mohammad javan nikkhah

Rice is the most important food for a significant portion of the world's population, especially in Asia. The infection of rice plants with Fusarium species is a major global problem. For the morphological and phylogenetic identification of Fusarium species associated with rice, a widespread sampling was conducted from the root, crown, leaf sheath, and stem of the rice plants in various rice-cultivating areas across Fars, Golestan, Guilan, Isfahan, Khuzestan and Mazandaran provinces. Species-specific primers were used to identify three closely related species: F. fujikuroi, F. proliferatum, and F. verticillioides, which could not be distinguished based on morphological criteria alone. By combining morphological characteristics and sequence data of TEF1-α and β-tubulin genomic regions, nine species including F. fujikuroi, F. verticillioides, F. proliferatum, F. globosum, F. andiyazi, F. anthophilum, F. incarnatum, F. culmorum and F. oxysporum were identified among 242 isolates examined. This study reports the first occurrence of Fusarium andiyazi on rice in Iran. In addition to the known causal agents of bakanae disease, the pathogenicity test revealed that two additional species F. culmorum and F. anthophilum can also be considered agents of bakanae disease on rice.

The vast land of Iran, with its diverse climate and geographical location, is known as a suitable habitat for different types of thyme. Thyme (Thymus spp.), a plant of great nutritional and medicinal value, holds a special place among pasture plants. Consequently, investigating the factors that threaten the survival of this plant is important. Two species of thyme, T. kotschyanus (Boiss. & Hohen) and T. pubescens (Boiss. & Kotschy ex Čelak) have significant habitats in the pastures of Alborz province. In this research, pests and diseases of these two species were investigated in 14 different stations in Alborz province. The samples were collected at three time points (May, July and September) and the collected insects were taken to the laboratory for identification. The results indicated the presence of six types of pests on thyme species: Aphis serpylli, Haplothrips reuteri, Tetranychus urticae, Aeolothrips mongolicus, a mite of the genus Bryobia, and also Cuscuta spp. as a parasitic plant. Alternaria leaf spot was also detected on thyme leaves. The results of this research are reported for the first time It can be used by regional managers and stakeholders and help in the protection of Thymus species in the rangelands of Alborz province.

Over the past two decades, biological control of wheat septoria tritici blotch (STB) caused by Zymoseptoria tritici has been evaluated as a reliable and safe alternative or complement to chemical control. The genus Trichoderma includes several biocontrol agents (BCAs) widely used worldwide against plant diseases. Besides direct control of plant pathogens, Trichoderma spp as BCAs provide numerous other beneficial features for their plant hosts. The current study aimed to assess the potential biocontrol ability of several antagonistic Trichoderma isolates with endophytic potential on STB reduction in wheat cultivar Tirgan under greenhouse conditions. The antagonists were used as a seed-coated treatment at the sowing time and sprayed on the plants 72 h before Z. tritici IPO323 inoculation. STB severity was calculated 21 days after the pathogen inoculation. The biocontrol isolates significantly (α = 0.01) reduced the disease severity on cv. Tirgan. T. harzianum AS12-2 provided the highest average percentage of STB inhibition in both application methods. Application of T. harzianum AS12-2 led to a reduction in the percentage of leaf area covered by lesions (PLACL) and the percentage of leaf area covered by pycnidia (PLACP) by 80.12% and 77.71%, respectively. Trichoderma biocontrol isolates could be considered safe and reliable alternatives/complements for sustainable agriculture in integrated management programs of wheat STB.

Some important herbs have been overly utilized in all over the world, which threatening them as endangered species. In this regard, in vitro culture has efficiently improved production and propagation rates of these plants. Accordingly, this research aimed to determine the best genotype, and explant for callus induction, as well as the effect of optimum concentration of hormones on traits including length, width, height and the callus volume of the explants derived from Ferula assa-foetida seedlings that have been grown in jars. Therefore, this experiment has been implemented as factorial in completely randomized design with at least 3 replications. Purposefully, the explants were placed in MS media supplemented with different levels of BAP and NAA. After 40 days, the traits including the length, width and volume of the callus were measured for induced callus. Given the mean comparison, the maximum size of target traits have been achieved for root explants cultured medium supplemented with 2 mg.L-1 BAP and 1 mg.L-1 NAA, while none of calluses derived from leaf and stem explants survived. Among the genotypes, the Kerman demonstrated higher callus growth rate than the Yazd, though this trait intensively increased through the time of culture. In addition, the calluses derived from Isfahan showed low growth rate, so it was removed from the present study.

Rice is one of the most important crops in Iran and the rice blast caused by Pyricularia oryzae causes significant losses in rice fields during outbreak season. At the beginning of the disease cycle, the primary inoculum imposes severe damage to plants as it leads to plant weakness. The damage will shortly become significant as the second wave of inoculum spreads. This study aimed to determine the occurrence and severity of blast disease on some native Iranian cultivars and estimate the number of conidia produced on second leaves. Pyricularia oryzae Cavara isolates were collected from rice leaves from the north of Iran. One out of 123 isolates was inoculated on seven Iranian native cultivars named Hashemi, Tarom Mahali, Sang Tarom, Binam, Champa, Alikazemi, and Domsiyah at the four-leaf stage. Lesions on leaves were scored and the percentage of the spotted area was calculated by software Image J1.52a. Various lesion types and a range of diseased areas varying from 30% recorded in Hashemi to 55% in Tarom Mahali were observed. Under optimum conditions, the sporulation potential was calculated. Isolate produced different amounts of spores on different cultivars; Hashemi showed the highest and Tarom Mahali showed the lowest sporulation with 63 and 11 spores per mm2 of spotted area. The result indicated that the resistance threshold of the native cultivars could be different but not sufficient. Additionally, to estimate the damage on each cultivar on farms, we need to investigate the sporulation potential of the pathogen through time in each cultivar

In this study, the effect of mycoendobiont fungi isolated from white button mushroom against Mycogone perniciosa, Lecanicillium fungicula and Trichoderma harzianum, as its important pathogens, was investigated for use in biological control and replacement of chemical compound. Isolates of mycoendobiont fungi were isolated from the cap, gills and stalk of healthy white button mushroom, collected from major mushroom growing farms in Iran. These isolates screened against aggressive strains mentioned above in dual culture tests and the most effective isolates were selected for greenhouse experiments. White button mushroom spawn was treated with the mycoendobiont fungi and disease incidence and growth parameters were measured. All mycoendobiont isolates tested were able to inhibit and compete with pathogens and controlled the incidence of wet bubble disease (M. perniciosa) from 21.42% to 97.61%, dry bubble disease (L. fungicula) from 60.63% to 96.80% and green mold disease (T. harzianum) from 64.28 to 96.93%. This test was performed in a completely randomized design with Three replications. Mycoendobiont isolates of Fusarium venenatum and Clonostachys rosea had the greatest inhibitory effect on the growth of all pathogens. In addition, Scedosporium apiospermum had a high growth inhibitory effect against M. perniciosa and T. harzianum. According to the results, mycoendobiont fungi of white button mushroom could be considered as the biocontrol agents and growth promoting with the aim of applying them in future.

Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris is one of the major yield-limiting factors in chickpea, particularly in west and northwest of Iran. Simple Sequence Repeats (SSR) were used to determine genetic structure of Fusarium oxysporum f. sp. ciceris populations from three infected areas located in west and northwest provinces of Iran during 2016-2017. A set of six microsatellite primer pairs revealed 35 alleles in each locus with an average of 5.83 per locus. The PIC value varied from 0.20 (FOAG11) to 0.39 (FODF7) with an average of 0.27 per locus. The average number of effective alleles (Ne), Number of private alleles (Na), allelic variability per locus (H), Shannon's Information Index (I) and percentage of Polymorphic Loci (%PL) in three populations were 1.33, 1.97, 0.2208, 0.357, and 74.29, respectively. The average genetic distance was calculated among the three populations. Nei’s pair-wise genetic distances between the populations varied from 0.0202 to 0.98. The total gene diversity (Ht) and gene diversities between subpopulations (Hs) were estimated to be 0.2208 and 0.2079, respectively. Gene diversity attributable to differentiation among populations (Gst) was 0.0585, while gene flow (Nm) was 8.0412. The AMOVA of genetic variation in Fusarium oxysporum f. sp. ciceris populations revealed that 5% of the variance occurred among populations and 95% within populations. PhiPT value was 0.054. The lowest genetic distance was found between Kurdistan and West Azerbaijan with Lorestan populations, while the highest genetic distance was observed between Lorestan and Kermanshah populations.

khamedj is one of the most important fungal diseases that in a favorable condition can lead to severe outbreaks and causing an 80% loss of the annual harvest. 30 isolates of Mauginiella scaettae, the causative agent of this disease including 27 isolates from different locations of the Iran, two isolates from Iraq and one isolate from Kuwait were collected from different palm cultivars during 1397-1397 and used in the present study. Seven EST-SSRs markers were applied to show the genetic diversity among these 30 isolates. Five loci including, SNOD1, SNOD26, SNOD22, SNOD17, and SNOD21 were polymorphic among the species and revealed a total of 16 alleles. Also, two microsatellite markers, including SNOD5 and SNOD16, did not amplify and showed no amplification. The dendrogram constructed based on the similarity index resulted in five major clusters so that two clusters were divided into sub-clusters. The highest similarity value was observed between isolates Ilam-sa and Behb-kh2 (0.75) and followed by isolates abad-ma1 and ramh-sa1 (0.57). On the other hand, two isolates Kheshbid-za and Abad-sa1 have the highest genetic distances with other isolates. Thirteen isolates were selected from the identified genetic groups to study pathogenic groups on the sayer cultivar. The results indicated that there were significant differences among isolates used. The results revealed that Behb-ma1 isolate and Mehr-sa isolate showed the lowest and highest pathogenicity, respectively.

Root and crown common rotdisease caused by Bipolaris sorokiniana, has caused severe losses of wheat products worldwide. A collection of 147 monoconidial isolates was made from cultivated wheat in West Azerbaijan (Miyandoab), Golestan (Gorgan), Alborz (Karaj), Kermanshah (Kermanshah) and Fars (Shiraz) provinces in 2017-2018. After morphological and molecular identification, mating type primers were designed based on a conserved DNA binding (alpha domain) in MAT-1 and the high mobility group (HMG) domain in MAT-2. Frequency of mating type idiomorphs of B. sorokiniana isolates were done by multiplex PCR. Among 147 isolates of five population, 86 isolates amplified by MAT-1 primer and 61 isolates by MAT-2 primer. Frequencies of MAT-1 alleles in Shiraz and Gorgan populations was more than the MAT-2 alleles. According to the results of the Chi-square test (X2), Karaj, Kermanshah and Miyandoab populations had not significant difference (P<0.05). Therefore, these three populations have the highest potential for sexual reproduction than the otherthreepopulations. In addition, isolates from all of the populations were studied to evaluate the sexual fertility in vitro. After two months, pseudothecia without mature asci and ascospores formed in media.

The purpose of this study was to estimate heritability, general and specific combining ability and the relationship between traits. Investigated traits were fruit number per plant, total yield per plant, average fruit weight, flesh thickness, flesh firmness, seed cavity size, total soluble solid, fruit length in melon. A complete diallel design was conducted for this purpose five parental genotypes were crossed and the hybrids were evaluated in a randomized complete block design with three replications. Analysis of variance of traits showed a significant differences between hybrids, therefore the first Griffing method was used to analyze the combining ability. The general combining ability effects were significant for all parents in the evaluated traits. The specific combining ability of yield per plant was higher than the general combining ability, which indicates the dominance and non-additive actions of genes. High narrow-sense heritability for yield per plant, flesh firmness and the number of fruit per plant (0.86, 0.72 and 0.70, respectively) indicated low environmental effect for these traits and the greater role of additive effects. The highest narrow-sense heritability was estimated for yield per plant (0.86). The role of non-additive effects of genes in controlling yield and flesh firmness traits was greater than additive effects. Therefore, the preparation of superior hybrids using breeding methods based on progeny tests will be effective in improving the mentioned traits in melon.

The members of Fusarium is one of the important causal egent of soil borne diseases which cause serious damage in the agricultural and horticultural crops in the world. In order to investigate Fusarium destributions in agricultural soils in the Bamyan and Daikundi with semi desert climates, Kandahar and Helmand with desert climates, sampling was performed in Afghanistan in 2018-2019. The PPA medium (Pepton PCNB Agar) was used for Fusarium isolations. For morphological characterisation, pure cultures were transferred to CLA (Carnation Leaf-piece Agar), SNA (Synthetic Nutrient Agar ) and PDA (Potato Dextrose Agar) mediums and gene sequencing of tef-lα regions were performed for molecular identification. Totally 134 Fusarium isolates were obtained and grouped in 11 different species. The presence of F. incarnatum-equiseti species complex (FIESC) (28%), F. solani (16%), F. acuminatum (6%), F.verticillioides (5%), F. oxysporum (3%), F. coeruleum (1%) and F. compactum (1%) with different frequency were detected in both regions of the sampling areas. F. sambucinum (1%) was detected in semi-desert climates (Bamyan and Daikundi provinces), While F. proliferatum (18%), F. culmorum (13%) and F. Pseudograminearum (1%) were isolated and identified in areas with desert climates (Helmand and Kandahar provinces). The results this study, geographical distribution of various species of Fusarium in Afghanistan, provide useful information about management of soil borne Fusarium pathogens.

Hyperspectral imaging technique as a new and efficient method is applied for detecting infection in agricultural products. It was used for classification of healthy and infected pistachio kernels by Aspergillus flavus fungus with and without considering infection stages. Two different fungus isolates, R5 and KK11 with and without capable of producing aflatoxin, respectively, were individually used to infect the pistachio kernel samples. From 960 to 1700 nm, three effective wavelengths of 1090, 1280, and 1700 nm were selected by principle component analysis method. After feature extraction, K-fold cross validation, support vector machine, and artificial neural network methods were used for classification. The results showed that the classification accuracy of the K-fold cross validation method was higher for classifying the healthy and infected pistachios without considering the infection stages and isolate type (99.71%). The maximum accuracy of the developed algorithms in classification of isolate type and infection stage was obtained as 69-91%.

Barley leaf stripe caused by the fungus Pyrenophora graminea, is one of the most important and devastating barley diseases in Iran. A total of 121 P. graminea isolates was recovered from cultivated barley in West Azerbaijan (Miyandoab region), East Azerbaijan (Miyaneh region), Ardebil (Ardebil region) and Zanjan (Khorramdareh region) provinces in 2016. After morphological and molecular identification, mating type primers were designed based on a conserved DNA binding (alpha domain) in MAT-1 and the high mobility group (HMG) domain in MAT-2. Frequency of mating type idiomorphs of P. graminea isolates were done by Multiplex PCR. Among 121 isolates, 56 isolates amplified by MAT-1 primer and 65 isolates by MAT-2 primer. Frequencies of two mating types in Khorramdareh (1) and Miyandoab populations had an excess of the MAT-1 and MAT-2 alleles, respectively. According to the results of the Chi-square test (X2), Miyaneh, Ardebil (1 and 2) and Khoramdareh (2) populations had not significant difference in compared with Khorramdareh (1) and Miyandoab populations. Therefore, these four populations have the highest potential for sexual reproduction than the two populations of Miyandoab and Khorramdareh (1). In addition, isolates from all of the populations were studied to evaluate the sexual fertility through directed crossing with opposite mating types. After six months, pseudothecia formed in media without mature asci and ascospores.

Western and north western provinces of Iran have the highest potential for chickpea cultivation in Iran. Vascular wilt caused by Fusarium oxysporum f. sp. ciceris (FOC) is one of the most important diseases of this crop which distributed in chickpea cultivation areas of these provinces. The purpose of this study was to identify the pathogenic races of FOC in the western and north western provinces. Sampling was conducted on three infected locations. After isolation of the fungal isolates, identification was performed using FOC specific primers by PCR method. Out of 116 isolates identified, 94 isolates belonging to three locations were selected to recognize the fungal races. Five races including R0, R1B/C, R1A, R6 and R5 showed the highest abundance in the studied areas, respectively. Presence of five races from the eight identified races of FOC in the world in western and north western provinces of Iran indicates the high genetic diversity of this fungus in the country and distribution of the races of this fungus is independent of environmental factors.

In order to study the Bipolaris and Curvularia species associated with corn, sorghum and sugarcane, sampeling were performed from different regions of Iran (including Alborz, Ardabil, Fars, Gilan, Golestan, Khuzestan, Mazandaran and Zanjan provinces) during the summer of 2011 and 2012. Fungal species were identified based on morphological features and molecular data of ITS regions of ribosomal DNA. Eleven species including Viz. Bipolaris cynodontis (48 isolates), B. sorghicola (16 isolates), B. maydis (17 isolates), B. sorokiniana (seven isolates), B. bicolor (four isolates) and B. oryzae (two isolates), Curvularia spicifera (19 isolates), C. papendorfii (ten isolates), C. ellisii (two isolates), and C. hawaiiensis (one isolate) were identified. Pathogenicity test of the species were performed on four to six leaf stage of corn, sorghum and sugarcane seedlings. Finally, pathogenicity of B. maydis, B. sorokiniana and B. oryzae on corn and B. sorghicola on sorghum seedlings were confirmed. This is the first report of occurrence and pathogenicity of B. sorokiniana and B. oryzae on corn leaves in Iran.

The objective of the present study was to isolate and characterize of fungal species from Persian oak trees rhizosphere in Zagros forests, Ilam province (Iran). For this purpose, sampling was performed from different regions of the above-mentioned forests during 2014–15. The serial dilutions of soil suspensions were cultured on Water agar (WA 2%), Potato dextrose agar (PDA), and Malt extract agar (MEA) media. Fungi were purified by transferring single hyphal tips on PDA medium.  At the first, the fungal isolates were identified using their morphological features. To confirm the morphological identification and taxonomic position of the isolates, molecular studied and phylogenetic analyses were then carried out using amplification and sequencing of ITS region of ribosomal DNA. Finally, the species Mortierella alpina, Penicillago nodositata,and Trichocladium jilongensis were identified based on morphological characters and molecular studies. These species were described and reported as new records for the mycobiota of Iran.

Single strand DNA or RNA known as aptamers, are able to detect the target molecule with specificity at the monoclonal antibody level. However, the placement of mycotoxins in small molecule groups and their high molecular weight differences with aptamers have become a serious challenge in introducing aptameric probes for them. In the present study, in order to achieve a new and shorter aptamers sequence for aflatoxin B1 (AFB1), an initial oligonucleotide library (Lib1) was designed based on the sequence of a known AFB1 aptamer (named Apt1, 50 bp) using the genetic algorithm. The best aptamer from the Lib1 library was selected based on the molecular docking results and has been modified to create a new library (Lib2) using the truncating strategy. Virtual screening of the Lib2 library in terms of their binding affinity over AFB1 molecule led to obtain the truncated aptamer, C52-T, with 19 bp in length. Type of interaction and stability of C52-T-AFB1 complex were investigated using molecular dynamics simulations (MD) and MM-PBSA method. The affinity constant of C52, C52-T and Apt1 aptamers over AFB1 were estimated through unmodified gold nanoparticle-based colorimetric assay. The experimentally founding and in silico results were completely consistent. It seems that the computational techniques have the great potential to introduce sensitive molecular probes to design specific mycotoxins aptasensors.

Polycyclic aromatic hydrocarbons (PAHs) are counted as important pollutants due to their ubiquitous occurrence and toxicity, mutagenic and carcinogenicity characteristics. They are counted as the main pollutants of soil. One of the most effective, environmentally friendly and cost-effective methods is bioremediation of contaminated soils. Fungi have been identified as the most successful microorganisms in the removal of these compounds. The purpose of this study was to identify endemic fungal strains compatible with PAH compounds. Contaminated dumping site of acidic sludge, produced from the recycling process of used oils in Eshtehard industrial complex was selected as the study area. Some endemic fungal species including Aspergillus fumigatus, Alternaria chlamydosporigena and Penicillium chrysogenum were isolated from contaminated soil and inoculated into contaminated soil. To examine the efficiency of fungal remediation of PAHs, after 21 and 50 days of inoculation, residual PAHs concentrations were measured by Gas chromatography mass spectrometry (GC/MS). The results interpreted by the mean and standard deviation of data using Excel software and comparison of means by Tukey HSD test. The results revealed that all three fungal species, identified in this study, were significantly effective in eliminating PAHs. The highest removal rate related to Naphthalene and Acenaphthene was due to the volatility of these compounds. Removal of Benzo [a] pyrene, which is the most cancerous PAH and has the highest molecular weight in this study, was 50%, 40% and 32% by Aspergillus fumigatus, Alternaria chlamydosporigena and Penicillium chrysogenum isolates, respectively. In addition, there was a significant difference between different species in reducing the concentration of different PAHs, at α = .05.

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.</span> 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.

During survey of rice fields in Fars and Khuzestan provinces at mature grain stage, 42 seed samples were collected. Fusarium isolates were obtained from these samples by planting seeds on selective Fusarium medium (Pepton-PCNB-Agar, PPA) by two methods i.e.; surface sterilization by sodium hypochlorite (NaClO 1%) and without disinfection. Cultivated samples were kept in incubator at 25±1° C and intermittent light (12 hours of light and 12 hours of darkness). Morphological identification of 26 purified Fusarium isolates was performed on the WA, PDA, CLA and SNA. Phylogenetic relationships of 18 isolates on the basis of TEF1-α gene region and ITS1-5.8S-ITS2 region sequences were investigated. The results revealed that Fusarium isolates belonged to two species complexes FIESC (Fusarium incarnatum-equiseti species complex, 50%) and GFSC (Gibberella fujikuroi species complex, 46%). Fusarium fujikuroi species in Fars and F. semitectum species in Khuzestan were most frequent. Two species F. fujikuroi and F. andiyazi belonging to the GFSC species complex were identified in two separate clades in phylogentic tree based on TEF1-α. On the basis of TEF1-α gene, F. merismoides was not identified, but the ITS region sequence was identified as Fusicolla acetilerea and differentiated this species from Fusarium species in the phylogenetic tree.

In order to assess the dynamic of Pyricularia oryzae at leaf and panicle neck blast stages, population genetic structures of the 142 isolates were investigated in a rice field and at three levels using SSR markers and five primer pair combinations. All the isolates were collected from the two-leaf and panicle stages at three levels. In the first level, isolates collected from leaf and panicle neck blast stages in the rice field were analyzed. In the second level, isolates belonged to the leaf and panicle neck populations were collected from tillers of the same rice hill and analyzed. In the third level, the ienvestigated isolates belonged to three populations and were obtained from three distinct leaf spots from the same rice leaf in a rice tiller. Based on constructed dendrogram for all of the 142 isolates, genetic similarity varied from 89 to 100% among the isolates. Population genetic estimations at the three studied levels showed that the amount of gene flow was 57.62, 10.68, and 4.783, respectively between the isolates. Detection of mating type idiomorphs of the studied isolates using multiplex PCR showed that all the isolates had Mat1-1 idiomorph. This indicates the absence of sexual reproduction within the M. oryzae populations in rice fields in Guilan province and the fungus reproduces only in an asexual way. Low genetic variation and high genetic similarity among the populations of the causal agent of rice blast disease in Guilan province confirm this result.

Venturia species include many important plant pathogenic species such as Venturiainaequalis and V. pyrina causing scab or black spot on apple and pear. The Venturia teleomorph is the overwintering structure developing in leaf and fruit litters. The immersed teleomorphicpseudothecia are produced in the host tissues. This Ascomycetous genus is characterized by small, solitary, scattered or gregarious, globose, sub-globose, papillateascomata with setae around the ostiole. Asci are bitunicate, oblong to obclavate with a short pedicel or not. Ascospores ellipsoidal with broadly rounded ends, pale brown, one septate with two unequal size cells. Venturia has some species like V. inopina as homothallic and some like V. populina as heterothallic species. Sexual reproduction with producing sexual air-born spores plays an important role in distribution of disease in early spring. Therefore, awareness of sexual cycle in plant pathogenic fungi is very important due to management and forecasting probability of incidence of new genotypes (races). Venturia and related anamorphic species were collected from different places of Iran (18 provinces including Alborz, Ardabil, Eastern Azarbaijan, Golestan, Guilan, Hamadan, Isfahan, Kermanshah, Kurdistan, Lorestan, Markazi, Mazandaran, North Khorasan, Qazvin, Razavi Khorasan, Tehran, Western Azarbaijan, and Zanjan) during years 2013 and 2014. The collected specimens (800 specimens) were identified based onmorphological features. Isolation of the fungi from the infected samples was conducted using single spore method by streaking out spores on 2% water agar (WA) and culturing of single germinated conidium on potato dextrose agar (PDA). V. inaequalis was determined as a dominant species in Iran. Ten isolates from each of V. inaequalis and V. pyrina were crossed together under in vitro condition. Some isolates were cultured as single isolate in the petri dishes for investigation of homothallism and heterothallism. Several isolates from two species were crossed together to survey the biological species definition. Mature leaves of Golab apple cultivar (Malusdomestica) (for V. inaequalis isolates) and pear (for V. pyrina isolates) were washed under running tap water to remove dust and debris, and cut into 2×2 cm squares avoiding the midrib of the leaf. The leaf pieces were autoclaved and placed in petri dishes with 2% water agar. A plug of mycelium of each isolate was scraped from the edge of three weeks old colonies on PDA and mixed onto the leaf pieces with a scalpel. The plates were sealed with parafilm and placed in a sealed plastic bag. The cultures were incubated at room temperature under 12 hours light / 12 hours dark condition for 2 weeks, after which they were maintained, inverted in the dark at 8°C. The cultures were checked for the production of pseudothecia for the first time after six months. Among crossed isolates, seven and five fertile crosses were obtained from V. inaequalis and V. pyrina isolates, respectively, after six to eight months. In single isolate cultures, no sexual fertility was observed which confirm the heterothallic nature of both species. Moreover, between isolates from two species, no fertile crosswasobtained, which is the evidence of two distinct biological species. This is the first study on sexual reproduction of thesetwo species under in vitro condition in Iran. Morphological features of obtained sexual organs in vitro were the same as pseudothecia, asci and ascospores features in vivo. All ascospores in an ascus were fertile and they germinated on water agar after 24 hours. Based on this study, isolates with two different mating types are available as tester isolates. These isolates will be useful to design the primers to detect MAT alleles. PCR based methods can be used to study population genetic diversity, phylogeny, disease epidemiology and population biology (fungal species overwintering in a region based on mating types frequencies and their proportion). Development of reliable methods of sexual reproduction in vitroseems to be a valuable toolfor studies e.g. classic genetic analysis, strains modification and a supplement for modern genetic manipulations and evolutionary analysis. Our results verified heterothallism in both of V. inaequalis and V. pyrinaspecies. There was nofertile cross between isolates from two species that is the evidence of two distinct biological species. In this study, some tester isolates for mating typewere obtained which appear to be useful for the next PCR-based molecular studies on MAT alleles.

Apple scab disease caused by Venturia inaequalis is the most important economic disease worldwide where the apple is grown and causes annual epidemics. It reduces the quality and quantity of the yields in different regions of Iran. In this study, diversity and population genetic structure of V. inaequalis were surveyed on different apple cultivars using 18 microsatellite markers. 51 isolates were obtained from infected leaf and fruit specimens from wild apple, Iranian endemic and commercial apples from Northern provinces of Iran (Mazandaran, Golestan and Guilan). 28 SSR primers were used to investigate the genetic diversity of this pathogen. Among them, 18 primers showed polymorphism between isolates and populations. AMOVA analyses revealed that 97% of the variation was distributed among individuals within populations, and 3% was attributable to the differences among populations. Gene diversity indexes including Nei’s gene diversity, Shannon index and allele numbers in every population showed that diversity within the population on endemic cultivars is more than the diversity in the population of wild and commercial cultivars. Also, the population of commercial cultivars has more diversity in comparison with the wild population. High genetic diversity within the populations is caused by annual sexual reproduction, gene flow between populations and probably the existence of this fungus for a long time in this region.

Pistachio die-back disease is an important disease in Iranian pistachio orchards. One of the most important fungal agents of the disease is Paecilomyces which has been recently identified as P. formosus based on phylogenetic and physiologic studies. In this study, four pairs of primers designed using mating type idiomorph sequences of P. variotii deposited in GenBank (NCBI). Designed primers were tested in some isolates of P. formosus and finally two primer pairs including; Mat1-1f224 and Mat1-1r224 for amplification of Mat1-1 and Mat1-2f165 and Mat1-2r165 primers for amplification of Mat1-2 idiomorphs were selected. Mating type idiomorphs were amplified in 124 isolates of P. formosus which have been obtained from pistachio trees and eight other species including; Pistacia mutica, Punica granatum, Prunus amygdalus, Caesalpinia gilliesii, Nerium oleander, Tamarix aphylla, Tamarix hispida, Haloxylon sp. and air of pistachio orchards. In 50 isolates (40.3%) Mat1-1, in 59 isolates (47.6%) Mat1-2 and in 15 isolates (12.1%) both idiomorphs were identified. 16 isolates from each mating type and three isolates which had both idiomorphs were selected and crossed in all of the possible combinations on PDA culture medium and cultures were incubated at 25 °C in continuous dark condition. After eight months, sexual reproduction was not observed in crosses.

Studying the effects of fungicides on rhizosphere and soil microbial communities is very important due to the critical role of microorganisms in soil and plant health. In this study, the effects of fungicide Rovral TS on dynamic and structure of cucumber rhizospheric bacteria was investigated by Next-Generation Sequencing based on 16S rRNA gene (V4 region) by means of Illumina MiSeq sequencing in three plant growth periods. Results showed that among 26 identified phyla and 352 identified bacterial genera for 18 samples, Proteobacteria and Pseudomonas were most dominate phylum and genus, respectively. Application of Rovral TS was resulted the changes in bacterial structure and dynamic. Decrease of relative abundance of Pseudomonas in middle stage of flowering in fungicide treatment and in the last stage of control treatment was observed. Moreover, the relative abundance of Sphingopyxis, Sphingobacterium and Chryseobacterim in fungicide-treated plants were more than control plant in all three stages. Analysis of diversity indices revealed that diversity in the rhizospheric soil of control plant was more than fungicide-treated plants (shanon=5.33), while species richness in control soil was greater than fungicide treated (Chao1=2324.17). In addition, the effect of plant developing stages on OUT number, diversity and richness indices (regardless type of treatment) was significant and rhizospheric soil belongs to before flowering stages benefit more diversity and richness.