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Entomopathogenic fungi produce a variety of degrading enzymes, including proteases, chitinases and lipases, to facilitate their entry through the massive barriers of insect cuticle. Isolates of the entomopathogenic fungi vary considerably in their proteolytic activity and virulence. The proteolytic activity of different isolates has been hypothesized to reflect their virulence toward the host. In this study, we evaluated the virulence and proteolytic activity of 17 Beauveria bassiana sensu lato isolates collected from different geographical regions in Iran. The selective medium D0C2 was used for isolating B. bassiana from soil samples. Casein substrate was used for protease assay. Total mortalities caused by different B. bassiana isolates through the dipping method, ranged from 25 to 60% with the highest and lowest rates for isolates BA and MITE, respectively. Our results revealed a wide variation in both proteolytic activity and virulence among the studied isolates. Additionally, we found a strong positive correlation between the proteolytic activity on Casein substrate and virulence of the isolates against the Khapra beetle, Trogoderma granarium. This finding will facilitate the screening and selection process of virulent fungal isolates as efficient agents for use in biological control programs of insect pests.  

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Genetic diversity relationships of 50 isolates of Cytospora schulzeri on apple from different parts of the Semirom region were analyzed using 15 polymerase chain reaction (PCR) based markers, 7 random amplified polymorphic DNAs (RAPDs) and 8 Microsatellite primed polymerase chain reaction (MP-PCR). Using 7 selected RAPD primers 113 bands were generated, of which 81 bands were polymorphic (71.7%), with an average of 11.57 polymorphic fragments per primer, and with 8 selected MP-PCR primers 107 amplified bands were observed with 78 polymorphic bands (72.3%), with an average of 9.75 polymorphic fragments per primer. In RAPD marker, number of polymorphic bands varied from 8 (241) to 15 (230, 238, OPA13) with an average of 11.57 per primer and which varied in size from 200 to 3750 bp. Percentage of polymorphism ranged from 64% (203 and 232) to a maximum of 83% (238). In MP-PCR marker, number of polymorphic bands varied from 6 (CAG) to 12 (GTG and ATG) with an average of 9.75 per primer and which varied in size from 200 to 3500 bp. Percentage of polymorphism ranged from 54% (CAG) to a maximum of 81% (ACTG). By combining markers, a total of 220 bands were detected, of which 159 bands (72%) were polymorphic and produced on an average 10.6 polymorphic bands per primer. The results showed that both markers were suitable for the detection of genetic polymorphism among apple C. schulzeri isolates. Estimated genetic relationship using similarity co-efficient (Jaccard’s) values between different pair of accessions varied from 0.54 to 0.89 in RAPD, 0.62 to 0.89 in MP-PCR and 0.62 to 0.87 with combined markers based similarities. High cophenetic correlation between the similarity matrix and corresponding dendrogram was obtained by RAPD + MP-PCR marker (r = 0.81). Cluster analysis of the data using UPGMA based on Jaccard´s similarity coefficient, divided the isolates into six groups, showing a high genetic diversity among populations of C. schulzeri.

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Ninety one monoconidial Bipolaris isolates were obtained from lesions on different parts of rice in different locations of Mazandaran province during the summer of 2009. Bipolaris species were identified using morphological features such as color and shape of colony and color and size of conidia and conidiophores. The isolates were separated into two species; 85 (93.4%) isolates belonged to Bipolaris oryzae and the remaining 6 (6.6%) isolates to Bipolaris cynodontis. Therefore B. oryzae is regarded as the major cause of rice brown spot disease in Mazandaran province. In order to analyze genetic diversity among B. oryzae isolates, 71 isolates were subjected to fingerprinting analysis by rep-PCR using BOX and REP primers. In cluster analysis, 15 clonal lineages and 54 haplotypes were identified. The largest clonal lineage contained with 36 haplotypes was the most common lineage. These results also indicate a relatively high level of genetic diversity among B. oryzae isolates. Also, pathogenicity test of a few B. oryzae isolates (12 isolates) was conducted under greenhouse condition and showed that those isolates were pathogenic to rice seedlings of cv. Tarom. All isolates produced some leaf spots 24 h after inoculation.

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The effects of water deficit on grain yield, yield components and the physiological char-acteristics of barley (Hordeum vulgare L.) varieties were studied in a split plot design dur-ing the 1999-2000 growing season in Karaj, Iran. Five irrigation levels [0, 25, 50, 75, and 100% crop water requirements (CWR)] and six barley varieties Karoon Kavir , Rei-hani(drought tolerant), Torkman , C-74-9 (intermediate) , Kavir Badia and Gorgan-4 (2 rowed type, drought sensitive) were arranged in the main plots and sub plots respectively. Results showed that water deficit induced stress and this decreased the grain yield and yield components. Water deficit increased the canopy temperature and those varieties showed a higher canopy temperature under non-stress conditions, performed better un-der drought conditions. Severe stress reduced the chlorophyll content (SPAD values) con-siderably, but the differences were not significant between the 50, 75 and 100% CWR treatments. In addition, no significant differences were observed in the chlorophyll con-tent of barley varieties under drought conditions. The effect of irrigation on the photo-chemical efficiency of photosystemII (Fv/Fm) and the other fluorescence parameters for all varieties were significant. Although the Fv/Fm values were not significant in barley varie-ties at any level of irrigation, in general Karoon Kavir and Reihani varieties showed a better performance under water deficit conditions. Proline content was significantly dif-ferent in various irrigation treatments, but its accumulation at any level of irrigation did not differ significantly in barley varieties. It was concluded that the higher canopy tem-perature (less negative T) under well irrigated conditions and higher grain yield, 1000-grain weight, Fv/Fm values under water stress conditions could possibly be the proper cri-teria for screening the drought tolerant barley genotypes under field or laboratory condi-tions.

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Anthracnose disease caused by Ophiognomonia leptostyla, is the most important and widespread fungal disease on Juglans regia. Walnut disease symptomatic samples were collected from different provinces of Iran, during 2015–2016. Fungal isolates were identified based on ITS-rDNA sequence data. Variance analysis of colony growth rate (mm/day) and acervulus density on medium, was significant. Acervulus density on medium was strongly correlated with colony growth rate. The Max acervulus density was 60% and > 80% for Hamedan and Mazandaran isolates respectively. The virulence of six selected isolates was examined on cv. Chandler. Virulence indices including spot diameter, disease severity, spot area average and logistic infection rate except spot number index, could successfully detect significant differences among isolates. SA-SE1 isolate from Mazandaran showed significantly the most virulence indices: disease severity (%), spot area and logistic infection rate. For the other five isolates, four significant levels in all virulence indices were observed. In summary after this isolate, other isolates including TA-ZY21, LA-SY21, U94-SR1, HA-GH22 and MA-K1 were placed in the next steps of virulence ranking. There was insignificant correlation between colony growth rate and disease severity. However, the acervulus density and disease severity were significantly correlated implying the importance of acervular conidial inoculum in secondary disease cycle progress. Disease severity was strongly correlated with number of spots, spot diameter and logistic infection rate. Disease severity was also negatively correlated with Mid-time (time to progress 50%). Moreover, there was positive relationship between logistic infection rate and three traits: number of spots, spot diameter and spot area average. This study was the first of the disease virulence components on cv. Chandler in Iran.
 

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In this study, 32 plant samples with leaf spot symptoms were collected from Chaharmahal and Bakhtiari province, Iran, in the spring and summer of 2018. Isolation and purification of 26 fungal isolates were performed on 2% water agar and potato dextrose agar to identify the causal agents. Fungal species were identified according to morphological characteristics and molecular data obtained from glyceraldehyde 3-phosphate dehydrogenase (gpdh) gene sequences. In this research, 11 species belonging to four genera of hyphomycetous fungi, including Alternaria cantlous, A. consortialis, A. multiformis, Bipolaris sorokiniana, B. zeicola, Curvularia spicifera, C. nicotiae, C. inaequalis, Stemphylium beticola, S. symphyti, and S. vesicarium are introduced. Among them, three species, including C. nicotiae from Salvia officinalis L., S. beticola from Plantago major L., and S. symphyti from Mentha pulegium L., are reported as new records for mycobiota of Iran. All collected plant species are reported as new hosts (matrix nova) for the identified fungal taxa.

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Some saprophytic fungi were isolated during fungi isolation causing black spot or scab-like symptoms (genus Venturia) from Iran. One saprophytic isolate from apple leaf was identified as a member of the family Phaeosphaericeae based on morphological characteristics of pseudothecia, asci, and ascospores on oatmeal agar (OA). Mature pseudothecia were produced on OA after one-month incubation at 24 °C and continuous dark conditions; however, the pseudothecia produced on PDA were immature. Pseudothecia were dark brown and globose in shape. Numerous bitunicate asci formed in a broad hymenium. Asci were clavate, apically rounded, short pedicellate, with eight overlapping biseriate fusiform ascospores. Ascospores with the smooth wall, straight or slightly curved and slightly constricted at the second septum. Phylogenetic analysis based on sequence data of ITS and LSU regions of ribosomal DNA confirmed the morphological identification and specified the isolate as Neosetophoma poaceicola. The N. poaceicola is the first report from Iran and apple leaf in the world.
 

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Delamination is one of the main defects in composite materials. Studying the initiation and propagation of delamination is useful for the design and production of high quality resistant materials. Therefore understanding the behavior of delamination damage and having enough knowledge about that will be helpful. In this study, generated acoustic emission signals from two types of glass/epoxy composite specimens at different layups, [0,90]5s and [0]10, is recorded. After that signals were analyzed by applying strain energy function. The results indicate that specimens at different layups have different stiffness behavior toward damage initiation and growth. Using strain energy function method initiation and growth of delamination can be evaluated. The obtained results provide useful information for the design and production of high quality resistant composite materials.

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Delamination is the most common failure mode in composite materials. It takes place in different modes, i.e. mode I, mode II or the combination of these modes. The present study is concerned with an investigation of mechanical and acoustic emission behavior of delamination. In this work, various lay-ups of glass/epoxy composite laminates have been used to study the delamination behavior when subjected to mode I, mode II and the mixed-mode I/II tests. First, the characterization of load-displacement curves of the specimens is done based on the AE parameters and mechanical responses and the curves were divided into three parts. The crack growth in the mode I was stable state and in the mixed-mode and mode II was unstable. In the next, interlaminar fracture toughness of the specimens, Gc, were measured using standard methodologies and acoustomechanical methodologies which is based on the mechanical behavior and AE information. It was found that the acoustomechanical method presents the lower limit of the interlaminar fracture toughness and agrees with the results that obtained from standard. The images were captured with Scanning electron microscope (SEM) from damage surfaces verifies the results that obtained from Acoustic emission.

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Objective: Influenza virus A (H1N1) is an important subtype of the influenza respiratory viruses, which has important worldwide implications. Hemagglutinin (HA), an important viral antigen, is responsible for binding to human cell receptors leading to an onset of the disease process. Considering the critical role of viral attachment, this study focuses on the extraction and cloning of HA and its large subunit HA1 genes to generate recombinant baculovirus shuttle vectors (bacmid) in order to produce recombinant proteins in insect cells. Methods: Human influenza virus A/New Caledonia 99/20/(H1N1) was propagated in MDCK cell culture. Total viral RNA was extracted using easy-red solution. The full-length HA genome and HA1 fragment were amplified by RT- PCR using specific primers, cloned into a pGEM®-TEasy vector, and then subcloned into a pFastBac HT plasmid. Finally, recombinant bacmids that contained the genes of interest were produced in E. coli DH10Bac™ cells. Results: Expected PCR products of HA genes were evaluated through gel electrophoresis and restriction enzyme analysis. Recombinant pGEM®-TEasy vectors and pFastBac HT donor plasmids were confirmed by PCR, digestion, and sequencing. Construction of recombinant bacmid DNA was verified by using blue-white colony screening, overnight electrophoresis, and PCR analysis that used either pUC/M13 or gene-specific primers. Conclusion: In this study, we have successfully constructed recombinant Bacmid DNA that encoded the full-length HA genome and its HA1 subunit. We intend to transfect sf9 insect cells with these constructs to generate recombinant baculovirus and produce large amounts of desired proteins for future studies.

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Sandwich composites are widely used in structural applications because of their appropriate mechanical properties and low strength/weight ratio. Delamination is common failure mode in these structures that lead to a reduction in strength and stiffness of composite. In this paper, using acoustic emission, initiation and propagation of delamination in sandwich composite specimens was investigated. The specimens were loaded under mode I loading. Then the characteristics of the signals related to different damage mechanisms were specified. The acoustic emission signals were classified based on their frequency ranges. Then the acoustic emission signals were recorded during the test specimens were processed using wavelet transform. Thus the percentage of energy in each components of the acoustic emission signal was specified. Each of these components has a certain frequency range corresponding to a damage mechanism. Thus the percentages of different damage mechanisms in each specimen were specified. The Scanning Electron Microscopy (SEM) was also employed to verify the results which were obtained from acoustic emission and wavelet transform method. The results showed acoustic emission is efficient tool for identification and separation of different damage mechanisms in sandwich structures.

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The AISI D2 steel is a high-chromium and high-carbon tool steel which has good mechanical properties such as high compressive strength and good through-hardening. Despite these advantages, fracture toughness of this steel is moderate. In this study, fracture toughness of AISI D2 steel was determined using Finite Element and Acoustic Emission methods. Selected steel (AISI D2 cold-work tool steel) was heat treated and tempered at different conditions. Then Compact testing specimens were prepared according to ASTM E399 standard and fracture toughness of the specimens was specified according to the standard method. The specimens were modeled in the commercial FE software (ABAQUS) and fracture toughness of the specimens was determined using FEM. Determination of fracture toughness using AE technique was carried out according to three methods: Acoustic Emission Energy Rate (AEER), Acoustic Emission Count Rate (AECR) and integral of sentry function. The results obtained from ASTM E399, Finite Element and Acoustic Emission methods were compared with each other. It was found that fracture toughness values which were obtained using AECR and integral of sentry function techniques are lower bound and the results obtained from FEM are upper bound values of the fracture toughness. Furthermore, fracture toughness values obtained using AEER were the most consistent with the results obtained from ASTM E399 standard method. Finally, it could be concluded that Acoustic Emission method can be used as a useful method for determination of fracture toughness of engineering materials.

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Structures during their lifetime experience plenty of static and dynamic loads. These loads cause failure or undermine the structures. So, reinforcement or repairing failed parts is one way to repair out of service structures. Composite materials have been used to reinforce structures. These materials enjoy advantages such as the proportion of their strength to their weight. As these structures get exposed to some load a number of failures get introduced. This research investigates the failure mechanisms of a notched 2024-T3 aluminum plate repaired with a composite patch using visual and acoustic emission methods. After constructing the specimens, tensile test has been conducted, and acoustic emission sensors have been stocked on the surface of the plate, so that they can record acoustic data. At the first stage, mechanical data obtained from the specimens in different states based on the number of layering have been analyzed. At the second stage, acoustic data, obtained from recording of acoustic emission signals, have been compared with the mechanical data. Also the images obtained from SEM were used to investigation of damages. According to this research, it is identified that a reasonable correspondence between the results obtained from mechanical and acoustic data and the desired functionality of the acoustic emission method in determining failure mechanism in those specimens that are repaired with composite patches.

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The influenza A virus is of global concern for the poultry industry, especially the H5 subtype as it has the potential to become highly pathogenic for poultry and mankind. Recently, plant expression systems have gained interest as an alternative for the production of vaccine antigens. The goal of the present study was to investigate the possibility of expressing the HA1 protein in Nicotiana tabacum via agroinfiltration. In this study, the Hemagglutinin type 1 (HA1) of a high pathogenic avian influenza virus of the H5N1 subtype was synthesized and transiently expressed in Nicotiana tabacum. To examine the possibility of expressing the HA1 protein in N. tabacum, a cDNA fragment encoding the HA1 gene was synthesized de novo, modified with a Kozak sequence, a C-terminal hexa-Histidine (6His) tag, and an endoplasmic retention signal (KDEL). The construct was cloned into vector and the resulting - HA1 plasmid was agro-infiltrated into N. tabacum. The relative gene expression of recombinant plant-produced HA1 was measured by quantitative real-time PCR. Guided by the gene expression profile, HA1 protein was extracted at 3 dpi and subsequently purified utilizing the 6His tag. A recombinant HA1 protein was immunogenically detected by conjugated polyhistidine antibody in western blot, dot blot and ELISA assay. In order to verify the right conformation of HA1 produced in plants, western blot was also done using mouse monoclonal anti-influenza A virus (H5N1/HA1) [2B7]. The results of Real Time PCR assay indicated that the foreign gene was transcribed in transfected leaves. Migration size of protein was detected at 45 kD by Western blotting and demonstrated no discrepancy compared to the positive control (HA1). ELISA results showed that the HA1 was expressed in the transfected leaves in high level as the yield of recombinant protein was 8.8 % of TSP and the yield of purified HA1 was 0.16 g purified protein per kg fresh weight of leaves. This is the first research about the transient expression of the tobacco-made HA1 protein where a synthetic sequence was used for its expression. Here, the efficacy of agro-infiltration for expression of HA1 antigen in tobacco was illustrated. Agro-infiltration expedites the process of recombinant antigens expression in plant tissues. Accordingly, our results provide great opportunity for the exploration of transiently plant-manufactured HA1 as vaccine candidate.

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Composite structures under impact loading are prone to a variety of damage mechanisms such as delamination, fiber breakage, or matrix cracking. It is proven that the impact-induced damage mechanisms of composite materials are dependent on scaling (in-plane and out-of-plane) and layup configurations.  The present study has investigated the effect of scaling and layup configurations on the failure mechanisms of composite materials under low-velocity impact force using acoustic emission, C-scan and CT-scan tools. For this purpose, four samples with quasi-isotropic configurations of [45_m/0_m/90_m/-45_m]_ns were manufactured, then they were loaded and acoustic signals were recorded. The three IS, PS, and SS samples were investigated based on D62624/D6264M ASTM standard test and the R sample had half of the in-plane dimension of them. The variables m and n vary according to the design plan. The obtained acoustic emission data were analyzed using sentry function, then C-Scan and CT-scan were utilized for damages’ size and location.   It was proven that scaling and layup configuration affect the type and intensity of damage mechanisms as well as mechanical behavior of the laminated composites. Furthermore, the acoustic emission method is shown as an indicator of scaling and layup configuration effects in glass/epoxy composite materials under the low-velocity impact.

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Ascorbic Acid (AsA) is a water-soluble antioxidant that makes plants resistant to environmental stresses by neutralizing free radicals. However, it is unknown to what extent this antioxidant may help Improve Irrigation Water Use Efficiency (IWUE) and reduce the adverse effects of water deficit on mung bean growth and yield. In an attempt to clarify whether exogenous application of this antioxidant could alleviate the adverse effects of water deficit on mung bean plants, two seasons (2019 and 2020) of field experiments were conducted using twelve combinations of three AsA levels (distilled water as a control and 10 and 20 mM of AsA) and four irrigation water amounts (25, 50, 75, and 100% of the plant water requirement). Based on the results, the maximum IWUE was obtained with W₅₀AsA₂₀ in the two seasons. The beneficial effect of AsA application on IWUE was determined under water stress conditions (W₅₀). High water deficit (W₅₀) plus applying 20 mM ascorbic acid, i.e. W₍₅₀₎AsA₍₂₀₎ treatment, improved seed yield about 43.7% as in the two seasons compared to high water deficit without ascorbic acid, i.e. W₍₅₀₎AsA₍₀₎. In 2019 and 2020, water saving in W₍₅₀₎AsA₍₂₀₎ compared to the control, was equal to 50% (2,550 and 2,500 m³ ha^-1, respectively). In W₍₅₀₎AsA₍₂₀₎ treatment, the increase of seed yield ranged between 79-107% in both seasons. Thus, the results reveal the potency of AsA to save water under low water supply and increase yield in mung bean fields.