Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Designing of polyoleosin-proinsulin fusion gene and its transformation to rapeseed (Brassica napus L.) Designing of polyoleosin-proinsulin fusion gene and its transformation to rapeseed (Brassica napus L.) 1 16 2204 10.22103/jab.2018.2204 FA Najmeh Jami M.Sc. Student, Department of Plant Production and Genetics, Agricultural Sciences and Natural Resources University of Khouzestan, Khouzestan, Iran. Esmael Ghasemi Assistant Professors, Department of Plant Production and Genetics, Agricultural Sciences and Natural Resources University of Khouzestan, Khouzestan, Iran. Alireza Shafei Assistant Professors, Department of Plant Production and Genetics, Agricultural Sciences and Natural Resources University of Khouzestan, Khouzestan, Iran. Payam Pourmohammadi Assistant Professors, Department of Plant Production and Genetics, Agricultural Sciences and Natural Resources University of Khouzestan, Khouzestan, Iran. 0000-0003-2292-6069 Journal Article 2018 06 19 Objective The use of plant Oleosin gene is a recommended method for large-scale, easier and cheaper production of recombinant protein. Attachment of oleosin gene to target gene cause to accumulate recombinant protein on seed oil bodies surface. Recent researches indicate that application of tandem Oleosin Genes (polyoleosin) fused to the target gene to facilitate recombinant protein purification is more efficient than single Oleosin. Materials and methods In present research, we designed a synthetic fusion fragment containing 5´- Kozak sequence - His-tags, three Oleosin Genes, a proteolytic site for a peptidase enzyme, Proinsulin protein, and a tetranucleotide stop codon. This synthetic sequence was cloned in binary vector pBI121by enzymatic digestion and ligation method. Then for transformation of cotyledon and hypocotyl explants of Canola, confirmed recombinant construct is transformed to Agrobacterium LBA4404 strain. Probable transgenic shoots were regenerated on selective medium containing kanamycin after 4-6 weeks. Results The analysis of transgenic shoots at DNA level was performed by PCR and amplification of a 120-bp fragment indicate integration of fusion gene in transgenic plant genome. Finally, the Proinsulin gene transcription was also confirmed by RT- PCR. Objective The use of plant Oleosin gene is a recommended method for large-scale, easier and cheaper production of recombinant protein. Attachment of oleosin gene to target gene cause to accumulate recombinant protein on seed oil bodies surface. Recent researches indicate that application of tandem Oleosin Genes (polyoleosin) fused to the target gene to facilitate recombinant protein purification is more efficient than single Oleosin. Materials and methods In present research, we designed a synthetic fusion fragment containing 5´- Kozak sequence - His-tags, three Oleosin Genes, a proteolytic site for a peptidase enzyme, Proinsulin protein, and a tetranucleotide stop codon. This synthetic sequence was cloned in binary vector pBI121by enzymatic digestion and ligation method. Then for transformation of cotyledon and hypocotyl explants of Canola, confirmed recombinant construct is transformed to Agrobacterium LBA4404 strain. Probable transgenic shoots were regenerated on selective medium containing kanamycin after 4-6 weeks. Results The analysis of transgenic shoots at DNA level was performed by PCR and amplification of a 120-bp fragment indicate integration of fusion gene in transgenic plant genome. Finally, the Proinsulin gene transcription was also confirmed by RT- PCR.

https://jab.uk.ac.ir/article_2204_4b4edc2630fe75800ddc29a7b4070add.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Optimization of Direct Regeneration of Astragalus verus in in vitro condition Optimization of Direct Regeneration of Astragalus verus in in vitro condition 17 30 2205 10.22103/jab.2018.2205 FA Saffieh Ebrahimi MSc, Plant Biotechnology, Complex Higher Education of Shirvan, North Khorasan, Iran. Fatemeh Zaker Tavallaie Assistant professor. Plant Biotechnology. Complex Higher education of Shirvan. North Khorasan, Iran 0000-0002-1934-6373 Mohammad Zarea Mehrjerdi Assistant Professor, Plant Biotechnology, Complex Higher Education of Shirvan, North Khorasan, Iran. Mhmood Ghorban-zadeh Associate Professor, Plant Biotechnology, Complex Higher Education of Shirvan, North Khorasan, Iran Journal Article 2018 06 13 Astragalus is one of the most valuable medicinal plants in Iran. The species of Astragalus verus is a native species of tragacant producer. Due to the importance of this species in the production of tragacant, exposed to extinction, the role of seed in its reproduction, as well as the possibility of its reproduction using micropropagation, this study was conducted with the aim of breaking seed dormancy, seed germination and direct regeneration of this plant. First, the effect of surface scratching on breaking of seed dormancy was investigated. Then, the effect of culture medium and sucrose concentration on seed germination and sterile seedling production was studied as factorial based on completely randomized design. The explants of hypocotyle and epicotyle were cultured on MS medium supplied by B5 vitamin containing BAP in 3 levels and IAA in 2 levels due to multiple shoot production. Rooting of multiple shoots was carried out on 1/2MS medium containing IAA in 2 levels and IBA in 3 levels based on completely randomized design with 6 treatments and 5 repeat.Scraping increased the germination rate by an average of 98%. Different media and sucrose concentration had no effect on germination percentage. With increasing sucrose, were increased the stem and root length, fresh weight and seedling dry weight. The highest percentage of shoots was obtained in MS medium containing 2 mg /L BAP without IAA with an average of 4.6 pcs. The highest percentage of rooting was obtained with 1.5 mg/l IAA with an average of 60%. The results of this study can be used in micropropagation, gene transformation and in vitro tragacant production. Astragalus is one of the most valuable medicinal plants in Iran. The species of Astragalus verus is a native species of tragacant producer. Due to the importance of this species in the production of tragacant, exposed to extinction, the role of seed in its reproduction, as well as the possibility of its reproduction using micropropagation, this study was conducted with the aim of breaking seed dormancy, seed germination and direct regeneration of this plant. First, the effect of surface scratching on breaking of seed dormancy was investigated. Then, the effect of culture medium and sucrose concentration on seed germination and sterile seedling production was studied as factorial based on completely randomized design. The explants of hypocotyle and epicotyle were cultured on MS medium supplied by B5 vitamin containing BAP in 3 levels and IAA in 2 levels due to multiple shoot production. Rooting of multiple shoots was carried out on 1/2MS medium containing IAA in 2 levels and IBA in 3 levels based on completely randomized design with 6 treatments and 5 repeat.Scraping increased the germination rate by an average of 98%. Different media and sucrose concentration had no effect on germination percentage. With increasing sucrose, were increased the stem and root length, fresh weight and seedling dry weight. The highest percentage of shoots was obtained in MS medium containing 2 mg /L BAP without IAA with an average of 4.6 pcs. The highest percentage of rooting was obtained with 1.5 mg/l IAA with an average of 60%. The results of this study can be used in micropropagation, gene transformation and in vitro tragacant production.

https://jab.uk.ac.ir/article_2205_06d5ae105ea1bea4d800bc96491876e9.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Study on antibacterial activity and cytotoxicity of recombinant peptide, Lasioglossin ɪɪɪ, against foodborne pathogens Study on antibacterial activity and cytotoxicity of recombinant peptide, Lasioglossin ɪɪɪ, against foodborne pathogens 31 44 2206 10.22103/jab.2018.2206 FA Mohammad B. Habibi Najafi Department of Food Science & Technology- Ferdowsi University of Mashhad 0000-0002-0498-1067 Abbas Tanhaeiyan Ph.D. Student of Agricultural Biotechnology Department, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran. Pariya Rahnama Ph.D. Student of Food Microbiology, Department of Food Science & Technology, Ferdowsi University of Mashhad, Mashhad, Iran. Marjan Azghandi Ph.D. Student of Animal Science Department, Ferdowsi University of Mashhad, Mashhad, Iran. Journal Article 2018 07 13 The increasing microbial resistance to existing antibiotics has increased the interest in novel antimicrobial compounds. Antimicrobial peptides (AMPs) represent an attractive alternative to classical antibiotics. Lasioglossins are a group of peptides with antimicrobial activity. The inhibitory effects of a recombinant synthetic Lasioglossin, Lasioglossin ɪɪɪ, on five food pathogens (Staphylococcus aureus, Salmonella typhimurium, Enterococcus faecalis, Listeria monocytogenes and Escherichia coli) and its cytotoxicity on the normal cell line was investigated in vitro. The findings showed great antimicrobial function of the peptide. Minimum inhibition concentration and minimum bactericidal concentration of Lasioglossin ɪɪɪ on the pathogens were the range of 3.851-8.625 and 7.703-15.406, respectively. Results indicated Staphylococcus aureus showed the highest sensitivity to Lasioglossin ɪɪɪ. The Lassioglossin demonstrated cytotoxic effect on human embryonic kidney cells in higher concentration (1652 µg/ml) in comparison to MIC and MBC values. The results indicate that this peptide can be competed with common antibiotics in terms of the bactericidal properties. The increasing microbial resistance to existing antibiotics has increased the interest in novel antimicrobial compounds. Antimicrobial peptides (AMPs) represent an attractive alternative to classical antibiotics. Lasioglossins are a group of peptides with antimicrobial activity. The inhibitory effects of a recombinant synthetic Lasioglossin, Lasioglossin ɪɪɪ, on five food pathogens (Staphylococcus aureus, Salmonella typhimurium, Enterococcus faecalis, Listeria monocytogenes and Escherichia coli) and its cytotoxicity on the normal cell line was investigated in vitro. The findings showed great antimicrobial function of the peptide. Minimum inhibition concentration and minimum bactericidal concentration of Lasioglossin ɪɪɪ on the pathogens were the range of 3.851-8.625 and 7.703-15.406, respectively. Results indicated Staphylococcus aureus showed the highest sensitivity to Lasioglossin ɪɪɪ. The Lassioglossin demonstrated cytotoxic effect on human embryonic kidney cells in higher concentration (1652 µg/ml) in comparison to MIC and MBC values. The results indicate that this peptide can be competed with common antibiotics in terms of the bactericidal properties.

https://jab.uk.ac.ir/article_2206_fea9c11c4ad9a395a636ed944a28b51a.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Identification of genetic variants in Arian line and investigation of their performance using whole genome sequencing Identification of genetic variants in Arian line and investigation of their performance using whole genome sequencing 45 60 2207 10.22103/jab.2018.2207 FA Hamed Kharrati Ph.D student, Institute of Biotechnology, School of Agriculture, Shiraz University, Shiraz, Iran. Esmael Ebrahimi Associate professor, Institute of Biotechnology, School of Agriculture, Shiraz University, Shiraz, Iran. Mohammad Dadpasand Associate professor, Department of Animal science, School of Agriculture, Shiraz University, Shiraz, Iran. Ali Niyazi Professor, Institute of Biotechnology, School of Agriculture, Shiraz University, Shiraz, Iran.. Ali Esmaelizadeh Professor, Department of Animal science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. Journal Article 2018 04 15 Objective This is the first study for discovering genetic variants in Arian line by whole genome sequencing data. The study of Arian line features at genomic level can unravel the genetic background of economic traits such as growth. Materials and methods In this study, the information of three whole genomes of Arian chickens was used. Results The results of this study indicated that, there were 63866 common variants among samples. 17604 variants Out of 63866 variants were reported as novel variants in this study. Also 604 variants are located in coding regions, which 204 out of them lead to change in the amino acid sequence of the proteins. Considering the importance of dietary protein quality for growth and weight gain for poultry, the results of gene ontology analysis indicated that the pathways of protein catabolism and post-translational modification of proteins, related to growth are considerable. Also candidate genes such as, SMURF2 and SUMO1 was suggested for these biological pathways. Another interesting result of gene ontology was the significance of response to stress pathways. Conclusions In the Arian line due to intense selection, the existence of stress is inevitable. Therefore, the significance of stress response can indicate the importance of these pathways in relation to the desired performance. Objective This is the first study for discovering genetic variants in Arian line by whole genome sequencing data. The study of Arian line features at genomic level can unravel the genetic background of economic traits such as growth. Materials and methods In this study, the information of three whole genomes of Arian chickens was used. Results The results of this study indicated that, there were 63866 common variants among samples. 17604 variants Out of 63866 variants were reported as novel variants in this study. Also 604 variants are located in coding regions, which 204 out of them lead to change in the amino acid sequence of the proteins. Considering the importance of dietary protein quality for growth and weight gain for poultry, the results of gene ontology analysis indicated that the pathways of protein catabolism and post-translational modification of proteins, related to growth are considerable. Also candidate genes such as, SMURF2 and SUMO1 was suggested for these biological pathways. Another interesting result of gene ontology was the significance of response to stress pathways. Conclusions In the Arian line due to intense selection, the existence of stress is inevitable. Therefore, the significance of stress response can indicate the importance of these pathways in relation to the desired performance.

https://jab.uk.ac.ir/article_2207_4aecfbe5d21e3f7912bf8eb29124423a.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Increasing expression of the PR1 gene in quince (Cydonia oblanga) by application of Bion elicitor Increasing expression of the PR1 gene in quince (Cydonia oblanga) by application of Bion elicitor 61 72 2208 10.22103/jab.2018.2208 FA Nasim Sarhangi Graduated M.Sc. Student, Payame Nour Univ., Karaj, Iran. Mansooreh Keshavarzi Associate Professor, Horticultural Research Institute, Cold and Temperate Center, Agricultural Research, Education and Extension Organization, Karaj, Iran. Ali Mohamad Shakib Associate Professor, Agricultural Biotechnology Research institute, Agricultural Research, Education and Extension Organization, Karaj Mohammadali Ebrahimi Associate Professor, Payame Nour Univiversity, Karaj, Iran. Journal Article 2018 02 17 One of the safe methods for fire blight disease management in quince plant (Cydonia oblanga) is through activating plant indigenous defense systems using defense elicitors such as Bion (Benzothiadiazole). Investigating the effect of PR1 defense genes group in induced defense mechanism by Bion in quince was the purpose of this study. For this, RNA was extracted from leaf tissues of bion-sprayed (400 mg/L) and water sprayed (control) plants was used for cDNA synthesis. First, a 750 bp fragment of the actin gene from quince, as a house keeping gene, was cloned and sequenced. The result of sequencing showed it has 99% homology with appleactin gene. Specific primers for quince actin gene were designed and used to study changes in PR1 gene expression in bion-treated comparing control plants using real-time PCR method. Based on the results, Bion treatment caused 10 times increase in the PR1 gene expression. Based of result, the effect of Bion in relative control of fire blight in quince can partly be related to PR1 defense proteins activities. One of the safe methods for fire blight disease management in quince plant (Cydonia oblanga) is through activating plant indigenous defense systems using defense elicitors such as Bion (Benzothiadiazole). Investigating the effect of PR1 defense genes group in induced defense mechanism by Bion in quince was the purpose of this study. For this, RNA was extracted from leaf tissues of bion-sprayed (400 mg/L) and water sprayed (control) plants was used for cDNA synthesis. First, a 750 bp fragment of the actin gene from quince, as a house keeping gene, was cloned and sequenced. The result of sequencing showed it has 99% homology with appleactin gene. Specific primers for quince actin gene were designed and used to study changes in PR1 gene expression in bion-treated comparing control plants using real-time PCR method. Based on the results, Bion treatment caused 10 times increase in the PR1 gene expression. Based of result, the effect of Bion in relative control of fire blight in quince can partly be related to PR1 defense proteins activities.

https://jab.uk.ac.ir/article_2208_cd3afef9b8b89558cd56638c3631868a.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Effect of zedo gum - Zataria multiflora essential oil nanoemulsion in the inhibition of free radical intercellular and expression of the NAD(P)H oxidase gene in wheat Effect of zedo gum - Zataria multiflora essential oil nanoemulsion in the inhibition of free radical intercellular and expression of the NAD(P)H oxidase gene in wheat 62 86 2209 10.22103/jab.2018.2209 FA Fatemeh Sohrabi Institute of Biotechnology, Shiraz University Gholamreza Kavoosi Associate Professor, Institute of Biotechnology, Faculty of Agriculture, Shiraz University, Shiraz, Iran. Journal Article 2018 04 16 The production of reactive oxygen species, such as hydrogen peroxide is inevitable in aerobic metabolism. Since, these compounds are effective at different concentration in a variety of processes in the cell and their excessive production lead to oxidative stresses, Therefore, their concentration must be controlled. In this study, Zataria multiflora essential oil with abundant phenolic compounds (thymol, carvacrol), and zedo gum polymers were used to encapsulate nanoemulsion production. The effect of zedo gum - Zataria multiflora essential oil nanoemulsion were investigated in the inhibition of ABTS (2,2-azinobis-3-ethylbenzothiazolin-6-sulfonic acid), hydrogen peroxide free radicals intercellular and expression of the (NOX:NAD(P)H oxidase) gene with accession number AY561153 (the largest producer of reactive oxygen species). Nanoemulsion with 88 mg/gr phenol content showed a high ability to inhibit the free radical ABTS and hydrogen peroxide. On the other hand, at concentration of 25 and 50 µg/ml increased the expression of NOX and at concentration of 100, 150 and 200 µg/ml decreased it. Our results showed that, different concentration of nanoemulsion have different effect on reactive oxygen species and, therefore, they have the ability to control oxidative stresses. The production of reactive oxygen species, such as hydrogen peroxide is inevitable in aerobic metabolism. Since, these compounds are effective at different concentration in a variety of processes in the cell and their excessive production lead to oxidative stresses, Therefore, their concentration must be controlled. In this study, Zataria multiflora essential oil with abundant phenolic compounds (thymol, carvacrol), and zedo gum polymers were used to encapsulate nanoemulsion production. The effect of zedo gum - Zataria multiflora essential oil nanoemulsion were investigated in the inhibition of ABTS (2,2-azinobis-3-ethylbenzothiazolin-6-sulfonic acid), hydrogen peroxide free radicals intercellular and expression of the (NOX:NAD(P)H oxidase) gene with accession number AY561153 (the largest producer of reactive oxygen species). Nanoemulsion with 88 mg/gr phenol content showed a high ability to inhibit the free radical ABTS and hydrogen peroxide. On the other hand, at concentration of 25 and 50 µg/ml increased the expression of NOX and at concentration of 100, 150 and 200 µg/ml decreased it. Our results showed that, different concentration of nanoemulsion have different effect on reactive oxygen species and, therefore, they have the ability to control oxidative stresses.

https://jab.uk.ac.ir/article_2209_db116b39f7a3ac5366079b1d9fe249a5.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Transcription activator-like effectors (TALEs) role in plant response to Xanthomonas Transcription activator-like effectors (TALEs) role in plant response to Xanthomonas 87 110 2210 10.22103/jab.2018.2210 FA Masood Shamsbakhsh Professor Associate, Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran Nargues Falahi Charkhabi Entomology and Plant Pathology, Aburaihan Campus, University of Tehran, Pakdasht, Tehran Heshmatollah Rahimian Professor, Department of Plant Protection, Sari University of Agricultural Sciences and Natural Resources, Sari, Iran. Journal Article 2018 08 30 Xanthomonas is an important plant pathogenic bacteria and cause economic losses in a wide range of crop plants. The type III secretion system as a crucial pathogenicity factor is used to inject effector proteins into the host cell. Transcription activator like effectors (TALEs), known in the most Xanthomonas species, are DNA-binding proteins which determine solely the outcome of plant-pathogen interaction. Because of determinative nature of TALE-target gene interaction, the knowledge of pathogen TALome diversity, forecasting susceptibility genes, genome editing using new developed methods have provided novel strategies for management of plant diseases. This article reviews TALEs structure, their role in pathogenicity and resistance, as well development of resistant plants to Xanthomonas using TALome derived results. Xanthomonas is an important plant pathogenic bacteria and cause economic losses in a wide range of crop plants. The type III secretion system as a crucial pathogenicity factor is used to inject effector proteins into the host cell. Transcription activator like effectors (TALEs), known in the most Xanthomonas species, are DNA-binding proteins which determine solely the outcome of plant-pathogen interaction. Because of determinative nature of TALE-target gene interaction, the knowledge of pathogen TALome diversity, forecasting susceptibility genes, genome editing using new developed methods have provided novel strategies for management of plant diseases. This article reviews TALEs structure, their role in pathogenicity and resistance, as well development of resistant plants to Xanthomonas using TALome derived results.

https://jab.uk.ac.ir/article_2210_1368ba1ab6ed38bb1f26f36673739d54.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Studying expression of leptin gene in different tissues of Kermani Sheep using Real Time PCR Studying expression of leptin gene in different tissues of Kermani Sheep using Real Time PCR 111 123 2211 FA Mohammadreza Mohammadabadi Department of Animal Science, Faculty of Agriculture, College of Agriculture, Shahid Bahonar University of Kerman 0000-0002-1268-3043 Mahboobeh Kord MSc Student, Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran. Mahmood Nazari Assistant Professor, Department of Animal Science, Ramin Agricultural & Natural Resources University of Khuzestan, Ahvaz, Iran. 0000-0003-4417-4654 Journal Article 2018 09 01 Objective Leptin is produced by white adipose tissue and plays an important role in regulation of feed intake, energy balance, fertility and immune functions. Thus, the aim of this research was to study leptin gene expression in adipose tissue, liver, kidney, lung and heart of Kermani sheep. Materials and methods Tissue samples obtained from 6 animals and RNA was extracted. Extracted RNA were immediately stored at -80°C. Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 1.5% agarose gel and were evaluated different levels of expression in studied different tissues. Results Results showed that the leptin gene was expressed in all the tested tissues and the highest level of expression was observed in adipose tissue and liver and the lowest level was detected in heart. Conclusions These results may show that leptin plays a particular role in fat metabolism. Further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions. Objective Leptin is produced by white adipose tissue and plays an important role in regulation of feed intake, energy balance, fertility and immune functions. Thus, the aim of this research was to study leptin gene expression in adipose tissue, liver, kidney, lung and heart of Kermani sheep. Materials and methods Tissue samples obtained from 6 animals and RNA was extracted. Extracted RNA were immediately stored at -80°C. Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 1.5% agarose gel and were evaluated different levels of expression in studied different tissues. Results Results showed that the leptin gene was expressed in all the tested tissues and the highest level of expression was observed in adipose tissue and liver and the lowest level was detected in heart. Conclusions These results may show that leptin plays a particular role in fat metabolism. Further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions.

https://jab.uk.ac.ir/article_2211_3323fe11e9595c09af38fe67567a9394.pdf

Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 10 3 2018 12 22 Identification and molecular characterization of the phytoplasma associated with leaf stunting and yellowing disease in Razavi and North Khorasan provinces Identification and molecular characterization of the phytoplasma associated with leaf stunting and yellowing disease in Razavi and North Khorasan provinces 124 131 2212 10.22103/jab.2018.2212 FA Saeed Tarighi Dept. Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad Mojtaba Dehghan Ph.D. Student of plant pathology, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran. Ebadollah Ebadi Ph.D. Student of plant pathology, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran. Journal Article 2018 07 03 In 2017, leaf stunting and yellowing were observed on twigs and branches of stone fruit trees in production regions in Razavi and North Khorasan provinces of Iran. For diagnosis of disease and identification of the causal agent, symptomatic leaf samples were collected in these orchards and were carried to the laboratory. The disease agent was successfully transmitted by grafting from naturally symptomatic samples to peach (Prunus persica) seedlings. Total DNA was extracted from plant samples and indexed by nested PCR using phytoplasma generic primers, P1/P7 and R16F2n/R2. PCR products were sequenced and the nucleotide sequences were compared with those of other phytoplasmas in GenBank and analyzed by virtual restriction fragment length polymorphism (RFLP). The phytoplasmas associated with leaf stunting and yellowing disease in Razavi and North Khorasan provinces were identiﬁed as members of the 16SrIX group or Pigeon pea witches broom (Candidatus Phytoplasma phoenicium). This is the first report of leaf stunting and yellowing disease and characterization of associated phytoplasma in these regions. In 2017, leaf stunting and yellowing were observed on twigs and branches of stone fruit trees in production regions in Razavi and North Khorasan provinces of Iran. For diagnosis of disease and identification of the causal agent, symptomatic leaf samples were collected in these orchards and were carried to the laboratory. The disease agent was successfully transmitted by grafting from naturally symptomatic samples to peach (Prunus persica) seedlings. Total DNA was extracted from plant samples and indexed by nested PCR using phytoplasma generic primers, P1/P7 and R16F2n/R2. PCR products were sequenced and the nucleotide sequences were compared with those of other phytoplasmas in GenBank and analyzed by virtual restriction fragment length polymorphism (RFLP). The phytoplasmas associated with leaf stunting and yellowing disease in Razavi and North Khorasan provinces were identiﬁed as members of the 16SrIX group or Pigeon pea witches broom (Candidatus Phytoplasma phoenicium). This is the first report of leaf stunting and yellowing disease and characterization of associated phytoplasma in these regions.

https://jab.uk.ac.ir/article_2212_05a70454516ecd9194c293b0e415777f.pdf