Virus-induced CRISPR-Cas9 system improved resistance against tomato yellow leaf curl virus.

Plant viruses are the most significant factors associated with massive economical losses in agricultural industries worldwide. Accordingly, many studies are dedicated to making virus-resistant crop varieties each year due to the ever-changing nature of viruses. Recently genome engineering methods have been used to confer interference against eukaryotic viruses. Research results on genome editing technics, in particular, CRISPR-Cas9, promises a feasible solution to make virus-resistant crops. In this research, we explored the possibility of utilizing CRISPR-Cas9 to obtain TYLCV resistant tomato varieties. Moreover, to overcome any potential off-target effects of Cas9, we used an inducible promoter to initiate Cas9 activity in case of the virus attack. Cas9 vector was transformed by the rgsCaM promoter, known as an endogenous silencer of RNAi and overexpressed after a virus attack. The golden gate cloning method was applied to construct sgRNAs. Intergenic region and coat protein-coding sequences of TYLCV were used to design sgRNAs. Infiltrated sensitive Money Maker varieties analyzed by real-time PCR, showed a significant reduction or delayed accumulation of viral DNA compared to the control plants. This result demonstrates the efficiency of using an inducible promoter in CRISPR-Cas9 constructs.

Genetic assessment of the internal transcribed spacer region (ITS1.2) in Mangifera indica L. landraces.

Mango (Mangifera indica) is one of the most important tropical fruits in the world. Twenty-two genotypes of native mangoes from different regions of southern Iran (Hormozgan and Kerman) were collected and analyzed for the ribosomal genes. GC content was found to be 55.5%. Fu and Li's D* test statistic (0.437), Fu and Li's F* test statistic (0.500) and Tajima's D (1.801) were positive and nonsignificant. A total of 769 positions were identified (319 with insertion or deletion including 250 polymorphic and 69 monomorphic loci; 450 loci without any insertion or deletion including 35 Singletons and 22 haplotypes). Nucleotide diversity of 0.309 and a high genetic differentiation including Chi square of 79.8; P value of 0.3605 and df value of 76 was observed among mango genotypes studied. The numerical value of the ratio dN/dS (0.45) indicated a pure selection in the examined gene and the absence of any key changes. Cluster analysis differentiated the mango used in this research (M. indica L.) into two genotypes but could not differentiate their geographical locations. The results of this study indicated that a high genetic distance exists between HajiGholam (Manojan) and Arbabi (Rodan) genotypes and showed higher genetic diversity in mango of Rodan region. Results of present study suggested that for successful breeding, the genotypes of Rodan region mango especially Arbabi mango can be used as a gene donor and ITS can be a suitable tool for genetic evaluations of inter and intra species.

Comparison the Effect of Ferutinin and 17ß-Estradiol on Bone Mineralization of Developing Zebrafish (Danio rerio) Larvae.

There is an urgent need to develop novel drugs for osteoporosis which occurs due to estrogen deficiency. Phytoestrogens derived from medicinal plants would be the best alternative to chemical drugs with harmful side effects. The main purpose of the present study was to investigate the effect of ferutinin compared to 17ß-estradiol (E2) on bone mineralization of zebrafish larvae. Regarding the lack of publications, the histology analysis was performed after exposure to E2 to find effective treatment on bone mineralization of developing zebrafish larvae. Then, the larvae were exposed to four concentrations of ferutinin at three time points to assess the mortality, the expression of some related genes and histology of the ceratohyal and hyomandibular of treated larvae. The RT-PCR result of the treatment groups demonstrated the similar expression pattern in the larvae which were exposed to 1.25 µg/mL of ferutinin and 2 µM of E2 at 2 dpf, which confirmed the result of histology analysis. In addition, RT-qPCR of high concentration of ferutinin and E2 demonstrated that bmp2a/b and esr1 were downregulated and upregulated when the larvae were exposed to 5 µg/mL of ferutinin and 10 µM of E2, respectively.

Fabrication of a Food Nano-Platform Sensor for Determination of Vanillin in Food Samples.

Herein, we describe the fabrication of NiO decorated single wall carbon nanotubes (NiO-SWCNTs) nanocomposites using the precipitation method. The synthesized NiO-SWCNTs nanocomposites were characterized by X-ray diffraction (XRD) and Transmission electron microscopy (TEM). Remarkably, NiO-SWCNTs and 1-butylpyridinium hexafluorophosphate modified carbon paste electrode (CPE/NiO-SWCNTs/BPrPF6) were employed for the electrochemical detection of vanillin. The vanillin sensor showed an ultra-high sensitivity of 0.3594 µA/µM and a low detection limit of 0.007 µM. In the final step, the NiO-SWCNTs/BPrPF6 was used as the suitable tool for food analysis.

The intraspecific variability of mitochondrial genes of Agaricus bisporus reveals an extensive group I intron mobility combined with low nucleotide substitution rates.

Intraspecific mitochondrial variability was studied in ten strains of A. bisporus var. bisporus, in a strain representative of A. bisporus var. eurotetrasporus and in a strain of the closely related species Agaricus devoniensis. In A. bisporus, the cox1 gene is the richest in group I introns harboring homing endonuclease genes (heg). This study led to identify group I introns as the main source of cox1 gene polymorphism. Among the studied introns, two groups were distinguished according to the heg they contained. One group harbored heg maintained putatively functional. The other group was composed of eroded heg sequences that appeared to evolve toward their elimination. Low nucleotide substitution rates were found in both types of intronic sequences. This feature was also shared by all types of studied mitochondrial sequences, not only intronic but also genic and intergenic ones, when compared with nuclear sequences. Hence, the intraspecific evolution of A. bisporus mitochondrial genome appears characterized by both an important mobility (presence/absence) of large group I introns and by low nt substitution rates. This stringent conservation of mitochondrial sequences, when compared with their nuclear counterparts, appears irrespective of their apparent functionality and contrasts to what is widely accepted in fungal sequence evolution. This strengthens the usefulness of mtDNA sequences to get clues on intraspecific evolution.

Copper increases laccase gene transcription and extracellular laccase activity in Pleurotus eryngii KS004.

The white-rot fungus Pleurotus eryngii secretes various laccases involved in the degradation of a wide range of chemical compounds. Since the laccase production is relatively low in fungi, many efforts have been focused on finding ways to increase it, so in this study, we investigated the effect of copper on the transcription of the pel3 laccase gene and extracellular laccase activity. The results indicate that adding 0.5 to 2 mM copper to liquid cultures of P. eryngii KS004 increased both pel3 gene transcription and extracellular laccase activity in a concentration-dependent manner. The most significant increase in enzyme activity occurred at 1 mM Cu2+, where the peak activity was 4.6 times higher than in control flasks. Copper also induced the transcription of the laccase gene pel3. The addition of 1.5 and 2 mM Cu2+ to fungal culture media elevated pel3 transcript levels to more than 13-fold, although the rate of induction slowed down at Cu2+ concentrations higher than 1.5 mM. Our findings suggest that copper acts as an inducer in the regulation of laccase gene expression in P. eryngii KS004. Despite its inhibitory effect on fungal growth, supplementing cultures with copper can lead to an increased extracellular laccase production in P. eryngii.

Effect of Bone Grafting on Hard and Soft Tissue Changes Following Immediate Implant Installation in Mandibular Molar Sites: A Double-Blind Randomized Clinical Trial.

This study investigated the effect of bone grafting on hard and soft tissue alterations after immediate implant insertion in mandibular molar sites. This randomized, double-blind clinical trial consisted of 30 healthy patients (17 women and 13 men aged 22-58 years) who required immediate implant installation to replace a first or second mandibular molar. Only subjects with a buccal gap between 2 and 4 mm were selected. The participants were randomly allocated to two groups. In the experimental group, the gap was augmented by an allograft, whereas in the control group no graft was applied. Marginal bone level, probing depth, keratinized gingival width, and bleeding on probing were assessed at the time of implant placement (T0), 1 month (T1), and 3 months (T2) after surgery. There was no significant difference in hard and soft tissue parameters between the grafted and nongrafted sites at any of the durations (P < 0.05). Bone level decreased significantly in both groups (P < 0.05). However, the amount of probing depth and the frequency of cases showing bleeding on probing did not alter over the experiment either in the test or in the control group (P > 0.05). Bone grafting simultaneously with immediate implant installation had no significant effect on hard and soft tissue outcomes when the buccal gap size was between 2 and 4 mm. Therefore, the use of a bone substitute is not mandatory up to the jumping distance of 4 mm in immediate implant surgery.

Yield performance of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.) P. Karst. (higher Basidiomycetes), using different waste materials as substrates.

In this research the effect of sawdust, malt extract, and wheat bran on yield, biological efficiency (BE), and mycelia growth of Ganoderma lucidum was investigated. Three kinds of sawdust (beech, poplar, and hornbeam) as basal medium were mixed with two levels of wheat bran (5% and 10% w/w) and malt extract (2.5% and 5% w/w) as medium supplement for production of G. lucidum in factorial experiments on the basis of completely randomized design with three replications. The results showed that various kinds of sawdust affect fruiting body yield, BE, and mycelia growth rate significantly. The highest fruiting body yield and BE (102.58 g/kg and 12.89%, respectively) were found using hornbeam sawdust. The beech sawdust promotes the mycelia growth rate more than other sawdust. Analysis of variance showed that there is a significant interaction between the sawdust type and wheat bran, sawdust type and malt extract, and wheat bran and malt extract as far as yield and BE of G. lucidum was concerned. A final comparison of the different formulae indicated that the best combinations for high yield (142.44 g/kg) and BE (18.68%) were obtained in a combination of poplar sawdust with 5% malt extract and 10% wheat bran. The highest mycelia growth rate (10.6 mm/day) was obtained in a combination of beech sawdust with 2.5% malt extract and 10% wheat bran.

Rheological properties of wood/bacterial cellulose and chitin nano-hydrogels as a function of concentration and their nano-films properties.

In this study, rheological properties of the Wood Cellulose NanoFibers (WCNF), Bacterial Cellulose NanoFibers (BCNF), and Chitin NanoFibers (ChNF) as well as physical properties of films prepared from each nano-hydrogel were investigated. Each nano-hydrogel was prepared in 2 concentrations of 0.5 and 1 wt% for rheological study. Rheological properties were measured using a rotational rheometer. The flow behaviour data were fitted with rheological models. Apparent viscosity was higher in higher concentrations of nano-hydrogels. Herschel-Bulkley model was the best model for flow behaviour data fitting. BCNF nano-hydrogels had the highest hysteresis loop while WCNF nano-hydrogels had the best structure recovery and lowest hysteresis loop. At LVE (Linear Viscoelastic Region), G' (storage modulus) and G″ (loss modulus) had a constant value, but as strain increased their values decreased. Storage modulus was found to be greater than loss modulus in all samples during frequency sweep test. BCNF nano-hydrogel showed the lowest frequency dependency. Chitin nanofilms had the highest elongation and stress value.

Modification of activated carbon by MIL-53(Al) MOF to develop a composite framework adsorbent for CO2 capturing.

In this research, a novel composite is synthesized based on activated carbon and MIL-53(Al) through the solution mixing method at different MOF weight fractions, and the CO2 loading of prepared samples are measured in the batch and continuous apparatus. The structure, crystallinity, surface area, and chemical functionality of activated carbon, MIL-53(Al), and developed composite are characterized through BET, X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR). The CO2 and N2 adsorption capacity of activated carbon, MIL-53(Al), and composites are examined in an isothermal batch reactor at the pressure range 0-110 kPa and equilibrium temperature 305 K. The adsorption isotherm of CO2 is correlated by the Langmuir and Toth models. Besides, the performance of composite is compared with MIL-53(Al) and activated carbon in a continuous packed bed at flow rate range 15-25 ml min-1 and temperature 32 °C, and the breakthrough curves are developed. The results show that increasing MOF content in the composite increases CO2 adsorption capacity, so the CO2 loading of synthesized composite containing 10%, 20%, and 30% MOF is 1.608, 1.704, and 1.792 mmol gr-1, respectively.

Evaluation of morphological traits of wheat varieties at germination stage under salinity stress.

Salinity stress is one of the major plant growth-limiting factors in agriculture. It causes ionic imbalance, thus decrease the growth and yield attributes of crops especially wheat. Seedling stage is considered as one of the most sensitive stages under salinity stress. Survival of seeds at seedling stage can overcome the adverse impacts of salinity stress to some extent. Selection of salt tolerant varieties in seedling stage is considered as an effective strategy. Hence, current study was conducted to examine the seed germination responses of four wheat varieties under different levels of salinity. The wheat varieties such as 'Rakhshan', 'Sirvan', 'Pishgam' and 'Heidari' were grown and four salinity levels of 0, 4, 8 and 12 dS/m were applied under completely randomized design. The varieties such as 'Sirvan', 'Rakhshan' and 'Heidari' showed significant response for germination compared to 'Pishgam' at 12 dS/m salinity. Furthermore, the variety 'Rakhshan' showed significantly higher germination rate (20.3%), higher root length (33.4%) and higher shoot length (84.3%) than 'Pishgam', 'Sirvan' and 'Sirvan' respectively. However, contrasting results were obtained for dry weight of seedlings where 12.2% increase was observed in 'Pishgam' over 'Rakhshan' at 12 dS/m salinity that might be due to higher capability to uptake of Na and Cl ions. In conclusion, 'Rakhshan' wheat variety proved to be the most salinity tolerant as it grew better under saline soil conditions. More investigations at field level are recommended to declare 'Rakhshan' as salinity tolerant cultivar.

Spotted hyena optimizer algorithm for capacitor allocation in radial distribution system with distributed generation and microgrid operation considering different load types.

In this paper, the optimal allocation of constant and switchable capacitors is presented simultaneously in two operation modes, grid-connected and islanded, for a microgrid. Different load levels are considered by employing non-dispatchable distributed generations. The objective function includes minimising the energy losses cost, the cost of peak power losses, and the cost of the capacitor. The optimization problem is solved using the spotted hyena optimizer (SHO) algorithm to determine the optimal size and location of capacitors, considering different loading levels and the two operation modes. In this study, a three-level load and various types of loads, including constant power, constant current, and constant impedance are considered. The proposed method is implemented on a 24-bus radial distribution network. To evaluate the performance of the SHO, the results are compared with GWO and the genetic algorithm (GA). The simulation results demonstrate the superior performance of the SHO in reducing the cost of losses and improving the voltage profile during injection and non-injection of reactive power by distributed generations in two operation modes. The total cost and net saving values for DGs only with the capability of active power injection is achieved 105,780 $ and 100,560.54 $, respectively and for DGs with the capability of active and reactive power injection is obtained 89,568 $ and 76,850.46 $, respectively using the SHO. The proposed method has achieved more annual net savings due to the lower cost of losses than other optimization methods.

MicroRNAs regulate the main events in rice drought stress response by manipulating the water supply to shoots.

MicroRNAs (miRNAs) are small endogenous regulatory RNAs that are involved in a variety of biological processes related to proliferation, development, and response to biotic and abiotic stresses. miRNA profiles of rice (Oryza sativa L. cv. IR64.) leaves in a partial root zone drying (PRD) system were analysed using a high-throughput sequencing approach to identify miRNAs associated with drought signalling. The treatments performed in this study were as follows: well-watered ("wet" roots, WW), wherein both halves of the pot were watered daily; drought ("dry" roots, DD), wherein water was withheld from both halves of the pot; and well-watered/drought ("wet" and "dry" roots, WD), wherein one half of each pot was watered daily, the same as in WW, and water was withheld from the other part, the same as in DD. High-throughput sequencing enabled us to detect novel miRNAs and study the differential expression of known miRNAs. A total of 209 novel miRNAs were detected in this study. Differential miRNA profiling of the DD, WD and WW conditions showed differential expression of 159 miRNAs, among which 83, 44 and 32 miRNAs showed differential expression under both DD and WD conditions. The detection of putative targets of the differentially expressed miRNAs and investigation of their functions showed that most of these genes encode transcription factors involved in growth and development, leaf morphology, regulation of hormonal homeostasis, and stress response. The most important differences between the DD and WD conditions involved regulation of the levels of hormones such as auxin, cytokinin, abscisic acid, and jasmonic acid and also regulation of phosphor homeostasis. Overall, differentially expressed miRNAs under WD conditions were found to differ from those under DD conditions, with such differences playing a role in adaptation and inducing the normal condition. The mechanisms involved in regulating hormonal homeostasis and involved in energy production and consumption were found to be the most important regulatory pathways distinguishing the DD and WD conditions.

Effects of therapeutic touch on blood hemoglobin and hematocrit level.

BACKGROUND: Therapeutic Touch (TT) is a widely used complementary therapy. This study investigated the effects of TT on hemoglobin and hematocrit level in students who were basically healthy. METHOD: The volunteers with a hemoglobin level less than 12 grams per deciliter (g/dl) were randomly assigned to three groups of TT, mimic therapeutic touch (MT), and control. Blood samples were collected before the first treatment and again a week after the last one and measurements were taken. RESULTS: TT increased the level of hemoglobin (.99 .13 g/dl) and hematocrit (2.82 .43%) significantly. MT also increased the level of hemoglobin (.55 .11 g/dl) and hematocrit (2.75 .44%) significantly. No significant changes were found in the control group. TT increased hemoglobin more effectively than MT (p< .05). CONCLUSIONS: Significant changes of both variables in TT and MTgroups suggest that more careful precision might be needed while selecting individuals as sham therapists in further experiments.

Antibacterial activity of Pseudonocardia sp. JB05, a rare salty soil actinomycete against Staphylococcus aureus.

Staphylococcus aureus is a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds against S. aureus and characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated as Pseudonocardia sp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL(-1) against S. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain of Pseudonocardia. The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

Biofield therapies: biophysical basis and biological regulations?

Complementary and alternative medicine (CAM) is increasingly popular in biomedical health care. One area of alternative medicine, biofield therapies, claims to manipulate individuals 'energy field' in order to enhance healing and wellbeing. This article reviews some recent studies addressing the characterization of endogenous energy fields and the way they affect the physiologic processes.