seyed alireza salami

The current research explored the biocontrol ability of two strains of Trichoderma fungi on infected Freesia corm by Fusarium disease. Enhancements in plant growth and performance were evaluated in response to the Trichoderma fungi in laboratory and greenhouse conditions. Laboratory experiments included five Trichoderma strains, i.e., T1, T20, and Th4, related to T. harzianum species, T2 related to T. atroviride species, and Tvi related to T. virens species. A dual culture test and investigation of the inhibition and colonization rate were carried out with 11 treatments in three replications. Results revealed that strains T1 and T2 completely covered the pathogen in 96 h of culture by 70.02% and 68.98% and successfully inhibited the colonization of the disease agent by 84.53% and 82.72%, respectively, compared to the other strains. Then, a greenhouse experiment was performed with eight treatments, using suspensions of Trichoderma strains separately or combined with the presence of Fusarium fungus isolate on Freesia cultivation. The results showed that the strains at different levels were able to increase the growth and yield of Freesia plants and control Fusarium disease. In general, the T2 strain was more effective in increasing plant growth and flower stem height in Freesia. The T1 strain specifically improved the corm diameter. Both T1 and T2 strains demonstrated effectiveness in controlling and reducing the severity of Fusarium disease and enhancing the growth quality of Freesia.

The present study was conducted to study the effect of different concentrations of Bacillus subtilis on morphophysiological traits of flowers and coriander in two saffron ecotypes in order to determine the appropriate concentration of bacteria.

A factorial experiment was conducted in a randomized complete block design with three replications in the research farm of Zanjan University. Experimental treatments: the first factor of five levels of different concentrations of bacteria(zero, 102, 105, 106, 108 ) and the second factor consisting of two ecotypes (Natanz, Zanjan) were considered.

Different concentrations of bacteria showed a significant effect on all morphophysiological traits measured. The main effect of ecotype also showed a significant effect on the traits at the level of one and five percent, except for the fresh weight traits of stigma, chlorophyll. Based on the results of analysis of variance, bilateral interaction (year × ecotype, ecotype × bacterium) and tripartite interaction (bacterium × year × ecotype) on flower yield and phosphorus content of saffron leaves were significant. Among the ecotypes, Natanz ecotype showed more than Zanjan ecotype in fresh and dry weight of pistachio and its diameter (16.29, 10.99 g and 19.71 mm, respectively).

Considering the effects of Bacillus subtilis on saffron ecotypes, it can be concluded that saffron ecotypes showed different reactions to biofertilizer due to their differences and different growth characteristics. According to the results, the greatest effect of Bacillus subtilis on morphophysiological traits was observed at a concentration of 108 cfu / ml.

In this study, we investigated important morphological traits of select populations of fiber Iranian hemp. The results of the analysis of variance showed significant diversity among the populations. Four of the eight genotypes showed large biomass suitable for use in the fiber industry. Kerman, Sirjan, Dezful, and Arak genotypes had the highest final height and the most prominent stem diameter. These populations also showed late flowering periods along with a short inflorescence length. Our classification study revealed that of the four high-fibrous candidates, three were classified in one group (Kerman, Sirjan, and Dezful), and the fourth one (Arak) was organized in the second group. The relationships among the traits showed a significant positive correlation between some characteristics, such as the final height, the footstalk diameter, and the number of days it takes for the first flower to appear. We also showed that there is a strong correlation between morphological traits and the geographical data of the genotypes. Our findings indicated that high elevation along with the low average annual temperature is suitable for the growth of medicinal cannabis, while warmer weather leads to taller plants with thicker stems andlate blooming periods, which are more relevant to be used in the fiber industry.

Milk thistle (Silybum marianum (L.) Gaertn) belonging to the Asteraceae family and known for its valuable secondary metabolite, silymarin. In order to get the cell suspension culture of Silybum marianum, the combination of 2,4-D and BAP hormones each with five different concentrations (0, 0.1, 1, 2 and 5 mg/L) and 3 different seedling explants (cotyledon, hypocotyl and root) was used to callus formation. High-quality callus was observed in eight different of hormone-explant combinations. Successful cell suspension culture was achieved only by using callus created from cotyledon explants treated with 0.1 mg/L 2,4-D and 5 mg/L BAP. Furthermore, the study examined the effects of 2% and 4% Serendipita indica cell wall extract as an elicitor on silymarin production in cell suspension culture at three different inoculation times (24, 48, and 72 h). After 48 h of inoculation with 2% fungal extract, the highest level of silymarin (199 ppm) was observed, which was significantly different from the control (46 ppm). The silymarin content of cells increased over time through elicitation with 4% fungal elicitor, while no similar pattern was observed with 2% fungal extract. Based on the results, the cell wall of S. indica can significantly enhance the amount of SLM in the cell suspension culture of S. marianum.

Agricultural saffron Crocus sativus L. is a medicinal, widely used and expensive plant and has important effective substances: crocin, picrocrocin and safranal, which gives it the characteristics of color, taste and smell. The search for new sources producing the aforementioned apocarotenoids makes the importance of research on it more obvious day by day. Since the country of Iran has diploid wild species with characteristics close to the sterile triploid cultivated species, in the present research, two species of Caspian saffron C. caspius and Zeiba C. Speciosus, along with the cultivated species, were investigated and 6 genes active in the production of the three mentioned effective substances , CCD2, ALDH, PSY, LCY, BCH, UGT74 (and a housekeeping gene TUB as an internal control gene with constant expression in all tissues) in two flower tissues, stigma and perianth (tepal) and in five stages (time) of flowering including: The stages of bud formation (immature bud), bud ready to bloom, newly opened flower, mature flower and old flower were investigated. Using of Real Time PCR, the expression ratio (fold change) to the internal control gene (with logarithm based on 2) was investigated in different genes and different and sometimes similar expression patterns were observed with the gene expression in the cultivated species in both the stigma and stigma tissues. Based on the results obtained in this study and the increase or decrease in the expression of some genes involved in the pathways of apocarotenoids production, at the same time as the growth of the stages from the bud to the senescence stage of the flower, it is expected from the three aforementioned substances or from other apocarotenoids in the metabolom of flowers, cases have been found in the non-triploid species and create approaches for the commercial use of Iranian wild saffron species and pave the way for its commercialization.

Cannabis (Cannabis sativa L.) is an herbaceous annual plant belongs to Cannabacea Family. (Ahmad et al., 2008). The resistance to water shortage, the ability to grow in different climatic conditions, and great genetic diversity are features of this plant (Amaducci et al., 2008). Drought is one of the most important environmental stresses limiting crop production worldwide and has adverse effects on plant growth, development, which may result in decreased chlorophyll a and b and increased proline content of leaf (Lum et al., 2014; Karimi et al., 2016). Plants generally adapt to drought stress by inducing a variety of physiological, biochemical, and morphological responses, and each of these factors can be effective in introducing drought tolerant cultivars. Among the physiological properties, leaf water condition, membrane stability, photosynthetic changes and related factors are of great importance (Farooq et al., 2009). Considering the pharmaceutical and industrial importance of cannabis, this study was conducted to identify the drought tolerant and sensitive ecotypes of cannabis based on physiological responses.

This study was done in research greenhouse of University of Tehran, Iran, from February to July 2017 on the base of factorial experiment in as a completely randomized design (CRD) with three replications. The first factor consisted of three soil moisture levels [100% (normal irrigation), 75% (mild drought stress), and 50% (serve drought stress)] of field capacity (FC). Also, the 12 Iranian ecotypes of cannabis were the second factor where collected from different geographical regions of Iran including Urmia, Tabriz, Sanandaj, Dasht-e-Moghan, Rasht, Khomein, Daran, Qom, Shahrood, Kerman, Tabas, and Saravan. The seedlings thinning was done at 3-4 leaf pairs stage and four plants were maintained in each pot. At the time point of sex determination of plants, one female plant was kept for future study. The irrigation was done uniformly to all pots until the emergence of fifth pair of leaves and afterwards, irrigation treatments were applied. During applying irrigation treatments, the soil humidity of the pots was measured before each irrigation cycle. Relative water content, electrolyte leakage, chlorophyll a, chlorophyll b, total chlorophyll content and carotenoid pigments, proline content, catalase and guaiacol peroxidase enzymes were measured at full flowering period - early fruiting. The analyses of variance of obtained data were down using SAS software (v.9.2) and Duncan's multiple ranges test was used for comparing the averages at the significance level of α = 0.05.

The results showed that the highest value of relative water content was obtained from the normal irrigation, which was 77.21% and was reduced to 16.70 and 31.13% under mild and serve drought stress, respectively. Interaction effect of Irrigation levels and ecotypes showed that Urmia ecotype had the highest value of relative water content in normal irrigation treatment, and Tabriz ecotype had lowest value of this parameter in severe drought stress. The electrolyte leakage Index was decreased by 10.54 and 24.11% at mild and severe drought stress, compared to normal irrigation, respectively. The highest value of electrolyte leakage was obtained from Tabriz ecotype in severe drought stress, and the lowest value of this parameter was obtained from Tabas and Saravan ecotypes in normal irrigation treatment. The highest values of chlorophyll b and total chlorophyll content were obtained for Tabas and Urmia ecotypes with 0.61 and 2.25 (mg.g-1 fw), respectively, at normal irrigation treatment. The lowest values of this parameters were obtained for Tabriz and Dasht-e-Moghan ecotypes with 0.17 and 0.84 (mg.g-1 fw), respectively, in severe drought stress. Water deficit decreased 28.12% of carotenoid pigments at severe drought stress compared to normal irrigation, and it increased values of proline, catalase and guaiacol peroxidase enzymes with 47.06, 29.18 and 22.78 (%) respectively, at severe drought stress compared to normal condition. The highest values of carotenoid pigments, proline, catalase and guaiacol peroxidase enzymes were observed in the ecotypes of Tabas, Urmia, Qom and Urmia [0.79 (mg.g-1 fw), 1.27 (mg.g-1fw), 0.0820 and 0.5800(Mc.min-1 mg-1 pro), respectively], and the lowest values of them were obtained for Tabriz, Dasht-e-Moghan, Khomein and Rasht ecotypes [0.34 (mg.g-1fw),0.48 (mg.g-1 fw), 0.0396 and 0.2744 (Mc.min-1 mg-1 pro), respectively].

The results of this study showed that Tabas ecotype had a significant advantage in maintaining relative water content, maintaining chlorophyll content and maintaining membrane stability. The Tabriz Ecotype is the most sensitive ecotype for drought conditions. Because it lost the most values of the relative water content, chlorophyll content and membrane stability in stress condition compared to other ecotypes. Therefore, it can be concluded that the physiological parameters measured under drought stress conditions can be used as a criterion for the identification of tolerant and sensitive ecotypes.

Effect of Bacillus subtilis (0, 102, 105, 106, 108 cfu/ml) was studied on saffron (Crocus sativus L.) ecotypes Natanz and Zanjan during 2017-2019 via a factorial experiment based on randomized complete blocks design in three replicates. Number of leaves per corm, average length of leaves, flowers, petals and sepals, stigma fresh and dry weights, fresh and dry flower yield, number of flowers per plot, total phenol and anthocyanin in petals and the amount of nitrogen, phosphorus and potassium were measured. All morphophysiological were significantly affected due to B. subtilis treatment. The highest and the lowest fresh and dry weight of stigma was observed in 108 cfu/ml (41.44 mg and 7.13 mg) and no treated samples (32.91 mg and 6.14 mg), respectively. The maximum stigma fresh and dry weight was observed in Natanz ecotype. The highest amount of nitrogen, phosphorus and potassium was recorded in 108 cfu / ml (5.55 %, 2061ppm 66.75 and 7540.8 ppm, respectively) and the lowest in control (3.25 %, 1314.42 ppm and 4339.8 ppm, respectively). The highest amount of nitrogen and phosphorus was observed in Zanjan ecotype. According to the results, utilizing the B. bacteria in a proper concentration as a bio fertilizer can be considered beneficial towards optimal growth indices in saffron.

Drought stress is one of the most important environmental factors, which limit the growth of plants. By the end of the 21st century, the incidence of drought stress is expected to increase because of the global warming phenomenon. As a consequence, trees growth and viability in the forests and urban greenspace will reduce. Thus, selection of plants that are more tolerant to severe drought stress and are able to cope with such environmental conditions needs to be considered in future silvicultural strategies. This study was carried out to identify candidate drought-tolerance proteins in Maclura pomifera. Therefore, we aimed to explore the performance of Maclura pomifera under a severe drought stress and analyse the proteome changes of Maclura pomifera leaf in response to drought.

The experiment was carried out on 4-year-old Maclura pomifera saplings genotypes cultivated on a flat field in the Botanical Garden of University of Tehran. Saplings were exposed to irrigation regimes of 100% and 25% field capacity in a completely randomized design. Leaf samples were collected and were frozen immediately in liquid nitrogen and then stored at −80◦C to be used for further analyses. Experiments were performed using the gradient pH 3-10 NL IPG strips for the isoelectric focusing. IEF was carried out using the PROTEAN IEF. Strips were then equilibrated first for 15 min in reducing solution and then 15 min in alkylating solution. Equilibrated IPG strips were then placed and fixed using hot agarose on the top of home-made 12 % SDS- polyacrylamide gels. Separation of proteins in the second dimension was carried out in Protean XL cell. The protein spots were visualized by staining with BioSafe Coomassie gel stains following manufacturer’s instructions.

After doing two-dimensional gel electrophoresis, 25 protein spots that had displayed significant protein level changes were identified. Differentially expressed, proteins were divided in three groups. The first group included stress and defense proteins such as lipoxygenase, two types of heat shock protein, Allergen, Convicilin and legumin A2 precursorm; the second group included oxidative stress proteins such as Catalase, Chloroplast stromal ascorbate Peroxidase, Cytosolic ascorbate peroxidase, Iron superoxide dismutase and Manganese superoxide dismutase. Thethird group included energy and metabolism proteins such as Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit, Ribulose -1,5- bisphosphate carboxylase /oxygenase small subuni, Translation elongation factor, Aldolase, Hydroxy-acid oxidase, Isopentenyl diphosphate isomerase, Dihydrolipoamide dehydrogenase and glyoxalase. The present results indicate that most proteins have been identified and their changes caused an increase in tolerance and adaptation of Maclura pomifera to drought stress. Also, our data suggest that drought tolerance of M. pomifera might be correlated with diminishing oxidative damage by activation of the antioxidant systems.

The present results indicate that most proteins have been identified and their changes caused an increase in tolerance and adaptation of Maclura pomifera to drought stress. Also, our data suggest that drought tolerance of M. pomifera might be correlated with diminishing oxidative damage by activation of the antioxidant systems.

The present study has been conducted to investigate the duration of growth period of different cannabis (Cannabis sativa L.) ecotypes and their responses to water stress based on a factorial completely randomized experiment design in the greenhouse condition at the University of Tehran in 2017. Irrigation levels include 50%, 75%, and 100% of field capacity, with the ecotypes being Urmia, Sanandaj, Tabriz, Dasht-e-Moghan, Rasht, Khomein, Daran, Qom, Shahroud, Kerman, Tabas, and Saravan. Results show that Rasht and Khomein ecotypes have had the highest and lowest duration of germination phase and growth degree-days (GDD1), respectively. The highest duration of vegetative, flowering and maturation phases belong to Dasht-e-Moghan, Rasht, and Dasht-e-Moghan ecotypes with 71.33, 30, and 46.66 (days), respectively, at 100% field capacity. The lowest durations of these phases could be seen in Tabas, Tabriz, and Saravan ecotypes with 42, 16, and 17 (days), respectively, at 50% field capacity. Also, the highest values of growth degree-days for vegetative (GDD2), flowering (GDD3), and maturation (GDD4) phases  for Dasht-e-Moghan, Kerman, and Dasht-e-Moghan ecotypes with 1788, 836, and 1169 (0C.d), respectively, at 100% field capacity, with their lowest values belonging to Tabas, Tabri,z and Saravan with 1039, 413, and 448 (0C.d), respectively, at 50% field capacity. Based on growth period duration and cumulative growth degree-days (total GDD), Tabas and Dasht-e-Moghan ecotypes are found as earliest and latest ecotypes, respectively.

Fire blight, the most important disease of pear tree causes necrosis by an oxidative stress in tissues. Therefore, identification of resistant cultivars and mechanisms of resistance to the oxidative stress of disease, that are mainly related to the chloroplasts, are important in breeding programs of this tree.  In order to study the role of chloroplasts in this interaction, expression of chloroplastic gene psbA that are under control of oxidation/reduction (redox), was evaluated in susceptible (Williams) and resistant (Harrow Sweet) cultivars during 48 h post inoculation by Erwinia amylovora in in vitro condition. In addition, expression of this gene was studied at presence of glutaraldehyde and rotenone as the inhibitors of the electron transport chain of chloroplast and mitochondria, respectively. The results showed higher necrosis progress rate in the in vitroshootlets of susceptible cultivar. Expression of psbA gene at presence of inhibitors in both presence and absence of E. amylovora was higher in cultivars Harrow Sweet. According to the results, the higher resistance level of cultivars Harrow Sweet to the disease could be due to the higher rapid responses and reaction of the chloroplasts of this cultivar in comparison to the cultivar Williams.

Cumin, Cuminum cyminum L., is the king of spices with a plethora of natural compounds with pharmacological features. Drought stress is a well-known factor that influences the production of some metabolites. We studied the impact of drought stress on gene expression and metabolite content in flower and leaf tissue organs of two ecotypes, Taybad and Ardakan populations. Plants were imposed into three water level conditions, control, moderate, and severe water deficit. Concerning the results, the expression of Limonene synthase in flower organ of the Ardakan genotype increased 2.2 times under 50% of field capacity, whereas, the expression of Flavone synthase in leaf tissue of the Ardakan ecotype, was the highest in this level of stress. Moreover, the majority of detected terpenoids were β-Acoradiene and γ-Terpinene in leaf and flower organs, respectively. Altogether, the monoterpenes content was decreased in both ecotypes, but sesquiterpenes increased only in the Ardakan population. Knowing the expression of key genes involved in the pathway of major metabolites in cumin under water stress conditions is important in the pharmaceutical industry and molecular researches.

Toxoplasma gondii is an obligate intracellular protozoan with worldwide distribution. Diagnosis of toxoplasmosis is a very critical issue, especially in pregnant women and immunocompromised patients. The aim of this study was rapid detection of T. gondii DNA in peripheral blood samples (PBS) employing HRM technique and using RE gene.

Totally, 242 samples from pregnant women and human immunodeficiency virus (HIV) patients were collected from different hospitals and medical centers of Tehran during Oct 2017 to Dec 2018. High resolution melting analysis (HRM) using partial sequences of repetitive element (RE) gene was done and compared with ELISA test.

Overall, 51 were positive for acute toxoplasmosis that among them, 12 and 20 reported as positive in pregnant women and HIV+ patients, respectively using HRM technique. Among 70 patients in chronic phase of disease, 10 and 3 samples were reported as positive for pregnant women and HIV+ patients respectively. From 121 negative control, 3 (4.62%) samples associated with HIV+ patients, showed positive real-time PCR and HRM analysis results.

For the first time, HRM technique via employing RE gene was used for detection of T. gondii infection in PBS. This method is suitable, helpful and in parallel with serological methods for early diagnosis of acute as well as active form of toxoplasmosis in pregnant women and HIV+ patients. The use of techniques based on melt curve and through employing next-generation dyes for diagnosis of T. gondii would be accessible for patients in developing countries.

In order to use alternative sources of fiber such as hemp to supply the paper and wood fiber, the exploration and utilization of abundant hemp variations seems to be necessary. In this study, some hemp populations collected from different part of Iran were evaluated with reference to wood anatomy and fiber biometry characteristics. Vessel characteristics of stems including average vessel lumen area, vessel density and porosity were measured using cross-sectioned tissues of stems. Fiber biometry characteristics of fibers including fiber length, fiber width, fiber cell wall thickness and fiber lumen width were measured and compared using macerated bast and core fibers. In conclusion based on vessel characteristics, Frs, San 02, Mahb, Zah and Mahl, populations, are expected to be putative high potential fiber populations, due to having the least vessel density and porosity leading to more woody core fiber content. Regarding fiber length, populations of Ban, Bsh, Frs and Kash, Bsh and Nhv, showed higher average for bast and woody cores, respectively. Hence these populations are expected to perform better in properties related to fiber length such as textile and paper strength and can be successfully used in paper making, fiberboard and textile industries as well as being used in selective breeding programs. Also all populations were evaluated based on their bast and wood fiber width, fiber cell wall thickness and fiber lumen width as effective factors on final product properties.

Cannabis sativa L. is an important plant with various uses in pharmaceutical and paper production industries. Due to the higher priority of female and male Cannabis plants for pharmaceutical uses and fibre industry, respectively, reduction of the quality of products after pollination, and also for detection of better genotypes before pollination for breeding purposes, early determination of sex in Cannabis is one of the major concerns of researchers and farmers. Seeds of 26 accessions from different regions of Iran along with one accession from Afghanistan were planted in the field based on randomized complete block design with 10 replications. Five female and five male plants were sampled from each accession for molecular and morphological analyses. Thirteen ISSR primers and two SCAR markers were used. Also, morphological differences, which possibly related to sex were measured in the Cannabis populations under study. The results showed significant differences among the accessions within female and male plants for plant height, number of days to flowering, days after full bloom and height of the first flowering node. Male plants were generally, but not always, taller than female plants and had a shorter life cycle. The highest expected heterozygosity was found in the ISSR3 primer and the lowest in the UBC825 primer. In total, 143 polymorphic bands and one polymorphic band were amplified using ISSR and SCAR primers, respectively. Out of 143 polymorphic ISSR bands, only 10 markers had significant relations with the gender of this plant. MADC6 SCAR marker was monomorph across the accessions. On the other hand, MADC5 was a polymorphic marker and showed a significant relation with gender. These markers, especially MADC5, have the potential to be used in sex determination of C. sativa.

Saffron (Crocus sativus L.) belonging to Iridaceae family as a source of apocarotenoids is one of the most valuable spices and medicinal plants in the world. Because of the large size and high complexity of saffron genome, its sequencing remains a challenge. The arrival of next-generation sequencing technologies (NGS) has allowed the rapid and efficient development for RNA sequencing. De novo assembly of transcriptome from short-read RNA-Seq data provides a great resource for the study of species without a reference genome. De novo assembly of the transcriptome has some unique challenges, particularly in the case of plants, which possess a large amount of paralogs, orthologs, homoeologs and isoforms. In this research, we attempted to compare the performance of de novo assembly tools including BinPacker, Bridger, Oases-Velvet and Trinity through consideration of a quality metrics such as N50 length, the total number of contigs and alignment scores. The results of these analyses revealed that assembly using Bridger had a superior performance for saffron transcriptome, Oases suffered from relatively high chimera rates and redundancies which causes genes family with high similarity assembled into one transcript, Trinity performs worse than Bridger in the increase of false positives. Our comparison study will assist researchers in selecting a well-suited assembler and offer essential information for the improvement of existing assemblers.

Medicinal Plants are almost reach sources of natural antioxidants such as terpenoids and phenolic compounds and have great potential as an alternative for synthetic antioxidants against oxidative stresses. Peppermint (Mentha piperita) infusion is a valuable worldwide source of antioxidants. The antioxidant property can be enhanced by inducing abiotic stresses. This experiment was conducted to evaluate prolong (4 months) water deficit stress (no stress, mild stress and severe stress as 100, 75 and 50% of field capacity, respectively) on cultivated peppermint on plant growth, secondary metabolite profile and antioxidant capacity of peppermint infusions. Both water deficit stress treatment decresed plant height, leaf wet and dry weight and leaf size significantly. Also, water deficit stress increased antioxidant capacity, total phenolic and flavonoid contents significantly. Some phenolic compounds such as quercetin, coumaric acid and luteolin were detected only in water-deficit conditions. Also, the amount of some identified amino acids and unsaturated fatty acids were changed under water deficit stress. Therefore, inducing water stress can enhance the biological value of peppermint and improve bioactive compounds and the antioxidant capacity of peppermint extract.

Cannabis sativa L. is an important economic plant for the production of medical, food, fiber and oils. Nevertheless, lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis genetic research. In this study, transcriptome sequences were used for identification and introduction of SSR markers in order to assess genetic diversity and separation of fiber and drug types. Based on the cannabis RNA-Seq data, 3383 SSR were identified from 1402 reads in fiber type whiles this number was 10381 for 4743 reads in drug type. Among these markers, trinucleotide repeat motifs were the most abundant, followed by mononucleotide and dinucleotide. Primers were successfully designed for amplification of 234 SSR markers from 152 individual sequences in fiber and 1543 SSR markers from 1372 individual sequences in drug using Primer3 with default parameters. This study outlines the first large-scale development of SSR markers for cannabis. The effectiveness of these molecular markers should be tested using different Iranian Marijuana populations and could be particularly useful for cannabis populations breeding.

Long intergenic non-coding RNA (lincRNA) has been introduced as key regulators of diverse biological processes, including transcription, chromatin organization, cell growth and tumorigenesis. With regard to the potential role of lincRNAs in cancer development, one may postulate that differential expression of lincRNAs could be employed as a tool in cancer diagnosis, prognosis, and targeted therapy. In this study, we aimed to explore the putative correlation between the expression levels of two lincRNAs: LINC00152 and LINC01082 in the bladder cancer (BC), in comparison with its adjacent non-cancerous tissue. Fifty Iranian subjects diagnosed with BC, representing in different stages and grades participated in this study.. The mRNA expression levels of the above mentioned lincRNAs were analyzed comparatively in cancerous and their adjacent non-cancerous counterpart tissues, of each subject by Real-Time PCR. The expression levels of LINC00152, and LINC01082 were significantly lower in tumor tissues in comparison with their adjacent normal tissues (P

To study effects of salicylic acid (SA) treatments (0, 0.5, 1 and 2 mM) in two forms of seed priming (for 24 h) and spraying on seedling at 4-6 leaf stage an experiment was conducted on periwinkle (Catharanthus roseus Don.) with and without exposure to a mild cold stress of 8 °C (for 32 h). Seeds were sown in trays containing peat-based substrate (with 20% sandy loam field soil). Transplants of both groups were treated with mild cold. Seedlings were grown in greenhouse with an average day/night temperature of 25/20± 2°C until their blooming. Seed emergence rate and percentage, activities of catalase (CAT), guaiacol peroxidase (GPX), polyphenol oxidase (PPO), CAT gene expression, height and number of nodes per plant, and days to flowering were evaluated. Results showed that mild cold stress together with salicylic acid at 0.5 mM (as either seed priming or spray on seedling) was the best treatment to accelerate the flowering, and improve growth parameters and antioxidant enzymatic activities. Thus, mild cold stress enhanced the positive effects of SA treatments on cold acclimation of periwinkle (particularly when associated with spraying, which is a simple administration method). Accordingly, it might be recommended for its seedling production and cultivation in temperate climates to prevent late spring frost damages.

Aberrant pre-mRNA alternative splicing is a common event in cancer cells. Many abnormally spliced RNA variants have been observed in tumor cells and they can be used as biomarkers or therapeutic targets in new drug design. Increasing our knowledge in understanding the mechanisms of alternative pre-mRNA splicing for cancer-related genes and determination of cancer specific isoforms are important for the development of new strategies in cancer therapy. The aim of this study was isoforms identification and expression of PIK3CA, FGFR3 and FGFR1 genes in bladder cancer by RNA Sequencing and Real-Time PCR.
This cross-sectional study was conducted at Urology Research Center of Sina Hospital, Tehran University of Medical Sciences, Tehran, from September 2014 to October 2016. Paired tumor and adjacent normal tissues samples were obtained from 30 bladder cancer subjects. Total RNAs were extracted from bladder tumor and normal tissues. Quantitative and qualitative examinations have been done. After quality control, fragmentation of RNAs and cDNA library construction, next-generation RNA sequencing was performed. Resulting raw data were analyzed with different bioinformatics software. Differential expression was confirmed by Real-Time PCR.
RNA sequencing results showed the number of PIK3CA (1 vs 3), FGFR3 (7 vs 6) and FGFR1 (9 vs 12) isoforms and their expression were different in bladder normal tissues in comparison to tumor tissues. Overexpression of PIK3CA gene have been observed in 42% of tumor samples but statistically was not significant. Increased FGFR3 gene (P=0.01) and decreased FGFR1 (P=0.01) expression were significant. There was an association with overexpression of FGFR3 and cigarette smoking ((P=0.037) and family history (P=0.004).
RNA sequencing make possible to do the accurate assessment of transcript abundance and identification of different isoforms resulted from aberrant pre-mRNA alternative splicing, which is a crucial process for the maturation of transcripts of multi-exon genes. Regarding the differences in isoforms expression in tumor and normal tissues of bladder cancer, they have potential to be used as biomarkers and sensitive targets for cancer therapy.

Streptococcus pneumoniae is a causative agent of morbidity and mortality worldwide. Diagnosis of pneumococcal infection includes conventional culture-based and molecular methods. Differentiation of S. pneumoniae from other mitis group streptococci is not reliable.
We aimed to evaluate the efficacy of lytB gene along with lytA gene for detection of S. pneumoniae in isolates and clinical samples using conventional and real-time PCR methods.
In this cross-sectional study, a total of 560 clinical specimens were collected from patients during February-September 2015. The samples were cultured on 5% sheep blood agar and suspected colonies were identified by biochemical assays. The antibiotic susceptibility test was performed by disk diffusion and serial microdilution methods. 46 culture-negative and 46 culture-positive samples were examined to evaluate the presence of lytA and lytB genes using conventional and real-time PCR methods.
A total of 46 (8.2%) isolates of S. pneumoniae were identified in suspected specimens. 52% (24/46) of isolates exhibited multiple drug resistance (MDR). All 46 isolates contained both lytA and lytB genes. Real-time PCR assay detected both genes with low CT values in culture-positive samples. Among culture negative specimens, one sample was positive for both the genes.
The lytB is similar to lytA in sensitivity for diagnosis of S. pneumoniae in isolates and clinical samples based on both the molecular methods. The results confirmed the applicability of real time PCR based on lytB genes for detection of S. pneumoniae.

For improvement asymbiotic seed germination media of temperate terrestrial orchid Epipactis veratrifolia , 12 different concentrations of the carbohydrates (fructose, glucose, sucrose and two combination of fructose with sucrose ) were assessed on the seed germination and protocorm development, in the presence (2g L−1) and absence of peptone in Fast medium. Results revealed significant differences between treatments on seed germination percentage and protocorm growth. Carbohydrate treatments; H12 (3.5g/l fructose g/l sucrose), H8(10 g/l sucrose), H2 (20 g/l glucose), and H11 (5 g/l fructose g/l sucrose) had significant effect on seed germination percentage. H11and H12 was the best medium for protocorm growth. H8P2(10 g/l sucrose g/l peptone), H2P2(20 g/l glucose g/l peptone) and H12P2(3.5g/l fructose g/l sucrose g/l peptone) were the best for seed germination respectively with 79/6%, 74/6% and 71/9% seed germination percentage. H10P2 (30 sucrose g/l g/l peptone) with 17/3mm growth, significantly was the best for protocorm growth. Therefore kind and concentration of carbohydrate and presence of organic nitrogen (peptone) influence asymbiotic seed germination percentage and protocorm growth and could improved both of them. This finding revealing that it is possible to improvement asymbiotic seed germination of this species orchid with combination of mono and disaccharides and organic nitrogen.

For evaluation of physiological and biochemical response of some grapevine cultivars to drought stress, an experiment was carried out in 2014 as a factorial arranged in a randomized complete block design with three replications. Three cultivars i. e. “Bidane Sefid”, “Chafte” and “Yaghooti” and four drought stress levels, including -0.3, -0.6, -1 and -1.5 MP soil water potential were employed. In this experiment, two years old grapes were cultured in pots with 28 cm in diameter and 60 cm in height, containing sandy-loam soil in field conditions. Results showed that in “Bidane sefid”, under drought stress (-0.3 to -1.5 MP), RWC, chlorophyll a and b content, and antioxidant enzymes CAT, APX, GPX activity along with DPPH scavenging capacity were declined, but electrolyte leakage, MDA content, LOX enzyme activity (responsible for membrane unsaturated fatty acid peroxidation), along with H2O2 accumulation were increased. Results indicated that ‘Bidane Sefid’ under drought stress had higher oxidative damage, led to lower membrane integrity. In comparison with “Bidane sefid”, “Chafte” and “Yaghooti” under drought stress, due to higher antioxidant enzymes (CAT, APX, GPX) with higher DPPH scavenging capacity, exhibited lower oxidative damage, led to higher membrane integrity. According to our results, it can be concluded that “Chafte” followed by “Yaghooti” had higher potential to tolerate drought stress compared with “Bidane sefid”.

The poultry industry occupies an important position in the provision of animal protein. Recently, next generation sequencing technology (RNA-Seq) has become available as a powerful tool to investigate transcriptional profiles for gene expression analysis of many organisms. The main use of RNA-Seq in agriculture species are focusing on finding the immune related genes or pathways by comparison of the whole transcriptome following pathogen challenge. Alternative splicing (AS) is the major fundamental mechanism generating the protein diversity and regulating the gene expression in eukaryotic organism. Identifying genes that are differentially spliced between two groups of RNA-sequencing samples is interesting subject in transcriptome with next-generation sequencing technology in this study used RNA sequencing to comparison isoforms of two breeds. A total of 64,819 transcripts were identified by aligning sequence reads to genome among the evaluated isoforms for expression analysis, 310 were significantly differentially expressed between two breeds, including 251 up-regulated and 59 down-regulated. The KEGG results of up regulated isoforms showed that that no pathway was found significantly different (FDR ≤ 0.05). However, enrichment analysis suggested that seven were over-represented (P-value ≤ 0.05) within the up regulated isoforms. Only one of them functionally related to immune system, natural killer cell mediated cytotoxicity. The results showed genes which are breed-specific expression and the comparative transcriptome analysis help to understand the difference of genetic mechanism.

Saffron (Crocus sativus L.) is one of the most valuable medicinal and spice herbs in the world. In spite of the ancient cultivation history in Iran, there are limited breeding studies on the plant due to its vegetative reproduction. In order to evaluation genetic diversity of Iranian saffron germplasm, sixty-five different saffron accessions from the main cultivation areas in Khorasan including Torbat heidarieh, Gonabad, Mahvelat, Ghaenat and Ferdows were collected and were studied by molecular markers. The used RAPD and ISSR primers produced 43 and 122 polymorphic markers loci, respectively, and totally 165 markers with average of 7.5 markers by each primer, totally. Diversity index ranged from 0.36 to 0.7 with average of 0.23. Also, marker index with the average of 0.16 varied in the range of 0.2 to 0.7. The accessions from Ghaenat and Mahvelat had the maximum (83.03%) and the minimum (52.73%) polymorphism, respectively. Grouping the studied saffron accessions using cluster analysis displayed four distinct groups which had little correspondence to their collection areas, while clustering for the main cultivation areas had relatively good correspondence to their geographical distances. So, it is expected to have nearly approaching improvements of qualitative and quantitative yields via the selection of superior clones of saffron.