During 2004–2007 various own rooted vineyards were inspected to study the fungi associated with vine trunk diseases in Iran. Samples from declining vines showing yellowing and reduced growth and different symptoms in wood, including browning of the wood, necrosis, brown and black streaking and white rot were collected. Fungal isolations were made from affected tissues onto Malt Extract Agar (MEA) supplemented with 1 g l–1 streptomycin sulphate (MEAS). Based on morphological and molecular characteristics, the following species were identified: Phaeoacremonium (Pm.) aleophilum, Phaeomoniella (Pa.) chlamydospora and less frequently Pm. parasiticum, Pm. inflatipes, Pm. cinereum, Cylindrocarpon liriodendri, Diplodia seriata and Neofusicoccum parvum. Results of the pathogenicity tests under field conditions showed that Pa. chlamydospora and Phaeoacremonium spp. caused large wood discoloration 10 months after inoculation without any external foliar symptoms. Phaeomoniella chlamydospora caused larger lesions than Phaeoacremonium spp. All inoculated species were re-isolated from the margin of the lesions completing Koch’s postulates. This study represents the first comprehensive work that investigates the molecular and morphological identification and pathogenicity of Phaeoacremonium spp. and Pa. chlamydospora associated with vine decline in Iran. This is also the first report of Pm. inflatipes, N. parvum and D. seriata associated with grapevine decline in this country.

The genus Phaeoacremonium is associated with decline disease of woody plants and with human infections. Members of this genus have broad host range and wide geographical distribution. During 2010, ten isolates of Phaeoacremonium were recovered from vascular tissues of persimmon (Diospyros kaki) trees, showing decline symptoms in Shiraz city. Anamorphic characteristics such as, conidiophore morphology, phialide type and shape, size of hyphal warts, conidial size and shape were investigated. Based on morphological characteristics the presence of Pm. rubrigenum in Iran was documented. To confirm morphological identification, DNA was extracted from isolates using a genomic DNA purification Kit. Region of internal transcribed spacers 1, 2 and 5.8S genes of rDNA were amplified using ITS4 and ITS1 universal primer set. Fragments of 630 bp were recovered from PCR, purified, sequenced, edited and deposited in GenBank. Pm. rubrigenum isolates had an average of 99 % identity with all P. rubrigenum sequences compared. This species is a new report from Iran.

To study of fungi and symptoms associated with vine trunk diseases, various vineyards in Kerman province were inspected during spring and summer of 2012. Samples showing decline symptoms were collected from trunk and branches of vines. Fungal isolation from affected tissues were done on malt extract agar medium (MEA) supplemented with 1 g l 1 streptomycin sulphate (MEAS). A total of 476 fungal isolates from diseased grapevines were obtained. Fungal isolates were identified based on their morphological and molecular characteristics. Morphological identifications of Phaeoacremonium spp. were confirmed by sequence analysis of partial β-tubulin gene (BT) sequences amplification using primers T1 and Bt2b. Botryosphaeria isolates were also confirmed by amplification and sequence of the internal transcribed spacer (ITS) region using the primers ITS4 and ITS5. The most common fungi isolated from vineyards were Phaeoacremonium aleophilum and Phaeomoniella chlamydospora with 103 and 30 isolates, respectively. This is the first report of molecular identification of Pm. aleophilum, Pa. chlamydospora, Pm. parasiticum and Botryosphaeria dothidea associated with different internal symptoms of vine decline in Kerman province.

An extensive survey on phylogeny and pathology of fungi associated with the oak decline in Zagros forests located in Western Azarbaijan, Kurdistan, Kermanshah, Lorestan, and Ilam provinces, a large number of collected fungal isolates seventeen isolates morphologically resembled the members of Phaeoacremonium. Similar DNA fingerprinting patterns were generated for all isolates using M13 primer. Thus, one representative isolate (IRAN 4348C) was analyzed based on tub2 sequence data using maximum parsimony and neighbor-joining algorithms and identified as Phaeoacremonium tuscanicum. Pathogenicity was confirmed following Koch’s postulates on two-year-old Quercus brantii seedlings under greenhouse conditions. To our knowledge, it is the first time Ph. tuscanicum is reported from oak trees and confirmed as a pathogenic fungal species on Q. brantii in the world.

Petri disease is responsible for grapevine decline and occurs wherever grapevines (Vitis vinifera) are cultivated. Phaeoacremonium species are among the principal hyphomycetes associated with Petri disease. During 2009, a field survey was conducted throughout different vineyards in the Fars province of Iran in order to determine the fungal pathogens associated with the vine decline observed in the region. Samples were taken from grapevines showing yellowing, interveinal chlorosis, leaf necrosis, reduced growth, wilting, wood necrosis and streaking, and xylem discoloration symptoms in cross section. Isolations were made from symptomatic wood tissues from cordons and trunks on malt extract agar supplemented with 1 mg ml-1 streptomycin sulphate (MEAS) and potato dextrose agar (PDA) media. Based on morphological and molecular characteristics two species of Phaeoacremonium, Phaeocremonium mortoniae and Pm. inflatipes, were isolated and identified from grapevines showing yellowing, slow dieback, stunted growth, and reduced foliage in Bavanat (Fars province, south-western Iran). Pathogenicity tests were conducted on rooted grapevine cuttings (cv. Askari) under greenhouse conditions. Based on the results of pathogenicity tests, both tested Phaeoacremonium species were pathogenic and caused significant vascular discoloration in inoculated cuttings four months after inoculation. The fungi were reisolated from the margins of the lesion and healthy tissue, completing Koch’s postulates. Based on our knowledge, this is the first report of Pm. mortoniae and Pm. inflatipes causing grapevine Petri disease in Iran.

One of the most important and elusive diseases of grapevine is Esca disease. Esca is a complex disease that comprises a combination of symptoms caused by various fungi including growth retardation, decrease in productivity and decline of the plant and usually affecting adult plants (8 to 10 years and older). The purpose of this research was to study the genetic variations of fungal agents associated with Esca disease in Khorasan province which was performed by using the molecular marker PAPD. Sampling carried out done from the grapevines of Bojnourd (North Khorasan) and Phaeoacremonium parasiticum, Phaeacremonium aleophilum, Phaeomoniella chlamydospora were isolated from wood brown decay while Fomitiporia mediterranea was isolated from wood white decay. The isolates with the most variability in morphology were selected and DNA polymorphism of the isolates was studied by RAPD-PCR. For this purpose after extracting DNA, RAPD _PCR was performed in 25 microliter reactions with 10 random primers. The results of RAPD analysis demonstrated that remarkable genetic variations existed in F. mediterranea while rather uniformity in Pa. chlamydosporare.. The results showed that RAPD-PCR technique have a great potential in determining genetic variations of the fungi associated with Esca disease. This is the first study of genetic variability in Iranian isolates of fungal agent of Esca using RAPD molecular markers in the populations of this species.

Togninia minima(anamorph: Phaeoacremonium aleophilum) is the most common species of Phaeoacremonium associated with Petriand esca di­sease in grapevines. A bipolar heterothallic mating strategy has been proposed for this species, which requires two isolates with opposite mating types for sexual reproduction. In the present study, the possibility for the induction of sexual stage among the Iranian isolates of Pm. aleophilum was investigated under laboratory conditions. For this purpose, different vineyards in East Azarbaijan province were inspected during the summer 2012 and samples were collected from grapevines with typical esca disease symptoms. Isolation and purification of the causal agent was carried out using routine plant pathology techniques. Pm. aleophilum isolates were identified based on morphological characteristics of the microscopic structures and previously designed Pm. aleophilum-specific primer set. In order to induce sexual stage, four isolates with different geographical origins were mated in all possible combinations on grapevine cane-water agar (GWA) plates. Perithecial formation was observed on wood segments and agar after 3-4 weeks of mating. The morphological characteristics of the sexual structures were in full agreement with those of T. minima. Two isolates with opposite mating type identity were selected as tester strains and mating identity of 34 T. minima isolates were determined by pairing with tester strains. The results showed that both mating types were present among the isolates examined in this study; however, skewed distribution of mating type alleles (2:1) was found among the tested isolates.

One of the most significant and destructive grapevine diseases is Esca which is caused by several fungal species. Trunk disease causes stunted growth، reduced yield، and ultimately، vine death. For this reason، during the summer 2011، infected samples were collected from Bojnourd (North Khorasan province) vineyards. Infected trunk samples were acquired that they were showing white and brown rots، dark brown spots and also samples without apparent symptoms. The aims of this work were، firstly، to apply species –specific primers for detecting fungal agents associated with Esca disease of grapevine; and comparing this method to the classical ones، and secondly، to detect frequency of each pathogenic agent using molecular and classical methods. In molecular method، species-specific primers which designed based on ITs region of rDNA were used to detect fungal agents of the Esca disease. They yielded a single amplicon of 550 bp، 360bp and 415 bp for F. mediterranea، Pa. chlamydospora and Phaeoacremonium spp. respectively. Results showed that the maximum frequency of incidence was related to P. chamydospora and Phaeoacremonium spp was in second. The lowest incidence frequency rate was related to F. mediterranea which was observed only in sample with white rot symptoms. Primers defined here can be used in the nursery sanitation program to produce plant free of Esca disease.

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.