Plant Disease and Biotechnology
Fatemeh Mohammadi; Aboozar Soorni; Rahim Mehrabi
Abstract
Two-spotted spider mite (Tetranychus urticae Koch) is one of the most important pests of beans (Phaseolus vulgaris L.). Since, complex gene networks are involved in creating sensitivity or resistance against the two-spotted spider mite; therefore, in this research we used biological system methods to ...
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Two-spotted spider mite (Tetranychus urticae Koch) is one of the most important pests of beans (Phaseolus vulgaris L.). Since, complex gene networks are involved in creating sensitivity or resistance against the two-spotted spider mite; therefore, in this research we used biological system methods to identify key networks. For this purpose, we used the RNA-Seq data related to the two-spotted spider mite stress on common bean plant. After providing the gene expression matrix, molecular networks were analyzed using weighted co-expression network analysis (WGCNA). After the modules identification, the gene functions in each module were investigated and analyzed. According to the results, a total of 699 genes were identified with differential expression in response to two-spotted spider mite stress, which were placed in 7 co-expression modules through hierarchical clustering. Gene ontology and interaction analysis of key genes using the String database showed that the response of common bean transcriptome to two-spotted spider mite infestation includes genes encoding protein kinases, catalysts, transcription factors, and metabolite production and pathways of hormonal message transmission. It is notable that among the most important genes that showed co-expression, WRKY and lipoxygenase were highlighted. The turquoise module had the higher number of genes involved in resistance, and this module and the yellow module had the highest correlation with the resistant variety after five and one day of contamination, respectively. Also, the black module had the highest correlation with the sensitive variety after five days of contamination. In conclusion, this study increases our knowledge of the molecular mechanisms involved in resistance to the two-spotted spider mite. Also, the genes examined in this research can be introduced as breeding targets to create resistance.
Plant Disease and Biotechnology
Abbas Saidi; Zahra Hajibrat; Ahmad Mosuapour Gorji; Rahim Ahmadvand
Abstract
Potato is one of the most important sources in the supply of food and industrial raw materials not only in Iran but also in the world which its yield is affected by various pathogens. Among these pathogens, PVY, PVX and nematode are the main factors of yield reduction. The molecular mechanism underlying ...
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Potato is one of the most important sources in the supply of food and industrial raw materials not only in Iran but also in the world which its yield is affected by various pathogens. Among these pathogens, PVY, PVX and nematode are the main factors of yield reduction. The molecular mechanism underlying disease resistance in potato remains largely unknown. In this study, analysis of gene expression profiles from the GEO data of three pathogen infections in potato was performed and morphological traits under four stresses were investigated. For this purpose, 501 common genes with different expression (DEGs) were studied in two experiments. Functional enrichment analysis showed that DEGs are more involved in nitrogen and primary metabolic cycle, GTP binding and GTPase binding, which are continuously up-regulated after inoculation with different pathogens. Based on the analysis of morphological traits under four stresses, PVY and PVX/PVY interaction left a significant difference with other stresses (PVX and nematode) on these traits. Potato microarray data extracted from GEO database were used for weighted gene co-expression network analysis (WGCNA). Based on the results of the network, 2 groups (modules) of genes were obtained whose expression profiles were highly correlated with each other in response to the above-mentioned four stresses. The results of this experiment provide valuable insight into the pathways and genes affected by PVY, PVX, PVX/PVY and potato nematode viruses, which may facilitate to identify genes resistant to many diseases in potato.
Plant Disease and Biotechnology
Son ay Baghdadi; Abdolhossein taheri; saeed nasrollahnejad; Farzad Aliramaji; leila fahmideh
Abstract
In order to detection and differentiation of two complications of disorder and lack of podding in soybeans, while visiting 185 soybean fields in Golestan and Mazandaran provinces, only from the summer cultivation of 17 fields from five different places in Golestan province, plants with signs of disorder ...
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In order to detection and differentiation of two complications of disorder and lack of podding in soybeans, while visiting 185 soybean fields in Golestan and Mazandaran provinces, only from the summer cultivation of 17 fields from five different places in Golestan province, plants with signs of disorder and the lack of acute podding were identified and 17 samples were selected from each field and sampling was done from the leaves or stems of the mentioned plants in order to extract RNA and DNA. Then, PCR was performed to detect nepovirus using the degenerate primer of nepovirus and to detect phytoplasma using a pair of general primers and a nested PCR test. The results of electrophoresis confirmed the amplification of the 1800 bp band in the general PCR, the 1250 bp fragment in the nested PCR related to phytoplasma, and also the amplification of the 640 bp band related to a nepovirus. Besides, no band of healthy plants was observed. at the same time, tissue grafting and mechanical inoculation were performed on GPX soybean benchmark plant using the mentioned samples and two types of symptoms appeared. From the sequencing of the disordered samples (production of a small number of seeds and small pods), Tomato ring spot virus strain ep31_63026 and from the sequencing of the samples with non-encapsulation (grassiness and no formation of pods and seeds), Aster yellows phytoplasma from the 16SrI-B group were identified, which confirmed the results of the reference plant. The phylogenetic analysis of sequencing results confirmed the presence of Phytoplasma and Nepovirus only in the summer culture of samples that had symptoms of disorder and lack of podding.
Plant Disease and Biotechnology
Mehrbanno Kazemi Alamuti; Mohammad Majdi; Ghasem Hosseini Salekdeh; Mohammad Reza Ghaffari
Abstract
Eurygaster integriceps is one of the known pests of wheat fields in Iran and West Asia. The role of neuropeptides in the stages of insect’s growth has led to a promising perspective for the production of a new generation of bio-insecticides based specific application. Insect’s neuropeptides along ...
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Eurygaster integriceps is one of the known pests of wheat fields in Iran and West Asia. The role of neuropeptides in the stages of insect’s growth has led to a promising perspective for the production of a new generation of bio-insecticides based specific application. Insect’s neuropeptides along with their specific receptors are one of the most diverse proteins that control physiological and behavioral activities in insects. Allatostatin is one of the important neuropeptides in insects which, by inhibiting the youth hormone, plays a role in regulating physiological processes such as feed and metabolism in some insects. In this study, using the information obtained from adult Eurygaster integriceps transcriptome, neuropeptides and specific receptors of the allatostatin family were investigated. Bioinformatics and phylogenetic studies of the data led to identify four neuropeptides A, B and C allatostatin family, as well as the neuropeptide receptors of A and B allatostatin. The results showed that the neuropeptides of the allatostatin family identified in Eurygaster integriceps are involved in various physiological processes. Considering the important role of neuropeptides in insects, these neuropeptides can be used to design specific insecticides compatible with the environment for managing control Eurygaster integriceps in future.
Plant Disease and Biotechnology
Behzad Naeimi Nezamabad; Fakhtak Taliei; Abbas Biabani; Hossein Sabouri; Masume Kheirgu
Abstract
Fungal leaf diseases are the most destructive factors to wheat (Triticum aestivum L.) yield. Therefore, identification of resistance sources to important fungal diseases is a great importance to increase wheat yield. In order to identify informative CBDP and SCoT markers for leaf rust, powdery mildew ...
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Fungal leaf diseases are the most destructive factors to wheat (Triticum aestivum L.) yield. Therefore, identification of resistance sources to important fungal diseases is a great importance to increase wheat yield. In order to identify informative CBDP and SCoT markers for leaf rust, powdery mildew and septoria leaf blotch disease, 63 wheat genotypes were planted through the augment design in agricultural research station of Araghimahaleh (Gorgan) during 2018-19. Clustering based on UPGMA, put the genotypes into 3 groups with 24, 17 and 22 individuals. Among the studied genotypes, genotypes No. 49 and 60 showed highest resistance to all three diseases. Association analysis showed that from 16 CBDP and SCoT primers, 99 bands produced in total, which 43 bands were polymorph. The maximum percent of polymorphism belonged to SCoT primers and the minimum polymorphism belonged to CBDP. SCoT11-2, CBDP 10-2 was shown significant correlation with all three diseases. SCoT21-3 was also significantly (P
Plant Disease and Biotechnology
Hanieh Abedinpour; NadAli Babaean Jeloudar; Ali Dehestani; GhorbanAli Nematzadeh
Abstract
Downy mildew is one of the most important diseases of cucumber plants reducing production in the world. Despite the availability of some fungicides to control the disease, demands for new and reliable natural products has been increased. In this study, the effects of salicylic acid (SA) and Azelaic acid ...
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Downy mildew is one of the most important diseases of cucumber plants reducing production in the world. Despite the availability of some fungicides to control the disease, demands for new and reliable natural products has been increased. In this study, the effects of salicylic acid (SA) and Azelaic acid (AZA) on some biochemical properties and expression of defense genes in cucumber plants were studied. Gene expression analysis indicated that expression of both β-1,3- glucanase and chitinase genes were increased after inoculation with Pseudoperonospora cubensis, but this increase was much higher for chitinase gene in SA-treated plants and for β-1,3- glucanase in AZA-treated plants. Also, chlorophyll a, b and total flavonoids contents in AZA and SA-treated plants were higher compared to control 6 hours after inoculation with pathogen. Total phenolic content was also increased in SA and AZA treated plants compared to the control 6 and 72 hours after treatment, respectively. These findings indicate that pretreatment of cucumber plants with SA and AZA prompts faster defense responses and increases the efficiency of the defense system against downy mildew.
Plant Disease and Biotechnology
Neda Asghari; Davoud Koolivand; Omid Eini
Abstract
Cucumber mosaic virus (CMV) is a type member of Cucumovirus genus in the family Bromoviridae. In this research, the expression profile of several genes related to development and methylation was tested in a sensitive variety of tomato infected by CMV-Fny. Tomato inoculation was carried out using a standard ...
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Cucumber mosaic virus (CMV) is a type member of Cucumovirus genus in the family Bromoviridae. In this research, the expression profile of several genes related to development and methylation was tested in a sensitive variety of tomato infected by CMV-Fny. Tomato inoculation was carried out using a standard isolates of CMV (Fny). To this aim, transcripts of RNA1, RNA2 and RNA3 of CMV were transcribed and then, inoculated using sap inoculation on tobacco plants at the four-leaf stage. After symptoms induction, leaf samples were collected at 7, 14 and 21 days post inoculation (dpi). The virus infection was confirmed by RT-PCR using CMV-CP specific primers corresponding coat protein gene. The expression of several genes related to development and methylation including Hsp90, AGO1 and AGO4 was tested by real-time PCR. The results showed that the expression of HSP90 was decreased after infection by CMV compared to healthy plans. In addition, the expression of AGO1 was increased at 7 and 14 dpi whereas the expression was decreased compared to healthy plans at 21 dpi. The expression of AGO4 which has the role in plant defense was increased in infected tomato plants at 14 and 21 dpi and AGO4 expression was decreased compared to healthy plant in 7 dpi. It seems, changes in AGOs expression refer to late induction of plant resistance against the virus infection.
Plant Disease and Biotechnology
Neda Khoshkhati; Omid Eini; Davoud KOOLIVAND
Abstract
Tomato bushy stunt virus (TBSV), a member of the genus Tombusvirus is one of the causal agents for curly disease in tomato plants. In this study, the interaction between a mycorrhizal fungus, Rhizoglomus irregular, and TBSV in tomato plants was investigated. In a completely randomized design experiment, ...
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Tomato bushy stunt virus (TBSV), a member of the genus Tombusvirus is one of the causal agents for curly disease in tomato plants. In this study, the interaction between a mycorrhizal fungus, Rhizoglomus irregular, and TBSV in tomato plants was investigated. In a completely randomized design experiment, tomato seedlings were inoculated with R. irregular and after four weeks they were inoculated with TBSV. Four treatments were included: control plants (C), TBSV -infected plants (V), mycorrhizal plants (M), TBSV -infected mycorrhizal plants (MV). Nineteen days after inoculation the infected plants were tested for symptom production and virus accumulation. Results of symptoms evaluation based on the disease severity index showed a lower disease severity in MV plants compared with V plants. Supporting this result, a lower level of virus accumulation was observed in V plants which was more significant at long-term infection. The expression of methylation-related genes including ADK, HEN1 and MET1 was tested by Real-time PCR. Results showed that the expression of these genes was significantly higher in MV plants as compared with V plants. An increase in the expression of methylation-related genes in MV plants indicates that resistance to the virus is likely to occur through methylation and also supports the lower level of virus accumulation in MV plants.
Plant Disease and Biotechnology
Zahra Sadeghi; Saeed Nasrollanejad; Milad Lagzian; Seyyed Esmaeil Razavi; Majid Jafari
Abstract
Yellowing diseases of field and greenhouse-grown cucurbits caused by whitefly-transmitted viruses are increasingly becoming important and cause economic losses in many cucurbits growing areas of the world. In this research, 195 samples with symptoms yellowing, stunting and deformation of leaves were ...
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Yellowing diseases of field and greenhouse-grown cucurbits caused by whitefly-transmitted viruses are increasingly becoming important and cause economic losses in many cucurbits growing areas of the world. In this research, 195 samples with symptoms yellowing, stunting and deformation of leaves were collected from fields of cucurbits in the South-East Iran (Sistan and Baluchestan). In order to detect CVYV and CYSDV, cDNAs were prepared using Random Hexamer primer and total RNAs extracted from the collected samples.Then, Polymerase Chain Reaction (PCR) was performed using exteracted RNA and specific primers (CYSCPf and CYSCPr for CYSDV, CV+ and CV- for CVYV). The results revealed that a 450 bp fragment from the CVYV-CP and a 700 bp fragment from CYSDV-CP were amplified from 55 and 17 samples, respectively. About 7% of the samples had simultaneous contamination with the two viruses. Based on the Phylogenetic studies CVYV were divided into two main groups I (subgroups IA and IB) and II: the first group belongs of isolates from Lebanon and Jordan, Tunisia and Spain, but the second group contained only isolates of Iran (Chabahar) which indicates the high diversity of this isolate. The results of the genetic diversity and selection indices showed that the gene flow from Europe to Asia is very small and the genetic difference is high between them. The CP region is under severe pressure a negative choice, and has caused variation and evolution of the CVYV. Generally, based on the results, the most diversity molecular and host are in Asia group.
Plant Disease and Biotechnology
zahra rezaloo; Samira Shahbazi; Hamed Askari
Abstract
Damping-off is one of the major diseases of Phaseolus vulgaris in different parts of the country. Because damping-off agents are soil-borne, the use of chemical methods had not satisfactory results, therefore, in recent years much attention has been paid to the biological struggle, especially the use ...
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Damping-off is one of the major diseases of Phaseolus vulgaris in different parts of the country. Because damping-off agents are soil-borne, the use of chemical methods had not satisfactory results, therefore, in recent years much attention has been paid to the biological struggle, especially the use of Trichoderma fungi. One of the Trichoderma mechanism is the induction of the plant's defense system. In this research, five spices of Trichoderma harzaianum, T. viride, T. Konigi, T. atroviride, T. virense were used to induce resistance of bean plant (Phaseolus vulgaris) against Rhizoctonia solani. In order to investigate the effect of mutation induction in the Trichoderma genome on the rate of plant resistance induction, five superior mutants (from each Trichoderma spices) were used for treatment of plants. The experiments were carried out in a completely randomized design with four replications. Yield, malondialdehyde, proline, chlorophyll a and b, carotenoids and protein concentration in treated plants in greenhouse condition were evaluated. The data from the evaluations showed that the mutants of these Trichoderma species decrease the diseases incidence more than the same wild type Trichoderma species, which lead to increase the bio control potential of mutant based bio-fungicides in competitive with wild type based or chemical compounds. The amount of soluble protein, enzymes activity, chlorophyll a and b and carotenoids increased in Trichoderma treatments compared to control (pathogen treatment). The results of these experiments showed that seed coating formulations (mutant and wild) had better efficiency than other formulations for controlling damping-off disease and resistance induction in bean plant.
Plant Disease and Biotechnology
Azam Badr Hadad; Farhad Nazarian Firouzabadi; Ahmad Ismaili; Hedayat Bagheri
Volume 7, Issue 19 , November 2017, , Pages 1-13
Abstract
Antimicrobial peptides are ancient and conserved molecules which are found in defense mechanisms of almost all living organisms from bacteria to animal and plant species. Identification and introduction of novel antimicrobial peptides, is a cost-effective way to improve crop plants resistance to pathogens ...
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Antimicrobial peptides are ancient and conserved molecules which are found in defense mechanisms of almost all living organisms from bacteria to animal and plant species. Identification and introduction of novel antimicrobial peptides, is a cost-effective way to improve crop plants resistance to pathogens by using recombinant DNA technology. Therefore, an expression construct containing omiganan antimicrobial encoding gene from the cytoplasmic granules of bovine neutrophils, was cloned and transferred to the tobacco leaf disk using Agrobacterium tumefaciens mediated-transformation. The presence of the antimicrobial peptide encoding gene in the genome of transgenic plants was confirmed by PCR analysis. Six putative transgenic lines and a non-transgenic control plant were selected for further molecular analysis. Total protein was extract from transgenic and non-transgenic control plants and used for antimicrobial activity assay against some human; E. coli, Salmonella, Staphylococcus, Bacillus, and plant: Xanthomonas and Pseudomonas pathogens by disc diffusion method. Results of this experiment showed that total protein extract from transgenic lines, as compared to non-transgenic plant, was significantly (P
Plant Disease and Biotechnology
Mohammad Amin Almasi; Galavizh Almasi
Volume 7, Issue 19 , November 2017, , Pages 51-63
Abstract
The most common virus affecting lettuce (Lactuca sativa L.) in the field worldwide is Lettuce mosaic virus (LMV), belonging to the family Potyviridea, genius Potyvirus. There are several techniques to detect LMV including serological and molecular assays, but these methods take a long time and requiring ...
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The most common virus affecting lettuce (Lactuca sativa L.) in the field worldwide is Lettuce mosaic virus (LMV), belonging to the family Potyviridea, genius Potyvirus. There are several techniques to detect LMV including serological and molecular assays, but these methods take a long time and requiring sophisticated tools. The aim of this study was to reduce the time required to detect LMV, using loop-mediated isothermal amplification (LAMP) technique. Leaf samples (38 samples) with symptoms similar to LMV were collected from Kurdestan province and were subjected to a serological (DAS-ELISA) test and lastly, four samples were detected as the positive samples. Then, total RNA was extracted and one-step reverse transcription (RT)-LAMP was carried out under isothermal conditions and results were evaluated by agarose gel electrophoresis and hydroxynaphthol blue (HNB) dye. A positive result using the HNB dye was a color change in master-mix from the violet to sky blue. In addition, a novel immunocapture (IC)-RT-LAMP assay for rapid and easy detection (without RNA extraction) of LMV was developed here and its potential compared with other assays. The advantages of RT-LAMP method in compare to other methods include the high specificity, high sensitivity, high rapidity, high efficiency, safety, fascinatingly, no requirement of expensive and tools for amplification, no need to RNA extraction (in IC-RT-LAMP assay), no post-amplification treatment of the amplicons, visual detection and user friendly.
Plant Disease and Biotechnology
Azadeh Ghaneie; Naser Safaie; Rahim Mehrabi; Forough Sanjarian
Volume 7, Issue 17 , July 2017, , Pages 43-54
Abstract
This study was carried out to evaluate the effect of salicylic acid on the reduction of disease severity of septoria tritici blotch (STB) in three tetraploeid and three hexaploeid wheat cultivars, under controlled conditions at seedling stage. The 10-day-old seedlings were initially treated with 0 (control), ...
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This study was carried out to evaluate the effect of salicylic acid on the reduction of disease severity of septoria tritici blotch (STB) in three tetraploeid and three hexaploeid wheat cultivars, under controlled conditions at seedling stage. The 10-day-old seedlings were initially treated with 0 (control), 1, 2 and 4 mM of salicylic acid with a handy sprayer. After 24 hrs, the seedlings were inoculated with fungal spore suspensions adjusted at 107 sopres/ml. The disease progress was evaluated 21 days post inoculation based on visual estimation of the percentage of leaf area with necrotic lesions bearing pycnidia. The results showed that there were significant differences on the reduction of disease severity among different concentrations of applied salicylic acid as well as among cultivar/ concentration interactions. In addition, the results showed that application of salicylic acid reduced disease severity of both tetraploid and hexaploid wheat cultivars and 1 mM salicylic acid was the most effective concentration on the reduction of disease severity. Although, the disease severity was significantly reduced by application of 2 and 4 mM salicylic acid, the effectiveness of these concentrations was less than application of 1mM. Overall, these results revealed that activation of plant defense systems through application of salicylic acid could play a significant role in systemic acquired resistance against STB disease.
Plant Disease and Biotechnology
Mohammad Amin Almasi
Volume 6, Issue 14 , August 2016, , Pages 73-84
Abstract
Alfalfa mosaic virus (AMV) causing a disease in Alfalfa (Medicago sativa) crop in Iran has been identified on the basis of determination of symptom expression and morphological properties. There are several techniques to detect the virus including serological test and molecular methods. Reverse transcription ...
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Alfalfa mosaic virus (AMV) causing a disease in Alfalfa (Medicago sativa) crop in Iran has been identified on the basis of determination of symptom expression and morphological properties. There are several techniques to detect the virus including serological test and molecular methods. Reverse transcription Loop-mediated isothermal amplification (RT-LAMP) assay is a novel technique for amplifying RNA under constant temperature, with high specificity, sensitivity, rapidity and efficiency that is used in this study for detection of Alfalfa mosaic virus. Leaf samples (100 samples) with symptoms similar to AMV were collected from Hamedan and Kurdestan provinces and were subjected to a serological test. All four RT-LAMP reaction primers (i.e. F3, B3, FIP and BIP) together with RT-PCR reaction primers were designed on the basis of the highly conserved sequence of coat protein (CP) gene. Total RNA was extracted and molecular reactions were carried out and finally was detected six positive sampled The advantages of this new method in compare to other methods include the high specificity, high sensitivity, high rapidity, high efficiency, safety, fascinatingly, no requirement of expensive and tools for amplification, no post-amplification treatment of the amplicons, visual detection and user friendly.
Plant Disease and Biotechnology
Elham Zamani; Forogh Sanjarian; Ebrahim Mohammadi-goltapeh; Naser Safaie
Volume 6, Issue 13 , May 2016, , Pages 1-12
Abstract
Induced resistance is one the ways by which plants cope with the biotic stresses. Pathogenesis-related proteins (PRs), including chitinase and β-1,3-glucanase are synthesized by plant in response to pathogenic infection. In present work, we have carried out inoculation experiment in wheat to investigate ...
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Induced resistance is one the ways by which plants cope with the biotic stresses. Pathogenesis-related proteins (PRs), including chitinase and β-1,3-glucanase are synthesized by plant in response to pathogenic infection. In present work, we have carried out inoculation experiment in wheat to investigate the effect of salicylic acid (SA) and Pseudomonas flourescens on Septoria tritici Bloch (STB) symptoms. Subsequently, the expression profiles of chitinase and β-1,3-glucanase were analyzed with semi-quantitative RT-PCR at 0, 4, 14 and 24 days after inoculation in both treated and control plants. Results showed that the expression of both chitinase and β-1,3-glucanase genes increased in interaction with Z. tritici and the application of SA and P. flourescens. On the other hand, the application together had no significant effect on gene expression. Briefly, it can be concluded that increment of chitinase and β-1,3-glucanase expression level with the application of SA and P. flourescens would play role induction of plant defense against STB.
Plant Disease and Biotechnology
Jalal Gholamnezhad; Forogh Sanjarian; Ebrahim Mohammadi Goltapeh; Naser Safaei; Khadijeh Razavi
Volume 5, Issue 12 , February 2016, , Pages 1-10
Abstract
As a rule the reference genes used in gene expression analysis have been selected for their housekeeping roles, but the variation observed in most of them is a major obstacle to their effective use. It is widely supported to identify and validate stable reference genes, since no single biological gene ...
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As a rule the reference genes used in gene expression analysis have been selected for their housekeeping roles, but the variation observed in most of them is a major obstacle to their effective use. It is widely supported to identify and validate stable reference genes, since no single biological gene is stably expressed between cell types or within cells under different conditions. In this study, suitability of seven wheat housekeeping genes for normalization of mRNA expression in wheat leaves infected by Mycosphaerella graminicola was investigated. Expression level of Actin, Rubisco, Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Translation Elongation Factor 1α (TEF-1α), α-Tubollin, eukaryotic release factors 1 and 3 (ERF1 and ERF3) genes were examined by reverse northern dot blot method. Expression stabilities of the reference genes were statistically analyzed by Ecxel and SAS softwares. α-Tubolin, TEF-1α and Actin were the three most stable genes whereas the expression of Rubisco and GAPDH had the least stability. The presented comprehensive data on changes in expression of various wheat housekeeping genes in wheat- M. graminicola phatosystem facilitate selection of reference genes for Reverse northern dot blot method.
Plant Disease and Biotechnology
Mohammad Ali Ebrahimi; Shiva Alahyar Parsa; Saeedeh Piraish
Volume 5, Issue 12 , February 2016, , Pages 55-66
Abstract
Molecular analysis is often required in many genome extracted from various tissues. In much molecular analysis of genetic studies of microbial communities, often require direct extraction of DNA from soil. In this study, after careful study of the various methods proposed three methods (chemical enzyme), ...
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Molecular analysis is often required in many genome extracted from various tissues. In much molecular analysis of genetic studies of microbial communities, often require direct extraction of DNA from soil. In this study, after careful study of the various methods proposed three methods (chemical enzyme), (chemi-mechanical) and (chemical mechanical enzymatic), in order to extract DNA from soil, a separate project was studied to most identify different methods of extraction. The comparison between the same conditions, a significant difference in the DNA product of the average concentration of 2/32 to 6/1 ng per µl DNA was diluted and also on the basis of information obtained from Nanodrop and electrophoresis device and review of the DNA extracted protein product DNA, methods (chemical, mechanical, enzymatic) better performance at higher concentrations, remove humic acid proteins and other contaminants was superior to other methods. The molecular detection of pathogens that extraction is important applications; the aim of this study is molecular detection of pathogen Fusarium oxysporum f. sp lycopersici and Fusarium oxysporum f. sp. radicis lycopersici of soil microbial community by molecular techniques to identify the special primer evaluated. Molecular detection results showed that there is tomato Fusarium infection in soil samples S1.
Molecular Plant Breeding
Arezo Mohamadian Farsani; Hamid Hatami Maleki; Reza Darvishzadeh; Faramarz Hoshyardel
Volume 4, Issue 7 , December 2015, , Pages 15-23
Abstract
Black stem disease is one of the most important fungi diseases of sunflower. The use of resistant genotypes is potentially one of the economical ways for its control. In this study, expression level of genes including phenylalanine ammonia-lyase 2 (PAL2) and thaumatin-like protein (TLP) were measured ...
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Black stem disease is one of the most important fungi diseases of sunflower. The use of resistant genotypes is potentially one of the economical ways for its control. In this study, expression level of genes including phenylalanine ammonia-lyase 2 (PAL2) and thaumatin-like protein (TLP) were measured in sunflower genotypes including ENSAT-B5, AS613 and mutant genotype M5-54-1 infected with MA6, MP8 and MP10 isolates of Phoma macdonaldii via quantitative RT-PCR technique. Results revealed that transcript levels of the both PAL2 and TLP genes were significantly affected by isolate and genotype and genotype-isolate interactions. In this study, the expression levels of genes PAL2 and TLP showed higher increase against the MP8 isolate. Among studied genotypes, genotype ENSAT-B5 possessed highest increasing in expression levels of genes PAL2 and TLP. The resistant and susceptible genotypes had possessed high and low transcript levels of PAL2 and TLP and disease symptoms were seen in susceptible genotypes.
Plant Disease and Biotechnology
Sahba Toosi; Farhad Shokouhifar; Saeed Malekzadeh Shafaroudi; Abdolreza Bagheri
Volume 4, Issue 7 , December 2015, , Pages 35-47
Abstract
Melon Vascular wilt caused by a soil-borne pathogen, Fusarium oxysporum f. sp melonis (Fom) is a distractive disease. The use of resistant cultivars is an effective way for controlling this fungus. Clear view about the interaction between virulence and resistant genes could provide applicable knowledge ...
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Melon Vascular wilt caused by a soil-borne pathogen, Fusarium oxysporum f. sp melonis (Fom) is a distractive disease. The use of resistant cultivars is an effective way for controlling this fungus. Clear view about the interaction between virulence and resistant genes could provide applicable knowledge to design breeding programs. Fungus effector genes play a critical role during the plant-pathogen interactions. Exploring for the homologous of effector genes in different formae speciales started since some effector genes have been reported from Fusarium oxysporum f. sp. lycopersici (Fol). Recently, using bioinformatics assisted approach a homologous Fol-SIX1 effector gene has been reported from Fom. Here we performed a functional analysis study to reveal interactions between Fol-SIX1 and the melon differential varieties harboring defined R genes. Coding sequence of Fol-SIX1 from the pTS1 construct was subcloned in the pCAMBIA3301 binary vector in three steps and confirmed by colony PCR and digestion analysis. The accuracy of the expression construct, pCaS1 was evaluated by bidirectional sequencing using the PSh4-F2 and PSh51-R primers. Agroinjection-mediated transient expression approach was used to express Fol-SIX1 in leaves of the melon varieties. Functional analysis of Fol-SIX1 was performed by agroinjection of LBA4404 carrying pCaS1 into the leaves of melon lines. the leaves of Ch-T and Ch-Fom2 varieties developed a green dried symptom in response to transiently expressed Fol-SIX1 at 24 hours post injection (hpi), while Ch-Fom1 and BG lines did not responsive to the infection until 48 hpi.
Plant Disease and Biotechnology
Farshad Bakhtiar; Ezatollah Farshadfar; Mostafa Aghaee Sarbarzeh; Habibollah Ghazvini; Farzad Afshari
Volume 5, Issue 10 , September 2015, , Pages 41-56
Abstract
In this research, 150 wheat doubled haploid lines were produced using chromosome elimination method by crossing between wheat and maize. Resistance of doubled haploid lines, their parents and check cultivars against strip and stem rust was evaluated at seedling and adult plant stages. Accordingly, eight ...
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In this research, 150 wheat doubled haploid lines were produced using chromosome elimination method by crossing between wheat and maize. Resistance of doubled haploid lines, their parents and check cultivars against strip and stem rust was evaluated at seedling and adult plant stages. Accordingly, eight known molecular markers which are tightly linked to resistance genes including Yr5, Sr31/Yr9/Lr26, Yr15, Sr38/Yr17/Lr37, Lr34/Yr18/Pm38, Yr27, Yr36 and Yr48 were screened in parents. Results showed that molecular markers for Yr5, Yr15, Yr27 and Yr36 couldn't detect polymorphism between parents as well as positive and negative controls. For gene block Sr38/Yr17/Lr37 and locus Yr48 allele sizes were not similar to those which were expected for these genes. Results also showed that MV17 and Flanders have gene block of Sr31/Yr9/Lr26, and only 3 lines of population DH-26: Ghods*3/MV17 had this gene block from which two doubled haploid lines showed resistance reaction to TTSTK and TTKSK of Puccinia graminis pers. f.sp tritici races. Genetic test for the presence or absence of gene block Lr34/Yr18/Pm38 on parents of doubled haploid lines showed that MV17 comprises this gene block. Evaluation of doubled haploid lines for this gene block showed that 6 doubled haploid lines of population DH-26:Ghods*3/MV17 have gene block Lr34/Yr18/Pm38 for which only one doubled haploid line showed resistance reaction to Puccinia striiformis Westend f.sp. tritici race of 7E158A+, Yr27 at both seedling and adult plant stages.
Plant Disease and Biotechnology
Mahsa Banaee; Forogh Sanjarian; Gholam Reza Bakhshi Khaniki
Volume 3, Issue 4 , September 2013, , Pages 25-32
Abstract
Acetyl transferases are enzymes responsible for enzymatic transfer of an acetyl group to suitable receptor molecule by using acetyl CoA as donor. Acetyltransferase reaction is involved in biosynthesis pathway of some important secondary metabolites such as antibiotics, as well as in their detoxification. ...
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Acetyl transferases are enzymes responsible for enzymatic transfer of an acetyl group to suitable receptor molecule by using acetyl CoA as donor. Acetyltransferase reaction is involved in biosynthesis pathway of some important secondary metabolites such as antibiotics, as well as in their detoxification. Trichothecenes are significant secondary metabolites produced by the plant fungal phatogens Fusarium ssp. Such as F. sporotrichioides and F. graminearums. These fungi possess specific genes in their genomes encodeinge acetyl transferase enzymes are affecting trichothecene. In this study, the gene encoding acetyltransferase from the fungus F. sporotrichioides, TRI 101, was cloned and transferred into tobacco plant as model plants and the effect of the enzyme on the detoxification of deoxynivalenol (DON), a well known trichthecene, was investigated. In addition it was shown that, in comparison whit the roots of wild type plants, transgenic roots grew normally in the deoxynivalenol-contained medium.
Plant Disease and Biotechnology
Fatemeh Hosseini; Reza Pourrahim; Mojdeh Maleki; Shirin Farzadfar
Volume 3, Issue 4 , September 2013, , Pages 119-131
Abstract
Potato virus A is one of the most important viruses infecting potato in the world. During spring and summer of the year 2011, a total of 280 symptomatic and 463 random leaf samples, and 1186 tuber samples were collected from potato fields located in Khorasan-e-razavi and Markazi provinces, Iran and tested ...
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Potato virus A is one of the most important viruses infecting potato in the world. During spring and summer of the year 2011, a total of 280 symptomatic and 463 random leaf samples, and 1186 tuber samples were collected from potato fields located in Khorasan-e-razavi and Markazi provinces, Iran and tested for PVA infection using DAS-ELISA method. Based on the results obtained, PVA infection was recorded in 39 and 11 % of symptomatic leaf samples, 23 and 6 % of random leaf samples and 17 and 13 % tuber samples in Markazi and Khorasan-e-Razavi provinces, respectively. Morever, biological properties of the selected PVA isolates were studied using host range studies on 10 plant species. Four PVA isolates were selected and their coat protein (CP) gene was amplified by specific primers using RT-PCR and their nucleotide sequences were determined (KF152950-3). The CP sequences of four Iranian and 37 Non-Iranian (from other parts of the world) PVA isolates were compared and a phylogenetic tree was reconstructed using Neighbor Joining-NJ method with 1000 bootstrap value. The PVA isolates in the phylogenic tree were clustered in two main groups (I and II) where group I had two subgroups (IA and IB). Three Iranian PVA isolates (Ar2, Kh4 and F2 all from Markazi province) were grouped in IA subgroup and showed highest similarity with two isolates from Germany and Netherlands (Y11427 and X91968) and the other Iranian PVA isolate (Gh4 from Khorasan-e-Razavi P.) was grouped in IB subgroup showing the highest similarity with an isolate from China (DQ309063). Comparison of the deduced amino acid (aa) sequences of CP among the four Iranian and the other PVA isolates revealed substitution of 2 to 7 aa mainly in N terminal of CP. This is the first phylogenetic study on Iranian PVA isolates and also the first report of PVA infection in Markazi province. This study showed variation among PVA isolates in Iran. These data could to be considered in potato breeding programs in order to achieve PVA resistant cultivars.
Plant Disease and Biotechnology
Samira Shahbazi; Hossein Ahari Mostafavi; Mohammad Ali Ebrahimi; Hamed Askari; Mahyar Mirmajlesi; Mahsa Karimi
Volume 3, Issue 5 , February 2013, , Pages 33-40
Abstract
Trichoderma spp. is considered as promising biological control agents. Antagonistic Trichoderma species control numerous of plant pathogenic fungi. The highest value of degrading enzymes production (chitinases and glucanases) between known micro- organisms belongs to this genus. Hence that chitinases ...
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Trichoderma spp. is considered as promising biological control agents. Antagonistic Trichoderma species control numerous of plant pathogenic fungi. The highest value of degrading enzymes production (chitinases and glucanases) between known micro- organisms belongs to this genus. Hence that chitinases have effective role in myco-parasitism mechanism of plant pathogenic fungi, therefore superior bio-control potential may then be found in isolates which produce a high capacity of these enzymes. Application of genetic engineering and direct mutation in fungal genome in order to increase T. harzianum antagonistic potential via enhancement in chitinase activity has some advantages. In this study, we use 250 Gry gamma irradiation dose to induce mutation in T. harzianum. Then, chitinase activity of 20 mutants have been evaluated and 15 mutants showed higher enzyme activity than wild type isolate. Thus, induced mutation by gamma irradiation in order to improve antagonistic capacity of native iranian Trichoderma spp. to obtain effective and useful bio- control agents is advisable.
Molecular Plant Breeding
maasoumeh Habibi; Asadollah Abiar Fini; Neda Mirakhorli; Mohsen Mardi
Volume 3, Issue 5 , February 2013, , Pages 85-92
Abstract
Bread wheat (Triticumaestivum) is one of the most important world food crops that are exposed to many pathogen. During the previous expression-profiling experiments, in addition to major resistant genes to disease in wheat, some defense-related genes such as S-Like RNase gene have been identified. Here ...
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Bread wheat (Triticumaestivum) is one of the most important world food crops that are exposed to many pathogen. During the previous expression-profiling experiments, in addition to major resistant genes to disease in wheat, some defense-related genes such as S-Like RNase gene have been identified. Here to study expression pattern of this gene in several fungal wheat diseases, some bioinformatics and laboratory studied were performed. In bioinformatics studies, several microarray libraries infected with Fusarium, Spike blight and Stripe rust were considered. In laboratory experiments Septoria tritici bloth was studied. So the level of expression was measured at 8 time interval, from 0h to 6 days after inoculation by Mycosphaerella graminicola in Wangshuibai as a resistant wheat cultivar and Falat as a susceptible wheat cultivar by semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The results show that the maximum expression of this gene, depending on types of disease and resistant cultivars, is obtained up to 24 hours after the inoculation. Thus, according to this results it can be concluded that this gene plays an important role in resistance to diseases and ,along with the main gene, increase and maintain resistance to many fungal diseases in wheat. Also this gene was confirmed by Nucleotide Blast.
Plant Disease and Biotechnology
M.R Safarnejad; M Basirat; M.A Ebrahimi; H Safarpour; S Nazari; B Mahmoudi; S Ataie Kachoie
Volume 2, Issue 3 , January 2013, , Pages 1-12
Abstract
Rhizomania is one of the most important Sugar beet diseases throughout the world. The disease is caused by beet necrotic yellow vein virus (BNYVV). The Polymyxa betae (Keskin) is the only natural transmitting agent of the disease between the plants. The fungus is an obligate parasite and could not be ...
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Rhizomania is one of the most important Sugar beet diseases throughout the world. The disease is caused by beet necrotic yellow vein virus (BNYVV). The Polymyxa betae (Keskin) is the only natural transmitting agent of the disease between the plants. The fungus is an obligate parasite and could not be cultured in the media, then detection of fungus is done normally by microscopic observation. In order to facilitate detection process, present study is done to develop specific antibodies against this fungus by applying recombinant protein. The Glutathione-S-transferase (GST) as a specific immunogenic protein is a critical enzyme expressed in zoospores, sporangia and resting spores and could be a good candidate to develop a serological test for P. betae. For this aim, the DNA region encoding fungal GST gene was isolated and cloned into pET28a bacterial expression vector. Large scale expression of the recombinant protein was performed in Escherichia coli (Migula). Purification was carried out by applying immobilized metal affinity chromatography under native conditions. The purified recombinant GST protein was used for immunization of rabbit. Purification of immunoglobulin was performed by affinity chromatography using protein A column. The purified antibodies were applied for efficient detection of infected plant in serological assays.
