Characterization of Cyclamen genotypes using morphological descriptors and DNA molecular markers in a multivariate analysis.

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.

Exogenously Applied GA3 Enhances Morphological Parameters of Tolerant and Sensitive Cyclamen persicum Genotypes under Ambient Temperature and Heat Stress Conditions.

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.

Anthocyanins and Carotenoids Characterization in Flowers and Leaves of Cyclamen Genotypes Linked with Bioactivities Using Multivariate Analysis Techniques.

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, ß-carotene, and cis-ß-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.

Assessment of Genetic Diversity and Population Structure of the Endangered Astragalus exscapus subsp. transsilvanicus through DNA-Based Molecular Markers.

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.

Assessment of Genetic Relationships between Streptocarpus x hybridus V. Parents and F1 Progenies Using SRAP Markers and FT-IR Spectroscopy.

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.

The evolutionarily conserved multifunctional glycine-rich RNA-binding proteins play key roles in development and stress adaptation.

The class IV glycine-rich RNA-binding proteins are a distinct subgroup within the heterogenous superfamily of glycine-rich proteins (GRPs). They are distinguished by the presence of an RNA-binding domain in the N-terminus; generally in the form of an RNA-recognition motif (RRM) or a cold-shock domain (CSD). These are followed by a C-terminal glycine-rich domain. Growing evidence suggests that these proteins play key roles in the adaptation of organisms to biotic and abiotic stresses including those resulting from pathogenesis, alterations in the osmotic, saline and oxidative environment and changes in temperature. Similar vertebrate proteins are also cold-induced and involved in, e.g. hibernation, suggesting evolutionarily conserved functions. The class IV RNA-binding GRPs are likely to operate as key molecular components of hormonally regulated development and to work by regulating gene expression at multiple levels by modifying alternative splicing, mRNA export, mRNA translation and mRNA degradation.

Identification of new adventitious rooting mutants amongst suppressors of the Arabidopsis thaliana superroot2 mutation.

The plant hormone auxin plays a central role in adventitious rooting and is routinely used with many economically important, vegetatively propagated plant species to promote adventitious root initiation and development on cuttings. Nevertheless the molecular mechanisms through which it acts are only starting to emerge. The Arabidopsis superroot2-1 (sur2-1) mutant overproduces auxin and, as a consequence, develops excessive adventitious roots in the hypocotyl. In order to increase the knowledge of adventitious rooting and of auxin signalling pathways and crosstalk, this study performed a screen for suppressors of superroot2-1 phenotype. These suppressors provide a new resource for discovery of genetic players involved in auxin signalling pathways or at the crosstalk of auxin and other hormones or environmental signals. This study reports the identification and characterization of 26 sur2-1 suppressor mutants, several of which were identified as mutations in candidate genes involved in either auxin biosynthesis or signalling. In addition to confirming the role of auxin as a central regulator of adventitious rooting, superroot2 suppressors indicated possible crosstalk with ethylene signalling in this process.

Association of atherosclerotic peripheral arterial disease with adiponectin genes SNP+45 and SNP+276: a case-control study.

OBJECTIVES: We hypothesized that adiponectin gene SNP+45 (rs2241766) and SNP+276 (rs1501299) would be associated with atherosclerotic peripheral arterial disease (PAD). Furthermore, the association between circulating adiponectin levels, fetuin-A, and tumoral necrosis factor-alpha (TNF-α) in patients with atherosclerotic peripheral arterial disease was investigated. METHOD: Several blood parameters (such as adiponectin, fetuin-A, and TNF-α) were measured in 346 patients, 226 with atherosclerotic peripheral arterial disease (PAD) and 120 without symptomatic PAD (non-PAD). Two common SNPs of the ADIPOQ gene represented by +45T/G 2 and +276G/T were also investigated. RESULTS: Adiponectin concentrations showed lower circulating levels in the PAD patients compared to non-PAD patients (P < 0.001). Decreasing adiponectin concentration was associated with increasing serum levels of fetuin-A in the PAD patients. None of the investigated adiponectin SNPs proved to be associated with the subjects' susceptibility to PAD (P > 0.05). CONCLUSION: The results of our study demonstrated that neither adiponectin SNP+45 nor SNP+276 is associated with the risk of PAD.

Fatty acid composition of lipids in pot marigold (Calendula officinalis L.) seed genotypes.

BACKGROUND: Calendula officinalis L. (pot marigold) is an annual aromatic herb with yellow or golden-orange flowers, native to the Mediterranean climate areas. Their seeds contain significant amounts of oil (around 20%), of which about 60% is calendic acid. For these reasons, in Europe concentrated research efforts have been directed towards the development of pot marigold as an oilseed crop for industrial purposes. RESULTS: The oil content and fatty acid composition of major lipid fractions in seeds from eleven genotypes of pot marigold (Calendula officinalis L.) were determined. The lipid content of seeds varied between 13.6 and 21.7 g oil/100 g seeds. The calendic and linoleic acids were the two dominant fatty acids in total lipid (51.4 to 57.6% and 28.5 to 31.9%) and triacylglycerol (45.7 to 54.7% and 22.6 to 29.2%) fractions. Polar lipids were also characterised by higher unsaturation ratios (with the PUFAs content between 60.4 and 66.4%), while saturates (consisted mainly of palmitic and very long-chain saturated fatty acids) were found in higher amounts in sterol esters (ranging between 49.3 and 55.7% of total fatty acids). CONCLUSIONS: All the pot marigold seed oils investigated contain high levels of calendic acid (more than 50% of total fatty acids), making them favorable for industrial use. The compositional differences between the genotypes should be considered when breeding and exploiting the pot marigold seeds for nutraceutical and pharmacological purposes.