m. a. tajick ghanbary

Endophytic fungi are a group of host-associated fungal communities that benefit their hosts. According to the conditions of their specific living environment, plant endophytic fungi produce many bioactive metabolites with different structural features. The bioactive compounds isolated from endophytic fungi, have significant effects on increasing the compatibility of both endophytic fungi and their host plants, such as the tolerance to biotic and abiotic stresses. In addition, some of these metabolites have indicated medicinal and ecological importance. In the present investigation, five fungal endophytes were isolated from the seeds of some medicinal plants. These endophytic isolates were characterized by sequencing of Internal Transcribed Spacer (ITS) regions as Acremonium sp. isolated from Echium amoenum, Epicoccum nigrum from Rosa canina, Fusarium sp. and Fusarium equiseti from Calendula officinalis and Lecanicillium aphanocladii from Physalis peruviana. To screen the phytochemical derivatives of ethyl acetate of these endophytic fungal isolates, the extracts were subjected to phytochemical analysis by gas chromatography-mass spectrometry (GC-MS). GC-MS spectrum of the compounds found in the extracts of the endophytic fungi was matched with the standard compounds present in the WILEY8 library and the National Institute of Standards and Technology (NIST14) library. The GC-MS analysis of extracts from these endophytic fungi revealed the presence of 49 phytocompounds such as 2,4-Di-tert-butylphenol, Hexadecane, Eicosane, Octadecane, Docosane, Nonadecane, Isopropyl myristate, Hexadecanoic acid, Undecane, Methyl stearate and so on. The results of the present study acknowledge that the endophytic fungi of these medicinal plants are the potential source of biologically active compounds and envisage the possible drug discovery using them. In addition, the compounds 14-Beta-H-Pregna and Cyclohexane,1,1'-(2-methyl-1,3-propanediyl) bis are reported here for the first time as fungal metabolites.

Fusarium wilt, root and crown rot caused by Fusarium oxysporum f. sp. ciceris, (FOC) is the highly significant soil-borne disease of chickpea in the Kurdistan province of Iran. The distribution of pathogenic races of FOC in Kurdistan province was determined during this research. Infected plant samples were collected from 42 fields in the chickpea production area of the Kurdistan province. The causative microorganism of the disease was isolated and purified from each sample, and then FOC isolates were identified by morphological characters. After the pathogenicity test and evaluation of pathogenic variability on the susceptible cultivar Kaka, the DNA extraction, the molecular identification of species, and races of pathogenic isolates were performed using FOC-specific linked primers. Among the collected isolates, 37 were identified as Fusarium oxysporum f. sp. ciceris. Molecular identification of races using SCAR-linked markers (1B/C, 0, 2,3,4,5, 6, and 1A) revealed that 28 out of 37 isolates belonged to race 0, and other isolates belonged to race 1B/C. There was no relationship between the prevalence of races and their geographical distribution. Identification of the races is crucial for the evaluation of resistance and the development of new commercial cultivars. The application of resistant cultivars is a fundamental approach for the integrated management of the Fusarium wilt, root, and crown rot for durable chickpea production

Black dot disease triggered by Dilophospora alopecuri causes considerable damage in some fields in Golestan province, North of Iran. D. alopecuri is its causal agent and carried in to wheat by wheat gall nematode. Extracellular appendages on the conidia adhere to the cuticule of the nematode juveniles. The disease was reported in united state of America, Canada, Germany, Ugoslavia, India and Pakistan.The fungi is classDothideomycetes and order Dothideales. The geographical distribution of disease was studied in South-Eastern Australia in the summer 1995.The disease in Iran has been reported by Bamdadian since 1973 from Baluchistan, Isfahan, Golestan, Khorasan, Kerman and Khuzestan. The disease agent causes considerable damage in some fields in Golestan province, North of Iran. Symptoms have been found during a survey of foliar disease of wheat in Golestan province. The disease incidence was very low but fungus interaction with seed gall nematode causes considerable damage. The symptoms start with yellow spindle-shaped flecks which develop and become tan brown with black border. This spots may occur on peduncle and heads. The pathogen survives as mycelium in host debris or as conidia on seeds. The purpose of this study was determining the geographical distribution, disease incidence (DI) and disease severity (DS). Then molecular identification of isolates with ITS rDNA in North of Iran, Golestan province.

 Wheat fields infected with the disease in the cities of Golestan province were visited and subjected to sampling during spring 2017-2018. 42 samples suspected to infection with the twist disease were gathered from seven cities of Golestan province. The rate of disease distribution from seven cities farms with disease symptoms, three farms were selected in each crop year and their geographical coordinates were recorded. In this research mapping the geographical distribution of disease was prepared by ArcGIS10.2 software. During 2018 and 2019, a survey was conducted to characterize the disease incidence (DI) and disease severity (DS). The D. alpoecuri leaf spot reading scales rate carried out in this study. No visible symptoms are observed and the leaf remain a health 0; A few chlorotic lesions are present and the infection site is a tan-spot colored 1: Necrosis and chlorosis both exist on the leaf 2: A few pycnidia are visible on the infected site and less than 30% of the leaf is occupied by pycnidia 3: The entire leaf is covered by pycnidial lesions scored 4. A significant difference was observed between the disease rates in the two cropping years. Seven D. alopecuri isolates were identified on the based on morphological and molecular parameters. After purification of fungal isolates, 50 conidia and pycnidia were selected in each isolate and the length and width of pycnidia, conidia cell number, conidia size (length and width), colony shape on the culture medium were also measured in the laboratory. After that identifying of the fungus isolates pathogenicity test was performed in the greenhouse. The D. alopecuri spore suspension with 1.75× 106 CFU concentration and for nematode population with 25000 larvae (L2) were used for inoculation to plants. After three months symptoms of leaf spot (twist) disease was appeared in tillering stage. Fungal DNA extraction from mycelium mass of selected isolates was performed by Murray and Thompson (1980) method and part of ITS region was amplified using ITS4 and ITS5 primers by polymerase chain reaction (PCR). The amplified PCR products of fragments were purified and sequenced at sequencing Microsense Company in Switzerland.

  Comparing of harvested fungal isolates which have been grown on PDA medium showed some variation in different characteristics such as the number of conidia walls, conidia appendages and pycnidia size. In both subsequent years, maximum disease incidence (DI) and disease severity (DS) was measured in Kalaleh by 3.6% to 3.9%, while for Bandar-Torkaman, the city with the lowest DS, were1.3% to 1.65%. The average size of conidia length was 4-8×1 μm in diameters.The pathogenicity test showed that the D.alopecuri is capable to produce twist  disease symptoms only  in the presence with seed gall nematode in the host. The nucleic acid sequences of Internal Spacer Transcribed (ITS) regions for Golestan isolates showed 100% similarity and had small genetic similarity with D. alopecuri MH859142.1 deposited in NCBI Genbank. The sequences belong to D.alopecuri from Gonbad, Aqala, Azadshahr, Maravehtapeh, Kalaleh, Minodasht cities were registered in NCBI Genbank with MW302360, MW291507, MW291561, MW303438, MW303517, MW303518 accession number, respectively.

This is the first study carried out on the morphological and molecular characteristics of twist disease agent isolates, disease severity and its distribution in Golestan province as a major wheat production area of Iran. Many of results especially molecular data and submitted sequences form Iranian isolates of D.alopecuri to databanks are new for Iran.

Grapevine trunk diseases (GTDs)are known as the most important factors in crop reduction and cause considerable economic problems in grapevines worldwide. During 2016 to 2018, several field surveys were conducted on numerous vineyards in Khorasan-Razavi province to study fungal species associated with grapevine trunk diseases. In this study, samples were collected from trunk and branches of trees showing yellowing, stunted growth, dieback and wood discoloration in cross sections. In this study, 258 fungal isolates were obtained and identified based on morphological characteristics and comparison of DNA sequence data (ITS-rDNA region and a part of β-tubulin gene). These isolates were identified as Phaeoacremonium minimum (75 isolates), P. parasiticum (19 isolates), P. iranianum (52 isolates), P. tuscanum (8 isolates), Fomitiporia mediterranea (56 isolates) and Seimatosporium vitis (48 isolates). Pathogenicity of the selected isolates was verified by inoculation of potted grapevines shoots under greenhouse conditions. Based on the mean length of wood discoloration in the wood, P. minimum and F. mediterranea were the most and least virulent species, respectively. Our findings indicated that known fungal trunk pathogens such as Phaeoacremonium species and F. mediterranea occur on grapevine in Khorasan-Razavi province. This study is the first report of S. vitis associated with grapevine decline in Iran.

The role of extracellular enzymes in mycoparasitism of Trichoderma spp. have been demonstrated in several cases. Trichoderma spp . produces several chitinolytic enzymes and this fungus is known as a powerful antagonist against Macrophomiona phaseolina, the causal agent of charcoal rot disease of soybean. In this study, two–dimensional protein pattern analysis and chitinaseactivity evaluation were performed for an Iranian Trichoderma koningii strain (NAS–K1) and its generated mutants were measured to indicate the potential role of endochitinases (N–acetylglucosam–inidase (NAG–I and NAG–II)) in its mycoparasitism. The results of chitinase activity assay using chitin and cell walls of M. phaseolina as substrates showed that the mutant isolates have significantly more enzyme activity compared to the wild type strain. The specific endochitinase enzyme activity in the mutant NAS–K1M25 was increased to approximately 2.5 times more and secreted three times more endochitinase than that of the wild type strain. This superior mutant showed up to 65% growth inhabitation against M. phaseolina in dual culture test (five times more than the wild type strain). Moreover, this strain showed sharper spots belong to endochitinase, and N–acetylglucosaminidase (I & II) presented in SDS–PAGE and 2D electrophoresis. Overall, induced mutation by gamma irradiation could be a useful method to develop such superior mutants, and the mutant NAS–K1M25 could be used as a potential biological control agent candidate for plant disease management programs of M. phaseolina. However, more detailed fermentation, formulation and field trial studies should be performed to finalize its biocontrol potentials.

To investigate the effect of Trichoderma harzianum, as a bioabsorbant to ameliorate the harmful effects of cadmium (Cd) on growth and yield of barley (Hordeum vulgare L. variety ‘Sahra’), a factorial pot experiment based on completely randomized design with three replicates was conducted. Trichoderma harzianum withtwo levels (with and without inoculation) and cadmium nitrate with four levels (0, 50, 100 and 150 mg.L-1) were the treatments. Results of ANOVA revealed that there was a significant interaction between Trichoderma and cadmium nitrate in terms of biological yield, straw yield, harvest index, spike number per plant and seed number per spike. Mean comparisons showed that Trichoderma inoculation at all Cd levels significantly improved both biological and straw yields. Trichoderma at 50 and 100 mg.L-1 of Cd also increased the spike number per plant (up to 120 and 66%, respectively) significantly. Increasing Cd levels decreased seed yield (19%), 1000 seed weight (18%), partitioning coefficient (57%) and tolerance index (23%) significantly. Inoculation of Trichoderma into growth medium had a significant effect on seed yield and tolerance index (up to 17 and 22%, respectively). In conclusion, Trichoderma harzianum inoculation at lower concentrations of Cd (50 and 100 mg.L-1) could be effective to improve growth parameters of barley plant.