b. sharifnabi

Apple (Malus domestica) is considered one of the most important economic products in Iran. Its cultivation is common in various regions and so far, more than 200 different domestic and foreign apple cultivars are identified, which Golden and Red Delicious cultivars are the most important cultivars in terms of cultivated area. Various fungal, bacterial, nematode, and viral agents cause weakness and decline of the apple tree and the quantitative and qualitative reduction of the crop. Apple root and crown rot is one of the important diseases of apple root, due to soil-born fungal pathogens, which lead to the decline of apple trees. The disease has a global spread and can occur at all ages of apple production. Different fungal species have been identified in infected regions so far, and it may be due to climate change. Sampling was performed from June to August 2019 from apple orchards with decline symptoms in Semirom and Padena in Isfahan province, Iran. Fungal species, including Fusarium solani, F. oxysporum, F. equiseti, F. acuminatum, and F. redolens, were morphologically identified. To confirm morphological identification, EF1/EF2 and ITS1/ITS4 primers were used in the PCR reaction. In vitro and greenhouse tests confirmed the pathogenicity of the identified species. As a result, it was proved that F. solani, F. oxysporum, F. equiseti, F. acuminatum, and F. redolens could be the causal agents of apple trees decline in Isfahan province, Iran, that F. solani, and F. oxysporum were more abundant.

Large flowered pelargonium (Pelargonium grandiflorum) is a perspective flowering crop which is distributed around the world. The area of its application is quite wide: from room floriculture to design the gardens and parks. Observation of leaf spot symptoms on this plant, which was collected from Alborz province (Karaj) motivated us to find the causal agent(s) of the disease. So, the symptomatic parts were cultured on the PDA medium after surface sterilization. Two fungal colonies were appeared on the culture medium. They were identified as Alternaria alternata and Botrytis cinerea according to the morphological characterizations. Molecular study using the gapdh for Alternaria and ITS regions and rpb2 gene for Botrytis confirmed the result of the morphological identification. In the pathogenicity tests, the same spots on inoculated plants with Alternaria and the same spots plus gray mold symptoms and fungal body on leaves, buds and stems of the inoculated plants with Botrytis were another confirmation. Based on our knowledge, this is the first report of these two fungal species on the Pelargonium grandiflorum in Iran.

Bean anthracnose disease caused by Colletotrichum lindemuthianum is one of the most important bean diseases in Iran. In this study, the presence of disease resistance genes was investigated using SCAR primers in 12 Iranian bean cultivars. Also, disease symptoms were evaluated based on the disease severity index in the greenhouse condition. Amplification of SCAR markers SAS13, SY20, SZ04 and SB12 indicated the presence of Co-42, Co-4, Co-6 and Co-9 resistant genes respectively. Cultivars Akhtar, Derakhshan, Almas, Talash and Koosha contained Co-42, Co-4 and Co-6 resistance gene revealed by amplification of SBB14, SC08 and SZ20 markers accordingly. Co-42 gene was detected in Goli and Naz cultivars and Co-4 and Co-42 genes were present in Pak, Sadri and Khomein cultivars. Co-42 gene that is linked to SH18 marker was observed in Dorsa and Shokofa cultivars. The presence of Co-2 resistance gene was observed only in Almas cultivar. Co-5 gene linked to SAB3 marker was only observed in the related differential cultivars. Phenotypic evaluation for disease symptoms showed that out of 12 cultivars studied, four (Naz, Almas, Dorsa and Shokofa) showed resistance reaction and two (Akhtar and Goli) were moderately resistant. Based on the disease severity index, these cultivars were considered relatively resistant. Simultaneous analysis of molecular and phenotypic data showed that only the two markers of SQ4 and SH18 succeeded in band amplification in resistant cultivars Diamond, Shokofa and Dorsa. The results of this study will be of help in producing resistant bean cultivars to anthracnose disease by resistance gene pyramiding.

Botrytis cinerea has a wide host range, the possibility of hiding plant infection from the initial stages, and causing severe damage (even up to 100%) to the host.  Due to the chance of growth and activity of the pathogen in field, greenhouse, in laboratory, and even in cold storage conditions, its importance is increasing. The presence of pathogen in seedlings, mature plants, ripe fruits, and even storage conditions in different parts of Iran and its importance, we decided to investigate the genetic diversity of the isolates using the ISSR marker. For this purpose, 21 isolates including 16 isolates from the collection of the University of Tehran (isolated from seedling and leaf of cauliflower, grizzly lettuce, needled lettuce, purple basil, onion, tomato, strawberry and pomegranate) and five isolates collected from two strawberry greenhouses in Alborz province, were used. Ten ISSR primers were used to determine the genetic diversity. Analyzing the results by NTSYS software version 2.02e (based on SM and UPGMA clustering method) showed that the isolates fall into four fingerprinting groups. It was also observed that the resulting bands have 100% polymorphism (93.33% in one case). The calculated cophenetic coefficient for the data (0.89) confirmed the accuracy of the obtained dendrogram. Evidence suggests that there is high genetic diversity in B. cinerea isolates and also there is no relationship between host and geographical region.

Pyrenophora lolii causes leaf spot on grasses including Festuca spp., Lolium spp., Dactylis spp., Avena sativa and wheat (Triticum aestivum). Infected oat leaves (Avena sativa) showing leaf spot symptoms were collected from the margin of barley fields in Golestan province of Iran during the spring of 2016. A morphological examination of the Pyrenophora specimen was carried out using light microscopy. Inoculation of oat leaves with the isolates of Pyrenophora lolii in greenhouse induced leaf spot on leaves. In order to confirm the morphological identification, sequences of glyceradehyde-3-phosphate dehydrogenase (gpd) gene and Internal transcribed spacer (ITS) regions were amplified using gpd1/2 and ITS1/4 primers, respectively. The phylogenetic analysis based on these sequences showed that the isolated Pyrenophora specimen clustered together with sequences of P. lolii. Based on result of morphological examination and phylogenetic analysis, it was concluded that the causal agent of leaf spot of A. sativa (oat) was P. lolii.

Root knot nematodes (Meloidogyne spp.) cause yield loss in all countries, of which, M. javanica, is the most widespread species in Iran. In order to identify M. javanica, 100 infected root and soil samples of root knot nematode were collected from different regions of Kerman province. After purification of populations and identification of M. javanica based on morphological and morphometerical characters of females and second stage juveniles (J2), total DNA was extracted from eggs, J2 and female adults. Specific 670 and 1600 bp bands were amplified in all M. javanica populations using species-specific primer pairs including OPARjav / OPAFjav and Mjavf / Mjavr These specific bands could not be amplified in other species such as M. incognita and M. arenaria. It seems that, application of these species specific primers in comparison with morphological characters would be more applicable, leading to easier identification of M. javanica.

Recent studies have shown the controlling effects of Trichoderma species and extract of a few plant species of the Brassicaceae family on some phytopathogenic fungi. In this study, using a completely randomized blocks design in the field, controlling effect of biological agents comprising mustard flour, Trichoderma koningii T18, T. virens T59, T. brevicompactum T30, T. harzianum T56, mixture of four Trichoderma isolates, mustard flour + mixture of four Trichoderma isolates and two commercial biological product of Trichodermin B and Subtilin were evaluated against wheat common bunt caused by Tilletia laevis. According to the infection index, all treatments were able to reduce infection percentage and showed significant differences (P<0.01) compared to control (infected with T. laevis without any biological agent) in which heads infection was 43.5%. Treatments of Mustard flour and mustard flour + mixture of four isolates, reduced the disease by 89.9% and 87.4% respectively. Consequently, it seems that wheat common bunt could be controlled by application of non-infected seeds and by treating seeds using mustard flour without using chemical fungicides.

Safflower (Carthamus tinctorius L.) is one of the multi-purpose oilseed crops which has a high adaptation to different conditions such as resistance to drought and it is suited to be grown in arid and semi-arid regions such as Isfahan province. Root rot disease is an important soil-borne disease of safflower in Isfahan, which can be caused by different pathogens. The objective of this study was to determine the causal agent of safflower root rot and to evaluate different genotypes for tolerance to the disease. Different species of Fusarium were isolated from sample collections. Laboratory and greenhouse inoculations indicated that F. solani was the only pathogenic species. In this experiment, 60 genotypes of safflower including breeding lines selected from various Iranian local populations and foreign cultivars were evaluated for reaction to the disease in a randomized complete block design with three replications in greenhouse. Artificial inoculation via injection of spore suspension of F. solani (106 spores/ml) was conducted on 8-week plants and then development of necrosis and death percentage were recorded. The results showed that there were significant differences among the genotypes in terms of reaction to the disease. The most resistant and susceptible genotypes were breeding lines of IUTE14310 and IUTC121 with mean necrosis of 9.67 and 28.33 mm, and death percentage of 32 and 74, respectively. Based on the means of necrosis and death percentage, the genotypes were significantly classified in 5 distinct groups including resistant (7 genotypes), moderately resistant (19 genotypes), tolerant (29 genotypes), moderately susceptible (3 genotypes), and susceptible (2 genotypes). The commercial foreign cultivars of AC Sunset, AC Sterling belonged to tolerant and moderately susceptible groups, respectively. However, Saffire was classified as a tolerant genotype. The local landrace of Kooseh which is widely grown in Isfahan province was classified as susceptible genotype. Phenotypic and genetic coefficients of variation (23.85 and 18.32 %, respectively) and a relatively high broad-sense heritability (59%) for necrosis and also the phenotypic and genetic coefficients of variation (25 and 21 %, respectively) and a high broad-sense heritability (73%) for death plants indicated that there was sufficient genetic variation for resistance and selection can be effective for producing resistant genotypes to Fusarium root rot disease.

Mutual relationship between endophytic fungi and some gramineuos plants is one of the recently known host-microbe interactions. Endophytic fungi are classified in class Ascomycetes, order Hypocreales, tribe Balansieae and mostly genus Neotyphodium. They are transferred into the progeny via maternal stock and have many benificial effects to their host plants, such as resistance to pests and diseases, environmental stresses such as pH fluctuation, drought, salinity and grazing. because Iran has a wide range of plant genomic resources and is known as one of the origins of gramineous plants, it is necessary to know more about endophytic fungi and their presence by using classical and molecular methods. Twenty isolates of endophytic fungi were obtained from seed and leaf sheaths of Bromus tomentellus, Lolium prenne, Festuca arundinacea and Festuca pratensis. Mycelia of endophytic fungi were observed red or pink in seed and leaf sheath when stained by Rose Bengal. The DNA of endophytic fungi was extracted by Murry and Thompson method with minor modification. In order to confirm the identity of endophytes, three universal and specific primer e. g. ITS1, ITS4, IS1, IS3, and 11-1, 11-2 were used. All isolates showed a 550-750 bp band with ITS1, ITS4 and eighteen of them showed a 444 bp band with IS1, IS3 primers, which is an indicator of Neotyphodium endophytes. when primers 11-1, 11-2 were used, a 1000 bp band was not observed in all isolates. Isolates FaSm and FpGon3 that did not have shown a 444 bp band, may probably differ from Neotyphodium endophytic fungi.

Mutual relationship between endophytic fungi and some gramineuos plants is one of the recently known host-microbe interactions. Endophytic fungi are classified in class Ascomycetes, order Hypocreales, tribe Balansieae and mostly genus Neotyphodium. They are transferred into the progeny via maternal stock and have many benificial effects to their host plants, such as resistance to pests and diseases, environmental stresses such as pH fluctuation, drought, salinity and grazing. because Iran has a wide range of plant genomic resources and is known as one of the origins of gramineous plants, it is necessary to know more about endophytic fungi and their presence by using classical and molecular methods. Twenty isolates of endophytic fungi were obtained from seed and leaf sheaths of Bromus tomentellus, Lolium prenne, Festuca arundinacea and Festuca pratensis. Mycelia of endophytic fungi were observed red or pink in seed and leaf sheath when stained by Rose Bengal. The DNA of endophytic fungi was extracted by Murry and Thompson method with minor modification. In order to confirm the identity of endophytes, three universal and specific primer e. g. ITS1, ITS4, IS1, IS3, and 11-1, 11-2 were used. All isolates showed a 550-750 bp band with ITS1, ITS4 and eighteen of them showed a 444 bp band with IS1, IS3 primers, which is an indicator of Neotyphodium endophytes. when primers 11-1, 11-2 were used, a 1000 bp band was not observed in all isolates. Isolates FaSm and FpGon3 that did not have shown a 444 bp band, may probably differ from Neotyphodium endophytic fungi.