Metabolomic and transcriptomic analyses of yellow-flowered crocuses to infer alternative sources of saffron metabolites.

BACKGROUND: The increasing demand for saffron metabolites in various commercial industries, including medicine, food, cosmetics, and dyeing, is driven by the discovery of their diverse applications. Saffron, derived from Crocus sativus stigmas, is the most expensive spice, and there is a need to explore additional sources to meet global consumption demands. In this study, we focused on yellow-flowering crocuses and examined their tepals to identify saffron-like compounds. RESULTS: Through metabolomic and transcriptomic approaches, our investigation provides valuable insights into the biosynthesis of compounds in yellow-tepal crocuses that are similar to those found in saffron. The results of our study support the potential use of yellow-tepal crocuses as a source of various crocins (crocetin glycosylated derivatives) and flavonoids. CONCLUSIONS: Our findings suggest that yellow-tepal crocuses have the potential to serve as a viable excessive source of some saffron metabolites. The identification of crocins and flavonoids in these crocuses highlights their suitability for meeting the demands of various industries that utilize saffron compounds. Further exploration and utilization of yellow-tepal crocuses could contribute to addressing the growing global demand for saffron-related products.

Prolonged exposure to freezing stress reduces the ability of chickpea seedlings to effectively tolerate extremely low temperatures.

The duration and intensity of freezing stress are the most critical factors determining injury in autumn chickpeas, limiting their production and development. To evaluate the effects of freezing temperature and duration on the survival rate (SU%), as well as the physiological and biochemical characteristics of autumn chickpea seedlings, a study was conducted using five different temperatures (0, -6, -8, -10, and -12°C) and five different durations (1 h, 2 h, 3 h, 4 h, and 5 h) of exposure to freezing stress. The SU% of chickpea seedlings decreased to zero after exposure to temperatures of -10°C and -12°C for 5 hours. As the temperature decreased from -8°C to -12°C and the duration of exposure to freezing stress increased from 1 to 5 hours, the leaf membrane stability index decreased by 33%, 48%, 46%, 57%, and 58%, respectively. The highest and lowest total pigment contents were observed after 1 hour at 0°C and 5 hours at -12°C, respectively. The maximum photochemical efficiency of photosystem II (Fv'/Fm') was not affected by temperatures as low as -8°C in any of the time treatments during the recovery period. However, this parameter's value decreased as the freezing stress duration increased. At -12°C, the activity of ascorbate peroxidase, catalase, and peroxidase increased by 44.6%, 38.3%, and 33.0%, respectively, as the duration of stress was increased from 1 hour to 5 hours. A positive and significant correlation was observed between plant dry weight, membrane stability index, photosynthetic pigment content, and Fv'/Fm' with SU% after exposure to freezing stress. The minimum temperature and the maximum duration of freezing stress tolerance in chickpea seedlings were observed at -12°C for two hours. Our findings confirm that prolonging the freezing duration disrupts the defense mechanisms of chickpea seedlings. Therefore, future studies on breeding chickpeas tolerant to freezing stress should concentrate on attributes strongly correlated with SU%.

Phylogenomic investigation of safflower (Carthamus tinctorius) and related species using genotyping-by-sequencing (GBS).

Safflower (Carthamus tinctorius, Asteraceae) is a source of high-quality edible oil growing in moisture-limited environments. Despite its economic importance, the relationships to close wild species in Carthamus and the presence and relationships of ecotypes within safflower are still not fully clarified. Here we use genotyping-by-sequencing to identify the wild progenitor of C. tinctorius, infer phylogenetic relationship within the series Carthamus and identify groups of closely related lineages within cultivated safflower. Phylogenetic and population genomic analyses found C. palaestinus to be the closest relative and single progenitor of C. tinctorius, which confirms the Levant as the area of domestication of the crop. Flow cytometry showed all analyzed samples of C. oxyacantha, C. palaestinus and C. tinctorius to be diploid (2n = 2x = 24) with 2C genome sizes of 2.4-2.7 pg. Analyses of a set of 114 worldwide distributed safflower accessions arrived at two to five genetic groups, which showed, however, no correlation with the geographic origins of these accessions. From this, we conclude that the trade of safflower seeds resulted in multiple introductions of genotypes from the Levant into other areas with suitable climate conditions for the plant, as well as exchange of genotypes among these areas.

Genetic Variation of Puccinia triticina Populations in Iran from 2010 to 2017 as Revealed by SSR and ISSR Markers.

Puccinia triticina is a major wheat pathogen worldwide. Although Iran is within the Fertile Crescent, which is supposed to be the center of origin of both wheat and P. triticina, the knowledge of the genetic variability of local populations of this basidiomycete is limited. We analyzed 12 inter simple sequence repeats (ISSRs) and 18 simple sequence repeats (SSRs) of 175 P. triticina isolates sampled between 2010 and 2017 from wheat and other Poaceae in 14 provinces of Iran. SSRs revealed more polymorphisms than ISSRs, indicating they were more effective in differentiating P. triticina populations. Based on a dissimilarity matrix with a variable mutation rate for SSRs and a Dice coefficient for ISSRs, the isolates were separated into three large groups, each including isolates from diverse geographic origins and hosts. The grouping of SSR genotypes in UPGMA dendrograms was consistent with the grouping inferred from the Bayesian approach. However, isolates with a common origin clustered into separate subgroups within each group. The high proportion of heterozygous alleles suggests that in Iran clonal reproduction prevails over sexual reproduction of the pathogen. A significant correlation was found between SSR and ISSR genotypes and the virulence phenotypes of the isolates, as determined in a previous study.

Does the Extension Torque Control Differ between Injured and Uninjured Knees of ACL-Deficient Individuals?

Background: Knee extension torque control decreases after anterior cruciate ligament (ACL) rupture. There is a controversy in neuromuscular control changes on the uninvolved side. We intended to evaluate the steadiness and accuracy of quadriceps muscle control in the healthy and deficient sides of people with acute ACL rupture. Methods: In this cross-sectional study, thirteen men with ACL rupture (age: 27.8±7.0, body mass index (BMI): 24.7±2.25: 24.7, days from injury: 48.1±21.3) participated in the study. We measured quadriceps force control, which is quantitatively assessed by the standard deviation (SD) of joint torque for a predefined submaximal target. The accuracy of muscular control or performance of quadriceps is commonly quantified by the root mean square of error (RMSE) was also measured. A two-way analysis of variance was conducted to assess SD and RMSE of two levels of quadriceps contraction (30% of muscle voluntary contraction (MVC), 50% of MVC) across both healthy and deficient knees. Results: There was a significant main effect for SD and RMSE of MVC percentage (p<0.001). SD of quadriceps torque in 50% of MVC (1.44 ± 0.13) was higher than 30% of MVC (0.88 ± 0.1). In contrast, there was no significant main effect for SD and RMSE of knee condition. Conclusion: After unilateral ACL rupture, the neuromuscular system becomes defected and quadriceps muscle control is then reduced in the healthy side. Therefore, the healthy side is also vulnerable to ligamentous damage. Besides, with the increasing intensity of physical activities, neuromuscular control decreases and the risk of re-injury rises.

Ancient Artworks and Crocus Genetics Both Support Saffron's Origin in Early Greece.

Saffron crocus (Crocus sativus) is a male-sterile, triploid flower crop, and source of the spice and colorant saffron. For over three millennia, it was cultivated across the Mediterranean, including ancient Greece, Persia, and other cultures, later spreading all over the world. Despite saffron crocus' early omnipresence, its origin has been the matter of a century-old debate, in terms of area and time as well as parental species contribution. While remnants of the ancient arts, crafts, and texts still provide hints on its origin, modern genetics has the potential to efficiently follow these leads, thus shedding light on new possible lines of descent. In this review, we follow ancient arts and recent genetics to trace the evolutionary origin of saffron crocus. We focus on the place and time of saffron domestication and cultivation, and address its presumed autopolyploid origin involving cytotypes of wild Crocus cartwrightianus. Both ancient arts from Greece, Iran, and Mesopotamia as well as recent cytogenetic and comparative next-generation sequencing approaches point to saffron's emergence and domestication in ancient Greece, showing how both disciplines converge in tracing its origin.

Genome Size and Chromosome Number Evaluation of Astragalus L. sect. Hymenostegis Bunge (Fabaceae).

Astragalus section Hymenostegis is one of the important characteristic elements of thorn-cushion formations in the Irano-Turanian floristic region. In this paper, we examined the chromosome number of 17 species (15 new reports) and provide estimates of genome size for 62 individuals belonging to 38 taxa of A. sect. Hymenostegis, some species outside this section, plus two Oxytropis species. Based on chromosome counts 11 species were found to be diploid (2n = 16), four species tetraploid (2n = 32) and two taxa hexaploid (2n = 48). From genome size measurements on silica-gel dried material, three ploidy levels (2x, 4x and 6x) were inferred, with a majority of species being diploid. The 2C values reach from 2.07 pg in diploid Astragalus zohrabi to 7.16 pg in hexaploid A. rubrostriatus. We found indications that species might occur with different cytotypes. A phylogenetic framework using nrDNA ITS sequences was constructed to understand the evolution of ploidy changes and genome sizes. It showed that genome size values among the studied taxa differ only slightly within ploidy levels and are nearly constant within most species and groups of closely related taxa within the genus Astragalus. The results of this study show that there is a rather strong correlation between genome sizes and chromosome numbers in sect. Hymenostegis. The resolution of the ITS-based phylogenetic tree is too low to infer evolutionary or environmental correlations of genome size differences. Polyploidization seems to contribute to the high species number in Astragalus, however, in sect. Hymenostegis it is not the main driver of speciation.

An ultra-high bandwidth nano-electronic interface to the interior of living cells with integrated fluorescence readout of metabolic activity.

We present the first ever broadband, calibrated electrical connection to the inside of a cell. The interior of a vital, living cell contains multiple dynamic and electrically active organelles such as mitochondria, chloroplasts, lysosomes, and the endoplasmic reticulum. However, little is known about the detailed electrical activity inside the cell. Here we show an ultra-high bandwidth nano-electronic interface to the interior of living cells with integrated fluorescence readout of metabolic activity. On-chip/on-petri dish nanoscale capacitance calibration standards are used to quantify the electronic coupling from bench to cell from DC to 26 GHz (with cell images at 22 GHz). The interaction of static to high frequency electromagnetic fields with the cell constituents induce currents of free charges and local reorganization of linked charges. As such, this enables a direct, calibrated, quantitative, nanoscale electronic interface to the interior of living cells. The interface could have a variety of applications in interfacing life sciences to nano-electronics, including electronic assays of membrane potential dynamics, nano-electronic actuation of cellular activity, and tomographic, nano-radar imaging of the morphology of vital organelles in the cytoplasm, during all phases of the cell life cycle (from development to senescence), under a variety of physiological environments, and under a broad suite of pharmacological manipulations.

Virulence of Leaf Rust Physiological Races in Iran From 2010 to 2017.

The wheat leaf rust fungus, Puccinia triticina, has widespread geographical distribution in Iran within the Fertile Crescent region of the Middle East where wheat was domesticated and P. triticina originated. Therefore, it is of great importance to identify the prevalence and distribution of P. triticina pathotypes in this area. From 2010 to 2017, 241 single-uredinium isolates of P. triticina were purified from 175 collections of P. triticina made from various hosts in 14 provinces of Iran, and they were tested on 20 Thatcher near-isogenic lines carrying single-leaf rust resistance genes. In total, 86 pathotypes were identified, of which the pathotypes FDTTQ, FDKPQ, FDKTQ, and FDTNQ were most prevalent. No virulence for Lr2a was detected, whereas virulence for Lr1 was found only on bread wheat in a few provinces in 2016. Only isolates from durum wheat and wild barley were virulent to Lr28. Although virulence for Lr9, Lr20, and Lr26 was observed in some years, the virulence frequency for these genes was lower than that of the other Lr genes. P. triticina collections from host plants with different ploidy levels or genetically dissimilar backgrounds were grouped individually according to genetic distance. Based on these results, collections from barley, durum wheat, oat, triticale, and wild barley were different from those of bread wheat.

Saffron (Crocus sativus) is an autotriploid that evolved in Attica (Greece) from wild Crocus cartwrightianus.

Crocus sativus, the saffron crocus, is the source of saffron, which is made from the dried stigmas of the plant. It is a male-sterile triploid lineage that ever since its origin has been propagated vegetatively. Its mode of evolution and area of origin are matters of long-lasting debates. Here we analyzed chloroplast genomes and genome-wide DNA polymorphisms obtained through genotyping-by-sequencing (GBS) to infer the parent and area of origin of C. sativus. These data were complemented by genome size measurements and analyses of nuclear single-copy genes. We could place 99.3% of saffron GBS alleles in Crocus cartwrightianus, a species occurring in southeastern mainland Greece and on Aegean islands, identifying it as the sole progenitor of the saffron crocus. Phylogenetic and population assignment analyses together with chloroplast polymorphisms indicated the C. cartwrightianus population in the vicinity of Athens as most similar to C. sativus. We conclude that the crop is an autotriploid that evolved in Attica by combining two different genotypes of C. cartwrightianus. Triploid sterility and vegetative propagation prevented afterwards segregation of the favorable traits of saffron, resulting in worldwide cultivation of a unique clonal lineage.

Phylogeny of the saffron-crocus species group, Crocus series Crocus (Iridaceae).

Phylogenetic relationships among the taxa of Crocus series Crocus are still unclear, preventing the understanding of species diversity and the evolution of the important spice saffron (Crocus sativus). Therefore, we analyzed sequences of two chloroplast (trnL-trnF, matK-trnK) and three nuclear (TOPO6, ribosomal DNA ETS and ITS) marker regions to infer phylogenetic relationships among all species belonging to series Crocus. Our phylogenetic analyses resolved the relationships among all taxa of the series. Crocus hadriaticus and the former C. pallasii subspecies appeared polyphyletic. The latter deserve elevating the subspecies to species rank, while for C. hadriaticus a detailed study of species boundaries is necessary. Multi-locus and also genome-wide single nucleotide polymorphism data obtained through genotyping-by-sequencing placed C. sativus within C. cartwrightianus with no indication that other Crocus species contributed to the evolution of the triploid. Our analyses thus made an autotriploid origin of C. sativus from C. cartwrightianus very likely.

Scanning Microwave Microscopy of Vital Mitochondria in Respiration Buffer.

We demonstrate imaging using scanning microwave microscopy (SMM) of vital mitochondria in respiration buffer. The mitochondria are isolated from cultured HeLa cells and tethered to a solid graphene support. The mitochondria are kept vital (alive) using a respiration buffer, which provides nutrients to sustain the Krebs cycle. We verify that the mitochondria are "alive" by measuring the membrane potential using a voltage sensitive fluorescent dye (TMRE). The organelles are measured capacitively at 7 GHz. Several technical advances are demonstrated which enable this work: 1) The SMM operates in an electrophysiologically relevant liquid (hence conducting) environment; 2) The SMM operates in tapping mode, averaging the microwave reflection measurement over many tapping periods; 3) A tuned reflectometer enables increased sensitivity; 4) Variable frequencies up to 18 GHz are used; 5) In contrast with traditional matching/resonant methods that exhibit high quality factor that fail in the presence of liquids, interferometric/tuned reflectometer gives the possibility to adjust the quality factor or sensitivity even in the presence of the liquid.

Effects of unstable footwear on gait characteristic: A systematic review.

BACKGROUND: Over the last three decades, several designs of unstable footwear have been developed in the forms of shoes, sandals and boots. There are marketing claims related to the positive effects of these shoes on the training of lower limb muscles and improving gait. Many studies have been performed on the effects of unstable footwear on muscle activity, balance, posture, energy expenditure, lower extremity disorders, and biomechanical changes. The analysis of the kinetics and kinematics characteristics of gait would provide objective representation of body movement. OBJECTIVE: To systematically review available evidence on the use of unstable footwear on kinetic and kinematic parameters to make specific recommendation for practice and future studies. METHOD: A computer-based search was undertaken through PubMed, Cochrane Library, Embase, PEDro, Web of Science and Google Scholar from 2005 to 2015. The included studies were appraised using McMaster Critical Review Form for Quantitative Studies. RESULT: Ten studies (quasi-experimental design) were included. CONCLUSION: Considering kinetic and kinematic interaction of variables in the included studies revealed that confounding factors may have high impact on biomechanical findings of unstable footwear. Then, more homogeneous studies, considering these factors, should be implemented in future studies to inform the best clinical practice.

The effect of new dynamic splint in pinch strength in De Quervain syndrome: a comparative study.

Objective For patients with De Quervain syndrome using thumb spica orthosis is restricting, so many patient are dissatisfied with using static model. The aim of this study is to compare the effect of modified dynamic and conventional static orthoses on pinch power and functional abilities of hand in De Quervain syndrome. Method In this quasi-experimental study, palmar and lateral pinch strength of the thumb, pain and functional abilities of hand, patient's satisfaction of orthoses, were evaluated after using modified dynamic and conventional static orthosis in two groups. Results Both orthoses improved palmar and lateral pinch strength of the thumb, pain level and functional abilities. In comparison of mentioned variables, there was no significant difference between two groups regarding to pain recovery and abilities improvement (p > 0.05). However, the satisfaction level of the patients who had used dynamic orthosis was higher (p < 0.05). Conclusion It seems, adding a joint to the static thumb spica leads to patient's relief and consequently to their satisfaction. Implications for Rehabilitation The dynamic thumb spica, is easier to use and patients satisfaction is higher than static thumb spica. Therefore, it can be used for conservative treatment of De Quervain syndrome.

Metabolic and Growth Rate Alterations in Lymphoblastic Cell Lines Discriminate Between Down Syndrome and Alzheimer's Disease.

BACKGROUND: Deficits in mitochondrial function and oxidative stress play pivotal roles in Down syndrome (DS) and Alzheimer's disease (AD) and these alterations in mitochondria occur systemically in both conditions. OBJECTIVE: We hypothesized that peripheral cells of elder subjects with DS exhibit disease-specific and dementia-specific metabolic features. To test this, we performed a comprehensive analysis of energy metabolism in lymphoblastic-cell-lines (LCLs) derived from subjects belonging to four groups: DS-with-dementia (DSAD), DS-without-dementia (DS), sporadic AD, and age-matched controls. METHODS: LCLs were studied under regular or minimal feeding regimes with galactose or glucose as primary carbohydrate sources. We assessed metabolism under glycolysis or oxidative phosphorylation by quantifying cell viability, oxidative stress, ATP levels, mitochondrial membrane potential (MMP), mitochondrial calcium uptake, and autophagy. RESULTS: DS and DSAD LCLs showed slower growth rates under minimal feeding. DS LCLs mainly dependent on mitochondrial respiration exhibited significantly slower growth and higher levels of oxidative stress compared to other groups. While ATP levels (under mitochondrial inhibitors) and mitochondrial calcium uptake were significantly reduced in DSAD and AD cells, MMP was decreased in DS, DSAD, and AD LCLs. Finally, DS LCLs showed markedly reduced levels of the autophagy marker LC3-II, underscoring the close association between metabolic dysfunction and impaired autophagy in DS. CONCLUSION: There are significant mitochondrial functional changes in LCLs derived from DS, DSAD, and AD patients. Several parameters analyzed were consistently different between DS, DSAD, and AD lines suggesting that metabolic indicators between LCL groups may be utilized as biomarkers of disease progression and/or treatment outcomes.

Isolation and characterization of a first set of polymorphic microsatellite markers in saffron, Crocus sativus (Iridaceae).

PREMISE OF THE STUDY: Twelve novel polymorphic microsatellite loci were developed and characterized from a repeat-enriched genomic library of Crocus sativus to study population and conservation genetics of this economically and medically important species. METHODS AND RESULTS: The microsatellite loci were isolated using a modified Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO) method. The average number of alleles per locus was 2.6. The observed and expected heterozygosities varied from 0.07 to 0.92 and 0.1 to 0.58, respectively. Polymorphic information content value ranged from 0.09 to 0.55 with an average of 0.34. Four out of twelve loci showed significant departures from Hardy-Weinberg equilibrium. CONCLUSIONS: The microsatellite markers reported here will be useful for evaluating genetic diversity and will likely serve as an ideal resource for use in marker-assisted breeding programs, germplasm analysis, and varietal identification.