منصوره میرابوالفتحی

Cupressus sempervirens L. var. horizontalis is one of the indigenous species of Iran, the natural habitats of which are regarded as significant forest reserves within Iran. In the northern of Iran, cypress trees have commonly been observed in both natural and artificially planted forests. The purpose of this study was to investigate the fungal factors affecting the common cypress tree die back in Hassanabad (Mazandaran province) during the summer of 2023. Study of the affected trees indicated drying in the terminal branches of the trees, blight and necrosis of the upper branches, as well as canker formation on the trunks of common cypress trees. The infected tissues of common cypress trees were cultured on potato dextrose agar medium. Fungal isolates were obtained. Based on the macroscopic and microscopic characters of the isolates they were identified as Diplodia cupressi in the laboratory. To confirm the morphological identification the rDNA sequence data (ITS1-5.8S-ITS2) were obtained using primers ITS1 and ITS4, and nBLAST. The pathogenicity of the fungus was approved on potted cypress seedlings. This study represents the major role of D. cupressi as the causal agent of Common cypress canker in North of Iran. It seems that the climate change and drought conditions provided the favorable environment to outbreak of the disease

Sphaerulina is a genus of fungi which belongs to the family Mycosphaerellaceae in the order Mycosphaerellales. In a survey of leaf-inhabiting fungi associated with trees in Hyrcanian forests, several isolates of the genus Sphaerulina were recovered from leaf spot symptoms of oak trees. Integration of multi-locus phylogenetic analyses (LSU, ITS, actA and tef1) and morphological identification were applied to identify the isolates. The results revealed that, the isolates belong to a species of the genus Sphaerulina, namely, S. quercicola. To the best of author’s knowledge, this is the first report of this species for the funga of Iran.

The Didymellaceae is one of the most species-rich families in the Pleosporales, and contains numerous plant pathogenic, saprobic and endophytic specie, inhabiting a wide range of ecosystems. To identify the fungal species of the Didymellaceae in Iran, we investigated nine unidentified phoma-like strains obtained from the Iranian Fungal Culture Collection (IRAN) and six additional strains obtained from various diseased plants in Khuzestan Province, during 2019–2021. The strains under study were recognized by combining the results obtained from multi-gene phylogenetic analyses (ITS, LSU, rpb2 and tub2), and morphological comparisons. Accordingly, six species belonging to four genera of the Didymellaceae were identified and described as follow: Didymella aquatica, D. segeticola, Ectophoma multirostrata, Epicoccum dendrobii, E. tobaicum and Neomicrosphaeropsis Juglandis. All these species are new records for the fungi of Iran. Furthermore, several new hosts are reported for these taxa.

Pistachio is one of the most important horticultural products in Iran which its production is always affected by biotic and abiotic limiting factors. Armillaria crown and root rot caused by Armillaria mellea, is an important disease in pistachio orchards, which causes the death of fertile trees. Control of the disease is difficult, costly and rarely successful. Symptoms of the disease in shoots of affected trees includes yellowing, early defoliation, sudden wilting, dieback of branches and reduction shoot growth. These symptoms, together with the decay and browning of woody tissues and the formation of white to creamy mycelial sheets in the roots and crown, as well as the observation of rhizomorphs of the fungus, are sufficient evidence for definitive diagnosis of Armillaria crown and root rot in pistachio trees. The infection process begins with the growth of hyphae or rhizomorphs and their firm attachment to the surface of the roots. The pathogen uses a combination of methods such as mechanical pressure, toxin production, and cell wall-degrading enzymes to penetrate the root tissues. Successful control of the disease is possible only by combining chemical, cultural, biological control methods and use of resistant rootstocks and cultivars due to delay in diagnosis of the disease before the spread of the pathogen in plant and soil and the appearance of symptoms, the presence of an extensive network of rhizomorphs at a distance from the tree and deep in the soil and formation of the mycelium beneath the plant bark or inside dead wood.

Iranian Pistachio is one of the most important horticultural product in export market. Crown and root rot caused by Phytophthora species is the most serious disease of plant, which annually destroys a considerable number of mature and young trees. This disease has been reported from all provinces of Iran. The pathogen is soil-borne and is distributed by sporangia or the released zoospores and infects the healthy trees. In the most Pistachio orchards, the key factors of disease development are the sensitivity of pistachio crown to Phytophthora and the flooding method of irrigation. The symptoms of the disease are include blight in early spring, drying of the green leaves during the growing season, gum exudation from the crown of tree  and the root rot. Gummosis can be successfully controlled by integrated disease management including orchard constructing in non-infected areas, using resistant or tolerant cultivars, using healthy rootstocks, improving the irrigation methods and avoiding the direct contact of water with tree crown, isolating the contaminated parts of the orchard from the healthy parts, using suitable fungicide, and biological control based on Trichoderma and Bacillus species.

In recent years, widespread tree dieback simultaneously with charcoal oak disease have occurred in Zagros forest especially in Ilam province. They dramatically caused damage to oak trees. This research was carried out to identify pathogenic fungi, to prove the pathogenicity and to investigate their pathogenic behavior during 2012-2015 in the forests of Ilam province. After several field observations across the forests of Ilam province, several samples were collected from infected parts of Persian Oak trees and transferred to the laboratory. Then, the samples were cultured to screen the pathogenic agents identified using morphological traits and validated by fungi keys. In order to prove the pathogenicity of the identified fungi, they were inoculated on two-year-old oak seedlings. In the final stage, the pathogenic behavior of fungi was studied. The results showed that fungi causing charcoal diseases in the forests of Ilam province were Biscogniauxia mediterranea and Obolarina persica. Furthermore, the fungi caused the disease on oak seedlings with dieback symptoms on the sections areas above the inoculum site. The study of the pathogenicity behavior of these fungi showed that their contamination began from inoculation site and developed in the wood, in addition to longitudinal extension, the hyphae colonized in transverse and radial directions towards the heartwood, so that it affects the xylem and phloem vessels. Considering the pathogenicity prove of these fungi and their contribution to the weakness and dieback of Persian oak trees, it would be possible to reduce the oak losses by eradicating the dead trees to prevent fungal spore spreading in the region.

Boxwood tree is one of the Iranian endemic trees expanded throughout Hyrcanian forests. They are of particular importance among the world's forest reserves. Recently, Calonectria pseudonaviculata has been considered as one of the most important causes of blight and leaf defoliation of the boxwood in Iran. For the first time, the present study was aimed to evaluate the genetic variation of Hyrcanian C. pseudonaviculata isolates using RAPD and ISSR markers.
Material & In this study, 75 fungal isolates were collected from the infected boxwoods throughout Hyrcanian forests in the North of Iran. The isolates were assessed based on morphology and colony color. Genetic diversity of the isolates was studied using RAPD and ISSR markers. A part of the beta-tubulin gene was sequenced and deposited at NCBI. Phylogenetic analysis was carried out using PAUP* v. 4.0b10.
The colony color of isolates on potato dextrose agar (PDA) medium was brown with pale hale, conidia were cylindrical, rounded at both ends, 1-septate, 48- 68 µm. Stipe was long (90-14 µm) and hyaline, with the extension terminating in a broadly ellipsoid papillate vesicle, the widest part above the middle. Phylogenetic tree based on the β-tubulin gene showed that all isolates are placed into the same group. Our results indicated that the population of this fungus has a low genetic diversity in Iran.
It seems that the variation of this pathogen is started to change genetically in some areas due to selection pressure phenomenon.

Thirty-seven isolates of Fusarium graminearum species complex obtained from wheat heads with Fusarium head blight symptoms were selected and used for phylogenetic studies. They were collected from different localities of Moghan plain (Ardebil province, Iran). Partial sequences of translation elongation factor 1-alpha (TEF), putative reductase (RED) and UTP-ammonia ligase (URA) genes were amplified using specific primers and sequences were analyzed using Maximum Parsimony method. Almost all strains of F. graminearum species complex belonged to F. graminearum sensu stricto. The results indicated homogeneity within F. graminearum species complex. Inter-simple sequence repeat markers (ISSR) were employed to study the genetic structure of F. graminearum s.s. populations collected from five localities in Moghan plain. The ISSR markers were generated by five primers and data analysed using UPGMA method with Jaccard's coefficient. Cluster analysis showed that all isolates were divided into four clades with 82 similarity level. The analysis of molecular variation indicated that most of the gene diversity (97%) was distributed within populations, whereas 3% of the variation was found among populations. Phylogenetic species identification and genetic diversity knowledge of major agent of FHB disease will be useful in defining the risk of pathogen evolution as well as benefiting disease management strategies.

In field surveys of charcoal disease of Persian oak (CDO) trees caused by Biscogniauxia mediterranea in Iran, the fungal stroma-bearing (apparent infected) oaks have been regarded as infected trees. However, this disease has a latent phase that its ignoring leads to wrong estimations. Therefore, this research was performed to estimate percentage of latently infected oak trees and dieback related to the disease. Oak trees were sampled randomly to isolate fungi from inner bark in two regions of Zagros forests. Also altitude, diameter at breast height (DBH), crown diameter, CDO sign (fungal stroma), dieback severity (DS), severity of leaf loss by insects (SLLI), fire damage (FD), sign of damage by bark beetles (BB) and wood-boring insects (WBI) were recorded during sampling. The isolates of B. mediterranea were identified by characteristics of anamorph and teleomorph states and species specific primers (MED1-MED2). The results reveal that all oak trees have more or less dieback symptoms. Latent infection of causal agent of CDO occures in 12% of sign free oaks equivalent to 10% of all examined oak trees. Approximately, 16.7% of evaluated oaks tress has sign of CDO. On the whole, 26.7% of oaks have latent or apparent infection of B. mediterranea. CDO correlates positively with DS, FD, DBH, SLLI, BB and WBI, and negatively with altitude and crown diameter. The dieback severity of infected oaks with B. mediterranea is more than that of non-infected ones. Moreover, dieback severity of oak trees having apparent infection is more than that with latent infection.

In order to determine the causal agents of Fusarium head blight, during 2015–16, wheat heads with disease symptom were collected from 10 and six main wheat production areas of Golestan and Mazandaran provinces (N Iran), respectively. A total of 431 Fusarium isolates were obtained belonging to nine Fusarium species based on their morphological characteristics. Fusarium graminearum species complex had the highest frequency among the species in both provinces, Golestan (52.0%) and Mazandaran (55.8%). Fusarium culmorum, F. equiseti, F. Acuminatum, and F. compactum had the highest mean of frequency in both provinces with 13.5, 9.7, 6.0 and 3.7%, respectively after F. graminearum. Also, F. cerealis, F. Avenaceum, and F. proliferatum and some unidentified isolates a total of 14% of the isolates were calculated. The lowest frequency was related to F. subglutinans (1%) that was isolated only from Golestan province. In order to determine the phylogeny of F. graminearum species complex in the North Iran, 53 out of 229 isolates were selected based on their distribution in the sampled areas. Partial genes of translation elongation factor 1-alpha (TEF) and putative reductase (RED) were amplified using specific primers. A commercial sequencing facility was used to generate fungal sequences. Almost all strains of F. graminearum species complex belonged to F. graminearum sensu stricto. The results indicated a homogeniety within F. graminearum species complex, however, there was a minor morphologically differences between some strains.

During recent years outbreak of box blight trees causing sudden leaf abscission have declined about 40000 hectares boxwood trees in Northern forests of Iran. Calonectria pseudonaviculata is the causal agent of the disease. Distribution of boxwood blight disease was monitored in north forests from June to September of 2014. Sampling locations was included the provinces: Mazandaran (Liresar, Sisangan, Chalandar, Toskatook, Kohnesara, Neka, Sorkhkola, Afrachal, Savadkooh, Chaeebagh, Vachat, Ghaemshahr, Amol, Behshahr), Giulan (Astara, Kalat, Gisoom, Roodsar, Siahkal, Bijarkenar) and Golestan (Bandargaz-Livan sharghi) provinces. The isolates were identified based on morphological characteristics at first, then molecular identification confirmed morphological identification using previously designed species-specific primer ITS113F and ITS4. The pathogenesis variation of Hyracanian isolates was tested in green houses using representative 35 isolates. Boxwood blight fungal agent was isolated from all surveyed areas. Morphological criteria distinguishing this species from close Cylindrocladium species including cylindrical, rounded at both ends, 1-septate, 48- 68 (47.3) ×4- 6 (4.8) µm of conidia along with the long stipe (90-14 µm) which was hyaline, extension terminating in a broadly ellipsoid papillated vesicle with the widest part at the middle resulted in to identify all isolates as Cylindocladium pseudonoviculatum. ITS113F and ITS4 primers amplified the expected 250 bp region in all Calonectria pseudonaviculata isolates specifically. Comparing the pathogenicity of 35 isolates on Buxus sempervirens subsp. Hyrcana saplings revealed pathogenicity degree variation among Hyricanian Cylindocladium pseudonoviculatum divided in 15 groups were scattered in three provinces.

Pistachio is the most important commercial product in Iran and root and crown rot (gummosis) is the most serious diseases of this crop. Causal agents of the disease were unknown in Sistan -va- Baluchistan province where 7000 ha are under cultivation of pistachio. During 2 years, thirty pistachio orchards including pistachio trees of different ages were inspected throughout different seasons. Eight Phytophthora isolates were isolated using citrus leaves pieces as baits on water-saturated soils collected from around the infected trees and cultured on PARP medium. Some isolates were isolated culturing the surface disinfected tissues of root and crown of infected trees directly on PARP as the semi-selective medium too. Two groups of isolates were obtained. Based on the morphological and physiological characteristics of the Sistan -va- Bluchistan pistachio isolates, the first group of isolates was identified as Phytophthora pistaciae and the second group identified as Phytophthora nicotianae. The two Phytophthora species were found pathogens using unripe pear and apple fruit inoculation method, detached pistachio twigs inoculation method and artificial inoculation of the crown area with a plug of cultured isolate medium. This is the first report of incidence of pistachio tree gummosis and its causal agents from Sistan -va- Baluchistan province.

Fusarium graminearum as the main cause of Fusarium head blight of wheat not only decreases the yield, but also causes mycotoxicosis in human and livestock because of producing dioxynivalenol and nivalenol. Sixty isolates of F. graminearum obtained from infected wheat farms of Iran were tested for the ability of deoxynivalenol, nivalenol and acetylen derivations production using specific primers of Tri13P1/Tri13P2 and Tri13F/Tri13DONR. A 583bp fragment responsible for the production of 15-Acetyl deoxynivalenol amplified in 36 isolates out of 60 tested ones, also in 24 isolates, an 859bp band responsible to product nivalenol were reproduced. None of the isolates produced 644bp fragment which is responsible for 3-Acetyl deoxynivalenol production. The data related to the distribution of Tri13 haplotypes by the PCR method showed that the majority of F. graminearum isolates were DON and 15-AcDON producer. Potential of trichothecenes B production of isolates possessing Trichothecenes-generating genes were evaluated using HPLC-SPE method. 77% of isolates produced 15-AcDON and 46% produced NIV. Also none of the DON-producing isolates was able to produce NIV, whereas the low levels of DON (10- 90 ppb) were evaluated in F. graminearum isolates producing high levels of NIV. Distribution of DON derivatives and NIV- producing F. graminarum isolates in different provinces showed that the most NIV- producing isolates were obtained from Golestan and Mazandaran provinces whereas DON chemotypes isolates were more than NIV-producing ones in Fars, Kerman, Hormozgan and Ardabil provinces.

Boxwood)Buxus sempervirens subsp. hyrcana) is one of the main forest evergreen plant species through the Caspian Sea provinces. Boxwood is endemic to Iran and it has particular importance among the forest reserves in the world. Boxwood blight disease was reported, from the forests of Mazandaran and Guilan province in 2013 (2, 3). Calonectria pseudonaviculata, the causal agent of buxus blight can complete its life cycle within one week under its favorite temperature and humidity (1). Development of the disease is as fast as that the boxwood blight symptoms were observed broadly in Guilan and Mazandaran provinces. To monitor the epidemiology of the disease, the infected leaves and twigs were collected from boxwood trees in Bandargaz area, located in Golestan province. The infected plant tissues were surface sterilized with hypochlorite sodium (%5) and cultured on PDA (potato dextrose agar) medium to isolate the causal agent. Fungal isolates were purified using hyphal type method on water agar medium (1.5%). For spore production, the colonies were grown on PCA (potato carrot agar) and incubated under near- ultraviolet light at 25°C. The colony color on PDA was brown with pale hale. No mycellial growth was observed at 30°C which had been reported (1). Conidia were produced frequently after 15 days on PCA medium. Conidia were cylindrical, rounded at both ends, 1-septate, 48- 68 (47.3) ×4- 6 (4.8) µm. Stipe was long (90-14 µm), hyaline, the extension terminating in a broadly ellipsoid papillate vesicle, and with the widest part above the middle. This is the first report of outbreak boxwood blight disease throughout all of Hyrcanian forests located in Iran.

Pistachio contamination to aflatoxin has been known as a serious problem for pistachio exportation. With regards to the increasing demand for Raman spectroscopy to detect and classify different materials and also the current experimental and technical problems for measuring toxin (such as being expensive and time-consuming), the main objective of this study was to detect aflatoxin contamination in pistachio by using Raman spectroscopy technique and artificial neural networks. Three sets of samples were prepared: non-contaminated (healthy) and contaminated samples with 20 and 100 ppb of the total aflatoxins (B1+B2+G1+G2). After spectral acquisition, considering to the results, spectral data were normalized and then principal components (PCs) were extracted to reduce the data dimensions. For classification of the samples spectra, an artificial neural network was used with a feed forward back propagation algorithm for 4 inputs and 3 neurons in hidden layer. Mean overall accuracy was achieved to be 98 percent; therefore, non-liner Raman spectra data modeling by ANN for samples classification was successful.

Ochratoxin A (OTA) is a mycotoxin which contaminates different plant products, including cereals, coffee, and dried grape. Aspergillus and penicillium species were identified as the main producer of OTA. Ochratoxin A is a potent nephrotoxin. The presence of OTA in raisin is an important barrier to raisin export. The maximum tolerable level of OTA in raisin is 10 µg /kg. During 2010- 2012 a research was carried out on the monitoring of ochratoxin A and ochratoxin A - producing fungi in raisin of Khorasan, Azarbaijan, and Qazvin provinces. The samples were collected from different processing stages of raisin, storages and factories. To study fungal contamination two methods of plating surface disinfected raisin and washing of raisin surface (suspension) on artificial media were used. Aspergillus section Nigri species were isolated and the growth of Aspergillus section Nigri species which were assumed to produce ochratoxin A were recorded after incubation period. To confirm morphological identification of A. carbonarius isolates, DNA of three A. carbonarius isolates were extracted and their ITS regions of ribomal DNA and β- tubulin genes were amplified by PCR and sequenced. Ochratoxin A contaminations of the raisin samples were estimated using HPLC- IAC method. To study ochratoxin A producing ability of Aspergillus section Nigri species isolated from raisin, 14 isolates selected as representative were cultured on PDB medium and OTA produced by each isolate was evaluated using HPLC- IAC method. All raisin samples were contaminated by Aspergillus section Nigri species. The mean of the assumed ochratoxin A producing -fungi contamination of raisin samples was 56. 31 (8-100%) but varied depending on the processing method, raisin variety and location. The highest contamination was found in injured not directly edible raisin. Twenty percent of the isolates belonged to A. carbonarus species and the rest to A. nigra. Ochratoxin A was found in five samples out of 44 (11%) raisin samples at the contamination levels of 0. 4 to 100 ng/g. 71% of tested A. carbonarius isolates produced ochratoxin between 0. 78- 108. 8 ppb. DNA sequencing data were generated for a part of 18S-ITS1-5. 8S-ITS2 and 28S region of the rDNA, and β- tubulin gene region. These sequences were compared to other sequences of A. carbonarus in GenBank and showed 100 % and 99% similarity, respectively. These data confirmed the morphological examinations. Sequences were submitted to GenBank under the accession numbers KF434631, KF434632, and KF434633 for ITS, and KF434634، KF434635 for β- tubulin gene. Since the highest contamination was found in injured raisins, they can be easily removed to reduce contamination by fungi and ochratoxin.

One of the most important diseases on wheat is Fusarium head blight (FHB) which causes high losses on wheat crop quantitatively and qualitatively. The main species caused FHB are Fusarium graminearum، F. culmorum and F. avenaceum. Fusarium graminearum isolates are concerned as the major agents to produce deoxynivalenol، 3-acetyldeoxynivalenol، 15-acetyldeoxynivalenol، nivalenol، and zeralenone in wheat. Regarding to high DON contamination of 2011 wheat crop production at Ardabil province، population analysis of the Fusarium graminearum species complex isolated from wheat crop of this area was considered as the purpose of this study. During Jun till July of 2011 the FHB wheat contaminated samples were collected from wheat fields and harvested wheat crop through Ardabil area and Fusarium isolates obtained from wheat samples using suitable laboratory methods. 321 F. graminearum isolates were identified based on morphological characters. The identification of all 321 isolates was confirmed molecularly using Nicholson''s specific F. graminearum primers. To resolve the different members of the F. graminearum clade، the species specific primers which designed for partial sequences of the ammonia ligase 2 (CTPS2) gene were used. F. asiaticum، F. meridionale، and other members of the F. graminearum clade can be distinguished based on differences in the length of the CTPS2-derived amplicons by these primers. The primers for the ammonia ligase 2 gene produce fragments of 162 and 3l1bp in isolates of F. asiaticum، while the 311bp fragments are observed in all other members of the F. graminearum clade To find the phylogenic relation four isolates were selected as the representative isolates and their ammonia ligase 2 gene amplicons were sequenced. The results of PCR products showed that 89. 4% of F. graminearum isolates (287 out of 321isolates) were F. asiaticum due to producing both 162 bp and 311 bp bands while in 10. 6 % of isolates only one band (311 bp) was produced which indicated this minority of isolates were not F. asiaticum but they were in the clade of F. graminearum. The sequences of the ammonia ligase 2 gene amplicons of four representative isolates confirmed the identification of F. asiaticum as the dominant species of F. graminearum clade on FHB contaminated wheat in Ardebil province. The results showed that F. asiaticum was the predominant species causing Fusarium head blight (FHB) of wheat suggested resulting in DON production throughout all regions of Ardabil province.

Early splitting Pistachio nuts in is one of the critical sources of aflatoxin contamination before and during harvest. In this research work physical characteristics,fungal, pest and aflatoxin contaminations were evaluated in early splitting pistachios.

Physical characteristics, fungal contamination and aflatoxin content were assessed in pistachio nuts employing advanced methods. This study was carried-out using three varieties of commercial nuts consisted of Ohadi, Kalleghoochi and Ahmadaghaii.The data was analyzed in a randomized complete block design using Mstatc statistical analysis software.

The highest levels of Aspergillus infection was observed in early splitting pistachios with dry and semi-dry hull. These pistachios also had the highest levels of carob moth infestation and shell staining as well as low kernel weights. In early splitting pistachios with soft hull the fungal infection, carob moth and shell staining were 0.75-10.3%, 1.5- 9.5% and 21.5- 44.1% respectively. This indicates the critical effects of early splitting pistachios as well as the risk of aflatoxin formation. The results showed that the levels of Aspergillus infection are increased by the presence of carob moth and ants, while there was low kernel weight. Aflatoxin was not detected in pistachio with healthy hulls. On the other hand, shell and kernel of early splitting pistachios contain different levels of aflatoxins (P<0.05). Aspergillus flavus isolates was alike to produce 67% aflatoxins B1 and B2 and 24% aflatoxins B1 and 9% were nontoxigenic.

Sorting out early splitting pistachios based on physical and appearance characteristics would reduce the levels of aflatoxins during or after processing. Early peaking is a critical factor to reduce fungal and aflatoxin contaminations. About 95% and 5% of aflatoxin contents were related to kernel and shell, respectively. Therefore, assessing the risk of contamination should be based on kernels.

Aflatoxins are highly toxic secondary metabolites produced by various Aspergillus species in food and feed. Nontoxigenic strains of A. flavus have been used for reducing aflatoxin of several crops. To determine the frequency of atoxigenic A. flavus population، samples were collected from different pistachio producing areas. A serial dilution method and AFPA medium was used to isolate Aspergillus section Flavi isolates. The ability of the isolates to produce aflatoxins was assayed using fluorescence detection (FD) around fungal colonies under UV 366 nm and color changing of colony after ammonia vapor (AV) exposure in the primary screening. First screening stage was validated by thin layer chromatography (TLC) after rice flour inoculation. The species of A. flavus were identified morphologically and molecularly based on species-specific primers. Overall، 524 isolates out of 681 isolates were identified as A. flavus. 53A. flavus isolates from nut and 10 isolates from soil did not produce any aflatoxins based on the methods used. Comparison of cultural and analytical methods showed FD methods overestimated the frequency of nontoxigenic strains while AV in combination with coconut agar medium produced similar results compare to TLC assays. The AV method can be used in the most laboratories as a practical، cheap، and rapid tool. Further efforts for assessment of competition potential of nontoxigenic strains versus toxigenic strains will be useful.

Turfgrasses have been recognized for their importance to our quality of life for many years. Today, they are cultured as a main component of cool-season grasses in the landscape and sport fields in Iran. Turfgrasses are susceptible to Fusarium species. To study the role of Fusarium species as the causal agents of foliar blight, crown and root rot of turfgrass, samples were collected from infected turfgrass plants from different areas in landscapes of Tehran, Ardebil, Khorasan and Khoozestan provinces during 2009 - 2010. Based on morphological and physiological characteristics and the diagnostic keys a total of 101 isolates seven species of Fusarium recovered. These include: F. solani (33%), F. chlamydosporum (8%), F. proliferatum (2%), F. graminarum (13%), F. oxysporum (8%), F. subglutinans (9%) and F. compactum (28%). Fusarium isolates were found pathogenic using their conidial suspensions in the soil of the one month- old turfgrass seedlings used in Iran as Baronburg trade mark which is mixed of Lolium perenne, Festuca rubra and Poa pratensis seeds in the greenhouse conditions. Based on the more pathogenic effect, the order of Fusarium species were: F. solani, F. subglutinan, F. chlamydosporum, F. oxysporum, F. compactum, F. graminearum, F. poliferatum, respectively. Pathogenicity test for above Fusarium isolates against five major genera of turfgrasses including: Poa pratensis (Kentucky bluegrass), F. rubra (Fescue), Agrostis stolonifera (Bentgrass), Lolium perenne (Rye grass) and Cynodon dactylon (Bermudagrass)– were also performed under greenhouse conditions. The results showed that Rye grass, Fescue, Kentucky bluegrass Bermuda grass and Bentgrass were more resistant to isolates of tested Fusarium species, respectively Pathogenicity test for any of five turfgrasses against above Fusarium species was performed on one month- old seedlings under greenhouse conditions. The virulence of the fungal isolates was evaluated.

During the last four years (2009- 2012) several reports of forest tree decline were received، complaining about serious damage and death of many trees. The disease has been spread throughout forests of Zagross mountains which extend from north west to south west of Iran covering approximately 4،000،000 ha. Also the disease has been spread throughout forests of Alborz mountains in the north of Iran. The infected tree species included Q. brantii، Q. castaneifolia and Zelkova carpinifolia. Q. brantii Lindl. (Persian oak) has dominated in all parts of Zagros mountains from north to south، especially on the southern side of mountain in Ilam، Lorestan، Kohgilouyeh va Boyer-Ahmad، Fars and Kermanshah provinces. Zelkova carpinifolia (Pall.) Dippel، and Quercus castaneifolia C. A. Mey are native to the Caucasus and Alborz mountains in northern parts of Iran. The decline began with browning of the leaves، viscous liquid exudatation on the branches and trunks resulting in a brown-black discoloration of bark and woody tissues. In the winter of the next year fungal growth induces a typical charcoal-black surface on diseased branches and trunks. The perithecia of B. mediterranea were observed in a black carbonaceous layer on the stem surface erupting from the declined trees and the ascospores were visible under light microscope. Perithecia were obovoid، containing، amyloid asci، with dark brown ellipsoid ascospores، with straight germ slits along the spore-length. On the basis of these morphological characteristics، the fungus was identified as Biscogniauxia mediterranea. Pathogenicity tests were conducted using an isolate of B. mediterranea on six-month-old Q. brantii seedlings using a mycelial plug of B. mediterranea colonized potato-dextrose agar and the symptoms were observed after two months and the same fungus was re-isolated. Based on previous studies infections occur in healthy living trees as endophyte and then become invasive under water stress conditions in most reports. B. mediterranea has been reported to be aggressive on drought stressed hosts. During the last ten years climate changes has occurred in Zagros forests and resulting drought stress extending desert areas followed by occurrence of the fine dust phenomenon has reduced the photosynthesis of the forest trees and making them more vulnerable to the disease. In spite of tolerance of Persian oak species to the range of temperatures from -31°C until +45، the incidence of charcoal disease has been increased dramatically on Q. brantii forests. On the basis of our current knowledge، Q. brantii represents a new host of B. mediterranea and this is the first report of Persian oak charcoal disease outbreaks throughout Zagros mountain forests of Lorestan، Ilam، Fars، Kohgiloye va Boyer – Ahmad and on Zelkova carpinfolia in Daland forests of Gorgan area.

Crown rot or gummosis is one of the most important diseases of pistachio trees in Qazvin province. For determination of Phytophthora species, their distribution and rate of damage in Qazvin province (Boeen Zahra region), samples from Crown tissue and soil were collected. In laboratory, isolation of fungus was performed by selective media (CMAPV and PARPH). The isolates were purificated by culturing tip of hypha and important morphological charactristics were studied to identify species. Two fungi, P. citrophthora and P. nicotianae were recognized as causal agents of gumosis in Boeen Zahra region and proved their pathogenecity. With repeated surveys from pistachio orchards in spring, summer and fall, observation of disease symptons and isolation of fungus from soil or tissue of infected trees, distribution of disease was determined in different area of Hosein Abad, Valad Abad, Arasang, Khonan, Jahan Abad and Eshrat Abad in Boeen Zahra region. Based on infection percentage of orchards, rate of gummosis damage in Boeen Zahra region was estimated very low and approximately 2-3 percent.