Comparison of phytochemical properties and expressional profiling of artemisinin synthesis-related genes in various Artemisia species.

The Artemisia genus belongs to the Asteraceae family and is used in the treatment of many different diseases such as hepatitis and cancer. So far, around 500 species of Artemisia have been found in different regions of the world. Artemisinin is one of the medicinal compounds found in Artemisia species. Hence, this medical feature encourages researchers to pay attention to various species of this genus to discover more genetic and phytochemical information. In the present study, five species of Artemisia including A. fragrans, A. annua, A. biennis, A. scoparia, and A. absinthium were compared to each other in terms of the artemisinin content and other phytochemical components. Moreover, the relative expression profiles of eight genes related to the accumulation and synthesis of artemisinin [including 4FPSF, DBR2, HMGR1, HMGR2, WIRKY, ADS, DXS, and SQS] were determined in investigated species. The result of high-performance liquid chromatography (HPLC) analysis showed that the content of artemisinin in various species was in the order of A. fragrans > A. annua > A. biennis > A. scoparia > A. absinthium. Based on the gas chromatography-mass spectrometry (GC-MS) analysis, 34, 26, 26, 24, and 20 phytochemical compounds were identified for A. scoparia, A. biennis, A. fragrans, A. absinthum, and A. annua species, respectively. Moreover, camphor (38.86%), ß-thujone (68.42%), spathulenol (48.33%), ß-farnesene (48.16%), and camphor (29.04%) were identified as the considerable compounds A. fragrans, A. absinthium, A. scoparia, A. biennis, and A. annua species, respectively. Considering the relative expression of the targeted genes, A. scoparia revealed higher expression for the 4FPSF gene. The highest relative expression of the DBR2, WIRKY, and SQS genes was found in A. absinthium species. Moreover, A. annua showed the highest expression of the ADS and DXS genes than the other species. In conclusion, our findings revealed that various species of Artemisia have interesting breeding potential for further investigation of different aspects such as medicinal properties and molecular studies.

Physiological and Molecular Responses of Barley Genotypes to Salinity Stress.

Among cereals, barley is tolerant to high levels of salinity stress; however, its performance and global production are still dramatically affected by salinity. In this study, we evaluated the behavior of a set of advanced genotypes of barley with aim of assessing the physiological and molecular mechanisms involved in salinity tolerance. The experiment was conducted using a hydroponic system at optimal growing temperature and photoperiod conditions. The results of the analysis of variance (ANOVA) showed significant effects for salinity treatments and genotypes in terms of all measured traits. Salinity stress significantly increased the root and shoot Na+ contents and root-to-shoot Na+ and K+ translocations. In contrast, other physiological features, gas exchange-related traits, as well as root and shoot biomasses were significantly decreased due to salinity stress. Based on the results of the multi-trait genotype ideotype distance index (MGIDI) as a multiple-traits method, G12 and G14 were identified as the superior salt-tolerant advanced genotypes. In the molecular analysis, salinity stress significantly increased the mean relative expression of HvSOS1, HvSOS3, HvHKT2, HvHKT3, HvNHX1, and HvNHX3 genes by 12.87-, 3.16-, 3.65-, 2.54-, 2.19-, and 3.18-fold more than the control conditions, respectively. The results of heatmap-based correlation and principal component analysis (PCA) revealed a clear association pattern among measured traits and expression data. Indeed, these associations confirmed relationships between tolerance pathways and physiological functions. In conclusion, the genotype G14 (D10*2/4/Productive/3/Roho//Alger/Ceres362-1-1) responded well to salinity stress and showed a better expression pattern of studied genes than other genotypes. Hence, this promising genotype can be a candidate for further assessments before commercial introduction.

An Analysis of Genetic Variability and Population Structure in Wheat Germplasm Using Microsatellite and Gene-Based Markers.

Knowledge of the natural patterns of genetic variation and their evolutionary basis is required for sustainable management and conservation of wheat germplasm. In the current study, the genetic diversity and population structure of 100 individuals from four Triticum and Aegilops species (including T. aestivum, Ae. tauschii, Ae. cylindrica, and Ae. crassa) were investigated using two gene-based markers (start codon targeted (SCoT) polymorphism and CAAT-box derived polymorphism (CBDP)) and simple-sequence repeats (SSRs). The SCoT, CBDP, and SSR markers yielded 76, 116, and 48 polymorphism fragments, respectively. The CBDP marker had greater efficiency than the SCoT and SSR markers due to its higher polymorphism content information (PIC), resolving power (Rp), and marker index (MI). Based on an analysis of molecular variance (AMOVA) performed using all marker systems and combined data, there was a higher distribution of genetic variation within species than among them. Ae. cylindrica and Ae. tauschii had the highest values for all genetic variation parameters. A cluster analysis using each marker system and combined data showed that the SSR marker had greater efficiency in grouping of tested accessions, such that the results of principal coordinate analysis (PCoA) and population structure confirmed the obtained clustering patterns. Hence, combining the SCoT and CBDP markers with polymorphic SSR markers may be useful in genetic fingerprinting and fine mapping and for association analysis in wheat and its germplasm for various agronomic traits or tolerance mechanisms to environmental stresses.

Stimulation of Secondary Metabolites and γ-Terpinene Synthase by Silver Nanoparticles in Callus Cultures of Carum carvi.

Biotechnology and nanotechnology are important tools for understanding biochemical pathways. They can be used efficiently for stimulating and increasing the production of secondary metabolites in medicinal plants. The present study aimed to identify the γ-terpinene synthase gene (CcTPS2) as an effective contributor to the biosynthetic pathway of monoterpenes. The effects of silver nanoparticles (AgNPs; 50 and 100 mg l- 1) and time (24 and 48 h) were examined on secondary metabolites in cell suspension cultures of Carum carvi. This involved the identification, isolation, and sequencing of a partial sequence in the CcTPS2 gene of C. carvi. The genomic sequence of CcTPS2 comprised 292 bp which were organized into two exons (110 and 82 bp) and one intron (100 bp), while the cDNA was 192 bp. In the scale of nucleotides, the CcTPS2 gene showed 96% similarity with the TPS2 gene of Oliveria decumbens. We generated sequence data of the CcTPS2 gene for the first time in this species, thereby enabling further developments in understanding the molecular mechanisms responsible for terpene biosynthesis and other chemical derivatives in C. carvi. The results of GC/MS and GC/FID showed that AgNPs strongly affected the secondary metabolites in cell suspension cultures of C. carvi. According to the results, the AgNPs (50 mg l- 1) increased p-cymene and carvone contents in comparison with the control. The exposure of plants to 100 mg l- 1 AgNPs induced the production of thymol and carvacrol. The results of real-time PCR revealed that the exposure of plants to 100 mg l- 1 AgNPs caused a significant upregulation of CcTPS2 expression for 24 h. These cell suspension cultures were elicited by AgNPs, the application of which proved as an effective method to improve the production of secondary metabolites in vitro.

Molecular and Physiological Variability in Bread Wheat and Its Wild Relative (Aegilops tauschii Coss.) Species under Water-Deficit Stress Conditions.

Aegilops and Triticum spp. are two ideal gene pools for the breeding purposes of wheat. In this study, a set of Iranian accessions of Aegilops tauschii Coss. and Triticum aestivum L. species were evaluated in terms of some physiological and biochemical features under control and water-deficit stress conditions. Moreover, several simple sequence repeat (SSR) markers were employed to identify marker loci associated with the measured traits. The results indicated that water-deficit stress significantly affected all measured traits and the highest reductions due to water-deficit were recorded for shoot fresh and dry biomasses (SFB and SDB), stomatal conductance (Gs), leaf relative water content (RWC), and chlorophyll b content (Chl b). In molecular analysis, 25 SSR markers generated 50 fragments, out of which 49 fragments (98%) were polymorphic. Furthermore, the genetic variation observed within species is more than between species. The results of cluster and Bayesian model analysis classified all evaluated accessions into three main clusters. Under control and water-deficit stress conditions, 28 and 27 significant marker-trait associations (MTAs) were identified, respectively. Furthermore, 10 MTAs showed sufficiently stable expression across both growth conditions. Of these, the markers Xgwm-111, Xgwm-44, Xgwm-455, Xgwm-272, and Xgwm-292 were associated with multiple traits. Hence, these markers could serve as useful molecular tools for population characterization, gene tagging, and other molecular breeding studies.

Presenting the spatial-temporal model for assessing and predicting qualitative changes of the groundwater resources in Shahriar, Tehran, Iran.

The quality of surface and groundwater are fundamental concerns. The World Health Organization (WHO) reports that 80% of diseases are waterborne. This percentage mostly lies in developing nations and underdeveloped countries. Determining the relationship between land-use patterns and groundwater quality changes is one of the methods to evaluate the impact of land-use on the hydrological condition of each region which ultimately assists in managing natural resources. In the present study, the spatial-temporal model via satellite images of Landsat TM, ETM, and OLI for2002, 2007, 2012, and 2017 are used to investigate all these modalities in Shahriar Plain, Tehran, Iran. Following image processing and analysis, the land-uses of the study area are classified into urban, farming, and bare lands. In order to assess the water quality of the study area, the periodic sampling data of piezometric wells in the plain were used from 2002 to 2017. Based on the US Salinity Laboratory (USSL) diagram, water qualities for agricultural uses are classified. Results demonstrated that land area has changed over time indicating an increase in urban land area and declines in agricultural and bare lands. Additionally, the results of the groundwater quality indicated C3S1 class in the second and third periods have increased to 3.58% (1025.93 ha) and 12.82% (9367.85 ha) while C2S1 class has been decreased in second and third periods to 3.58 and 12.82%. C1S1 and C4S1 classes during the three periods have been 0.

Genetic diversity analysis in a mini core collection of Damask rose (Rosa damascena Mill.) germplasm from Iran using URP and SCoT markers.

BACKGROUND: Rosa damascena Mill is a well-known species of the rose family. It is famous for its essential oil content. The aim of the present study was to assess the genetic diversity and population structure of a mini core collection of the Iranian Damask rose germplasm. This involved the use of universal rice primers (URP) and start codon targeted (SCoT) molecular markers. RESULTS: Fourteen URP and twelve SCoT primers amplified 268 and 216 loci, with an average of 19.21 and 18.18 polymorphic fragments per primer, respectively. The polymorphic information content for URR and SCoT primers ranged from 0.38 to 0.48 and 0.11 to 0.45, with the resolving power ranging from 8.75 to 13.05 and 9.9 to 14.59, respectively. Clustering was based on neighbor-joining (NJ). The mini core collection contained 40 accessions and was divided into three distinct clusters, centered on both markers and on the combination of data. CONCLUSION: Cluster analysis and principal coordinate analysis were consistent with genetic relationships derived by STRUCTURE analysis. The findings showed that patterns of grouping did not correlate with geographical origin. Both molecular markers demonstrated that the accessions were not genetically diverse as expected, thereby highlighting the possibility that gene flow occurred between populations.

Molecular diversity analysis in hexaploid wheat (Triticum aestivum L.) and two Aegilops species (Aegilops crassa and Aegilops cylindrica) using CBDP and SCoT markers.

BACKGROUND: Evaluation of genetic diversity and relationships among crop wild relatives is an important task in crop improvement. The main objective of the current study was to estimate molecular variability within the set of 91 samples from Triticum aestivum, Aegilops cylindrica, and Aegilops crassa species using 30 CAAT box-derived polymorphism (CBDP) and start codon targeted (SCoT) markers. RESULTS: Fifteen SCoT and Fifteen CBDP primers produced 262 and 298 fragments which all of them were polymorphic, respectively. The number of polymorphic bands (NPB), polymorphic information content (PIC), resolving power (Rp), and marker index (MI) for SCoT primers ranged from 14 to 23, 0.31 to 0.39, 2.55 to 7.49, and 7.56 to 14.46 with an average of 17.47, 0.34, 10.44, and 5.69, respectively, whereas these values for CBDP primers were 15 to 26, 0.28 to 0.36, 3.82 to 6.94, and 4.74 to 7.96 with a mean of 19.87, 0.31, 5.35, and 6.24, respectively. Based on both marker systems, analysis of molecular variance (AMOVA) indicated that the portion of genetic diversity within species was more than among them. In both analyses, the highest values of the number of observed (Na) and effective alleles (Ne), Nei's gene diversity (He), and Shannon's information index (I) were estimated for Ae. cylindrica species. CONCLUSION: The results of cluster analysis and population structure showed that SCoT and CBDP markers grouped all samples based on their genomic constitutions. In conclusion, the used markers are very effective techniques for the evaluation of the genetic diversity in wild relatives of wheat.

Applications of CRISPR-Cas9 as an Advanced Genome Editing System in Life Sciences.

Targeted nucleases are powerful genomic tools to precisely change the target genome of living cells, controlling functional genes with high exactness. The clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) genome editing system has been identified as one of the most useful biological tools in genetic engineering that is taken from adaptive immune strategies for bacteria. In recent years, this system has made significant progress and it has been widely used in genome editing to create gene knock-ins, knock-outs, and point mutations. This paper summarizes the application of this system in various biological sciences, including medicine, plant science, and animal breeding.

Effects of methyl jasmonate and phloroglucinol on thebaine and sanguinarine production in cell suspension culture of Persian poppy (Papaver bracteatum Lindl.).

The biosynthesis path engineering could be very promising for mass production of alkaloids by applying elicitors in the cell suspension culture of Persian poppy (Papaver bracteatum Lindl.). In this work, the effects of different concentrations of methyl jasmonate (MJ) and phloroglucinol (PG) on thebaine and sanguinarine productions in vitro were investigated. Roots as explant and supplementing 3 mg L-1 2,4-Dichlorophenoxyacetic acid with 0.5 mg L-1 Benzyl amino purine to modified MS medium were selected to achieve the most efficient combination for callus induction and production of callus fresh and dry weights. At 48 h after treatment, the addition of PG and MJ individually and in combination together significantly increased both thebaine and sanguinarine contents than the control. The results of high-performance liquid chromatography (HPLC) detection indicated that the highest production rate has been achieved through a synergic effect of two elicitors after 48 h. Results revealed that adding 200 µM of MJ and 100 mg L-1 PG increased thebaine and sanguinarine contents by 56.36 and 107.71-fold than control cells, respectively.

Stabilization of soybean oil during accelerated storage by essential oil of ferulago angulata boiss.

This study has been considered effect of Ferulago angulata essential oil on stabilizing soybean oil during accelerated storage. The essential oil was extracted by Clevenger-type apparatus. For analysis of the essential oil, GC/MS was used. Main components of the essential oil were monoterpene and sesquiterpene hydrocarbons. The essential oil of F. angulata at four concentrations, i.e. 125 (SBO-125), 250 (SBO-250), 500 (SBO-500) and SBO-Mixture (60 ppm TBHQ +60 ppm essential oil) were added to preheated refined soybean oil. TBHQ was used at 120 ppm as standard besides the control. Antioxidant activity index (AAI), free fatty acid (FFA) content, peroxide value (PV) and p-anisidine value (p-AnV) were served for appreciation of efficacy of F. angulata in stabilization of soybean oil. Results from different tests showed that SBO-mixture had highest effect and followed by SBO-TBHQ, SBO-250, SBO-125, SBO-500 and Ctrl. These results reveal F. angulata is a strong antioxidant and can be used instead of synthetic antioxidant.