S.leila Akbari kiarood; kamran Rahnama; morteza Golmohammadi; Saeed Nasrollanejad
Abstract
This research was conducted to identify endophytic bacteria over five-year old citrus trees in the east of Guilan province. A total of 63 endophytic bacteria were isolated from symptomless leaves of Citrus aurantium, Citrus sinensis, Citrus sinensis var. thomson, Citrus unshiu, Citrus limon, Fortunella ...
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This research was conducted to identify endophytic bacteria over five-year old citrus trees in the east of Guilan province. A total of 63 endophytic bacteria were isolated from symptomless leaves of Citrus aurantium, Citrus sinensis, Citrus sinensis var. thomson, Citrus unshiu, Citrus limon, Fortunella margarita and Citrus medica. The isolates were grouped based on colony type and color. Then the morphological and biochemical characteristics of the six isolates were assessed as candidate of different groups. According to the molecular studies and phylogenetic analyses of 16srRNA region these isolates were identified as Bacillus megaterium, Bacillus cereus, Arthrobacter agilis, Acinetobacter junii and Pseudomonas azotoformans species. . Among them, three species including A. agilis, A. junii and P. azotoformans are reported as citrus endophytic bacteria from Iran and the world for the first time. The morphological, biochemical, molecular and phylogenetical analyses are able to differentiate genus and species of the endophyte isolates.
Saadi Karami; Mohammad Javan Nikkhah; Khalil-Berdi Fotouhifar; Vahid Rahjoo; Abdollah Ahmadpour; Amin Alidadi
Abstract
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 ...
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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.
Ali Baradar; Ahmad Hosseini; somaye abdani babaki; Samin Alsadat Hosseini Farhangi
Abstract
Bean yellow mosaic virus, a species of the genus Potivirus, has a wide host range and a broad geographical distribution. BYMV causes high annual economic damage in various legumes such as faba beans in Iran. In this study, 13 BYMV isolates were collected from faba bean fields of different provinces of ...
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Bean yellow mosaic virus, a species of the genus Potivirus, has a wide host range and a broad geographical distribution. BYMV causes high annual economic damage in various legumes such as faba beans in Iran. In this study, 13 BYMV isolates were collected from faba bean fields of different provinces of Iran (Sistan and Baluchestan, Hormozgan, Kerman, Khuzestan, Fars, Lorestan, Ilam, Hamadan, Ghazvin, Zanjan, Ardabil, East Azerbaijan). The coat protein (CP) region of the collected isolates was sequenced and then compared with the CP sequence of 178 isolates available in GenBank. The selected Iranian sequences showed 86-99% nucleotide sequence identities with other BYMV isolates. Phylogenetic relationships based on CP nucleotide sequences were estimated using the Maximum Likelihood method, after removing all recombinant sequences. Accordingly, all isolates excluding three isolates, AI38, PAC-1, BYMV-W were placed in eight monophyletic groups. Iranian isolates were located in two distinct groups, along with broadbean, lentil, bean, gladiolus and sunflower isolates from Japan, Australia, Iraq and Spain. According to the results there is no significant relation among clustering of BYMV isolates based on phylogenetic analysis of CP sequences and original host and country. The CP structure analysis of Iranian isolates and other selected isolates from GenBank revealed conservation of the C-terminus and the central region of the coat protein and the variation of the N-terminus.
Mohammad Morad Moradi; Ahmad Hosseini; Samin Hosseini Farhangi
Abstract
Bean yellow mosaic virus (BYMV) is one of the most important members of the genus Potyvirus that is distributed worldwide. In the present study, the complete genome sequence and molecular characteristics of a BYMV sunflower isolate (BYSun) from Iran has been reported. The complete nucleotide sequence ...
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Bean yellow mosaic virus (BYMV) is one of the most important members of the genus Potyvirus that is distributed worldwide. In the present study, the complete genome sequence and molecular characteristics of a BYMV sunflower isolate (BYSun) from Iran has been reported. The complete nucleotide sequence of BYSun was determined using degenerate and specific primers and 5`-RACE. The viral genome comprises 9547 nucleotides, excluding a 3´-terminal poly (A) sequence. The genome of BYSun has a 206 nt 5´-non coding and a 171 nt 3´-non coding region. The RNA encodes a single polyprotein of 3056 amino acid residues and has a deduced genome organization typical for the members of the genus Potyvirus and nine cleavage sites of the polyprotein were predicted. The small overlapping ORF (PIPO) in the P3 gene was also deduced. When analyzed using the RDP4 program, at least one recombination breakpoint was identified in BYSun genome. BYSun along with 23 full-length sequences of BYMV isolates from Genbank were subjected to phylogenetic analysis that showed the highest nucleotide and amino acid sequence identitywith Australian isolate of BYMV-S isolated from Broad bean. Phylogenetic analysis of different BYMV isolates sequences revealed existence of six to eight phylogenetic groupings depending on parts of investigated genome. BYSun and BYMV-S grouped apart from other isolates and were placed within a distinct group, currently designated “S”. This is the first report of a full-length sequence of a BYMV isolate from Iran.
Vahid Khosravi; Mohammad Javan Nikkhah; Hossein Saremi; Shahram Naeimi; Rasoul Zare
Abstract
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 ...
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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.
Bahram Sharifnabi; Fariba Ghaderi; Nahid Javadi
Abstract
Almond (Prunus amygdalus L.) is a deciduous tree in the Rosaceae family and is grown mostly for its edible seed. It is one of the most popular nuts on a worldwide basis and widely grown in Iran in Isfahan and Chaharmahal and Bakhtiari provinces. Root and crown rot causal agents which are caused by several ...
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Almond (Prunus amygdalus L.) is a deciduous tree in the Rosaceae family and is grown mostly for its edible seed. It is one of the most popular nuts on a worldwide basis and widely grown in Iran in Isfahan and Chaharmahal and Bakhtiari provinces. Root and crown rot causal agents which are caused by several pathogens were studied. According to morphological characteristics, 53 isolates were identified as Phytopythium litorale (13 isolates), Phytophthora cactorum (19 isolates), and P. crytogea (21 isolates). Out of 53 isolates, 25 isolates were selected for molecular studies. Specific primer pairs of BPhycacL87FRG (F)/BPhycacR87RRG and Cryp1(F)/Cryp2 were used to confirm morphological identification of species. The reaction of crown and root of 8-month-old almondcultivars Sangi, Rabee, Shahrood12 and GF677 to virulent isolates of P. cactorum, P. cryptogea,and Pp. litorale was evaluated under greenhouse conditions. Comparison of pathogenicity means revealed that there are significant differences among P. cactorum, P. cryptogea,and Pp. litorale isolates.The results alsoshowedthat Pp. litorale was more aggressive than P. cactorum. On all tested almond cultivars, disease severity percentage showed that Sangi is the most susceptible cultivar and GF677 showed the least susceptibility. In this study, phylogenetic relationships of some Phytophthora and Phytopythium species based on sequencing of the rDNA-ITS region is provided. This is the first report of isolation and pathogenicity of Phytopythium litorale from almond orchards in Iran.
Parisa Sharifi Nezam Abad; Mina Koohi Habibi; Akbar Dizadji; Siamak Kalantari; Masoumeh Ranjbar Aghdam
Abstract
During 2008-2009, a total of 154 gladiolus (Gladiolus grandiflorus) corms were collected from the distribution centers in Karaj (Iran). After cultivation, fresh leaf samples of all germinated corms were tested by DAS-ELISA, which revealed the infection of 74.02% of plants with Bean yellow mosaic virus ...
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During 2008-2009, a total of 154 gladiolus (Gladiolus grandiflorus) corms were collected from the distribution centers in Karaj (Iran). After cultivation, fresh leaf samples of all germinated corms were tested by DAS-ELISA, which revealed the infection of 74.02% of plants with Bean yellow mosaic virus (BYMV). Following biological purification of an isolate, designated as BYMV-GPK, its host range was determined. The molecular mass of BYMV-GPK coat protein was estimated as 34 KDa by SDS-PAGE, which was confirmed by western blot analysis. DAS-ELISA and tissue print immunoassay (TPIA) techniques showed similar sensitivity in detection of BYMV in leaf tissue of infected gladiolus plants, but none of them was able to detect BYMV in corm tissue of the same infected plants. Two fragments of GPK genome, corresponding to HC-Pro and 3´ end regions with 600 and 1100 bp in length, respectively, were amplified by RT-PCR. Phylogenetic analysis based on the nucleotide and deduced amino acid sequence of HC-Pro and genomic 3´ end region showed that BYMV isolates clustered into separate groups according to their host origin and GPK isolate was closely related to a trifolium and other gladiolus isolates.
Bahar Karimian; Mohammad Javan Nikkhah; Doostmorad Zafari; Khalil Fotoohifar; Shahram Naiemi
Abstract
Strains of Trichoderma harzianum s. l. have a great importance because of affecting mankind by being used as biological control agents against plant pathogens. In the present study, we planned to characterize the phylogenetic structure of T. harzianum recovered from different geographical origins and ...
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Strains of Trichoderma harzianum s. l. have a great importance because of affecting mankind by being used as biological control agents against plant pathogens. In the present study, we planned to characterize the phylogenetic structure of T. harzianum recovered from different geographical origins and various habitats in some climates of Iran (NC, SCD, CSW). In order to screen 239 collected Trichoderma isolates, the strains were pre-screened by RAPD-PCR, and representatives were selected for sequencing of ITS1-5.8 s-ITS2 genomic region. Totally, 176 T. harzianum isolates were obtained. Fourteen phylotypes were found as a result of maximum parsimony analysis of 76 ITS1 and ITS2 sequences related to T. harzianum, including one single-member phylotype representing a novel ITS allele for T. harzianum, endemic to Iran. Finding a unique ITS allele of T. harzainum in climate CSW may be an indicative of the effective role of this kind of climatic conditions on arising this new ITS allele.
