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The genetic variability of five populations of Quercus petraea originating from the Transylvania and Maramureș regions of Romania was investigated in this study to provide insights into the species’ adaptability, population dynamics, and potential for preservation in the face of environmental challenges. To achieve this, sequence-related amplified polymorphism (SRAP) markers, in conjunction with a set of 18 primer combinations, were employed. The outcomes of the analysis revealed a range of polymorphisms spanning from 69.78% to 85.75%. Additionally, the assessment of genetic diversity using Shannon’s information index (I) yielded values ranging between 0.2887 and 0.3955, while Nei’s gene diversity (He) exhibited a spectrum from 0.1833 to 0.2582. The analysis of genetic variability, conducted via molecular variance (AMOVA), unveiled that 9% of the genetic variation was attributable to differences among the populations, while a substantial 91% resided within the populations. A further investigation of the population structure revealed that the construction of a UPGMA dendrogram based on Nei’s genetic distances elucidated the presence of two principal genetic clusters, a finding that was reinforced by a Principal Coordinate Analysis (PCoA). The genetic diversity revealed by Quercus petraea using SRAP molecular markers offers promising potential for upcoming breeding programs to identify optimal genitors, facilitating the development of well-adapted oak populations in the Transylvania and Maramureș regions.

Morphological and molecular characterization of germplasm is essential for the improvement of cultivated plants efforts. This study investigated the genetic diversity of 32 Cyclamen genotypes comprising 16 C. persicum varieties and 16 Cyclamen species using multivariate analysis for 36 morphological traits (19 quantitative and 17 qualitative) and molecular characterization (SRAP and SCoT markers). The color CIELab parameters were collected via PCE-CSM7 that separately measured the leaves dark and silvery patterns and the flower’s slip (region of the petal top), eye (the region of the petal base) and sepal. Genetic diversity was also evaluated using Shannon Wiener (H′) and Simpson’s (λ) Indices, and Pilou evenness (J) using the library vegan from R software. According to the principal component analysis, the variables that contributed the most were leaf pattern color, leaf abaxial surface color, pedicel coiling, leaf and stem number. The color indicators of Cyclamen leaves showed decreased L* values in darker colored genotypes, whereas increased a* values were noticed in flower eye and lower in slip. Molecular characterization was based on 26 SRAP and 12 SCoT markers that produced clearly repeatable DNA bands and exhibited significant levels of polymorphism. Based on the morphological traits and molecular markers data, the UPGMA method for hierarchical clustering technique was used to generate the dendrograms, and their entanglement was obtained using the Tanglegram algorithm from the dendextend package with the R software. Entanglement analysis (0.30) between dendrograms obtained from the morphological and genetic analysis using SRAP markers showed a high association. Comparison between color measurements of flowers (entanglement=0.45) and leaves (entanglement=0.47) with SCoT analysis revealed differences at species level, discriminating between similar genotypes. Combined phenotypic and molecular analysis improved the comprehensive estimation of real diversity in the investigated Cyclamen genotypes. The findings of the present study are useful for quantifying diversity and genetic variability in Cyclamen breeding and genetic investigations.

Micropropagation has an important role in the large-scale production of blackberry plant material, given the high proliferation rates of this species. The aim of the present study was to evaluate the proliferative capacity of blackberry grown in vitro on wheat starch-gelled culture medium compared to classical agar-gelled medium and to assess the genetic fidelity between the proliferated shoots in starch-gelled culture medium and their mother plants. Six blackberry varieties (‘Čačanska Bestrna’, ‘Chester Thornless’, ‘Driscoll’s Victoria’, ‘Loch Ness’, ‘Polar’, and ‘Karaka Black’) were tested. For the in vitro shoots proliferation, Murashige and Skoog (MS) medium supplemented with 0.5 mg dm−3 6-benzyladenine (BA) was used. The conventional medium was gelled with 0.5% plant agar, and wheat starch was used as an alternative gelling agent in a concentration of 5%. The results showed that for all blackberry cultivars, the highest number of shoots/inoculum was obtained in wheat starch-gelled culture medium, with a maximum value of 54.42 ± 4.18 presented by ‘Karaka Black’. Considering the length of the proliferated shoots, all tested cultivars presented outstanding results on the culture medium gelled with 5% wheat starch. The highest values regarding shoots length were observed on the ‘Chester Thornless’ followed by ‘Čačanska Bestrna’, and ‘Loch Ness’ with values of 5.55 ± 0.04 cm, 5.46 ± 0.06 cm, and 5.37 ± 0.09 cm, respectively. The genetic uniformity of the micropropagated shoots in relation to their mother plants was confirmed by sequence-related amplified polymorphism (SRAP) and start codon targeted (SCoT) molecular markers.

In this work we aimed to develop an effcient micropropagation protocol for highbush blueberry (Vaccinium corymbosum L. ‘Duke’) and hazelnut (Corylus avellana L. ‘Tonda di Giffoni’) using the Plantform bioreactor. The influence of different quantities of the culture media (300 ml, 400 ml and 500 ml/bioreactor) was tested for both species. The immersion time was one at every hour lasting for 1 minute and aeration was once/hour lasting for 4 min each. Our results show that highbush blueberry had the highest proliferation rate (6.20±0.81) in the bioreactor containing 500 ml of the culture medium while the longest shoots (4.19±0.25 cm) were recorded in the bioreactor with 400 ml of the culture medium. Hazelnut had the highest proliferation rate (0.93±0.07) in the bioreactor containing 400 ml while the longest shoots (3.13±0.49 cm) were measured in the bioreactor containing 500 ml medium. The highest rooting percentage (77.78%) in hazelnut was recorded in the bioreactor containing 400 ml medium.

The aim of this work was to determine the effect of zeatin (Z), dihydrozeatin (DHZ) and 2-isopentenyl-adenine (2 iP) on shoot length and proliferation rate (PR) in three Vaccinium corymbosum L. cultivars: ‘Blueray’, ‘Duke’ and ‘Patriot’. The culture medium used in this research was prepared from stock solution: Woody Plant Medium (WPM) supplemented with 5 mg/l2iP, 0.25 mg/l Z, 0.5 mg/lZ, 0.25 mg/l dihydrozeatin (DHZ), 0.5 mg/l DHZ and a combination of 0.5 mg/lZ + 0.25 mg/l DHZ. The results show that the lowest PR (1.26±0.03) was recorded in ‘Duke’ on the medium containing 0.25 mg/l DHZ, while the highest PR (4.26±0.10) of this variety was recorded on the medium supplemented with a combination of 0.5 mg/l zeatin and 0.25 mg/lDHZ. The highest average length of the shoots (3.82±0.07) recorded was also on this medium. Among the treatments, the highest PR (3.66±0.07) induced by a single cytokinin was obtained on WPM+5 mg/l2iP. Similar results were obtained in ‘Blueray’ undergoing the same treatment with hormone combinations (0.5 mg/l zeatin and 0.25 mg/lDHZ); the highest PR recorded was 4.1±0.23. On the contrary, ‘Patriot’ showed the highest proliferation rate (5.06±0.11) on the medium supplemented with 5 mg/l 2iP, but the average length of the shoots was slightly lower (4.21%) than that on the medium with hormone combinations.

Nowadays, the field of entirely artificial hybrids raises ethical problems in the animal world and to a lesser amount in plants. Throughout the years, yellow Cyclamen has been particularly important for both breeders and passionate growers as being a peculiar color for this species. The possibility to artificially induce hybrids between species that can never normally cross it’s now achievable. This paper describes the possibility of obtaining high ornamental yellow flowered cyclamen, through chromosome doubling. The pollen and seed sterility can be overcome by doubling the chromosomes. In this sense, there are two full sets from each parent, resulting in a fertile hybrid, by introducing the in vitro culture into colchicine supplemented medium.

Cyclamen genus is part of the Primulaceae family consisting of 24 species widely cultivated as ornamental and medicinal plants. They also possess high plasticity in terms of adaptability to alternating environmental conditions. In this regard, the present study investigates the germination and morphological parameters of heat-tolerant and heat-sensitive Cyclamen persicum accessions in the presence of different GA3 solutions (0, 30, 70 and 90 mg/L) under ambient temperature and heat stress conditions. Heat-tolerant genotypes, mainly C3-Smartiz Victoria (6.42%), C15-Merengue magenta (6.47%) and C16-Metis silverleaf (5.12%) had the highest germination rate with 90 mg/L GA3 treatment compared with control. Regarding heat-sensitive genotypes, C11-Verano (5.11%) and C13-Metis Origami (4.28%) had the lowest values in mean germination time, along with the Petticoat genotypes C1 (73.3%) and C2 (80.0%) with a high germination percentage. Heat-tolerant genotypes positively responded to GA3 (70 and 90 mg/L) even under heat stress conditions, by their higher values in plant height, an ascending trend also seen in heat-sensitive genotypes under GA3 treatment (70 and 90 mg/L). According to the hierarchical clustering, several heat-tolerant genotypes showed peculiar behavior under heat stress conditions, namely C3 (Smartiz Victoria), C7 (Halios falbala) and C8 (Latinia pipoca) which proved to be susceptible to heat stress even under GA3 application, compared with the other genotypes which showed tolerance to higher temperatures. In the case of heat-sensitive genotypes, C4 (Smartiz violet fonce), C6 (Metis blank pur), C11 (Verano) and C13 (Metis origami) possessed higher positive or negative values compared with the other heat-sensitive genotypes with increased doses of GA3. These genotypes were shown to be less affected by heat stress, suggesting their positive response to hormone treatment. In conclusion, the above-mentioned genotypes, particularly heat-tolerant C15 and heat-sensitive C2 with the highest germination capacity and development can be selected as heat-resistant genotypes to be deposited in gene banks and used in further amelioration programs under biotic and/or abiotic stresses to develop resistant genotypes.

The present study was carried out to evaluate and compare in vitro antioxidant (2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC), and ferric reducing antioxidant power (FRAP)), antimicrobial, anticancer activities, and the individual carotenoids and anthocyanins content of methanol extracts of the Cyclamen genotypes: Persian cyclamen accessions (Cyclamen persicum Mill.), sowbread (C. mirabile Hildebr.), and ivy-leaved cyclamen (C. hederifolium Mill.) aerial parts. The HPLC-PDA analysis revealed the presence of five individual carotenoids (i.e., neoxanthin, violaxanthin, lutein, β-carotene, and cis-β-carotene) as the main compounds in Cyclamen leaves, and the presence of seven individual anthocycanins (i.e., cyanidin 3,5-di-O-glucoside, peonidin-rutinoside, peonidin 3,5-di-O-glucoside, peonidin 3-O-glucoside, malvidin 3-O-glucoside, malvidin 3,5-di-O-glucoside, and malvidin-rutinoside) in Cyclamen flowers reported, hereby, for the first time. The highest phenolic content was found in the leaves of LC6, C. mirabile (46.32 ± 0.14 mg/g gallic acid equivalents [GAE]), and in the flowers of C. persicum Merengue Magenta (FC15) (58.63 ± 0.17 mg/g GAE), whereas the highest flavonoid content was reported in C. persicum Halios Falbala leaves, namely LC9 (54.90 ± 0.27 mg/g quercetin equivalents [QE]) and in flowers of C. persicum Victora (FC2) (77.87 ± 0.25 mg/g QE). The highest antioxidant activity in DPPH and FRAP assays was reported in C. persicum Dark Violet (LC1) and Victoria (LC2), whereas C. mirabile (LC6) had the highest activity in the TEAC assay. In flowers, high antioxidant activities in DPPH and TEAC were noticed in C. persicum Superserie Red (FC7) and Dark Violet (FC1), respectively, and Halios Falbala (FC9) exhibited the highest activity in the TEAC assay. Additionally, FC9 exhibited the highest antibacterial activity in almost all tested bacteria compared with the leaves extracts. Furthermore, the highest in vitro citotoxicity in MDA-MB-231 cells was noticed in C. hederifolium LC18 (56.71–69.35%) and FC18 (40.07–41.43%), with a lower effect against BJ cells demonstrating selective toxicity. The above findings, highlight the potential use of the Cyclamen flower and leaf extracts as significant anticancer agents along with their antioxidant and antimicrobial properties.

Nowadays, the interest to essential oil-bearing plants has increased significantly due to their therapeutic and industrial potential. Therefore, the main aim of this study was to examine and confirm the presence of essential oils in seven different Ranunculaceae species from Romania by hydrodistillation. The results show that five (Aconitum toxicum, Aconitum variegatum, Hepatica nobilis, Ranunculus carpaticus, and Ranunculus polyanthemos) out of the seven investigated species contain volatile oils isolated for the first time from aerial parts. The hydrodistillation of the above-mentioned species resulted in a yellow to cloudy white colored essential oil with a waxy texture and yields ranging from 0.013 to 0.245 %. Our results are in accordance with some previous studies that have revealed similar quantities of essential oils in other taxa belonging to the Ranunculaceae family.

The climatic data of the last years presents a significant growth of temperatures even in the cultivation areas of two-rowed spring barley. These intervals of drought appear especially in the period of grain formation and filling, bearing a negative impact on production, both from the qualitative and quantitative points of view. Thus, in order to simulate the drought phenomenon, the method proposed by Blum (1983), cited by Elena Petcu and colab., (2014), was used. This method consists in treating the plants with desiccants (e.g. Sodium chlorate) in a 2% concentration, 14 days after the anthesis. Thus, the cultivars that manage to translocate a greater quantity of reserve carbohydrates from the vegetative organs towards the grains, present a better stability of grain weight under conditions of hydric stress.

Astragalus exscapus L. subsp. transsilvanicus (Schur) Nyár. (Fabaceae) is a rare plant endemic to the Transylvanian Plateau, represented by 24 identified populations. Limited information on the genetic variation and population structure is available, which obstructs efficient measures for conservation strategy. The present study aimed to analyze the genetic diversity and population structure of eight populations of A. exscapus subsp. transsilvanicus revealed by sequence-related amplified polymorphism (SRAP) markers. A total of 164 bands were amplified, 96.7% of which (159) were polymorphic. Nei’s gene diversity index (He) was estimated to be 0.228 at the population level and 0.272 at the subspecies level. The genetic differentiation among populations (Gst) was 0.165, which indicated a low proportion of total genetic diversity. The analysis of molecular variance (AMOVA) indicated that 17% of the total variation of A. exscapus subsp. transsilvanicus is found among the populations, while 83% was found within the populations. A UPGMA dendrogram, principal coordinate analysis, and the STRUCTURE software grouped the populations into two clusters uncorrelated with the provenience of the 125 individuals, which might be attributed to fragmentation processes, insect pollination, population size, and specific environmental conditions of the habitats.

The main aim of this study was to investigate the genetic diversity and phylogenetic relationships of six Ranunculus species using SRAP markers. The genotyping of 73 Ranunculus accessions showed that 12 primer pairs generated a total number of 212 polymorphic fragments with a 99.53 % level of polymorphism. Nei’s gene diversity ranged between 0.2237 and 0.3436 with an average of 0.3001. The UPGMA method with Nei72’s similarity coefficient ranging from 0.23 to 1.11 separated 73 accessions into two distinct groups (A and B). Cluster-A included R. acris members, while cluster-B included the rest of the tested species. Cluster-B was subsequently divided into two sub-groups (I and II). Subgroup-I consisted of R. carpaticus and R. serpens subsp. nemorosus accessions, while subgroup-II included the members of R. platanifolius, R. polyanthemos, and R. repens. R. polyanthemos and R. repens were founded the most closely related species. These findings provide a brand-new understanding of the evolutionary relationships of above mentioned Ranunculus species and may be useful tools for plant breeding and conservation purposes.

A micropropagation protocol via direct shoot organogenesis from Streptocarpus x hybridus Voss. leaf explants was established in this study. The shoot induction of three Streptocarpus cultivars (‘Snow White’, ‘Black Panther’ and ‘Slumber Song’) was successfully achieved on Murashige and Skoog (MS) medium supplemented with 0.2 mg L-1 -indole-3-acetic acid (IAA) and 0.2 mg L-1 thidiazuron (TDZ). In proliferation stage, the effects of two combinations of plant growth regulators -PGR- (V1-0.2 mg/L-1 IAA + 0.5 mg/L-1 BAP and V2-1.0 mg L-1 NAA + 0.2 mg L-1 TDZ) on shoot number and length were examined. The results suggest that PGRs combinations significantly influenced shoot proliferation and root induction in all Streptocarpus cultivars. Among the treatments, 0.2 mg L-1 (IAA) in combination with 0.5 mg L-1 6-benzylaminopurine (BAP) were the most effective for in vitro shoot multiplication and rooting. The in vitro rooting percentage was also determined before subjecting the plantlets to the acclimatization process. Due to acclimatization, Streptocarpus plantlets showed a very high rate of survival (90%). The generated PCR-RAPD profiles for the selected in vitro-raised plants and donor plants were similar which indicates the clonal or true-to-type nature of the progenies.

In Romania, Quercus is represented by five to nine species, depending on their taxonomic classification. The purpose of this study was to determine the seed germination rate and seedling growth of Quercus robur and Quercus petraea species in different soil conditions. The results obtained show that there are differences between trees growing and development. Our results show that seed quality and lithological substrate have an influence on seed germination of Quercus robur and Quercus petraea species. This fact has an important role in forest resource management.

Micropropagation of Lycium barbarum cv. 'Ningxia N1' was achieved. The cultures were by initiated by axenical seed germination. The highest shoot proliferation was obtained on the MS media with 1.33 or 2.22 µM benzyl adenine, gelled with wheat starch as an agar alternative. The treatments with 2.22 µM benzyl adenine ensured proliferation rates superior to the ones with 1.33 μM benzyl adenine, but the latter provided longer and more robust shoots. Use of large microcuttings as an explant onto the multiplication media ensured higher in vitro explant survival, higher number of shoots regeneration and more vigorous plantlets. The microcuttings inserted vertically into the media yielded superior growth and multiplication as compared to the microcuttings placed horizontally. The non-rooted, elongated shoots from the treatment 1.33 μM benzyl adenine were either rooted in vitro on a hormone-free MS medium with starch or used for direct ex vitro rooting and acclimatization. The optimal number of microcuttings/vessel for in vitro rooting was 40 and the rooted plantlets were efficiently acclimatized ex vitro by three methods: float hydroculture in floating cell trays, floating perlite, and in Jiffy7 pellets.

The chemical composition of Romanian table grape varieties was determined to evaluate their phenolic and aroma profiles because they are factors useful for the variety characterization and consumer acceptance. Two white table grapes (‘Aromat de Iaşi’ and ‘Timpuriu de Cluj’) and two red table grapes (‘Napoca’ and ‘Someşan’) were studied. The phenolic composition of berry skins and seeds was determined using spectrophotometric methods. The profile of anthocyanins and hydroxycinnamoyl tartaric acids (HCTs), and the content of trans-resveratrol were determined in the skins using high-performance liquid chromatographic (HPLC) methods. Free volatile compounds were quantified by gas chromatography/mass spectrometry (GC/MS). A different phenolic and aromatic composition was found in the varieties studied. Among white grapes, ‘Aromat de Iaşi’ stands out for the great number of aroma compounds, and ‘Timpuriu de Cluj’ showed high contents of polyphenols, particularly oligomeric flavanols in skins and seeds (1,171 and 1,189 mg kg-1, respectively) and total skin HCTs (181.6 mg kg-1). The red variety ‘Napoca’ had high contents of total anthocyanins (380 mg kg-1) and total skin HCTs (183.2 mg kg-1), and it is characterized by high percentages of acylated anthocyanin derivatives (28.5%) and trans-caffeoyltartaric acid forms (58.8%). ‘Someşan’ grapes showed high contents of free volatile compounds, particularly of 1-hexanol, (Z)-3-hexen-1-ol and (E)-2-hexenoic acid (167.0, 59.4 and 167.0 µg kg-1). These results may contribute to the knowledge of Romanian table grapes perspective and to a better exploitation of these varieties.

Cyclamen species (swinebread spp.) are perennial herbaceous plants which belong to the Primulaceae family, widely used as medicinal or ornamental plants. Currently, gibberellic acid (GA3), an environmentally friendly bioregulator is widely used to enhance the productivity and phenotypic characteristics of multiple ornamental crops. A dual experiment was conducted to evaluate the influence of gibberellic acid (50, 100 or 150 mg/L), on germination earliness of seeds, along with growth and ornamental quality of selected Cyclamen species. Additionally, a greenhouse experiment with long and short sunlight photoperiod was conducted to evaluate its importance in the development of Cyclamen plants. Our results indicated that under short sunlight exposure, seeds germinated earlier than those kept in long sunlight photoperiod. Plants treated with foliar applications of GA3 exhibited higher plant height, leaf area, number of leaves, root length and tuber development. Significant differences were found in germination percentage (GP), mean germination time (MGT) and seedling vigor index (SVI), within different GA3 supplementation and greenhouse sunlight intensity (short and long days). In conclusion, our study suggests that seedling vigor and ornamental quality can be increased by GA3 application, an economic strategy for enhancing germination and growth of selected Cyclamen species.

The genetic relationship among three Streptocarpus parents and twelve F1 hybrids was assessed using sequence-related amplified polymorphism (SRAP) molecular markers and Fourier-transform infrared (FT-IR) spectroscopy. Both methods were able to discriminate F1 hybrids and parents as revealed by cluster analysis. For hybrid identification, the type III SRAP marker was the most effective due to the presence of male-specific bands in the hybrids. Different behaviors in the biochemical variability of DNA samples have been observed by FT-IR spectral analysis, which might be attributed to the inherent nature of the genomic DNA from parents and their F1 progenies. Mantel test was also carried out to compare morphological, SRAP, and FT-IR results based on genetic distances. The highest correlation coefficient was found between morphological and SRAP marker distances (R = 0.607; p ≤ 0.022). A lower correlation was observed between the morphological and FT-IR distance matrix (R = 0.231; p ≤0.008). Moreover, a positive correlation was found between the distances generated with SRAP and FT-IR analyses (R = 0.026) but was not statistically significant. These findings show that both SRAP and FT-IR techniques combined with morphological descriptions can be used effectively for nonconventional breeding programs for Streptocarpus to obtain new and valuable varieties.

It is reported that probiotics provide several health benefits as they help in maintaining a good balance and composition of intestinal flora, and increase the resistance against invasion of pathogens. Ensuring adequate dosages of probiotics at the time of consumption is a challenge, because several factors during processing and storage affect the viability of probiotic organisms. Major emphasis has been given to protect the microorganisms with the help of encapsulation technique, by addition of different protectants. In this study, probiotic cells (Bifidobacterium lactis 300B) were entrapped in alginate/pullulan microspheres. In the encapsulation formula glycerol was used as cryoprotectant in the freeze drying process for long time storage. It was observed that the survival of Bifidobacterium lactis 300B when encapsulated without cryoprotectant was higher than the formula with glycerol in the fresh obtained microspheres. The addition of glycerol was in order to reduce the deep freezing and freeze drying damages. In the chosen formulations, glycerol did not proved protection for the entrapped probiotic cells in the freeze drying process, for which the use of glycerol as cryoprotectant for alginate/pullulan Bifidobacterium lactis 300B entrapment is not recommended.

In order to assess the effect of gelling agents upon lilac in vitro proliferation rate, eight types of gelling agents were tested, at concentrations considered to be optimal, based on preliminary tests as follow: 3 g/L Fibrous Agar, 6g/L Daishin Agar, 4 g/L Gelcarin, 2 g/L Gelrite, 15 g/L Isubgol, 4 g/L Micro Agar, 6 g/L Phyto Agar and 4 g/L Plant Agar. The culture media was Murashige & Skoog (MS) supplemented with 4 mg/L 6-benzylaminopurine (BAP). The plant material consisted of shoots picked from in vitro cultures on MS+0.5 mg/L BAP gelled with Plant Agar. The highest multiplication rate was 3.76, obtained in the experimental treatment with the media gelled with 15 g/L Isubgol, while the lowest proliferation rate was 1.4, on the media gelled with Daishin Agar. The shoots obtained in these eight experimental treatments were rooted ex vitro and acclimatized using the floating perlite method, in two experimental treatments: one with hormone-free floating perlite and one supplemented with 1mg/L indole-3-butyric acid (IBA). The highest rooting and acclimatization percentage (90%) was obtained in the hormone-free floating perlite experimental treatment, from shoots obtained in the media gelled with 6 g/L Daishin Agar.

The aim of our research was to investigate the way in which the agar types used for culture media preparation influence damask rose in vitro proliferation rates, as well as the quality of the shoots obtained and their ability for ex vitro rooting and acclimatization. For this purpose, seven types of gelling agents were tested: 6 g/L Daishin Agar, 4 g/L Gelcarin, 2 g/L Gelrite, 15 g/L Isubgol, 4 g/L Micro Agar, 6 g/L Phyto Agar and 4 g/L Plant Agar. The culture media consisted of Murashige & Skoog supplemented with 4 mg/L 6-benzyladenine. The highest multiplication rate, 5.1, was obtained on the culture media supplemented with 6g/L Daishin Agar and 4 g/L Plant Agar and the lowest one was 2.8, obtained on the media gelled with 4 g/L Gelcarin. Although the multiplication rate on the media gelled with Isubgol was lower (3.2), the shoots obtained in this experimental treatment got acclimatized with 90 % survival percentage and developed the best root system, regarding the number of roots/shoot and root length.

Conservation agriculture is an alternative to conventional agriculture and one of the most efficient systems for sustainable agricultural development, stimulating soil biological activity, increasing organic matter and humus content. In order to evaluate the impact of the conservation agriculture system compared to the conventional agriculture system on wheat yield, a bi-factorial experiment on wheat crop based on split-plot model was organized in the Transylvania Plain, Romania, where there are a few research results in the field and the area has a high potential for the implementation of conservation agriculture system. First factor (‘A’) was agriculture system, having two levels: ‘A1’ tillage (classic) system and ‘A2’ no-tillage system, and the second, ‘B’ factor was fertilization, with three levels: ‘B1’ no fertilization, ‘B2’- N80P40 kg/ha, which is the average level practiced by farmers in the area and ‘B3’- N150P75 kg/ha, which is the recommended level for obtaining a high production performance specific to ‘Ariesan’ wheat cultivar used in this experiment. The results pointed out that, in comparison with conventional agriculture, the application of no-tillage system determined a lower wheat yield by 353 kg per surface unit. Fertilization assured a significant increase by 610 kg wheat grains per ha. In the case of the conservation agriculture system, the highest production gain was 1,260 kg/ha for ‘B3A2’ level compared to ‘B1A2’ level, where the highest fertilization level was applied. The lowest production gain was 410 kg/ha registered by ‘B3A1-B1A1’ which was the highest fertilizer dose in case of conventional agriculture. In conclusion, the application of no-tillage system increased the efficiency of fertilization in terms of wheat yield compared to the classical agriculture. As a result, it is recommended as farmers dealing with wheat cropping in the area and also in other regions with similar soil and climate conditions to implement conservation agriculture.

The paper aimed at making a comparison between conservation agriculture and conventional agriculture on wheat yield, in a three crop rotation (wheat, maize and soybean) system. A three factorial experiment based on the split-plot model and variance analysis was set in the Transylvania area of Romania, as follows: A factor-agriculture system: A1-tillage, A2-no-tillage, B factor-fertilization: B1-N40P40 kg/ha. B2-N40P40 kg/ha+N50P30 kg/ha; C factor- treatments: C1-4 treatments, C2-3 treatments, C3-2 treatments at heading, C4-2 treatments at blooming. Fertilization applied in its B2 variant in conservation agriculture brought 460 kg/ha production gain, significantly and statistically ensured. Under conservation agriculture, two treatments applied at blooming determined 318 kg/ha production gain, statistically and significantly covered. The interaction fertilization-treatments produced the highest wheat yield 5001 kg/ha in case of the B2C3 variant, in conventional agriculture, and 5272 kg for the same variant in conservation agriculture, the difference of 953 kg compared to the central variant B1C1 being statistically ensured. Under the conditions of Transylvania, farmers could adopt conservation agriculture as a solution for increasing wheat production, reducing the number of treatments and applying more fertilizer upon blooming. The advantage of the implementation of such a system resides in its beneficial effect on soil structure, water reserve and biodiversity, as well.

The potato (Solanum tuberosum) is the 3rd most important crop worldwide, and Phytophthora infestans is the most devastating pathogen to potato crops. In this study, it has identified markers for resistance and susceptibility to late blight using potato genotypes that differ in their resistance/susceptibility to Phytophthora infestans. Using Fourier transform infrared spectroscopy, many absorbance bands have been identified as specific to resistant potato plants, and others were specific to susceptible potato plants. For each case (resistance/susceptibility), three bands were identified: 941-1180 cm-1, 1336-1483 cm-1, and 1483-1703 cm-1 and 1056-1294 cm-1, 1442-1585 cm-1, and 1585-1832 cm-1, respectively. As it is a simple, rapid, and inexpensive technology, FTIR (Fourier Transform Infrared) spectroscopy offers an excellent opportunity for studying potato resistance/susceptibility to late blight.

The somatic embryogenesis is an advanced method for clonal propagation and a useful tool for ex situ conservation of genetic resources. In this paper, the results of an experiment to investigate the influence of development stage of explants and culture medium on the germination percentage in two oak species (three provenances of Quercus robur and two provenances of Q. frainetto), are presented. A high significant influence of the development stage of explants and a significant influence of the interaction provenance x stage on the germination percentage were recorded for Q. robur explants, whilst no significant differences between the germination percentages against the nutritive media used were fould for both oak species.

Adventitious rooting is a complex process and a key step in the vegetative propagation of economically important woody, horticultural and agricultural species, playing an important role in the successful production of elite clones. The formation of adventitious roots is a quantitative genetic trait regulated by both environmental and endogenous factors. Among phytohormones, auxin plays an essential role in regulating roots development and it has been shown to be intimately involved in the process of adventitious rooting. Great progress has been made in elucidating the auxin-induced genes and auxin signaling pathway, especially in auxin response Aux/IAA and Auxin Response Factor gene families. Although some important aspects of adventitious and lateral rooting signaling have been revealed, the intricate signaling network remains poorly understood. This review summarizes some of the current knowledge on the physiological aspects of adventitious root formation and highlights the recent progress made in the identification of putative molecular players involved in the control of adventitious rooting. Despite much has been discovered regarding the effects and regulation of auxins on plant growth since the Darwin experiments, there is much that remains unknown.

In this study we used the Fourier-Transformed Infrared (FT-IR) technique to examine the compatible reaction of potato (Solanum tuberosum) to infection by the late blight agent Phytophthora infestans. Three virulent isolates have been used, different by their level of pathogenicity on R2 potato. The response was dependent on the pathogenicity of the isolate. The Infrared spectra in the middle infrared region (MIR) of infested versus healthy (control) leaves showed that controls absorb (intensity) mostly in the range 650-950 cm-1.The absorbance was higher for the most virulent isolate in the range 950-1650 cm-1. In the domain 3000-4000 cm-1 for the control and less pathogenic interactions, have absorbed more than the highly infested samples. The shape of spectra has generally been the same shape. However, the number of peaks suppressed/induced after inoculation and in function of the isolate differs indicating a perturbation of plant metabolsim. FTIR technique provides important information about the events occurring during the interaction.

Conference Information: 12th International BIOVEG Symposium Plant Biodiversity and Food Diversification Place: University of Agricultural Sciences and Veterinary Medicine, September 27-29, 2010 Cluj-Napoca, Romania The aim of the BIOVEG Symposium is to present the latest scientific results on all aspects of research in plant biotechnology. Various tools are now available to describe the biological diversity of plant species useful for food diversification. They allow fine analysis of basic components, but also their interactions, using different and complementary disciplines such as genetics, microbiology, biochemistry and physiology. The involvement of Plant biodiversity and Food diversification in one symposium is not a common process. However, the FAO data (2008) relevant to the state of the food worldwide are clear on at least two main points: I) there is a well defined relationship between quality of diet and dietary diversification; II) the most affected countries are India, countries in sub-Saharan Africa, the Pacific area of Asia and China as they have a high potential of biodiversity. If food is a vital necessity, the overall diet is different between rich and poor countries. In poor countries, cereals, roots and tubers cover 66% of eaten plants. In rich countries, fruits and vegetables play a much larger role (19% vs. 11%). In rich countries, attention is increasingly paid to the nutritional quality and food intakes for a better life. In poor countries, it is foremost eat to survive. Nearly 900 million peoples suffering from hunger in the world but 90% of them live in developing countries. India (231 MH) is at the head, followed by sub-Saharan Africa (212 MH), Asia - Pacific (189 MH) and China (123 MH). The rest of the world has barely 100 MH malnourished peoples. However, the Pacific area of Asia provided the world in many domesticated plant species (Citrus, numerous fruits and vegetables). Tools to describe biodiversity of plant species for food diversification have made many advances in recent years. New technologies allow deep and detailed analysis of the biodiversity and quality of the food supply. Several different and complementary disciplines such as genetics, microbiology, biochemistry and physiology are involved. Beyond the simple description step, it is to improve products or value-added varieties and also to maintain traditional pools of genetic diversity for tomorrow. We must also appeal to disciplines such as plant breeding or the wide range of plant biotechnologies to generate new diversity aspects and to preserve existing genetic resources. This requires of course a broad collaboration of local people, the best able to defend their specificities. In this context, 2010 Conference of Scientific Journeys of the BIOVEG network was organized around five main sessions: 1. Genetic resources, local domestication, plant breeding, cytogenetics. 2. Plants - micro-organisms interaction which include phytopathology, symbiotic relationship, toxins impacts. 3. Final biochemical composition, antioxidants analysis, lipid contents and composition, metabolic pathways regulations. 4. Non-conventional diversity creation methods such as induced mutations. 5. Ex situ and in situ genetic resources conservation.

Seabuckthorn is a spinescent, deciduous and dioecious berry-producing shrub, with a high economical and ecological potential. It is frequently used as a pioneer species in anthropic and eroded soils due to its low pedoclimatic demands, strong rooting system and ability to fix atmospheric nitrogen. Seabuckthorn berries, leaves and bark have a high content of nutritive and active substances which promote this species for use in both food and medical industries. One of the most requested therapeutical products on the market is the seabuckthorn oil, extracted from both pulp and seeds. Two important parameters in analyzing seabuckthorn oil quality are fatty acids and tocopherols. In Transylvania region most of seabuckthorn orchards are established with local, low productive and less uniform planting material, randomly collected from wild flora. In order to assess the opportunity of introducing new seabuckthorn varieties in Transylvania, a selection process was initiated. In this context, five elites were selected from wild populations in the Danube Delta, using biometrical criteria. They were later compared to a representative individual from a local population and to a number of homologated cultivars, with respect to morphology and some lipophilic components (oil content, fatty acids and tocopherols). For both pulp and seeds, total lipids were extracted using a modified Folch method. Fatty acids were analyzed by gas chromatography using flame ionization detection. Tocopherols were analysed using a Shimadzu VP Series liquid chromatograph with a fluorescence detector FR-10 AXL.

Romanian hazelnut (Corylus avellana) germplasm is held in a national collection at SCDP Valcea. A clear situation of the held accessions is necessary for an efficient management of the germplasm collection. In order to achieve this, the genetic variability of 43 accessions was assessed using 23 RAPD primers. The RAPD analysis was carried out as a screening test to confirm the genetic identity of some accessions. Based on the screening results, 12 accessions were selected for analysis using nine SSR primers. A high level of genetic diversity was observed (He=0.75, Ho=0.81, F=-0.061) among the analyzed samples. A genetic similarity matrix was constructed and the resulting UPGMA dendrogram revealed three major groups, corresponding to the geographical origin of the accessions. In order to increase the effectiveness of genebank management, the identification of duplicate and mislabeled accessions with the aid of molecular markers is of high interest, especially being the first one of this kind in a Romanian hazelnut germplasm collection.

In order to use markers for the selection of wheat resistance to the common bunt, there were tested certain wheat microsatellite markers (SSRs) on certain local accessions of bread wheat. A number of eleven SSR markers were tested on parental forms (five lines of bunt resistant wheat - Bt5, Bt8, Bt10, Bt11 genes and a number of eight cultivars and lines susceptible to common bunt. From the eleven tested markers, the Xgwm633 marker and Xgwm114 marker identified polymorphism on parental forms. Relying on co-segregation method, the Xgwm633 marker revealed a polymorphic fragment of 200 bp on the resistant wheat line 99419G4-1A/1-1 and a 230 bp fragment on the susceptible cultivar Glosa. So the fragment of 200 bp obtained with this marker could be assigned to some common bunt resistant genes. The Xgwm114 marker revealed a polymorphic fragment of 120 bp on resistant wheat line 00274G2-31, harboring the Bt11 resistant gene, and two fragments of 160 bp and 180 bp on susceptible cultivar Glosa. So the fragment of 120 bp obtained with this marker could be assigned to Bt11 resistant gene. That result was also confirmed by BSA (Bulked Segregant Analysis) method. The linkage relationship between markers and resistant genes was evaluated on the F2 populations resulted from the cross between 99419G4-1A/1-1 and 00274G2-31 the wheat resistant lines with the susceptible cultivar (Glosa). The value of recombination between the Xgwm633 marker and a common bunt resistant gene was evaluated at 32 cM. A smaller value of recombination has been estimated between the marker Xgwm114 and the resistant gene (8 cM).

Interspecific hybridizations represent one of the apple breeding methods by which a wide variability can be achieved, useful for creating new cultivars. The study of 2190 interspecific hybrids, obtained from 25 combinations among crab apple species (Malus coronaria, M. floribunda, M. niedzwetzkyana, M. zumi, M. prunifolia) and different apple cultivars, points out a large variability of the F1 seedlings for several traits, with significant importance in apple breeding programs. The first year of fructification, as mean per hybrid combination, varied from 6 (M. zumi x ‘Jonathan’) to 9.3 years; the average hybrid’s age for fructification was 7.4 years. The size of fruits among families varied from 1.5 (‘Starkrimson’ x M. prunifolia) to 4.0 (‘Reinette Baumann’ x M. zumi), the mean being settled at 2.8 (therefore below mark 3, meaning ‘small fruits’). The lowest infection rate both for apple scab and powdery mildew attack was noticed at hybrids from M. coronaria x ‘Reinette Baumann’. On the whole, the hybrids with genitors of ‘species x cultivars’ type have had an early fruiting and a better resistance to scab and powdery mildew, compared to the ones from ‘cultivars x species’ combinations. The large variability of the studied traits gave the possibility to identify offsprings with desirable characteristics on nine hybrid combinations. Among these, 53 elite plants were selected, with a strength of selection of 2.42%, therefore a relative low value, correlated with the peculiarities of the interspecific population and the selection criteria for dessert apple.

Plum pox virus (PPV) is considered the most destructive viral pathogen of stone fruits. Although PPV is widespread in all plum growing areas from Romania and causes serious yield losses, little is known about the virus variability. For this reason we investigated 100 PPV isolates collected from 13 different plum orchards in the Transylvania - Romania. PPV detection was made by DAS-ELISA and by IC-RT-PCR. PPV strains were serologically determined by TAS-ELISA using PPV-D and PPV-M specific monoclonal antibodies. Molecular strain typing was done by RT-PCR targeting three different genomic segments corresponding to (Cter)CP, (Cter)NIb/(Nter)CP and CI. RFLP analysis was used to distinguish the two major strains, D and M based on a RsaI polymorphism located in (Cter)CP. All PCR products targeting (Cter)CP and 10 PCR products spanning the (Cter)NIb/(Nter)CP were sequenced. All PPV isolates typed as PPV-M by serological analysis and by molecular differentiation in the genomic region corresponding to (C-ter)CP proved to be PPV recombinant (PPV-Rec) when the molecular analysis were performed in the region corresponding to NIb/CP. Sequencing confirmed a high similarity with different sequences of PPV-Rec previously reported. All these recombinant isolates share the same recombination breakpoint and conserve the DAG motif, which is considered essential for aphid transmission. Overall results provided that in Transylvania the predominant strain is PPV-D, followed by PPV-Rec which shares the CP gene with M strain and, therefore, it is serologically detected as PPV-M with M-specific monoclonal antibodies. In this big plum growing area, mixed infections (PPV-D+PPV-Rec), which might generate additional variation by recombination, are also frequent.

Phenotypic selection in apple seedling populations (F1), derived from semidiallel hybridizations between genitors with different peculiarities (Florina and Liberty, resistant to apple scab attack; Starkrimson and Golden spur, with spur ideotype) was completed with Marker Assisted Selection (MAS), based on Bulk Segregant Analysis (BSA) technique. Molecular analyses performing by Bulk Segregant Analysis were intended to indentifying markers for scab resistance, powdery mildew resistance and architectural tree structure (ideotype) with bulks of five to six plants per groups, and testing the bulks with RAPD polymorphic primers. The polymorphic bands between the bulks were mostly associated to apple scab resistance genes. Concerning powdery mildew resistance and architectural tree structure analysis the results are inconclusive. Although it seems that the primer 54 showed a polymorphic band in a resistant to powdery mildew hybrid, which is missing in the sensitive hybrids, the result is only apparently favourable since this band is present in both parental forms (Starkrimson, tolerant to powdery mildew and Liberty, relative susceptible). Molecular profile of the standard ideotype and spur ideotype, resulted in hybrids originated in standard parental form (Florina) and spur parental form (Golden spur), were usually monomorphic. Genetic association among the parental molecular polymorphism for scab resistance and the other two important traits (powdery mildew resistance and ideotype), was not conclusive.

In ornamental apple breeding process, the vigour of the tree is an important characteristic, this trait being analyzed in selection of seminal descendants obtained by hybridization. The vigour of the apple trees was studied in 25 interspecific hybrids F1 belonging to different Malus ornamental species, used as maternal or paternal genitors, crossed with several cultivars and selections. The seedling vigour was noted when hybrids were four to seven years old, in accordance with UPOV guideline standard: 3 - weak; 5 - medium; 7 - strong. The mean of marks for tree vigour of F1 hybrids in the 25 combinations ranged between 4.00 and 6.13. Compared with the mean of experiment as control (5.20), the most vigorous seedlings were obtained in the families ‘Cluj 218/2’ x M. niedzwetzkyana and ‘Reinette Baumann’ x M. prunifolia, both with significant differences of growth versus the mean growth of all combinations. The weakest growth was noted in progenies belonging to ‘Rosu de Cluj’ x M. coronaria, ‘Starkrimson’ x M. prunifolia, ‘Frumos de Voinesti’ x M. floribunda, ‘Cluj II-1-2’ x M. floribunda, M. zumi x ‘Reinette Baumann’, M. coronaria x ‘Reinette Baumann’. No significant differences were found between the mean of marks for tree vigour in the group of combinations in which the Malus species were used as maternal genitors (5.18) and paternal genitors (5.20). The coefficient of variability ranged in the experiment between 2.6% and 27.4%; in ten families a lower variability for seedlings vigour (under 10%) was identified, in 13 a medium one (between 10-20%) and in two a large one (more than 20%). Even if in some combinations the variability of tree vigour was not very large, there seems to be fair possibilities to identify by selection the seedlings with a desired vigour in each family.

Apple scab caused by Venturia inaequalis, is an important disease, spread worldwide, having a major negative impact in apple production. In this study, there were analyzed four apple cultivars (Liberty, Florina, Starkrimson, Golden Spur), used as genitors in a diallel cross, and their seedlings. In the field, plants selection was done after presence or absence of the scab on leaves, and afterwards, phenotipically selection was completed by marker assisted selection. The molecular analysis conducted with the markers AL-07, AM-19 and U1400, detects the presence of the Vf gene, introgressed from Malus floribunda 821, which confers resistance to the pathogen, in the parental cultivars Liberty and Florina, and in all the F1 hybrids belonging these two genitors. Starkrimson and Golden Spur were susceptible scab parents with recessive homozygous genotype (vfvf), while Liberty and Florina were identified as heterozygous scab resistant parents with genotype (Vfvf). From the 60 apple seedlings analyzed, grouped in six combinations after their genitors (in each combinations were selected ten hybrids, among them five resistant and five susceptible to apple scab attack) 28 were resistant and heterozygous for Vf gene (Vfvf) and 32 were susceptible to scab with recessive homozygous genotypes (vfvf), being obtained almost Mendelian ratio for response to the pathogen, 50% resistant: 50% susceptible. Seedlings belonging to Starkrimson x Golden Spur were only recessive homozygous genotype (vfvf), and in combination resistant x resistant (Liberty x Florina) were identified susceptible and resistant offspring, but at last, only with heterozygous for Vf gene (Vfvf) and no one with dominant homozygous genotypes (VfVf).

Several traits of growth on greenhouse carnation were analysed at seven varieties, growing both in vivo and in vitro. In vitro culture, proliferation and root formation were more intense in Pink Sol, Caribe and Tanga than in Pink Sim, Obor and Domingo cultivars. The highest intervarietal variability, under in vivo conditions, was recorded for the number of flowers per plant and number of petals per flower, while in vitro all analysed characters showed an extremely large span of variability values (s% = 26.5 - 109.1). The heritability in broad sense showed high values for the analysed characters, both in vivo (H2 = 0.548 - 0.992) and in vitro (H2 = 0.688 -0.953) conditions. There has been noted a tight positive and negative correlation among a lot of the traits of plants, both in vivo and in vitro. Correlations among the desirable traits can be used as selection indices, in carnation breeding.

In Romania, Prunus genetic resources are kept in collections of varieties, populations and biotypes, mainly located in research and development institutes or fruit growing stations and, in the last years, by some private enterprises. Creating the experimental model for the Germplum database based on phenotypic descriptors and SSR molecular markers analysis is an important and topical objective for the efficient characterization of genetic resources and also for establishing a public-private partnership for the effective management of plum germplasm resources in Romania. The technical development of the Germplum database was completed and data will be added continuously after characterizing each new accession.

The selection with co-dominant SSR markers existent on the market, accomplished in dihaploid wheat lines, offers the opportunity to rapidly stabilize the selected resistant individuals into homozygous estate. As biological material there were used 21 resistant and two sensitive dihaploid lines. All dihaploid lines tested, with field resistance to Septoria tritici leaf blotch, expressed at least once such a resistant gene. The most frequently there were encountered in the tested material the Stb 7 and Stb 8 leaf blotch resistant genes. The size of amplification products specific for SSR markers linked with Stb 7 and Stb 8 resistant genes were similar to those obtained by Adhikari et al. (2004) but the size of amplification products linked with Stb 2 resistant genes were quite different, probably due to the polymorphism that characterizes microsatellite markers at the allelic level. The Stb3, Stb 4 and Stb 6 resistant genes were totally absent. The results obtained for Stb 1 and Stb 5 were inconclusive.