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Many items affect the yields of every recombinant protein production. Promoters are one of the key regulatory elements which control the level of recombinant protein expression in the host. Although in some studies (Hepatitis B Surface Antigen) HBsAg was cloned in E. coli, in many of them common promoters were used. In this study two co-vectors (PHK and PHGK) were designed and used. Both of them were shuttle types based on phycocyanin-specific promoter and each was a combination of two types of vectors: a plasmid and a transposon. ELISA results, in line with Western blot results, showed the ability of phycocyanin promoter as well as revealed the expression level of HBsAg in transgenes from the Top10 PHK vector is higher than those of Top10 PHGK. The expression of HBsAg under the phycocyanin promoter in the present study is 7.5 µg/lit. In the current study as a pilot step, our attitude and objective of bacteria expression are to evaluate the function of the phycocyanin promoter based on the HBsAg gene. Due to the nature of the shuttle vector, in continuation of the present study to reduce the non-responsive populations, high antigen expression, meet the need for renewal and, other benefits, expression of HBsAg in other than bacterial host is under investigation.

Drought stress is one of the most significant limiting factors affecting crop products. Foxtail millet (Setaria italica) is one of the superior drought-tolerant crop plants, comprising high collinearity with other staple cereals. The present study used a meta-analysis approach to identify genomic regions and candidate genes associated with drought tolerance and yield-related traits in foxtail millet. Meta-analysis using all 448 collected original quantitative trait loci (QTLs) lead to detection of 41 meta-QTLs (MQTLs) on the nine chromosomes of foxtail millet. The confidence interval (CI) of the identified MQTLs was determined to be 0.31–14.47 cM (5.23 cM average), which was 3.5 times narrower than the mean CI of the original QTLs. Based on the available RNA-seq and microarray data, 1631 differentially expressed genes (DEGs) were detected in 41 MQTLs. Through synteny analysis, 8, 4, and 2 ortho-MQTLs were recognized within co-linear regions of foxtail millet with rice, barley, and maize, respectively. To detect the most important genome regions involved in the genetic control of yield maintenance and drought tolerance of foxtail millet, 10 MQTLs with physical intervals of below 1 Mb and seven hotspot regions with high-value QTL-overview index were distinguished. Several candidate genes involved in sensing and signaling, transcription regulation, ROS inhibition, and adaptation to abiotic stress of foxtail millet were detected by seeking drought-responsive genes in MQTL regions with CI

Chickpea is an important food legume cultivated in several countries. A sudden drop in autumn temperature, freezing winter temperature, and late spring cold events result in significant losses in chickpea production. The current study used RNA sequencing of two cold tolerant (Saral) and sensitive (ILC533) Kabuli chickpea genotypes to identify cold tolerance-associated genes/pathways. A total of 200.85 million raw reads were acquired from the leaf samples by Illumina sequencing, and around 86% of the clean reads (199 million) were mapped to the chickpea reference genome. The results indicated that 3710 (1980 up- and 1730 down-regulated) and 3473 (1972 up- and 1501 down-regulated) genes were expressed differentially under cold stress in the tolerant and sensitive genotypes, respectively. According to the GO enrichment analysis of uniquely down-regulated genes under cold stress in ILC533, photosynthetic membrane, photosystem II, chloroplast part, and photosystem processes were enriched, revealing that the photosynthesis is severely sensitive to cold stress in this sensitive genotype. Many remarkable transcription factors (CaDREB1E, CaMYB4, CaNAC47, CaTCP4, and CaWRKY33), signaling/regulatory genes (CaCDPK4, CaPP2C6, CaMKK2, and CaHSFA3), and protective genes (CaCOR47, CaLEA3, and CaGST) were identified among the cold-responsive genes of the tolerant genotype. These findings would help improve cold tolerance across chickpea genotypes by molecular breeding or genetic engineering.

Global warming is posing a serious threat to crop production that can cause various types of environmental stresses. The genetic information of lentil in response to environmental stresses is severely lacking. Heat shock proteins (HSPs) can help plants deal with the adverse conditions caused by abiotic stress. Therefore, the identification of key HSPs can help provide comprehensive insights into molecular mechanisms of abiotic stress. In this study, RNA sequencing data analysis was used to identify, classify, and study the changes in expression of HSPs in lentil under cold, heat, drought, and salt stresses. Finally, the expression changes of the top HSPs under all treatments and control condition were validated by qRT-PCR. The results showed that among the six identified HSP classes, HSP40 and HSP60 have the highest and lowest percentage of transcription members, respectively. Our findings also show that the expression of HSPs is opposite under heat and cold stress. In addition, the results showed that the highest levels of HSPs’ expression occurred in response to temperature stresses, especially heat stress, while this change was not significant in drought and salt stresses. In general, our findings show that there is variation in the expression of HSPs in response to abiotic stresses, which can play an important role in better understanding the molecular mechanism of lentil tolerance to abiotic stresses.

Background Seeds of Bunium persicum, a member of the Apiaceae family, are traditionally consumed in the Iranian diet. Although many of its biological properties have been fully studied, no reliable information about the anticancer/cytotoxicity of this herb is available currently. Methods Herein, we first determined the major bioactive compounds of B. persicum seeds extract (BPSE) using GC-Mass analysis. Then, the cytotoxicity of the extract alone and in combination with vincristine (VCR), a common chemotherapy drug, was investigated on Breast cancer cell lines, MCF-7 and MDA-MB-231, and a normal cell line, MCF-10A, via MTT assay. Furthermore, the anticancer activity of these compounds was evaluated on MCF-7 cells through ROS assay, Real-Time PCR, Western blot, flow cytometry, and cell cycle assay. Results Our results showed that the extract considerably reduced the viability of cancerous cells while was harmless to normal ones. Besides, the combination of BPSE with VCR resulted in synergistic activity. The MCF-7 cells’ intracellular ROS increased more than 2 times after treatment with BPSE and/or BPSE-VCR. Moreover, the IC30 of BPSE (100 µg/ml) significantly increased the BAX/BCL-2 and P53 genes expression, while reducing the expression of the MYC gene. Also, the cell cycle was arrested in the G2 phase, in treated cells. The BPSE-VCR combination synergistically reduced the NF-κB and increased the Caspase-3 proteins’ expression. The percent of apoptosis in the cell treated with the extract, VCR, and their combination was 27, 11, and 50, respectively. Conclusion The present study demonstrated the anticancer activity of the BPSE and its potential for application in combination therapy with VCR.

Lentil cultivation could be challenged by combined heat and drought stress in semi-arid regions. We used RNA-seq approach to profile transcriptome changes of Lens culinaris exposed to individual and combined heat and drought stresses. It was determined that most of the differentially expressed genes observed in response to combined stress, could not be identified by analysis of transcriptome exposed to corresponding individual stresses. Interestingly, this study results revealed that the expression of ribosome generation and protein biosynthesis and starch degradation pathways related genes were uniquely up-regulated under the combined stress. Although multiple genes related to antioxidant activity were up-regulated in response to all stresses, variation in types and expression levels of these genes under the combined stress were higher than that of individual stresses. Using this comparative approach, for the first time, we reported up-regulation of several TF, CDPK, CYP, and antioxidant genes in response to combined stress in plants.

Selection of transgenic plants by using genes encoding screenable markers of plant origin with health benefit properties, such as anthocyanin is an important aim in plant genetic engineering. In this study, Malus domestica MYB10 (MdMYB10) gene, was used for Agrobacterium tumefaciens-mediated transformation of two SBS-02 and SBS-04 sugar beet lines. The impact of different light regimes on plant tissue culture from a combination of light, dark/light and dark was investigated. The results of this study showed that the MdMYB10 gene was successfully integrated into the selected purple transgenic lines, suggesting that the expression of MdMYB10 gene in sugar beet shoots can be used as a screenable markers for transformation, possibly replacing antibiotic resistant genes. Furthermore, the results of the antibacterial activity of transgenic plants extracts showed that the total extract obtained from transgenic lines significantly (P

Introduction Nowadays, increasing environmental risks to human health have caused increasing attention to use chemical nitrogen sources efficiently or nitrogen supplying from organic amendments. In this respect, chicken manure seems to be a good alternative to chemical nitrogen fertilizers. The purpose of this experiment is to investigate the role of urea and poultry manure and ecological factors (climate) on the quantity of some chemical compositions of the potato tuber. Materials and Methods To study the effects of urea and poultry manure on some potato tuber compositions, the factorial experiment was conducted in 2015-2016 during the growing season in 2 regions; Azna and Khorramabad which is located in Lorestan province in the west of Iran. The factors were 2 different sources of nitrogen supply including urea fertilizer and poultry manure. Results Results show that the highest activity of nitrate reductase (NR) in both sites was observed using 10304 kg of poultry manure ha-1. The results also showed that the tuber nitrate content was only affected by the fertilizer source. In both studied places, the highest tuber nitrate was associated with high levels of urea fertilizer. Based on the results, the vitamin C of potato tubers grown by poultry manure were also superior to the relative vitamin C content. According to the results of this experiment, the accumulation of more than 2 times the nitrate in the tubers was obtained from urea fertilizer (244.2 versus 100 ppm). Conclusions In order to achieve food security and an efficient and sustainable food chain, chemical fertilizers must be replaced with organic fertilizers. Thus, the belief that the use of organic fertilizers, at any level, would not cause any problem for the health of products is definitely incorrect.

The present study was conducted to determine the changes which occurred in the water-stressed fenugreek (Trigonella foenum-gracum L.) in response to melatonin treatment. Fenugreek is a valuable med...

Expression of strong antimicrobial peptides in plants is of great interest to combat a wide range of plant pathogens. To bring the Dermaseptin B1 (DrsB1) peptide to the intimate contact of the plant pathogens cell wall surface, the DrsB1 encoding sequence was fused to the C-terminal part of the two copies of the chitin-binding domain (CBD) of the Avr4 effector protein and used for Agrobacterium rhizogenes-mediated transformation. The expression of the recombinant protein in the tobacco hairy roots (HRs) was confirmed by molecular analysis. Antimicrobial activity analysis of the recombinant protein purified from the transgenic HRs showed that the (CBD)2-DrsB1 recombinant protein had a significant (p

Breeding new rice varieties with high Eating and Cooking Quality (ECQ) characteristics is an important goal in rice breeding programs. To this end, 242 Recombinant Inbred Lines (RILs) of two populations derived from Alikazemi/IR67017-180-2-1-2 and Alikazemi/Saleh crosses were evaluated in two environments. The main and epistatic QTLs and QTL × Environment (Q × E) interactions were analyzed using the Mixed-Model-based Composite Interval Mapping method. A total of 17 main QTLs with additive effects were mapped on 6 rice chromosomes, including 6 QTLs for Amylose Content, 5 QTLs for Gelatinization Temperature and 6 QTLs for Gel Consistency. Seven QTLs were found on chromosome 6 associated with a common RM4128 marker (close to the Alk locus) influencing the ECQ characteristics with favorable alleles from the common female parent. Five QTL clusters were identified on chromosome 6, 7, 8, and 9, suggesting that the co-located QTLs with pleiotropic effects may control ECQ traits. A major QTL cluster was found for AC and GC between RM4128 and RM549 markers close to the Alk locus on chromosome 6, which is in the vicinity of another QTL located in RM587-RM4128 interval. 13 QTL pairs were identified to have a significant epistatic, Additive × Additive effects involved in controlling ECQ traits. A small phenotypic variation was found for the epistatic QTLs and Q × E interactions, suggesting the complexity of ECQ inheritance traits in rice. Further comprehensive studies and validation of the detected QTLs could be useful for marker-assisted selection to improve grain quality in rice.

Extreme temperature is one of the serious threats to crop production in present and future scenarios of global climate changes. Lentil (Lens culinaris) is an important crop, and there is a serious lack of genetic information regarding environmental and temperature stresses responses. This study is the first report of evaluation of key genes and molecular mechanisms related to temperature stresses in lentil using the RNA sequencing technique. De novo transcriptome assembly created 44,673 contigs and differential gene expression analysis revealed 7494 differentially expressed genes between the temperature stresses and control group. Basic annotation of generated transcriptome assembly in our study led to the identification of 2765 novel transcripts that have not been identified yet in lentil genome draft v1.2. In addition, several unigenes involved in mechanisms of temperature sensing, calcium and hormone signaling and DNA-binding transcription factor activity were identified. Also, common mechanisms in response to temperature stresses, including the proline biosynthesis, the photosynthetic light reactions balancing, chaperone activity and circadian rhythms, are determined by the hub genes through the protein-protein interaction networks analysis. Deciphering the mechanisms of extreme temperature tolerance would be a new way for developing crops with enhanced plasticity against climate change. In general, this study has identified set of mechanisms and various genes related to cold and heat stresses which will be useful in better understanding of the lentil's reaction to temperature stresses.

Structural mutations providing herbicide resistance may cause a modification of the three dimensional structure of a protein which will lead to a decrease in the herbicide efficacy. Wild oat (Avena ludoviciana Durieu.) is an increasingly disruptive weed in areas of intensive cereal production, thus the aim of this research was to identify mutations conferring resistance to ACCase-inhibitor herbicides at greenhouse, laboratory and in silico scales. Among the selected biotypes, No. 3 in the position 1781 (Ile1781-Leu) and No. 14 in the position 2041 (Ile2041-Asn), showed resistance to ACCase-inhibitor. The above mutations were confirmed using the specific primers and PCR-based methods. Analysis of molecular docking indicated that residues of Trp1948 and Pro2001 are important in the binding site and showed remarkable variation in the mutation types. Using molecular dynamic simulation analysis, we demonstrated that mutation types changed the conformation of the enzyme. These changes resulted in compressed conformation in the active site, which limited the availability of binding herbicide-enzyme. In present, no crystallography molecular structure and modeling reported on the ACCase of plants and this study investigated interactions of clodinafop propargyl and ACCase CT domain in A. ludoviciana by modeling, docking and simulations for the first time. Totally, bioinformatics analysis as well as PCR-based method confirmed that herbicide resistance conferred by nucleotide mutations in the gene sequence.

Opium poppy (Papaver somniferum L.) remains the only commercial source for several pharmaceutical alkaloids. In present study, opium poppy plants were genetically manipulated via VIGS technique using pTRV2-BBE (to silencing of BBE1 gene), pTRV2-COM (to simultaneous silencing of T6ODM and CODM genes) and pTRV2-BBE-COM (to simultaneous silencing of T6ODM, CODM and BBE1 genes) constructs. Also, via Agrobacterium-mediated transient expression technique, poppy plants were genetically manipulated using ACS (to simultaneous silencing of T6ODM, CODM and BBE1) and ACS-4ꞌOMT2 (to simultaneous silencing of T6ODM, CODM and BBE1 and over-expression of 4ꞌOMT2) constructs. HPLC analysis showed lower sanguinarine and noscapine levels and elevated levels of morphine in plants infiltrated with pTRV2-BBE construct. In plants infiltrated with pTRV2-COM construct, reduction in the morphine and codeine content and a substantial increase in accumulation of thebaine and papaverine were observed. A substantial increase in the accumulation of thebaine and papaverine and lower levels of morphine, codeine, sanguinarine and noscapine were observed in plants infiltrated with pTRV2-BBE-COM and ACS constructs. Plants infiltrated with ACS-4ꞌOMT2 showed lower levels of codeine, sanguinarine and noscapine and elevated levels of morphine, thebaine and papaverine. Although, previous studies reported the role of our selected genes in benzylisoquinoline alkaloids (BIAs) biosynthetic pathway, but we simultaneously engineered these genes to alter the levels of specific BIAs and take advantage of all genes at the same time. For the first time, we showed different pattern in BIAs accumulation in simultaneous gene manipulation compared with single gene manipulation in previous reports, especially for higher levels of thebaine (about 1550%) and papaverine (about 155%). We also determined potential role of BBE1 gene in noscapine biosynthesis. This is the first report of simultaneous silencing and over-expression of biosynthetic genes in opium poppy and our finding could establish vast potential for metabolic engineering in opium poppy.

Salinity is one of the main abiotic stresses limiting crop productivity. In the current study, the transcriptome of wheat leaves in an Iranian salt-tolerant cultivar (Arg) was investigated in response to salinity stress to identify salinity stress-responsive genes and mechanisms. More than 114 million reads were generated from leaf tissues by the Illumina HiSeq 2500 platform. An amount of 81.9% to 85.7% of reads could be mapped to the wheat reference genome for different samples. The data analysis led to the identification of 98819 genes, including 26700 novel transcripts. A total of 4290 differentially expressed genes (DEGs) were recognized, comprising 2346 up-regulated genes and 1944 down-regulated genes. Clustering of the DEGs utilizing Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated that transcripts associated with phenylpropanoid biosynthesis, transporters, transcription factors, hormone signal transduction, glycosyltransferases, exosome, and MAPK signaling might be involved in salt tolerance. The expression patterns of nine DEGs were investigated by quantitative real-time PCR in Arg and Moghan3 as the salt-tolerant and susceptible cultivars, respectively. The obtained results were consistent with changes in transcript abundance found by RNA-sequencing in the tolerant cultivar. The results presented here could be utilized for salt tolerance enhancement in wheat through genetic engineering or molecular breeding.

Vermicompost can have an effective role in plant growth and nutrition. It can also reduce the harmful effects of stress caused by environmental factors. This study was a factorial experiment with a completely randomized plot design and three replications. The results showed that the calcium concentration (+40% +39%) of root and calcium (+71% +67%) and potassium contents (+49% +59%) of leaf under conditions of moderate and severe water stress increased under vermicompost treatments at the ratio of 30%. Sodium absorption (−37% −21%) in moderate and severe conditions was significantly reduced by vermicompost treatment. In moderate and severe water stress, the growth medium of 30% vermicompost significantly enhanced proline (+39%) and soluble protein content (+28%) compared to the control. In conclusion, our results proved vermicompost fertilizer due to its structural properties and components, served to reduce the negative effects of water stress by increasing the absorption of calcium (Ca) and potas...

One goal in the face of deficit water conditions is to increase growth and yield. Agro-industrial production frequently causes environmental pollution by using chemical fertilizers. In recent decad...

Late-season heat stress (LSH) is a limiting factor for wheat production. Besides, low zinc and poor protein diet usually is a problem in low-income countries. The primary calorie source in such countries is prepared from bread. This study aims to mitigate heat stress by zinc and nitrogen application and improve zinc and protein content in wheat grain. We did the field experiments as a split-split-plot based on a randomized complete block design with four replications to assess zinc and nitrogen’s possible mitigation effect on LSH and protein and zinc enrichment of wheat grain during two years. Factors included LSH by delay in planting date (optimum, late, and very late) as the main plot, nitrogen (0, 75, 150, and 225 kg ha–1) in subplots, and zinc (0, 10, and 20 kg ha–1) as sub-subplots. We measured yield, yield components, physiological traits, zinc, and protein contents in the grain. Results showed that the highest relative water content and cell-membrane thermal stability were attained at the optimum planting date, 150 kg N ha–1 and 20 kg Zn ha–1. The maximum chlorophyll a and carotenoids contents in wheat cells were recorded in the optimum planting date, 225 kg N ha–1, and 20 kg Zn ha–1. Heat stress reduced the grain yield. In the second year of the experiment, the grain number per unit area was more than that of the first year; however, the highest grain yield was achieved in the first year owing to the higher mean grain weight. Nitrogen application decreased the adverse effects of heat stress on grain yield by increasing the grain number. Zinc application diminished the adverse effects of heat stress by increasing the mean grain weight. The adverse impact of the LSH on grain yield was more than that of biological yield. Heat stress reduced the hectolitre weight and zinc content of the grain. Meanwhile, it increased grain protein. In general, under LSH, the application of 225 kg N ha–1 and 20 kg Zn ha–1 can reduce the adverse effects of heat on the grain quality and quantity.

Water availability is the main factor limiting crop growth and productivity in dry regions. This study was carried out in order to determine the effect of spraying methanol solution on the photosynthetic characteristics of bean plants. The main aim of our experiment was to improve plant performance under stress caused by water shortage. Two factors were involved: water-deficit stress, such as severe stress (25% of field capacity), mild stress (75% of field capacity), and no stress (100% of field capacity), and application of methanol solution spray at four concentrations: control (without spraying), 10, 20, and 30%. Methanol was applied three times at different growth stages (seedling, flowering, and podding stage) in 10-d intervals. The treatment with 20% methanol at the seedling stage resulted in increased net photosynthesis (P N), intercellular CO2 concentration (C i), and decreased transpiration rate (E) under no stress and mild stress conditions. Under severe stress, 10 and 20%-methanol treatments resulted in increased C i, maximal quantum yield of PSII photochemistry, and decreased E. At the flowering stage, methanol treatments resulted in decreased E and increased C i under mild and severe stress. At the podding stage, 10 and 20%-methanol treatments resulted in increased P N, C i, and total chlorophyll content under mild stress. In conclusion, we suggested that foliar application of methanol had a positive role in enhancing photosynthetic performance.

In estimation of genetic parameters in perennial tree species on the basis of analysis of variance (ANOVA), heterogeneity of years and genotype × environment interaction for data sets during the juvenility to maturity life period is ignored. Therefore, a linear mixed model based on restricted maximum likelihood (REML) approximation for modeling of covariance structure of longitudinal data can improve our ability to analyze repeated measures data. In the present research, a modeling of variance-covariance structure by mixed model based on the REML approach has been used for characteristics of 26 apricot genotypes recorded during three years. Fitting unstructured covariance (UN) models for all traits indicated a great heterogeneity of variances among repeated years and the trends of response variables in the genotypes (except for RWC) was due to imperfect correlation of subjects measured in different years. Based on the same structure, positive correlations were estimated among fruit set, potassium content, and yield of pistil in repetitive years, and most traits showed high heritability estimation. To our knowledge, this is the first report in plant that genotypic correlation and heritability and their standard errors are estimated in a repeated measures data over years using REML approximation.

Water availability is an important factor for plant growth in arid environments. In recent decades, vermicompost (VC) fertilizer has been used in agriculture as a safe and effective fertilizer with high water-holding capacity. The aim of the present study was to characterize effects of VC fertilizer on photosynthetic activity of chickpea (Cicer arietinum L. cv. Karaj) under drought conditions at three different growth stages. Tests were carried out with four volumetric ratios of VC to soil, i.e., 0:100, 10:90, 20:80, and 30:70, and three levels of drought stress, i.e., no stress (NS), moderate drought (MS), and severe drought (SS) (100, 75, and 25% of field capacity, respectively). Evaluations were performed at the seedling, flowering, and podding stage. We found that the VC treatment under NS conditions significantly increased total chlorophyll content [Chl (a+b)], intercellular CO2 concentration (C i), net photosynthetic rate (P N), transpiration rate (E), and maximal quantum yield of PSII photochemistry (Fv/Fm) at all three stages. The VC addition of 10 and 20% significantly enhanced the Chl content and Fv/Fm under MS and Fv/Fm, C i, and P N under SS at the flowering stage. In conclusion, our results proved a positive effect of the VC fertilizer on photosynthesis of chickpea under NS conditions, but it was not found under MS and SS.

Due to its high productivity and sucrose content, sugarcane (Saccharum officinarum) is becoming the source of high-value bioproducts. Expression of bacterial extracellular polysaccharide genes in non-biopolymer accumulating plants is an excellent resource for production of added-value products. To this end, an expression cassette containing a full-length glucosyltransferase (gtfI) gene from Streptococcus downei driven by a CaMV promoter was expressed in a commercial sugarcane cultivar (CP48-103) using a biolistic approach. Copy number was assessed for a number of selected transgenic sugarcane lines by DNA blot analysis, where it was corroborated that each transgenic line contained at least two gtfI copies. The southern blot analysis of gtfI-expressing lines showed that the number of integrated copies ranged from two to four. The expression of gtfI in transgenic sugarcane plants was confirmed by mRNA blot analysis and qRT-PCR analysis. The expression of gtfI in transgenic sugarcane plants resulted in an approximate 30% reduction in sucrose accumulation, suggesting that mutansucrase actively converted sucrose to mutan polymer. In internodal stalk tissues, mutan polymer accumulated up to 55.9 mg/g FW, which apparent through glucan staining. The levels of glucose and fructose increased nearly by twofold, suggesting that mutansucrase may also have hydrolyzing activity.

Nano and bulk-forms of zinc oxide (ZnO) are used extensively in industry and consequently may accumulate in the environment. However, there is little information available on the comparative effects of these forms during the critical early stages of plant life. Furthermore, the role of chelating agents, which affect the bioavailability of metals, in ameliorating plant stress due to exposure to nano and bulk-forms of ZnO is not well characterised. In this study, the effects of different concentrations (0.5, 2.5, 5, 10, 50 and 100 ppm) of nano ZnO (22 nm) and bulk ZnO (natural form, 1000 nm) with and without organic (citrate) and inorganic (EDTA) chelators on germination and seedling growth, and oxidative stress markers of Nicotiana tabacum L. were compared. Chelators (without ZnO) enhanced root growth, whilst ZnO negatively affected seedling growth. ZnO toxicity was often mitigated by adding chelators, especially citrate, although at the highest levels (50 and 100 ppm) of ZnO, toxicity was more pronounced when chelated with EDTA, but was decreased with citrate. Collectively, our findings provide important information regarding the different morpho-physiological and biochemical effects of bulk and nano ZnO and organic and inorganic chelators (citrate and EDTA), which are all prevalent in the environment.

Background: Gastric cancer as one of the most important diseases affecting health in all worldwide. Current studies have confirmed associations of cytokine gene polymorphisms with the risk of gastric cancer development. The current research aimed to assess the association of IL-1B+3954 genotypes with the risk of gastric cancer in the Iranian population. Materials and Methods: This case-control study covered 49 gastric cancer patients compared to 53 cancer free individuals as a control group. Genomic-DNA extraction was carried out from bioptic samples of patients and peripheral blood of healthy volunteers. Polymorphism of IL-1B +3954 genotypes were analysed with a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: The frequencies of IL-1B +3954 A1A1, A1A2 and A2A2 genotypes in healthy individuals were 26.4, 66 and 7.6 %, respectively. However, in gastric cancer patients, A1A1, A1A2 and A2A2 with 4.1, 51 and 44.9% were observed (p

High temperature stress, especially on the early season of plant growth stages, is an agricultural problem in many areas in the world. A temporary or continually high temperature leads to a set of morphological, biochemical and physiological changes in plants, which consequently reduces the plant growth and development and finally may cause a severe reduction in economic yield. The main goal of this study was to assess plant response to high temperature stress (HTS) in early seedling of canola. This study is the first experiment on the effect of heat stress on proteome of canola. In the present research, a proteomics approach was used to evaluate the effects of high temperature stress, including 45 °C day/34 °C night for 2, 6 and 12 hour, on early seedling stage (2-day old) of canola. Proteins were isolated from hypocotyl and separated by two-dimensional polyacrylamide gel electrophoresis. Out of 381 protein spots, 28 and 34 proteins were significantly down- and up-regulated, respectively. The trend of mRNA expression for sucrose binding protein, a scorbate peroxidase and triosephosphateisomerase, was in accordance with their trend at translation level. Results of this study suggest that the up-regulation of proteins involved in cellular traffic, energy and metabolism, and down-regulation of some proteins involved in disease and defense, protein synthesis and signal transduction could be the main reason of physiological and morphological responses to high temperature stress. The observed increases in the level of ascorbate peroxidase protein and mRNA expression in canola hypocotyl in response to HTS suggests that ascorbate peroxidase is a short term high temperature stress response protein and is thus a candidate for gene modification strategies aimed at producing high temperature canola varieties. These results also suggest that the up regulation of protein involved in energy and metabolism in response to the heat stress can use most of nutritive reserves in seedling of canola and might explain the reduced growth of canola in heat stress conditions.

A simple, rapid and efficient ultrasound and salt-assisted liquid–liquid extraction (USALLE) method coupled with high-performance liquid chromatography (HPLC) has been introduced for extraction, clean-up and pre-concentration of oleuropein from olive leaves as a model analyte. In this technique, the plant sample was transferred into the extraction solvent (consisting of phosphate buffer and miscible organic solvents) and the mixture was exposed to ultrasonic waves. After ultrasonic-assisted extraction (UAE), phase separation was performed by addition of salt to the liquid phase. During salt-assisted liquid–liquid extraction (SALLE) the analyte was transferred into the supernatant organic phase. Various parameters that affect the extraction efficiency such as ultrasonic time and temperature, sample amount, type and volume of miscible organic solvent, type and concentration of salt and pH were evaluated and optimized. The calibration curve shows good linearity (r2 = 0.9934) and precision (RSD < 5.5%) in the range of 2.5–50 μg mL−1. The limit of detection (LOD) and limit of quantitation were 0.5 and 2.5 μg mL−1, respectively. The recoveries were in the range of 90.0–97.0% with RSD values ranging from 4.0 to 6.5%. Unlike the conventional extraction methods for plant extracts no evaporation and re-solubilization operations were needed in the proposed technique.

Weeds are serious problem in crop production and wild oat is a grass weed of economic and agronomic significance. We need to extend our basic knowledge of weeds especially in molecular genetics and gene expression. For study of gene expression by semi-quantitative and quantitative PCR, it is recommended that normalization of reference genes be carried out in order to select the most stable reference gene for a precise gene expression study. The purpose of this research was evaluation of four reference genes in response to treated and untreated (control) by herbicide in two tissues (stem and leaf) of non-target site resistance wild oat (A. ludoviciana). Four candidate reference genes including Actin, Ef1α (elongation factor 1 alpha), GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and TBP (TATA-box-binding protein) were used to determine stable reference gene exposed to the herbicide using the statistical methods of NormFinder, BestKeeper and delta-Ct. NormFinder indicated that TBP and Actin genes are the best combination of two genes for normalizing calculations (with a combined gene stability value of 0.012) for qPCR analysis under herbicide stress in different tissues of non-target site resistance wild oat. Based on the statistical results, the Ef1α gene was identified as the unstable reference gene. Totally, according to results of this study, TBP gene is the most stable reference gene and therefore, this gene can be used as a reference gene for future studies of quantitative PCR analysis of herbicide stress-responsive gene expression in wild oat and potentially in other grass weed species.

To identify miRNAs and assess the expression of genes involved in drought stress tolerance, leaf and root ESTs were analyzed in barley. To this end, the EGassembler bioinformatics service and IDEG6 were used for the pre-processing and identify differentially expressed genes among EST libraries, respectively. Furthermore, root contigs were analyzed by C-mii software for miRNAs identification. The expression profile of two drought tolerance candidate genes was studied, using Real time-PCR in a 2 (Nimruz and Spontaneum) × 2 (control and 50% FC) factorial experiment in a completely randomized design with 3 replications. Results of this study showed that N-butyl-N-methylpiperidinium (Pip1;4) and a non-specific lipid transport protein (nsLTP) were found to express differently under drought stress. The expression level of barley HvPip1;4 (HvPiP1;4) and non-specific lipid transport (HvnsLTP) genes increased by 95.98 and 54.53 fold after 72 h of drought stress, respectively. There was a significant difference (P

The original version of this article unfortunately contained a mistake in the unit “μg/l”. The unit “μg/l” should be corrected to “μg/ml” throughout the paper.

Zinc oxide nanoparticles (NPs) are the third highest in terms of global production among the various inorganic nanoparticles, and there are concerns because of their worldwide availability and accumulation in the environment. In contrast, zinc is an essential element in plant growth and metabolism, and ZnO NPs (nano-ZnO) may have unknown interactions with plants due to their small sizes as well as their particular chemical and physical characteristics. The present study examined the effect of nano-ZnO (25nm) and bulk or natural form (

BACKGROUND Using proper growing medium is known to be an effective way to improve crop growth and yield. However, the effects of growing media on geranium essential oil have scarcely ever been examined in detail. In this research, the effects of different growing media (soil, sand, pumice, perlite and perlite + cocopeat) on growth, oil yield and composition of geranium were studied. RESULTS Growth was significantly improved in soilless-grown plants compared with soil-grown plants. Oil yield of soilless-grown plants (except for pumice) was about threefold higher than that of soil-grown plants. The increase in oil yield was correlated with higher leaf dry weight (r2 = 0.96), as oil content was not affected. The citronellol/geranium ratio of oil was clearly affected by growing media, ranging from 5:1 in soil culture to 3:1 in soilless culture. The latter is acceptable for perfumery. CONCLUSION Compared with soil, soilless media could produce higher yields of high-quality geranium oil that fits market requirements. Growth, oil yield and composition of plants grown in sand (a cheap and abundant growing medium) were not significantly different from those of plants grown in perlite and perlite + cocopeat. © 2013 Society of Chemical Industry

An experiment was conducted to investigate the effect of methanol solution spray treatment on growth, biomass, and functional properties of bean plant under drought stress. The experiment was set up as a fully-randomized design with three replications. The first factor was four concentration levels of spray solution (Control, 10, 20, and 30%) and the spray was applied three times during the plant growth season at 10-day intervals. The second factor was three levels of drought stress; severe drought stress (25% field capacity), moderate drought stress (75% field capacity), and non-stress (100% field capacity). Test results showed that under 20% of methanol and conditions of non-stress there was significant growth of protein content of root and leaf in comparison with the control. Antioxidant enzyme activity was not affected by application of methanol solution spray but leaf antioxidant enzymes activity declined. Under non-stress and methanol concentrations of 10 and 20%, results showed a significant increase in all morphological properties compared with the control treatment. Under conditions of severe and moderate drought stress, level of methanol solution spray did not mitigate the negative effects of drought stress on the studied properties.

This study conducted two experiments involving in vitro anther culture of Zea mays L. The first experiment tested 46 maize genotypes, including inbred lines, single and three-way cross hybrids, and line A188 as control, in three different induction basal media (IMSS, N6 and YPm) for their androgenic responses. The results showed that the embryos were established 2–3 weeks after the anthers of the few responsive genotypes were cultured. Most responsive genotypes produced embryos in at least one of the three basal media; therefore, genotype is more important than the type of medium for androgenesis in maize. The mean number of anthers that developed to embryo ranged from 19 embryos per Petri dish in YPm medium for the cross (DH5 × DH7) genotype to 0 for some maize genotypes. In the second experiment, this research reports for the first time the effect of carbohydrates and polyethylene glycol (PEG) as a non-metabolized osmoticum on the embryogenesis anther culture of maize. The genotype DH5 × DH7 was used for this experiment, and the media were varied by altering sucrose, maltose, and PEG concentrations. Results showed that the maximum embryogenesis (32 embryos per Petri dish) was obtained by YPm basal medium supplemented with 60 gl−1 sucrose + 0.0125 M PEG and 30 gl−1 sucrose + 30 gl−1 maltose + 0.0125 M PEG. The lowest rate of embryogenesis was observed in YPm basal medium with 60 gl−1 maltose and 0.0125 or 0.025 M PEG. Sucrose or a high concentration of maltose was found to be necessary for embryogenesis in anther culture of maize. Therefore, the addition of low levels of PEG and/or different sugars in the experimental design appeared to improve the protocol currently available in the world, especially for anther embryo yield and haploid plant regeneration in maize.

A novel and rapid ultrasound- and salt-assisted liquid-liquid extraction coupled with high-performance liquid chromatography has been optimized by response surface methodology for the determination of oleuropein from olive leaves. Box-Behnken design was used for optimizing the main parameters including ultrasound time (A), pH (B), salt concentration (C), and volume of miscible organic solvent (D). In this technique, a mixture of plant sample and extraction solvent was subjected to ultrasound waves. After ultrasound-assisted extraction, phase separation was performed by the addition of salt to the liquid phase. The optimal conditions for the highest extraction yield of oleuropein were ultrasound time, 30 min; volume of organic solvent, 2.5 mL; salt concentration, 25% w/v; and sample pH, 4. Experimental data were fitted with a quadratic model. Analysis of variance results show that BC interaction, A(2) , B(2) , C(2) , and D(2) are significant model terms. Unlike the conventional extraction methods for plant extracts, no evaporation and reconstitution operations were needed in the proposed technique.

This study was a factorial experiment with a completely randomized design and three replications. The four levels of methanol spraying were used. Spraying was carried out three times during the growing season at 10-day intervals beginning at 4 weeks after sowing. The spraying of solution continued until saturation of droplets on the leaves was achieved. The levels of water-deficit stress applied were nonstress, moderate water stress and severe water stress. The results showed that there was a significant difference (P ≤ 0.05) between the methanol and water-deficit stress treatments for chlorophyll (Chl) a and Chl b, carotenoid, total chlorophyll, net photosynthesis (PN ), intercellular CO2 (Ci ), maximal quantum yield of photosystem II photochemistry (Fv /Fm ), leaf moisture, water use efficiency and relative water content. The application of foliar methanol at all levels of water-deficit stress significantly decreased the catalase activity of the roots. Under all levels of water-deficit stress, the 30% (v/v) methanol treatment significantly decreased peroxidase activity in the roots over that for the control. The results suggest that foliar application of methanol can decrease the negative effects of water-deficit stress on Phaseolus vulgaris L. cv. Sadry.

Using proper growing medium is known to be an effective way to improve crop growth and yield. However, the effects of growing media on geranium essential oil have scarcely ever been examined in detail. In this research, the effects of different growing media (soil, sand, pumice, perlite and perlite + cocopeat) on growth, oil yield and composition of geranium were studied.Growth was significantly improved in soilless-grown plants compared with soil-grown plants. Oil yield of soilless-grown plants (except for pumice) was about threefold higher than that of soil-grown plants. The increase in oil yield was correlated with higher leaf dry weight (r² = 0.96), as oil content was not affected. The citronellol/geranium ratio of oil was clearly affected by growing media, ranging from 5:1 in soil culture to 3:1 in soilless culture. The latter is acceptable for perfumery.Compared with soil, soilless media could produce higher yields of high-quality geranium oil that fits market requirements. Growth, oil yield and composition of plants grown in sand (a cheap and abundant growing medium) were not significantly different from those of plants grown in perlite and perlite + cocopeat.

Members of the Camelidae (camels, dromedaries, llamas, alpacas, guanacos and vicunas) are known to produce Igs (immunoglobulins) devoid of light chains and CH1s (constant heavy-chain domains). The antigen-specific binding fragments of these heavy-chain antibodies therefore comprise one single domain (the so-called 'VHH') and are of great importance in biotechnological applications. To evaluate the expression and biological activity of sdAbs (single-domain antibodies) in plants, which, on account of their small size and antigen-recognition properties, would have a major impact on antibody-engineering strategies, we constructed a pBI121-VHH gene encoding the recombinant sdAb fragments with specificity for a cancer-associated mucin, MUC1. Analysis of transgenic tobacco (Nicotiana tabacum cultivar Xanthi) plants by PCR and Western blotting demonstrated the expression of sdAb, while ELISA results with various MUC1 antigens and immunocytochemistry with cancerous cell lines confirmed that the activity of these molecules compared favourably with that of the parent recombinant antibodies. Protein purification was achieved by using sequential (NH4)2SO4 precipitation, gel filtration and immunoaffinity chromatography. Analysis of the purified VHH by ELISA indicated that the purified antibody fragments were able to react successfully with a MUC1-related peptide. These results reaffirm that the tobacco plant is a suitable host for the production of correctly folded VHH antibody fragments with diagnostic and therapeutic applications.