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Potato tubers are rich sources of various nutrients and unique sources of starch. Many genes play major roles in different pathways, including carbohydrate metabolism during the potato tuber's life cycle. Despite substantial scientific evidence about the physiological and morphological development of potato tubers, the molecular genetic aspects of mechanisms underlying tuber formation have not yet been fully understood. In this study, for the first time, RNA-seq analysis was performed to shed light on the expression of genes involved in starch biosynthesis during potato tuber development. To this end, samples were collected at the hook-like stolon (Stage I), swollen tips stolon (Stage II), and tuber initiation (Stage III) stages of tuber formation. Overall, 23 GB of raw data were generated and assembled. There were more than 20000 differentially expressed genes (DEGs); the expression of 73 genes involved in starch metabolism was further studied. Moreover, qRT-PCR analysis revealed that the expression profile of the starch biosynthesis DEGs was consistent with that of the RNA-seq data, which further supported the role of the DEGs in starch biosynthesis. This study provides substantial resources on potato tuber development and several starch synthesis isoforms associated with starch biosynthesis.

With the development of genome editing technologies, editing susceptible genes is a promising method to modify plants for resistance to stress. NPH3/RPT2-LIKE1 protein (NRL1) interacts with effector Pi02860 of Phytophthora infestans and creates a protein complex, promoting the proteasome-mediated degradation of the guanine nucleotide exchange factor SWAP70. SWAP70, as a positive regulator, enhances cell death triggered by the perception of the P. infestans pathogen-associated molecular pattern (PAMP) INF1. Using a clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, a construct was made to introduce four guide RNAs into the potato cultivar Agria. A total of 60 putative transgenic lines were regenerated, in which 10 transgenic lines with deletions were selected and analyzed. A mutant line with a four-allelic knockdown of StNRL1 gene was obtained, showing an ~90% reduction in StNRL1 expression level, resulting in enhanced resistance to P. infestans. Surprisingly, mutant lines were susceptible to Alternaria alternata, suggesting that StNRL1 may play a role as a resistance gene; hence, silencing StNRL1 enhances resistance to P. infestans.

Abstract 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.

Satureja is rich in phenolic monoterpenoids, mainly carvacrol, that is of interest due to diverse biological activities including antifungal and antibacterial. However, limited information is available regarding the molecular mechanisms underlying carvacrol biosynthesis and its regulation for this wonderful medicinal herb. To identify the putative genes involved in carvacrol and other monoterpene biosynthesis pathway, we generated a reference transcriptome in two endemic Satureja species of Iran, containing different yields (Satureja khuzistanica and Satureja rechingeri). Cross-species differential expression analysis was conducted between two species of Satureja. 210 and 186 transcripts related to terpenoid backbone biosynthesis were identified for S. khuzistanica and S. rechingeri, respectively. 29 differentially expressed genes (DEGs) involved in terpenoid biosynthesis were identified, and these DEGs were significantly enriched in monoterpenoid biosynthesis, diterpenoid biosynthesis, sesquiterpenoid and triterpenoid biosynthesis, carotenoid biosynthesis and ubiquinone and other terpenoid-quinone biosynthesis pathways. Expression patterns of S. khuzistanica and S. rechingeri transcripts involved in the terpenoid biosynthetic pathway were evaluated. In addition, we identified 19 differentially expressed transcription factors (such as MYC4, bHLH, and ARF18) that may control terpenoid biosynthesis. We confirmed the altered expression levels of DEGs that encode carvacrol biosynthetic enzymes using quantitative real-time PCR (qRT-PCR). This study is the first report on de novo assembly and transcriptome data analysis in Satureja which could be useful for an understanding of the main constituents of Satureja essential oil and future research in this genus.

To investigate the effect of soluble salicylic acid and micronutrient elements on some of the biochemical and agronomic characteristics of rapeseed under water deficit conditions studied in a split factorial experiment based on a randomized complete block design with three replications at the Faculty of Agriculture, Lorestan University during 2016-2017 growing season. Water deficit was considered as the main factor with two levels (irrigation at 20 (control) and 70% of field capacity depletion), and combined treatments of two factors of foliar application of micronutrient elements (non-consumption and spraying at a concentration of 2 per thousand) and salicylic acid concentration of 0, 0.5, 1 and 1.5 mM) as the subplot. The results showed that the effects of water deficit and salicylic acid on all traits under study were significant. The results showed that triple interaction was also significant on catalase, peroxidase. proline, oil percent, seed yield and oil yield. The highest average oil percentage, seed and oil yield were 46.02%, 4424 and 2036 kg.ha-1 of the treatment combination (no stress+ solubility of 1.5 mm salicylic acid and concentration of 2 per thousand micronutrient fertilizer). The results of this study, based on the changes in the mean, showed that salicylic acid and micronutrient elements reduced the effect of water deficit in canola seedlings. In general, Based on the results of this study, it is recommended to reduce the negative effects of water deficit stress and produce proper seed yield, by using Ferti Mix Trio fertilizer and 1.5mM salicylic acid under water deficit conditions.

Objective Lactic acid bacteria (LAB) synthesize valuable industrial and pharmaceutical important biopolymers, such as Alternan, by expressing glycosyltransferase enzymes by utilizing extracellular sucrose. In this study, due to the importance of such versatile biopolymers in the industry and medicine, a gene encoding an Altranansucrase (Asr) was introduced to sugar beet plants. Materials and methods A gene encoding the Asr gene was isolated from Leuconostoc mesenteroides bacterium and introduced to sugar beet plants by using Agrobacterium-mediated transformation. Molecular techniques were used to analyse transgenic plants and sugar content of sugar beet lines. Results Out of 131 transformed explants, only three transgenic plants were produced, showing a transformation efficiency of 2.3%. The Asr gene integration in transgenic plants genome and expression were confirmed by specific PCR and semi-quantitative RT-PCR analysis, respectively. Sugar analysis of sugar beet transgenic plants showed that control plants with a 19.6% brix value (Sucrose) had more sucrose than transgenic plants with an average brix value of 14.4%. Brix in transgenic plants was lower than that of control plants. The amount of sugar (sucrose) in the transgenic asr-expressing plants reduced by 36.1% relative to the untransformed control plants. Conclusions The results of this study showed that Altrenansucrase can convert substantial amount of sugar beet sucrose into Alternan biopolymer, producing 36.6 mg/g FW alternan biopolymer with pharmaceutical and industrial applications.

Plant nutrition has a vital role in crop production. This study was performed to investigate the effects of different application methods of some nutrients (nano Fe, Zn, and Ti), and gibberellin on yield, some morphophysiological and grain protein of white beans in 2018 as a factorial experiment in a randomized complete block design with four replications. Experimental factors included seed priming (hydropriming, gibberellin priming, titanium nano dioxide, and nano-Zn priming) and micronutrient foliar spraying (zinc, iron, and zinc + iron). The results illustrated that seed priming and foliar application significantly affected yield, yield components and chlorophyll content. Plant height increased in seed priming treatment with gibberellin and foliar application of zinc + iron by 13% compared to the control. Furthermore, this treatment enhanced the number of sub-branches per plant by 32% compared to the control. Grain yield components such as the number of pods per plant and 100-grain weight were also affected by seed priming with nano-Zn, and the simultaneous spraying of iron and zinc that grain yield by 18%, so that grain yield by 2649 kg ha–1 in hydropriming treatment reached to 3211 kg ha–1 in nano-Zn priming with simultaneous application of zinc and iron. Nano-Zn priming with iron foliar application caused the highest biological yield (9011 kg ha–1), which increased by 19% compared to control. Nano-Zn priming increased grain protein percentage by 21%. This treatment along with the foliar application of zinc + iron, significantly enhanced leaf chlorophyll content compared to other treatments. Therefore, to increase the yield of white beans, priming treatment with nano-Zn as well as foliar application of zinc + iron can be used.

Background and purpose: Rapid emergence of traditional antibiotic-resistant pathogens is one of the most important global challenges in medical sciences. To this end, substitution of current antibiotics with strong antimicrobial peptides could be of great benefit. Materials and methods: In this study, the DNA sequence encoding dermaseptin B1 (DrsB1) antimicrobial peptide derived from Phyllomedusa bicolor frog species was fused to either N or C terminal end of the sequence encoding the chitin-binding domain of the Avr4 gene from Cladosporium fulvum. The recombinant expression vectors containing two separate structures were transferred to Agrobacterium rhizogenes bacterium and then used to produce Hairy Roots (HRs) in tobacco plants. Recombinant dermaseptin B1 peptides were extracted from HRs and their antimicrobial activity was evaluated against some important human pathogens. Results: Transgene integration and expression of recombinant DrsB1 in hairy roots were confirmed by PCR and semi-quantitative RT-PCR analysis, respectively. Antimicrobial activity of the protein extracts from transgenic HRs showed that both recombinant proteins had significant inhibitory effects on the bacterial pathogens growth (P>0.01). CBD-DrsB1 recombinant protein had the highest inhibitory activity against Enterococcus faecalis and Bacillus subtilis, whereas DrsB1-CBD recombinant protein showed the least antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Conclusion: To the best of our knowledge, this study for the first time showed that the new recombinant peptides possess a high antibacterial activity. Acquiring resistance to antimicrobial peptides in bacteria is not readily feasible, therefore, present findings may find application as suitable alternative for current antibiotic drugs.

Background and Aim: Enterococcus species are opportunistic pathogens and their pathogenicity seems to be related to the presence of a number of pathogenicity genes. Since donkey’s milk is a new non-allergenic source of nutrition, this study was performed to assess the antibiogram and detection of pathogenicity genes in some Enterococcus faecalis isolates from donkey’s milk. Materials and Method: In this experimental study, several Enterococcus faecalis strains were isolated from donkey’s milk. Resistance patterns of the isolates to 10 antibiotics including vancomycin were investigated based on CLSI protocol. Statistical comparison was made by Fisherchr('39')s exact test. Polymerase chain reaction (PCR) amplification was used to study gel E, esp, ace, as and efaA pathogenicity related genes. Results: Enterococcus faecalis isolates showed a different antibiogram pattern. Isolates were resistant to azithromycin, and erythromycin, but were susceptible to ampicillin and penicillin. Previously isolated LUB93929 and LUB93101 isolates were found to be susceptible and resistance to vancomycin, respectively. GelE, ace and efaA genes were detected in both Enterococcus faecalis isolates and also in E. faecalis in the control strains. The aggregation substance gene (as) was only amplified in LUB93101 isolate. Interestingly, esp gene was not detected in any of the isolates. Conclusion: Despite resistance to vancomycin and presence of some pathogenicity related genes in this study, E. faecalis isolates may not be human pathogens due to lack of pathogenic factors. The esp gene is crucial for biofilm formation and rise of nosocomial infections. Donkey’s milk Enterococcus faecalis isolates are not able to form biofilm and seem not to bring any problem.

Objective Cold is one of the important stresses in cold and temperate regions and improving cold tolerance is in the top priority of the lentil breeding programs. NAC transcription factors play an important role in response to various biological stresses, including cold stress. Materials and Methods: In this study, to evaluate the expression of MfNAC, MtNAC57 and GmNAC2 genes, the plants of Gachsaran lentil cultivar were subjected to five temprature treatments in a completely randomized design with three replications. Twenty-nine old plants were subjected to duration of cold stress treatments including 6h, 12h, 24h and 48h at 2°-4°c and plants grown under greenhouse conditions (23°C) were considered as controls. Total RNA extraction from tissues was performed using lithium chloride method and cDNA was synthesized using TaKara kit. For each treatment, real-time PCR was performed in both shoot and root in three biological and two technical replications. Actin gene was used as internal control. Results The results of analysis of variance of all studied genes showed that there was a significant difference between the different duration of cold treatments in both shoot and root. GmNAC2 gene expression decreased significantly in the shoot, 6 hours after stress, while in the root, the expression decreased 24 hours after stress. The expression of MtNAC57 gene in 6h, 12h, and 24h treatments decreased compared to control, but in the root in 12h treatment, a significant expression was observed. MfNAC gene in shoot showed a significant reduction in expression in all treatments, while in root a significant increase was only observed during long-term treatment of 48h. Conclusions In the present study, for first time, the expression pattern of some members of NAC gene family was examined in lentils under cold stress and the results could be useful for studies on this plant and other legumes.

Introduction: Lactic acid bacteria )LAB) are a group of gram-positive, non-spore forming, cocci or rod shaped, catalase-negative organisms, which are generally recognized as safe (GRAS). The LAB can cause changes in the food flavor and texture. Enterococci are a group of LAB capable of producing antimicrobial peptides. Materials & Methods: This experimental study was conducted to identify and isolate genes encoding antimicrobial peptides from an E. faecium LUB950217 strains isolated from oak tree. In doing so, genomic DNA was extracted from E. faecium strains and polymerase chain reaction (PCR) analysis was performed using specific primers. Amplified PCR products were sequenced by Sanger sequencing. Bioinformatic analysis was carried out and the peptide was modeled. Findings: Both DNA and protein Blast search confirmed that the PCR product encoded an antimicrobial peptide, known as LUB950217 enterocin with 36 amino acids of 4658.28 Da molecular mass. The isolated enterocin peptide had 86-100% similarity to other known enterocins in the peptide data bases. The LUB950217 enterocin was almost water insoluble and it was found to be cationic (2.2, pH=7). Discussion & Conclusions: The LUB950217 enterocin seems to inhibit both gram-positive and gram-negative bacteria growth in vitro, suggesting LUB950217 enterocin can be used in food stuff and animal feed. Furthermore, isolation, cloning, and expression of this peptide may render resistance to fungi and bacteria pathogens in crop plants.

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.

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.

Despite the increasing demand for production of recombinant proteins, only plant cell cultures are considered as the efficient systems for producing eukaryotic valuable products. Although plant cells are grown in the straightforward and simple conditions, optimization of culture conditions is crucial for a higher yield. In the present study, the establishment of transgenic tobacco hairy roots (HRs) expressing a Dermaseptin B1 recombinant antimicrobial peptide (C2-B1) is examined. Direct and indirect transformation (DT and IT) methods were applied to compare the amount of the recombinant peptide production. The integration, transcription and expression of the transgene in the HRs were confirmed by PCR, semi-quantitative RT-PCR and western blot analysis, respectively. The C2-B1 level was quantified by ELISA analysis of HRs extracts. The results of this study showed that the longer the inoculation and co-culture period, the higher the transformation efficiency. Antimicrobial activity by of crude protein extract from transgenic HR clones showed a significant (P ≤ 0.01) difference between HRs derived from DT and IT methodes methodsmethodes . Transgenic clones derived from both DT and IT methodes and grown in liquid medium showed a difference in terms of total protein content significantly. HRs derived from IT methodmethod produced a significantly higher amount of C2-B1 recombinant peptide in liquid B5 medium supplemented with sucrose.

Antimicrobial peptides have been long known to confer resistance to plant pathogens. In this study, new recombinant peptides constructed from a dermaseptin B1 (DrsB1) peptide fused to a chitin-binding domain (CBD) from Avr4 protein, were used for Agrobacterium tumefaciens-mediated transformation of tobacco plants. Polymerase chain reaction (PCR), semi‐quantitative RT‐PCR, and western blotting analysis demonstrated the incorporation and expression of transgenes in tobacco genome and transgenic plants, respectively. In vitro experiments with recombinant peptides extracted from transgenic plants demonstrated a significant (P

In order to study the effect of bio-fertilizers, planting method and superabsorbent polymer on quantitative yield of saffron under rainfed condition, a factorial experiment based on randomized complete block design with three replications was conducted at the Agricultural Research Station of Khorramabad, Iran during 2015-2016 growing seasons. In this study, Nitroxin bio-fertilizer (consumption 5 Lit/ha and non-consumption), Stockosorb superabsorbent polymer (consumption 240 kg/ha and non-consumption) and planting method (streaking and cluster) was tested. Results showed that triple interactions was significant for number of corm/m2 in first and second year and combined treatment including non-Nitroxin in cluster culture method using superabsorbent with 80.67 corm/m2 for first year and with 183.67 corm/m2 for second year was the superior treatment. Results showed significant differences for number of produced flowers and for fresh and dry weight of stigma in both years; and in the first year, treatment included cluster method cultivation, consumption of superabsorbent and no consumption of Nitroxin was superior; while in the second year, superior treatment was use of superabsorbent, Nitroxin and cluster method. For stigma dry weight of saffron, triple interactions was significant and in the first year, cluster method and consumption of superabsorbent and no usage of Nitroxin was superior (with 0.6 kg/ha), and in the second year, use of Nitroxin and superabsorbent and cluster method was superior (with 0.719 kg/ha). In general, results of the present study in dry-farming conditions over the two years, cluster method cultivation with usage of superabsorbent and Nitroxin could be recommended. Reasons of this recommendation and introduction of the mentioned method are dry climate conditions in area of study and reduction in consumption of chemical fertilizers for development of sustainable agriculture.

One goal in the face of drought stress conditions is to increase growth and yield through the reduction of negative effects of stress. Vermicompost can play an effective role in plant growth and development and in reducing harmful effects of various environmental stresses on plants due to its porous structure, high water storage capacity, having hormone-like substances, plant growth regulators, and high levels of macro and micro nutrients. This study considered the physiological, biochemical, and photosynthetic responses of the chickpea to different combinations of vermicompost and water stress in a greenhouse environment. Two factors were involved, addition of vermicompost to soil at four ratios: control (100 wt% (weight percentage) soil); 10 wt% vermicompost+90% soil; 20 wt% vermicompost+80 wt% soil; 30 wt% vermicompost+70 wt% soil weight percentage, and treatment of water stress at three levels including 75, 50, and 25% of field capacity. The results showed that vermicompost had a significant effect on all traits under stress and non-stress conditions. Application of vermicompost in soil, especially at the levels of 20 and 30 wt% significantly increased all studied traits under non-stress conditions. Under moderate stress conditions, vermicompost at 30 wt% treatment resulted in a significant increase in the photosynthetic pigments, CO2 assimilation rate, internal leaf CO2 concentration, transpiration, the maximal quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), concentrations of Ca and K in root and leaf tissues, proline and soluble protein contents in root tissues. Peroxidase (POX) and catalase (CAT) enzyme activities decreased significantly with increasing proportions of vermicompost, but the activity of superoxide dismutase was not significantly different. In conclusion, the above results showed that vermicompost fertilizer had a positive effect on physiological, biochemical, and photosynthetic responses of chickpea under non-stress and moderate stress conditions, but no positive effect was determined under severe water stress.

Background and Aim: The use of bioactive compounds of medicinal plants in prevention, control and treatment of human diseases has a long history. Most of plant bioactive compounds have highly complex chemical structures, and their chemical syntheses are often uneconomical. Moreover, their production is still dependent on plants. Plants of Papaveraceae family contain a variety of bioactive compounds that have many uses in traditional and modern medicine. In the present study, we evaluated the antimicrobial activity of alkaloid extracts of four plant species of Papaveraceae family against some human pathogens. Materials and Methods: Crude alkaloid compounds of Papaver macrostomum, Roemeria refracta, Papaver somniferum and Glaucium grandiflorum plants were extracted using cain method. The antimicrobial activity of plant alkaloid extracts against Pseudomonas aeroginosa PTCC 1310, Listeria monocytogenes PTCC 1297, Staphylococcus aureus PTCC 1189, Klebsiella pneumoniae PTCC 1290 and Candida albicans PTCC 5027 pathogens were assessed using agar diffusion method. Results: The results of this study indicated that the alkaloid extracts of tested plants were effective against fungal and bacterial pathogens. All plant alkaloid extracts exhibited more inhibitory effect against fungal pathogen than the bacterial pathogens. Comparison of MIC and MBC values for bacterial pathogens revealed that all plant alkaloid extracts showed more inhibitory effects against the gram-positive bacteria than gram-negative bacterial pathogens. The results also showed that the alkaloid extracts of R. refracta have stronger inhibitory effect against fungal and bacterial pathogens compared with other plants. Conclusion: The antimicrobial compounds present in the plant species of Papaveraceae family are suitable candidates to produce new antibiotics. The results of this study demonstrated that antimicrobial compound in R. refracta plant could be used in the treatment of infectious diseases caused by C. albicans and S. aureus pathogens.

Background and purpose: Lactic acid bacteria (LAB) play an important role in human and other organisms life. Some of LABs kill pathogenic and other harmful microorganisms. Enterococcus are, gram-positive, catalase-negative, cocci forming, non-sporogenesis, and facultative anaerobic bacteria. This study was done to identify and isolate possible probiotic bacteria with benefits to human health from Persian oak sap (Quercus brantii var. persica). The aim was to identify beneficial bacteria for plant pathogenic biological control and industrial applications. Materials and methods: LABs were identified using conventional methods, including culture dependent methods and 16S rRNA sequencing method. Antibiogram analysis was performed and the presence of virulence genes, including efaA (endocarditis antigen), as, ace, esp and gelE was examined by PCR. Results: It was found that out of 285 colonies, 160 (56.1%) were catalase-negative and gram-positive. Results of 16S rRNA sequencing showed that the bacteria isolated bacteria belong to the Enterococcus Faecium species. E. faecium strains of this study were sensitive to a number of clinically important antibiotics such as vancomycin, ampicillin, ciprofloxacin, and nitrofurantoin. Two strains of E. faecium bacteria, one with the ability to produce Co2 from glucose (KX185054) and one with no Co2 production ability (KX185055) were identified. The 16S rRNA sequence of identified strains were deposited in NCBI database. PCR amplification did not amplify virulence genes except efaA (endocarditis antigen). Conclusion: In this study, two different E. faecium strains were isolated from the oak which can be good candidates for probiotics and biological control.

Abstract Dermaseptin B1 (DrsB1), an antimicrobial cationic 31 amino acid peptide, is produced by Phyllomedusa bicolor. In an attempt to enhance the antimicrobial efficacy of DrsB1, the DrsB1 encoding 93 bp sequence was either fused to the N or C terminus of sequence encoding chitin‐binding domain (CBD) of Avr4 gene from Cladosporium fulvum. Tobacco leaf disk explants were inoculated with Agrobacterium rhizogenes harboring pGSA/CBD‐DrsB1 and pGSA/DrsB1‐CBD expression vectors to produce hairy roots (HRs). Polymerase chain reaction (PCR) was employed to screen putative transgenic tobacco lines. Semi‐quantitative RT‐PCR and western blotting analysis indicated that the expression of recombinant genes were significantly higher, and recombinant proteins were produced in transgenic HRs. The recombinant proteins were extracted from the tobacco HRs and used against Pectobacterium carotovorum, Agrobacterium tumefaciens, Ralstonia solanacearum, and Xanthomonas campestris pathogenic bacteria and Alternaria alternata and Pythium sp. fungi. Two recombinant proteins had a statistically significant (p

Salt stress is one of the major adverse environmental factors limiting crop productivity. Considering Iran as one of the bread wheat origins, we sequenced root transcriptome of an Iranian salt tolerant cultivar, Arg, under salt stress to extend our knowledge of the molecular basis of salinity tolerance in Triticum aestivum. RNA sequencing resulted in more than 113 million reads and about 104013 genes were obtained, among which 26171 novel transcripts were identified. A comparison of abundances showed that 5128 genes were differentially expressed due to salt stress. The differentially expressed genes (DEGs) were annotated with Gene Ontology terms, and the key pathways were identified using Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway mapping. The DEGs could be classified into 227 KEGG pathways among which transporters, phenylpropanoid biosynthesis, transcription factors, glycosyltransferases, glutathione metabolism and plant hormone signal transduction represented the most significant pathways. Furthermore, the expression pattern of nine genes involved in salt stress response was compared between the salt tolerant (Arg) and susceptible (Moghan3) cultivars. A panel of novel genes and transcripts is found in this research to be differentially expressed under salinity in Arg cultivar and a model is proposed for salt stress response in this salt tolerant cultivar of wheat employing the DEGs. The achieved results can be beneficial for better understanding and improvement of salt tolerance in wheat.

Opposition to destructive and side effects of salt stress, have limited suitable water and soil sources for plant culturing, is possible by increasing the tolerance level of seeds and seedlings. In present research, the effect of priming technique was studied with Na2SiO3 or KNO3 in 3, 6, 9 and 12 hours on Medicago sativa cv. Hamadani in response to 200 mM salt stress. Salinity caused a decrease of most of germination and growth indices from seeds and seedlings, whereas 3h-Na2SiO3 priming increased final germination percentage (FGP) and (3/6/9h)-KNO3 increased FGP, coefficient velocity of germination (CVG), compared to control. An improvement in mean germination rate (MGR) and mean germination time (MGT) was shown by 6h-Na2SiO3 and all KNO3 treatment times and increase of Vigor index (I) was observed at 3h-KNO3. Pretreatment with (3/6/9h)-Na2SiO3 and all h-KNO3 decreased T50 at stress and in non stress conditions, 9h-Na2SiO3 and 9h-KNO3 caused a decrease in mean daily germination (MDG) and an increase in Timson index and improvement of coefficient uniformity of germination (CUG). 12 hour treatments from both substances exacerbated the conditions and decreased the FGP, Vig (I) and (II) and decreased CVG. All-Na2SiO3 and (9/12)-KNO3 decreased the dry weight of shoots at stress. Proline was raised only with KNO3 and salt affected pigments negatively and priming increased them. In conclusion, priming elevated tolerance to salt stress and treatment compositions with the lower times were more efficient.

In order to evaluate the effects of methanol on germination percentage, germination speed index, plumule and radicle length, plumule and radicle dry weigh, vigor index and consumed endosperm of bean in condition of water deficit, a factorial experiment done based on completely randomized design with three replications. The first factor was different levels of methanol included 0 (control), 5, 10 and 15 volumetric percentage, and second factor was negative water deficit in four levels of 0, -3, -6 and -9 bar. Results of this study showed that there was a significant difference among different methanol concentrations regarding germination percentage, germination speed index, plumule and radicle length, plumule and radicle dry weight, vigor index and consumed endosperm (p 0.01). Different levels of methanol caused significant decrease on germination characteristics compared with control. Drought stress at -9 bar level significantly decreased germination percentage, germination speed index, radicle length, radicle dry weigh and consumed endosperm compared with other levels. Results showed that in non-stress condition, methanol levels led to significant decrease on germination characteristics compared with control. In drought stress condition, methanol levels caused significant decrease on germination percentage, plumule dry weigh, radicle length and radicle dry weight compared with control.

Introduction Despite the fact that Iran is one of the major chickpea (Cicer arietinum L.) center of origins, limited information is available regarding chickpea genetic variation and diversity. Genetic diversity information is crucial for the choice of proper parents to establish new breeding programs. Chickpea germplasm is poor, suggesting the need for gaining enough knowledge of genetic diversity among available chickpea genotypes. A number of molecular techniques have been developed to unveil the genetic potentials of plant materials. Random Amplified Polymorpic DNA (RAPD) seems to be a reliable molecular marker to investigate the genetic diversity of chickpea genotypes in IRAN. The objectives of present research was: (1) to investigate the power of RAPD markers for estimation of genetic diversity among chickpea genotypes in west of Iran, (2) to investigate the genetic relationships between chickpea genotypes, and (3) to determine whether chickpea genotypes could be distinguished by RAPD marker data. Materials and Methods Random amplified polymorphic DNA markers (RAPD) were used to assess the genetic relationship between 18 different chickpea genotypes representing the cultivated chickpea cultivars in west of Iran. Genomic DNA was isolated according to Murray & Thompson (1980). Eighteen oligonucleotide primers were selected according to the number of literature published with the highest number of polymorphic bands. Polymerase chain reaction (PCR) was carried out in a total volume of 25 μl including 2 units of Taq DNA polymerase, 30 ng of genomic DNA template, 10 pmol of primers, 0.2 mM of dNTPs, and 2.5 μl of 10 × reaction buffer. DNA amplifications were performed in a thermocycler. The thermal profile was as follow: One time denaturation at 94◦C (5 min), followed by 40 cycles of denaturation at 94◦C (3 min), annealing at each primer proper Tm (1 min) and extension at 72◦C (2 min) and one time final extension at 72◦C (5 min). PCR products were analyzed on 1.5 % agarose gels in TBE buffer running at 100 V for 2h. The gels were stained using ethidium bromide and visualized with UV light. The reproducibility of the DNA band patterns was evaluated duplicate gel electrophoresis analysis. Only clear and repeatedly amplified RAPD DNA bands were scored as (1) for present bands and (0) for absent ones. Results and Discussion Out of 18 random RAPD primers used in this study, 17 primers amplified genomic DNA across all the genotypes. In total, 201 polymorphic bands (96.63%) out of 208 reproducibly scoreable RAPD markers were generated (OPM-05 primer did not produce any band). On average, 12.2 bands per primer were observed in RAPD analysis. Cluster analysis using Dice coefficient of similarity and UPGMA (r=0.98) method based on polymorphic fragments, grouped all eighteen genotypes into 6 groups with 77% accuracy. Based on dendrogram obtained, Bivanij (Landrace genotype) showed the least similarity with the 6th cluster. Although there was no redundancy among the genotypes tested, the majority of genotypes were clustered together. ICARDA genotypes may have been improved from an Iranian landrace. Conclusions Genetic diversity among chickpea cultivars using RAPD markers have been studied by a number of researchers. Although in most cases a low level of polymorphism with RAPD markers have been reported, this study showed a considerable amount of polymorphism. Furthermore, our results showed that cultivated chickpea cultivars in west of Iran have many genes in common. We recommend further studies to be conducted by using more number of chickpea genotypes as well as more robust molecular markers. Results of this study can be used in germplasm management/conservation practices, developing core collections and as guidance to plant geneticist and breeders for planning future explorations, and crop improvement purposes. These findings may help to avoid genetic vulnerability and erosion, keeping chickpea genetic diversity and germplasm.

Analysis of genetic diversity is a major step for understanding evolution and breeding applications. Recent advances in the application of the polymerase chain reaction make it possible to score individuals at a large number of loci. The RAPD technique has been successfully used in a variety of taxonomic and genetic diversity studies. The genetic diversity of 18 accessions of Thymus kotschyanus collected from different districts of Iran has been reported in this study, using 30 random amplified polymorphic DNA primers. Multivariate statistical analyses including principal coordinate analysis (PCOA) and cluster analysis were used to group the accessions. From 29 primers, 385 bands were scored corresponding to an average of 13.27 bands per primer with 298 bands showing polymorphism (77.40%). A dendrogram constructed based on the UPGMA clustering method revealed three major clusters. The obtained results from grouping 18 accessions of T. kotschyanus with two studied methods indicated that in the most cases the applied methods produced similar grouping results. This study revealed nearly rich genetic diversity among T. kotschyanus accessions from different regions of Iran. The results showed RAPD marker was a useful marker for genetic diversity studies of T. kotschyanus and it was indicative of geographica variations.

Study of genetic diversity in medicinal plant is very important for improvement and evolutionary variations. In this study, assessment of genetic diversity in two subspecies of Thymus daenensis was evaluated, using intron-exonic markers. Thirty primers produced 633 polymorphic bands (98% polymorphism). The highest polymorphic information content (PIC) included ISJ5 and ISJ9 primers and the lowest PIC also included IT15-32 primer. The highest marker index (MI) produced by IT10-6 primer. Results of Analysis of Molecular Variance (AMOVA) showed that intra-sub specific variation was more than inter-sub specific variation. Dendrogram obtained from Cluster analysis, using NTSYS-pc software and UPGMA method based on Dice's similarity matrix, divided accessions into 4 groups. The maximum range of genetic similarity was observed between two accessions of sub-species daenensis. Two accessions of Fars and Semnan formed a separate group. Results showed that clustering based on molecular data and principal coordinate analysis had a medium alignment. Grouping based on cluster analysis also could separate two subspecies of Thymus daenensis. Results obtained from this study showed that intron-exonic markers had an effective potential in assessment of genetic relationships between the two sub-species of daenensis.