotype. Results of F2 plant population analysis suggest these markers are to be linked to the Y chromosome.

AFLP analysis and improved phytoextraction capacity of transgenic gshI-poplar clones (Populus x canescens L.) for copper in vitro

Abstract Clone stability and in vitro phytoextraction capacity of vegetative clones of P. x canescens (2n = 4x = 38) including two transgenic clones (ggs11 and lgl6) were studied as in vitro leaf disc cultures. Presence of the gshI-transgene in the transformed clones was detected in PCR reactions using gshI-specific primers. Clone stability was determined by fAFLP (fluorescent amplified DNA fragment length polymorphism) analysis. In total, 682 AFLP fragments were identified generated by twelve selective primer pairs after EcoRIDMseI digestion. Four fragments generated by EcoAGTDMseCCC were different (99.4% genetic similarity) which proves an unexpectedly low bud mutation frequency in P. \ canescens. For the study of phytoextraction capacity leaf discs (8 mm) were exposed to a concentration series of ZnSO4 (10-1 to 10-5 ᴍ) incubated for 21 days on aseptic tissue culture media WPM containing 1 μᴍ Cu. Zn2+ caused phytotoxicity only at high concentrations (10-1 to 10-2 ᴍ). The transgenic poplar cyt-ECS (ggs11) clone, as stimulated by the presence of Zn, showed elevated heavy metal (Cu) uptake as compared to the non-transformed clone. These results suggest that gshI-transgenic poplars may be suitable for phytoremediation of soils contaminated with zinc and copper.

Morphological and RAPD analysis of poplar trees of anther culture origin

Our objectives were to improve the rate of haploid plant regeneration through increasing the rate of callus initiation on the anthers and sustaining shoot regeneration frequency, and to analyze the field population of anther culture origin by morphological and molecular methods. Regarding the callus initiation, the most responsive clones were ‘N-90’(59%) in P. nigra and ‘D-29’ (75%) in P. deltoides. The rate of shoot regeneration and number of shoots/calli ranged from 4%–79% and 1–9, respectively. From the 208 rooted plants 8 haploid, 179 diploid, 4 tetraploid and 17 aneuploid plants were found. In the field population, the haploid plants could be easily identified by their retarded development and morphological characteristics (size and shape of the leaves, strong branching, etc). Several diploid plants showed depressed developmental and morphological traits similar to the haploid ones. Three traits (growth rate, leaf blade length and shape of leaf base) of the 6 different morphological characteristics measured were in correlation with the ploidy level within the poplar field population. Six primers of the 48 primers tested were able to detect polymorphism among the field plants.

Characterization of FaSPT ,a SPATULA gene encoding a bHLH transcriptional factor from the non-climacteric strawberry fruit

The involvement of basic-helix-loop-helix (bHLH) transcription factors in essential physiological and developmental processes is well established. Although a lot of animal bHLH proteins were characterized functionally, much less bHLHs of plant origin have been studied so far. Using a cDNA-AFLP approach, a ripening-related SPATULA gene was identified from strawberry fruit (Fragaria × ananassa Duch.), which encodes a bHLH protein. It is an orthologue of an Arabidopsis SPATULA protein, which has an important role in carpel and fruit development. Our experiments revealed that FaSPT is repressed by auxin in green fruits, and shows different expression patterns in receptacles at various stages of fruit ripening by ethylene treatment. Moreover, we applied a reverse genetic tool to elucidate the in planta function of FaSPT in early fruit development. To our knowledge, this work is the first report for the characterization of a SPATULA gene from a non-climacteric fruit.

DNA methylation and gene expression differences during alternative in vitro morphogenetic processes in eggplant (Solanum melongena L.)

SummaryIn vitro-cultured cotyledon explants of eggplant (Solanum melongena L.) are capable of producing somatic embryos or shoots, depending on which growth regulators are exogenously applied (auxins versus cytokinins). Rapidly growing suspension cultures were established to model these alternative morphogenetic processes. Clear differences in DNA methylation and gene expression patterns could be detected when suspension cultures grown under different growth regulator regimes were compared by restriction fragment length polymorphism (RFLP) or Differential Display analyses. Some of the random genomic clones showing methylation pattern differences, as well as some differentially expressed transcripts were analyzed in detail. One of the genomic clones, SMG56, proved to be the first identified 1-aminocyclopropane-1-carboxylate (ACC) synthase gene in eggplant. From 15 cloned. Differential Display fragments two somatic embryogenesis (SME39, SME50) and one organogenesis (SME88) related transcripts were identified.

Assessment of morphological and molecular similarity of Hungarian white grape varieties

The aim of this paper was to find possible link between molecular and morphological similarities of 38 Hungarian white grape varieties. Three aspects of morphological and molecular similarity were assessed in the study: comparison of the ordered variety pairs, assessment of molecular and morphological mean similarity differences and separation of varieties into similar groups by divisive cluster analysis to define (DIANA). Molecular similarity was calculated from binary data based on allele sizes obtained in DNA analysis. DNA fingerprints were determined at 9 SSR loci recommended by the European GrapeGen06 project. Morphological similarity was calculated on the basis of quantitative morphological descriptors. Morphological and molecular similarity values were ordered and categorized after pairwise comparison. Overall correlation was found to be weak but case by case assessment of the variety pairs confirmed some coincidence of molecular and morphological similarity. General similarity position of each variety was characterized by Mean Similarity Index (MSI). It was calculated as the mean of n-1 pair similarity values of the variety concerned. Varieties were ordered and compared by the difference of the index. Five varieties had low morphological and high molecular MSI meaning that they share several SSR marker alleles with the others but seems relatively distinct according to the expression of their morphological traits. Divisive cluster analysis was carried out to find similar groups. Eight and twelve cluster solutions proved to be sufficient to distinct varieties. Morphological and molecular similarity groups partly coincided according to the results. Several clusters reflected parent offspring relations but molecular clustering gave more realistic results concerning pedigree.

Grain quality and baking value of perennial rye (cv. ‘Perenne’) of interspecific origin ( Secale cereale × S. montanum )

The aim of the project was to determine the grain quality, technological properties and baking values of ‘Perenne’ (registered perennial rye cultivar), obtained through the interspecific crossing of Secale cereale L. and S. montanum Guss., and to compare with annual rye varieties. The crude protein content of ‘Perenne’ grains was higher (‘Perenne’: 18%, annual varieties: 12%) and contained more crude fibre, crude fat and ash than the annual varieties. Regarding the quantity and composition of amino acids, ‘Perenne’ showed values between S. montanum (high amino acid content) and S. cereale (lower amino acid content). While its farinograph softening value was inferior to those of the annual varieties, its flour mixed with wheat flour outperformed them. In terms of other properties (falling number, farinograph water absorption capacity, baking test) of ‘Perenne’ flour, whether in mixtures or in pure form, was not left behind the annual varieties. Perennial rye can also be used for bread making since it has g...