Payame Noor UniversityCrop Biotechnology2252-07834120141221Proteome Analysis of Wheat Flag Leaf Response to Drought StressProteome Analysis of Wheat Flag Leaf Response to Drought Stress1131412FAMohammad RezaAzimiDepartment of Agronomy and Plant Breeding, University of Tehran, Karaj, Iran.GhasemHosseini SalekdehَAssociate Professor, Department of Genomics, Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran.Journal Article20140909<span>Water shortage is one of the most important environmental factors in limiting plant production worldwide. Molecular breeding may help to develop drought tolerant plants. Proteomics approach can help in comprehensive analysis of stress responsive genes and identification of drought signaling pathways. These drought tolerance candidate genes or their regulatory genes may be further analyzed for their possible implication in increase plant tolerance to drought stress. In this studied we analyzed the changes in proteome of wheat flag leaf in response to drought. We compared a drought tolerant with drought susceptible genotypes under normal and stress well-watered and stress conditions. Stressed plants were exposed to 50% field capacity before anthesis. Out of 900 proteins analyzed across two dimensional gels, 57 protein spots showed significant differences in response to stress. Of these, 42 protein spots could be identified using mass spectrometry analysis. Differentially expressed proteins and enzymes could be grouped in different functional groups. Our results showed that tolerant genotype may use various mechanisms particularly the up-regulation of genes involved in oxidative stress defense in flag leaf. This may help the tolerant genotypes to better remove reactive oxygen species generate by stress in flag leaf and maintain its physiological and photosynthetic activities.</span><span>Water shortage is one of the most important environmental factors in limiting plant production worldwide. Molecular breeding may help to develop drought tolerant plants. Proteomics approach can help in comprehensive analysis of stress responsive genes and identification of drought signaling pathways. These drought tolerance candidate genes or their regulatory genes may be further analyzed for their possible implication in increase plant tolerance to drought stress. In this studied we analyzed the changes in proteome of wheat flag leaf in response to drought. We compared a drought tolerant with drought susceptible genotypes under normal and stress well-watered and stress conditions. Stressed plants were exposed to 50% field capacity before anthesis. Out of 900 proteins analyzed across two dimensional gels, 57 protein spots showed significant differences in response to stress. Of these, 42 protein spots could be identified using mass spectrometry analysis. Differentially expressed proteins and enzymes could be grouped in different functional groups. Our results showed that tolerant genotype may use various mechanisms particularly the up-regulation of genes involved in oxidative stress defense in flag leaf. This may help the tolerant genotypes to better remove reactive oxygen species generate by stress in flag leaf and maintain its physiological and photosynthetic activities.</span>https://cropbiotech.journals.pnu.ac.ir/article_1412_494d59dd4486499d79087fcd62c48083.pdfPayame Noor UniversityCrop Biotechnology2252-07834120141223Study of changes in the phenylalanine ammonia-lyase 2 (PAL2) and thaumatin-like protein (TLP) genes transcripts in sunflower genotypes in response to fungi isolates of Black stem diseaseStudy of changes in the phenylalanine ammonia-lyase 2 (PAL2) and thaumatin-like protein (TLP) genes transcripts in sunflower genotypes in response to fungi isolates of Black stem disease15231413FAArezoMohamadian FarsaniDepartment of Plant Breeding and Biotechnology, Faculty of Agriculture, Urmia University, Urmia, Iran.HamidHatami MalekiDepartment of Agronomy and Plant Breeding, Faculty of Agriculture, University of Maragheh, Maragheh, IranRezaDarvishzadehDepartment of Plant Breeding and Biotechnology, Faculty of Agriculture, Urmia University, Urmia, Iran.FaramarzHoshyardelDepartment of Plant Breeding and Biotechnology, Faculty of Agriculture, Urmia University, Urmia, Iran.Journal Article20141023Black 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.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.https://cropbiotech.journals.pnu.ac.ir/article_1413_efdf74b8219ce41fe08835ded56339bd.pdfPayame Noor UniversityCrop Biotechnology2252-07834120141226Cytogenetical study of different ecotypes of Cichorium intybus L.Cytogenetical study of different ecotypes of Cichorium intybus L.25341414FASadeghImaniM.Sc. student of Plant Breeding, Payame Noor University, Tehran, Iran.RaheleRahbarianAssistant Professor, Department of Biology, Payame Noor University, Tehran, IranAliMasoumiAssistant Professor, Department of Agriculture, Payame Noor University, Tehran, Iran,SaeedKhavari KhorasaniKhorasan Razavi Agricultural and Natural Resources Research CenterJournal Article20140926Cytogenetical studies and karyotypes can be used for many purposes such as , to study chromosomal characteristics, identifying diploid and poly ploid species and hybridization process , to study of genetic diversity and finding taxonomic relationships, and to gather information about past evolutionary events. In this study, 8 ecotypes of Cichorium intybus L. Included Karaj, Taft, Khomein, Talesh, Semirom, Mollasani and Baft investigated based on cytogenetic characteristics. Results showed that all the studied ecotypes are diploid (2n=2x=18). But the length of the chromosomes is very diverse (from 0.97 to 5.56 µm for the short arm and from 1.02 to 7.14 µm for the long arm). There was no satellite in all chromosomes. In classifying based on cluster analysis method, Baft and Talesh ecotypes were in a same cluster and Tafresh and khomein ecotypes were in another cluster. Also Taft, Semirom and Mollasani ecotypes were in the third cluster. Because of the differences and diversity of traits, is expected that different clusters show more heterosis in crosses and it can be used in breeding programs. According to stebbins table, Tafresh and khomain ecotypes were in 1A class and other ecotypes were in 2B class.Cytogenetical studies and karyotypes can be used for many purposes such as , to study chromosomal characteristics, identifying diploid and poly ploid species and hybridization process , to study of genetic diversity and finding taxonomic relationships, and to gather information about past evolutionary events. In this study, 8 ecotypes of Cichorium intybus L. Included Karaj, Taft, Khomein, Talesh, Semirom, Mollasani and Baft investigated based on cytogenetic characteristics. Results showed that all the studied ecotypes are diploid (2n=2x=18). But the length of the chromosomes is very diverse (from 0.97 to 5.56 µm for the short arm and from 1.02 to 7.14 µm for the long arm). There was no satellite in all chromosomes. In classifying based on cluster analysis method, Baft and Talesh ecotypes were in a same cluster and Tafresh and khomein ecotypes were in another cluster. Also Taft, Semirom and Mollasani ecotypes were in the third cluster. Because of the differences and diversity of traits, is expected that different clusters show more heterosis in crosses and it can be used in breeding programs. According to stebbins table, Tafresh and khomain ecotypes were in 1A class and other ecotypes were in 2B class.https://cropbiotech.journals.pnu.ac.ir/article_1414_7c3b5c5218d4fc3f050e468986aed6a8.pdfPayame Noor UniversityCrop Biotechnology2252-07834120140923Functional analysis of Fol-SIX1 in resistant and susceptible Cucumis meloFunctional analysis of Fol-SIX1 in resistant and susceptible Cucumis melo35471415FASahbaToosiM.Sc. student, Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, IranFarhadShokouhifarAssistant Professor, Research Center for Plant Sciences, Ferdowsi University of Mashhad, , Mashhad, IranSaeedMalekzadeh ShafaroudiAssistant Professor, Department of Plant Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, IranAbdolrezaBagheriProfessor, Department of Plant Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, IranJournal Article20140905Melon 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.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.https://cropbiotech.journals.pnu.ac.ir/article_1415_8e0ea2ce80cfb0bf610cc2998286a9d4.pdfPayame Noor UniversityCrop Biotechnology2252-07834120141222Study on the presence of resistance gene loci to yellow, stem and leaf rust diseases using molecular markers in pre-released wheat linesStudy on the presence of resistance gene loci to yellow, stem and leaf rust diseases using molecular markers in pre-released wheat lines49581416FARahimMehrabiAssistant Professor, Cereal Research Department, Seed and Plant Improvement Institute, Karaj, IranMohsenSarhangiLaboratory Technicians, Cereal Research Department, Seed and Plant Improvement Institute, Karaj, IranElhamAla-HassaniLaboratory Technicians, Cereal Research Department, Seed and Plant Improvement Institute, Karaj, IranHabibolahGhazviniAssistant Professor, Cereal Research Department, Seed and Plant Improvement Institute, Karaj, IranFarzadAfshariProfessor, Cereal Research Department, Seed and Plant Improvement Institute, Karaj, IranJournal Article20141009Yellow, leaf and stem rusts are among the most devastating diseases of wheat in Iran and worldwide. The use of resistant cultivars is the most effective and economic approach to control these diseases. To date, a significant number of molecular markers linked to the resistance loci to these diseases have been identified. In this study, pre-released wheat lines of four major wheat climate zones of Iran were evaluated for the presence or absence of molecular markers linked to 11 resistance loci including Lr46/Yr29/Pm39, Lr34/Yr18/Pm38, Lr67/Yr46, Sr2/PBC, Sr24/Lr24, Sr26, Sr31/Yr9/Lr26, L21, Sr38/Yr17/Lr37, SrCad and Lr29. The results showed that Lr46/Yr29/Pm39, Lr34/Yr18/Pm38 and Lr67/Yr46 loci existed in four and five genotypes, respectively, while Sr31/Yr9/Lr26 and Sr38/Yr17/Lr37 loci existed only in one genotype. The remaining genotypes did not possess markers linked to resistance loci tested in this study. The results showed that the frequency of presence of these loci in pre-released lines is low and, thus, lines possessing the resistance loci should be incorporated in wheat breeding programs in order to increase the frequency of the resistance genes in new cultivars.Yellow, leaf and stem rusts are among the most devastating diseases of wheat in Iran and worldwide. The use of resistant cultivars is the most effective and economic approach to control these diseases. To date, a significant number of molecular markers linked to the resistance loci to these diseases have been identified. In this study, pre-released wheat lines of four major wheat climate zones of Iran were evaluated for the presence or absence of molecular markers linked to 11 resistance loci including Lr46/Yr29/Pm39, Lr34/Yr18/Pm38, Lr67/Yr46, Sr2/PBC, Sr24/Lr24, Sr26, Sr31/Yr9/Lr26, L21, Sr38/Yr17/Lr37, SrCad and Lr29. The results showed that Lr46/Yr29/Pm39, Lr34/Yr18/Pm38 and Lr67/Yr46 loci existed in four and five genotypes, respectively, while Sr31/Yr9/Lr26 and Sr38/Yr17/Lr37 loci existed only in one genotype. The remaining genotypes did not possess markers linked to resistance loci tested in this study. The results showed that the frequency of presence of these loci in pre-released lines is low and, thus, lines possessing the resistance loci should be incorporated in wheat breeding programs in order to increase the frequency of the resistance genes in new cultivars.https://cropbiotech.journals.pnu.ac.ir/article_1416_7e72c95cf29ac05ed7856638607e5d78.pdfPayame Noor UniversityCrop Biotechnology2252-07834120141222Construction and functional analysis of pBI105, a plant expression vector to facilitate cloning and recombinant protein purificationConstruction and functional analysis of pBI105, a plant expression vector to facilitate cloning and recombinant protein purification59691430FAKasraEsfahaniAssistant Professor, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.0000-0003-3079-7009Ali HatefSalmanianAssociated Professor, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.Journal Article20140918Binary vectors such as pBI121 are widely used for introducing genes of interest into plants via Agrobacteriun-mediated transformation method. These vectors usually have low number of unique recognition sites of restriction enzymes in their multiple cloning sites. They also lack elements such as sequences necessary for protein purification. In this article, a new plant expression vectorwith a developed multiple cloning site including recognition sites of 13 restriction enzymes, a sequence for coding 8xHis-Tag to purify recombinant proteins and the sequence of M13 reverse sequencing primer, is introduced. Construction of new vector was confirmed by PCR, digestion pattern analysis, and sequencing. The T-DNA regions of the new vector and pBI121 as a control sample were transformed to Nicotiana tabbacum L. cv. Samsun by using Agrobacterium tumefaciensstrain LBA4404. Integration of T-DNA of the vector to the genome of plantlets, which were regenerated on selective culture, was analyzed by PCR. GUS assay also confirmed expression of the transgene in transgenic seedlings. These evidences indicated that the new plant expression vector, pBI105, is efficient for plant transformation.Binary vectors such as pBI121 are widely used for introducing genes of interest into plants via Agrobacteriun-mediated transformation method. These vectors usually have low number of unique recognition sites of restriction enzymes in their multiple cloning sites. They also lack elements such as sequences necessary for protein purification. In this article, a new plant expression vectorwith a developed multiple cloning site including recognition sites of 13 restriction enzymes, a sequence for coding 8xHis-Tag to purify recombinant proteins and the sequence of M13 reverse sequencing primer, is introduced. Construction of new vector was confirmed by PCR, digestion pattern analysis, and sequencing. The T-DNA regions of the new vector and pBI121 as a control sample were transformed to Nicotiana tabbacum L. cv. Samsun by using Agrobacterium tumefaciensstrain LBA4404. Integration of T-DNA of the vector to the genome of plantlets, which were regenerated on selective culture, was analyzed by PCR. GUS assay also confirmed expression of the transgene in transgenic seedlings. These evidences indicated that the new plant expression vector, pBI105, is efficient for plant transformation.https://cropbiotech.journals.pnu.ac.ir/article_1430_736941c582272df950fce136229683ac.pdf