Hyaluronic Acid-Targeted Niosomes for Effective Breast Cancer Chemostarvation Therapy.

Chemotherapy is widely used for cancer therapy; however, its efficacy is limited due to poor targeting specificity and severe side effects. Currently, the next generations of delivery systems with multitasking potential have attracted significant attention for cancer therapy. This study reports on the design and synthesis of a multifunctional nanoplatform based on niosomes (NIO) coloaded with paclitaxel (PTX), a chemotherapeutic drug commonly used to treat breast cancer, and sodium oxamate (SO), a glycolytic inhibitor to enhance the cytotoxicity of anticancer drug, along with quantum dots (QD) as bioimaging agents, and hyaluronic acid (HA) coating for active targeting. HN@QPS nanoparticles with a size of ∼150 nm and a surface charge of -39.9 mV with more than 90% EE for PTX were synthesized. Codelivery of SO with PTX remarkably boosted the anticancer effects of PTX, achieving IC50 values of 1-5 and >0.5 ppm for HN@QP and HN@QPS, respectively. Further, HN@QPS treatment enhanced the apoptosis rate by more than 70% in MCF-7 breast cancer cells without significant cytotoxicity on HHF-2 normal cells. Also, quantification of mitochondrial fluorescence showed efficient toxicity against MCF-7 cells. Moreover, the cellular uptake evaluation demonstrated an improved uptake of HN@Q in MCF-7 cells. Taken together, this preliminary research indicated the potential of HN@QPS as an efficient targeted-dual drug delivery nanotheranostic against breast cancer cells.

Response of seed yield and biochemical traits of Eruca sativa Mill. to drought stress in a collection study.

Drought tolerance is a complex trait in plants that involves different biochemical mechanisms. During two years of study (2019-2020), the responses of 64 arugula genotypes to drought stress were evaluated in a randomized complete block design with three replications under field conditions. Several metabolic traits were evaluated, i.e. relative water content, photosynthetic pigments (chlorophyll and carotenoids), proline, malondialdehyde, enzymatic antioxidants (catalase, ascorbate peroxidase, and peroxidase), total phenolic and flavonoid contents and seed yield. On average, the drought stress significantly increased the proline content (24%), catalase (42%), peroxidase (60%) and malondialdehyde activities (116%) over the two years of study. As a result of the drought stress, the seed yield (18%), relative water content (19.5%) and amount of photosynthetic pigments (chlorophyll and carotenoids) dropped significantly. However, the total phenolic and flavonoid contents showed no significant changes. Under drought stress, the highest seed yields were seen in the G50, G57, G54, G55 and G60 genotypes, while the lowest value was observed in the G16 genotype (94 g plant-1). According to the findings, when compared to the drought-sensitive genotypes, the drought-tolerant arugula genotypes were marked with higher levels of proline accumulation and antioxidant enzyme activity. Correlation analysis indicated the positive effects of peroxidase, catalase and proline on seed yield under drought conditions. These traits can be considered for the selection of drought-tolerant genotypes in breeding programs.

Seedling nanopriming with selenium-chitosan nanoparticles mitigates the adverse effects of salt stress by inducing multiple defence pathways in bitter melon plants.

Advances in the nanotechnology fields provided crucial applications in plant sciences, contributing to the plant performance and health under stress and stress-free conditions. Amid the applications, selenium (Se), chitosan and their conjugated forms as nanoparticles (Se-CS NPs) have been revealed to have potential of alleviating the harmful effects of the stress on several crops and subsequently enhancing the growth and productivity. The present study was addressed to assay the potential effects of Se-CS NPs in reversing or buffering the harmful effects of salt stress on growth, photosynthesis, nutrient concentration, antioxidant system and defence transcript levels in bitter melon )Momordica charantia(. In addition, some secondary metabolite-related genes were explicitly examined. In this regard, the transcriptional levels of WRKY1, SOS1, PM H+-ATPase, SKOR, Mc5PTase7, SOAR1, MAP30, α-MMC, polypeptide-P and PAL were quantified. Our results demonstrated that Se-CS NPs increased growth parameters, photosynthesis parameters (SPAD, Fv/Fm, Y(II)), antioxidant enzymatic activity (POD, SOD, CAT) and nutrient homeostasis (Na+/K+, Ca2+, and Cl-) and induced the expression of genes in bitter melon plants under salt stress (p ≤ 0.05). Therefore, applying Se-CS NPs might be a simple and effective way of improving crop plants' overall health and yield under salt stress conditions.

High-Density Linkage Mapping of Agronomic Trait QTLs in Wheat under Water Deficit Condition using Genotyping by Sequencing (GBS).

Improvement of grain yield is the ultimate goal for wheat breeding under water-limited environments. In the present study, a high-density linkage map was developed by using genotyping-by-sequencing (GBS) of a recombinant inbred line (RIL) population derived from the cross between Iranian landrace #49 and cultivar Yecora Rojo. The population was evaluated in three locations in Iran during two years under irrigated and water deficit conditions for the agronomic traits grain yield (GY), plant height (PH), spike number per square meter (SM), 1000 kernel weight (TKW), grain number per spike (GNS), spike length (SL), biomass (BIO) and harvest index (HI). A linkage map was constructed using 5831 SNPs assigned to 21 chromosomes, spanning 3642.14 cM of the hexaploid wheat genome with an average marker density of 0.62 (markers/cM). In total, 85 QTLs were identified on 19 chromosomes (all except 5D and 6D) explaining 6.06-19.25% of the traits phenotypic variance. We could identify 20 novel QTLs explaining 8.87-19.18% of phenotypic variance on chromosomes 1A, 1B, 1D, 2B, 3A, 3B, 6A, 6B and 7A. For 35 out of 85 mapped QTLs functionally annotated genes were identified which could be related to a potential role in drought stress.

Exogenous melatonin increases salt tolerance in bitter melon by regulating ionic balance, antioxidant system and secondary metabolism-related genes.

BACKGROUND: Melatonin is a multi-functional molecule widely employed in order to mitigate abiotic stress factors, in general and salt stress in particular. Even though previous reports revealed that melatonin could exhibit roles in promoting seed germination and protecting plants during various developmental stages of several plant species under salt stress, no reports are available with respect to the regulatory acts of melatonin on the physiological and biochemical status as well as the expression levels of defense- and secondary metabolism-related related transcripts in bitter melon subjected to the salt stress. RESULTS: Herewith the present study, we performed a comprehensive analysis of the physiological and ion balance, antioxidant system, as well as transcript analysis of defense-related genes (WRKY1, SOS1, PM H+-ATPase, SKOR, Mc5PTase7, and SOAR1) and secondary metabolism-related gene expression (MAP30, α-MMC, polypeptide-P, and PAL) in salt-stressed bitter melon (Momordica charantia L.) plants in response to melatonin treatment. In this regard, different levels of melatonin (0, 75 and 150 µM) were applied to mitigate salinity stress (0, 50 and 100 mM NaCl) in bitter melon. Accordingly, present findings revealed that 100 mM salinity stress decreased growth and photosynthesis parameters (SPAD, Fv/Fo, Y(II)), RWC, and some nutrient elements (K+, Ca2+, and P), while it increased Y(NO), Y(NPQ), proline, Na+, Cl-, H2O2, MDA, antioxidant enzyme activity, and lead to the induction of the examined genes. However, prsiming with 150 µM melatonin increased SPAD, Fv/Fo, Y(II)), RWC, and K+, Ca2+, and P concentration while decreased Y(NO), Y(NPQ), Na+, Cl-, H2O2, and MDA under salt stress. In addition, the antioxidant system and gene expression levels were increased by melatonin (150 µM). CONCLUSIONS: Overall, it can be postulated that the application of melatonin (150 µM) has effective roles in alleviating the adverse impacts of salinity through critical modifications in plant metabolism.

Changes in root microbiome during wheat evolution.

BACKGROUND: Although coevolutionary signatures of host-microbe interactions are considered to engineer the healthy microbiome of humans, little is known about the changes in root-microbiome during plant evolution. To understand how the composition of the wheat and its ancestral species microbiome have changed over the evolutionary processes, we performed a 16S rRNA metagenomic analysis on rhizobacterial communities associated with a phylogenetic framework of four Triticum species T. urartu, T. turgidum, T. durum, and T. aestivum along with their ancestral species Aegilops speltoides, and Ae. tauschii during vegetative and reproductive stages. RESULTS: In this study, we illustrated that the genome contents of wild species Aegilops speltoides and Ae. tauschii can be significant factors determining the composition of root-associated bacterial communities in domesticated bread wheat. Although it was found that domestication and modern breeding practices might have had a significant impact on microbiome-plant interactions especially at the reproductive stage, we observed an extensive and selective control by wheat genotypes on associated rhizobacterial communities at the same time. Our data also showed a strong genotypic variation within species of T. aestivum and Ae. tauschii, suggesting potential breeding targets for plants surveyed. CONCLUSIONS: This study performed with different genotypes of Triticum and Aegilops species is the first study showing that the genome contents of Ae. speltoides and Ae. tauschii along with domestication-related changes can be significant factors determining the composition of root-associated bacterial communities in bread wheat. It is also indirect evidence that shows a very extensive range of host traits and genes are probably involved in host-microbe interactions. Therefore, understanding the wheat root-associated microbiome needs to take into consideration of its polygenetic mosaic nature.

Molecular mechanisms of fat deposition: IL-6 is a hub gene in fat lipolysis, comparing thin-tailed with fat-tailed sheep breeds.

Tail fat content affects meat quality and varies significantly among different breeds of sheep. Ghezel (fat-tailed) and Zel (thin-tailed) are two important Iranian local sheep breeds with different patterns of fat storage. The current study presents the transcriptome characterization of tail fat using RNA sequencing in order to get a better comprehension of the molecular mechanism of lipid storage in the two mentioned sheep breeds. Seven (Zel  =  4 and Ghezel  =  3) 7-month-old male lambs were used for this experiment. The results of sequencing were analyzed with bioinformatics methods, including differentially expressed genes (DEGs) identification, functional enrichment analysis, structural classification of proteins, protein-protein interaction (PPI) and network and module analyses. Some of the DEGs, such as LIPG, SAA1, SOCS3, HIF-1 α , and especially IL-6, had a close association with lipid metabolism. Furthermore, functional enrichment analysis revealed pathways associated with fat deposition, including "fatty acid metabolism", "fatty acid biosynthesis" and "HIF-1 signaling pathway". The structural classification of proteins showed that major down-regulated DEGs in the Zel (thin-tailed) breed were classified under transporter class and that most of them belonged to the solute carrier transporter (SLC) families. In addition, DEGs under the transcription factor class with an important role in lipolysis were up-regulated in the Zel (thin-tailed) breed. Also, network analysis revealed that IL-6 and JUNB were hub genes for up-regulated PPI networks, and HMGCS1, VPS35 and VPS26A were hub genes for down-regulated PPI networks. Among the up-regulated DEGs, the IL-6 gene seems to play an important role in lipolysis of tail fat in thin-tailed sheep breeds via various pathways such as tumor necrosis factor (TNF) signaling and mitogen-activated protein kinase (MAPK) signaling pathways. Due to the probable role of the IL-6 gene in fat lipolysis and also due to the strong interaction of IL-6 with the other up-regulated DEGs, it seems that IL-6 accelerates the degradation of lipids in tail fat cells.

Evaluation of drought tolerance indices for barley landraces under irrigated and dry conditions / Avaliação de índices de tolerância seca para terraços de cevada em condições irrigadas e secas

Barley cultivation for drought areas requires a reliable assessment of drought tolerance variability among the breeding germplasms. Hence, 121 barley landraces, advanced breeding lines, and varieties were evaluated under both moisture non-stress and stress field conditions using a lattice square (11×11) design with two replications for each set of the trials. Twelve drought tolerance indices (SSI, TOL, MP, GMP, STI, YI, YSI, HM, SDI, DI, RDI, and SSPI) were used based on grain yield under normal (Yp) and drought (Ys) conditions. Analysis of variance showed a significant genetic variation among genotypes for all indices except for TOL and SSPI indices. Yp had a very strong association with Ys (r = 0.92**) that indicates high yield potential under non-stress can predict better yield under stress conditions. Yp and Ys were positively and significantly correlated with MP, GMP, STI, YI, HM, and DI indices, whereas they were negatively correlated with SSI and SDI. In principal component analysis (PCA), the first PC explained 64% of total variation with Yp, Ys, MP, GMP, STI, YI, HM, and DI. The second PC explained 35.6% of the total variation and had a positive correlation with SSI, TOL, SDI, and SSPI. It can be concluded that MP, GMP, STI, YI, HM and DI indices with the most positive and significant correlation with the yield at both non-stress and stress environments would be better indices to screen barley genotypes, although none of the indices could undoubtedly identify high yield genotypes under both conditions.

O cultivo de cevada para áreas secas exige uma avaliação confiável da variabilidade da tolerância à seca entre os germoplasmas reprodutores. Assim, 121 linhagens crioulas de cevada (linhas de reprodução avançada e variedades) foram avaliadas em campo sob condições sem estresse e com estresse de umidade do solo, utilizando-se para isso um arranjo experimental de malha quadrada (11×11), com duas repetições para cada conjunto de ensaios. Foram utilizados 12 índices de tolerância à seca (SSI, TOL, MP, GMP, STI, YI, YSI, HM, SDI, DI, RDI e SSPI), com base no rendimento de grãos sob condições normais sem estresse (Yp) e com estresse de seca (Ys). A análise de variância mostrou uma variação genética significativa entre os genótipos para todos os índices, com exceção dos índices TOL e SSPI. Yp teve uma associação muito forte com Ys (r = 0,92**), o que indica que o potencial de alto rendimento sob condições sem estresse pode prever melhor rendimento sob condições de estresse. Yp e Ys foram positivamente e significativamente correlacionados com os índices MP, GMP, STI, YI, HM e DI, enquanto, foram correlacionados negativamente com os índices SSI e SDI. Na análise de componentes principais (PCA), o primeiro PC explicou 64% da variação total com Yp, Ys, MP, GMP, STI, YI, HM e DI. O segundo PC explicou 35,6% da variação total e apresentou correlação positiva com SSI, TOL, SDI e SSPI. Pode-se concluir que, os índices MP, GMP, STI, YI, HM e DI com a correlação mais positiva e significativa com a produção nos ambientes sem estresse e com estresse seriam melhores índices para a seleção de genótipos de cevada, embora nenhum dos índices pudesse concretamente identificar genótipos de alto rendimento sob ambas as condições.

Column-free purification and coating of SpyCatcher protein on ELISA wells generates universal solid support for capturing of SpyTag-fusion protein from the non-purified condition.

•Spy Tag-Protein covalent interaction is rapid and specific method for protein immobilization.•Column free purification of SpyCatcher protein enables develop a universal solid support for SpyTag protein purification.•This method is highly simple and applicable to other proteins.

Insecticide Susceptibility in Tuta absoluta (Lepidoptera: Gelechiidae) and Metabolic Characterization of Resistance to Diazinon.

The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a globally important economic insect pest of tomatoes that has rapidly expanded in range. The pest is generally controlled by insecticides; however, reduced susceptibility of T. absoluta to many recommended insecticides has been reported. The aims of this study were to determine the efficacy of diazinon, spinosad, abamectin, and Bt on second instar larvae of Iranian greenhouse populations of T. absoluta from Soufian (SF) and Qom (QM) and a susceptible (SS) population originally from Bonab, and assess the possibility of resistance in the populations. The results showed that spinosad was the most effective insecticide, whereas in contrast, Bt was ineffective against the populations. Abamectin was effective, but susceptibility in SF population was lower than in QM and SS. Susceptibility to diazinon was low in SF and QM populations. Mediation of diazinon resistance by metabolic mechanisms and activity of detoxifying enzymes and acetylcholinesterase on resistance were assessed. The results showed that enhanced activity of monooxygenases and esterases may have played a role in resistance of T. absoluta to diazinon. Our results will be useful in resistance management in this injurious pest and reduce the negative impacts of high rates of insecticide use on the environment.

Watercress-based electrospun nanofibrous scaffolds enhance proliferation and stemness preservation of human adipose-derived stem cells.

The present study describes the effects of Watercress extract (WE) based electrospun nanofibrous mats on the regulation of adhesion, proliferation, cytoprotection and stemness preservation of adipose-derived stem cells (ADSCs). Watercress (Nasturtium officinale) is one of the most important medicinal plant with a board spectrum of biological functions. For this purpose, WE-loaded PCL-PEG nanofibers were fabricated by electrospinning and characterized using FE-SEM and FTIR. Adhesion, proliferation and cytoprotection of ADSCs on the nanofibers was investigated using FE-SEM and MTT assays. Analysis of cell cycle was carried out by flow-cytometry. Finally, qPCR was applied to measure the expression levels of cell cycle-regulated genes and stemness markers of ADSCs grown on the nanofibers. In this study, we found that WE-loaded PCL-PEG nanofibers had great antioxidant potential and exhibited higher cytoprotection, better adhesion, and significantly increased proliferation of ADSCs. The greater proliferation and preserving stemness ability of ADSCs on WE based nanofibers was further confirmed by higher expression levels of cell cycle-regulated genes and stemness markers. These results demonstrate that WE-loaded PCL-PEG electrospun nanofibrous mats appear suitable to support ADSCs adhesion and proliferation while concurrently preserving the cell stemness, therefore representing a hopeful approach for applying in stem cell based regenerative medicine.

The effect of ketorolac and triamcinolone acetonide on adipogenic and hepatogenic differentiation through miRNAs 16/15/195: Possible clinical application in regenerative medicine.

OBJECTIVE: The role of miR-16/15b/195 and complementary compounds in the commitment of mesenchymal stem cells (MSCs) to different lineages is unknown. The aim of this work was to investigate the effect of ketorolac and triamcinolone acetonide on adipogenic and hepatogenic processes, respectively. Also, miR-16/15 and miR-195 expression levels were evaluated under different induction conditions. METHODS: MSCs were isolated, expanded and directed using adipogenesis medium or medium supplemented with ketorolac, and also subjected to hepatogenic differentiation using a cocktail of hepatocyte growth factor (HGF) and Oncostatin M (OSM) either with or without triamcinolone acetonide. Periodic acid-Schiff (PAS) staining, Oil red O staining, albumin and adiponectin protein secretion were evaluated. MiR-16 family expression level was assessed using qRT-PCR in four induced groups as compared to non-induced cells. RESULTS: The expression levels of miR-16 and miR-15 increased significantly from adipose derived stem cells to adipocytes (p<0.01). Positive stimulatory effects of ketorolac and triamcinolone on adipogenic and hepatogenic induction, respectively, were observed. Ketorolac stimulated upregulation of miR-16/15b in the adipogenic induced stem cells. Interestingly, triamcinolone caused upregulation of miR-15b and miR-195 in hepatic commitment. CONCLUSIONS: Two highly effective stimulators were identified. Ketorolac is similar to indomethacin and affects adipogenic differentiation. Triamcinolone is involved in hepatogenic commitment of stem cells like differentiation process of macrophages, adipocytes and osteocytes. The alteration of miR-16 family members' expression indicates that this family may play a possible role in directing stem cell fate.

An Integrative Genetic Study of Rice Metabolism, Growth and Stochastic Variation Reveals Potential C/N Partitioning Loci.

Studying the genetic basis of variation in plant metabolism has been greatly facilitated by genomic and metabolic profiling advances. In this study, we use metabolomics and growth measurements to map QTL in rice, a major staple crop. Previous rice metabolism studies have largely focused on identifying genes controlling major effect loci. To complement these studies, we conducted a replicated metabolomics analysis on a japonica (Lemont) by indica (Teqing) rice recombinant inbred line population and focused on the genetic variation for primary metabolism. Using independent replicated studies, we show that in contrast to other rice studies, the heritability of primary metabolism is similar to Arabidopsis. The vast majority of metabolic QTLs had small to moderate effects with significant polygenic epistasis. Two metabolomics QTL hotspots had opposing effects on carbon and nitrogen rich metabolites suggesting that they may influence carbon and nitrogen partitioning, with one locus co-localizing with SUSIBA2 (WRKY78). Comparing QTLs for metabolomic and a variety of growth related traits identified few overlaps. Interestingly, the rice population displayed fewer loci controlling stochastic variation for metabolism than was found in Arabidopsis. Thus, it is possible that domestication has differentially impacted stochastic metabolite variation more than average metabolite variation.

The effect of dimethyl sulfoxide on hepatic differentiation of mesenchymal stem cells.

INTRODUCTION: Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are suitable choices in autologous stem cell treatment of liver-associated diseases due to their hepatic differentiation potential. Dimethyl sulfoxide (DMSO) is an amphipathic molecule with potential of delivering both lipophilic and hydrophilic agents into cells, also a common cryoprotectant for freezing of the cells. DMSO was used in some protocols for induction of AT-MSCs towards hepatocyte like cells. However, the effect of DMSO on hepatogenic differentiation of AT-MSCs were not surveyed, previously. In the present study, we aimed at evaluation of the effect of DMSO on differentiation of AT-MSCs into hepatic lineage. METHODS: We isolated mesenchymal stem cells (MSCs) from adipose tissue, and then verifies multi-potency and surface markers of AT-MSCs . Isolated AT-MSCs randomly dispensed in four groups including Group 1: HGF treated, 2: HGF+ DMSO treated, 3: HGF+ DMSO+ OSM treated, and group control for a period of 3 weeks in the expansion medium without serum; EGF and bFGF were also included in the first days of inductions. The morphologic changes during induction period was observed with microscopy. The secretion of albumin (ALB) of the differentiating MSCs was investigated using ELISA, and urea production was evaluated using colorimetric assay. The qRT-PCR was performed for quantitation of hepatocyte marker genes including AFP, ALB, CK18, HNF4a, and HNF6. The glycogen storage of differentiated cells was visualized by periodic-acid Schiff‘s staining. RESULTS: The results demonstrate that DMSO speeds up hepatic differentiation of AT-MSCs characterized by rapid changes in morphology; higher expression of hepatic marker gene (ALB) in both mRNA and protein level (P < 0.05); also increased transcriptional levels of other liver genes including CK18, HNF4a, and HNF6 (P < 0.01); and moreover, greater percentage of glycogen storage(p < 0.05) in DMSO-treated groups. CONCLUSION: DMSO catalyzes hepatic differentiation; therefore, using DMSO for acceleration of the hepatogenic protocols of AT-MSCs appears advantageous.

Genome-wide analysis of alternative splicing events in Hordeum vulgare: Highlighting retention of intron-based splicing and its possible function through network analysis.

In this study, using homology mapping of assembled expressed sequence tags against the genomic data, we identified alternative splicing events in barley. Results demonstrated that intron retention is frequently associated with specific abiotic stresses. Network analysis resulted in discovery of some specific sub-networks between miRNAs and transcription factors in genes with high number of alternative splicing, such as cross talk between SPL2, SPL10 and SPL11 regulated by miR156 and miR157 families. To confirm the alternative splicing events, elongation factor protein (MLOC_3412) was selected followed by experimental verification of the predicted splice variants by Semi quantitative Reverse Transcription PCR (qRT-PCR). Our novel integrative approach opens a new avenue for functional annotation of alternative splicing through regulatory-based network discovery.

Validation of EST-derived STS markers localized on Qfhs.ndsu-3BS for Fusarium head blight resistance in wheat using a 'Wangshuibai' derived population.

A few EST-derived STS markers localized on Qfhs.ndsu-3BS, a major QTL for resistance to Fusarium head blight (FHB) in wheat, have been previously identified in the 'Sumai 3'/'Stoa' population. In this study, we used a 'Wangshuibai' (resistant) /'Seri82' (susceptible) derived population, linkage group, QTL, and quantitative gene expression analysis to assess the genetic background dependence and stability of the EST-derived STS markers for use in marker aided selection to improve FHB resistance in wheat. Based on our results, a QTL in the map interval of Xsts3B-138_1-Xgwm493 on chromosome 3BS was detected for FHB resistance, which accounted for up to 16% of the phenotypic variation. BLASTN analysis indicated that Xsts3B-138_1 sequence had significant similarity with the resistance gene analogue. Real-time quantitative PCR showed that the relative expression of Xsts3B-138_1 in 'Wangshuibai' at 96 h after inoculation was 2.6 times higher than 'Seri82'. Our results underlined that EST-derived STS3B-138 markers could be predominantly used in marker aided selection to improve FHB resistance in wheat.

Genetic properties of milk thistle ecotypes from Iran for morphological and flavonolignans characters.

The aim of present study was to investigate the genetic variation within and between 32 milk thistle ecotypes collected from northern (23 accessions) and southern (9 accessions) regions of Iran along with two introduced varieties, CN seeds and Budakalaszi, for morphological and flavonolignans properties. The two collections were assessed at separate field experiments. MANOVA for all the morphological traits showed significant difference between ecotypes. Univariate ANOVA verified these differences for most of the traits in the northern ecotypes (first collection) while for southern ecotypes no significant differences were obtained for the studied traits except seed yield. Among and within ecotypes genotypic coefficient of variation indicated higher level of variation among ecotypes than within ecotypes. In both of the experiments, there was a large genetic variation for silybin and silymarin quality and quantity. Cluster analysis of 34 accessions was performed for morphological traits and silymarin and silybin characteristics, separately. The resulting dendrogram based on silybin and silymarin characteristics revealed that the native accessions such as Dezfoul, Fereydounkenar and Nour, had highest flavonolignans and they were better than the foreign varieties. Also, there was no clear relationship between clustering based on morphological traits and flavonolignan compounds.