Esmailzadeh Hosseini, S., Khodakaramian, G., Salehi, M., Bertaccini, A. (2016). Characterization of 16SrII group phytoplasmas associated with alfalfa (Medicago sativa) witches’ broom disease in diverse areas of Iran. Journal of Crop Protection, 5(4), 581-590. doi: http://doi.org/10.18869/modares.jcp.5.4.581
Seyyed Alireza Esmailzadeh Hosseini; Gholam Khodakaramian; Mohammad Salehi; Assunta Bertaccini. "Characterization of 16SrII group phytoplasmas associated with alfalfa (Medicago sativa) witches’ broom disease in diverse areas of Iran". Journal of Crop Protection, 5, 4, 2016, 581-590. doi: http://doi.org/10.18869/modares.jcp.5.4.581
Esmailzadeh Hosseini, S., Khodakaramian, G., Salehi, M., Bertaccini, A. (2016). 'Characterization of 16SrII group phytoplasmas associated with alfalfa (Medicago sativa) witches’ broom disease in diverse areas of Iran', Journal of Crop Protection, 5(4), pp. 581-590. doi: http://doi.org/10.18869/modares.jcp.5.4.581
Esmailzadeh Hosseini, S., Khodakaramian, G., Salehi, M., Bertaccini, A. Characterization of 16SrII group phytoplasmas associated with alfalfa (Medicago sativa) witches’ broom disease in diverse areas of Iran. Journal of Crop Protection, 2016; 5(4): 581-590. doi: http://doi.org/10.18869/modares.jcp.5.4.581
Characterization of 16SrII group phytoplasmas associated with alfalfa (Medicago sativa) witches’ broom disease in diverse areas of Iran
1Plant Protection Department, Bu-Ali Sina University, Hamedan, Iran, Plant Protection Research Department, Yazd Agricultural and Natural Resources Research and Education Center, AREEO, Yazd, Iran.
2Plant Protection Department, Bu-Ali Sina University, Hamedan, Iran.
3Plant Protection Research Department, Fars Agricultural and Natural Resources Research and Education Center, AREEO, Zarghan, Iran.
4Department of Agricultural Sciences, Alma Mater Studiorum, University of Bologna, Italy.
Abstract
Alfalfa witches’ broom (AWB) is one of the most important alfalfa diseases in Iran. To characterize 16SrII group phytoplasmas associated with this disease, symptomatic and asymptomatic plants were collected during 2013-2015 and subjected to direct and nested polymerase chain reaction (PCR) using P1/P7, R16mF2/R16mR2 and R16F2n/R16R2. PCR amplicons of ~1.8, ~1.4 and ~1.25 kb respectively, were obtained only from all symptomatic plants. Restriction fragment length polymorphism (RFLP) analysis of R16F2n/R2 amplicons showed that the phytoplasma associated with AWB disease were members of 16SrII group subgroups 16SrII-D and -C. Blast analysis of these amplicon sequences and sequence homology of collected strains and strain sequences retrived from GenBank (AWB strains Chahgeer, Juyom and Bushehr) confirmed that AWB phytoplasmas collected from Bafg, Ardakan, Bahabad and Herat (Yazd province), Nikshahr (Sistan-Baluchestan), Bam, Zarand, Jiroft (Kerman province), Bushehr (Bushehr province), Tabas (South Khorasan province), Jowkar (Hamedan province) and Zardenjan (Esfahan province) cluster with phytoplasma strains enclosed in the 16SrII-D subgroup, while AWB strains from Chahgeer (Yazd province) and Juyom (Fars province) cluster with phytoplasma strains in the 16SrII-C subgroup. Based on these results the predominant strains of 16SrII phytoplasmas associated with AWB disease in Iran were classified in the 16SrII-D subgroup. In Ashkezar and Abarkouh in Yazd province entire alfalfa farm was infected with witches’ broom disease. In 3 year alfalfa stands in Ashkezar alfalfa farms were plowed due to high incidence of the disease.
Conci, L., Meneguzzi, N., Galdeano, E., Torres, L., Nome, C., Nome, S. 2005. Detection and molecular characterization of an alfalfa phytoplasma in Argentina that represents a new subgroup in the 16Sr RNA Ash yellows group (‘Candidatus phytoplasma fraxini’). European Journal of Plant Pathology, 113: 255-265.
Deng, S. J. and Hiruki, C. 1991. Amplification of 16S ribosomal RNA genes from culturable and nonculturable mollicutes. Journal of Microbiological Methods, 14: 53-61.
Esmailzadeh Hosseini S. A., Salehi, M., Khanchezar, A. and Shamszadeh, M. 2011a. The first report of a phytoplasma associated with pot marigold phyllody in Iran. Bulletin of Insectology, 64 (Suppl): S109-S110.
Esmailzadeh Hosseini, S. A., Khodakaramian, G., Salehi, M. and Bertaccini, A. 2016a. First report of 16SrVI-A and 16SrXII-A phytoplasmas associated with alfalfa witches’ broom disease in Iran. Journal of Plant Pathology 98 (2): 174.
Esmailzadeh Hosseini, S. A., Khodakaramian, G., Salehi, M. and Bertaccini, A. 2016b. Molecular identification and phylogenetic analysis of phytoplasmas associated with alfalfa witches’ broom diseases in the western areas of Iran. Phytopathogenic Mollicutes 6 (1): 16-22.
Esmailzadeh Hosseini, S. A., Khodakaramian, G., Salehi, M., Fani S. R., Mirchenari, S. M., Salehi, E. and Bertaccini, A. 2015b. Incidence, distribution and economic importance of alfalfa witches’ broom disease in Sistan-Baluchestan (Iran) and characterization of associated phytoplasma. Phytopathogenic Mollicutes, 5 (2): 84-90.
Esmailzadeh Hosseini, S. A., Khodakaramian, G., Salehi, M., Fani, S. R., Bolok Yazdi, H. R., Raoufi, D., Jadidi, O. and Bertaccini, A. 2015a. Status of alfalfa witches’ broom phytoplasma disease in Iran. Phytopathogenic Mollicutes, 5 (1-Suppl): 65-66.
Esmailzadeh Hosseini, S. A., Mirzaie, A., Jafari-Nodooshan, A. and Rahimian, H. 2007. The first report of transmission of a phytoplasma associated with sesame phyllody by Orosius albicinctus in Iran. Australsian Plant Diseasae Notes, 2 (1): 33-34.
Esmailzadeh Hosseini, S. A., Salehi, M. and Mirzaie, A. 2011b. Alternate hosts of alfalfa witches’ broom phytoplasma and winter hosts of its vector Orosius albicinctus in Yazd-Iran. Bulletin of Insectology, 64 (Suppl): S247-S248.
Esmailzadeh Hosseini, S. A., Salehi, M., Khodakaramian, G., Mirchenary, S. M. and Bertaccini, A. 2015c. An up to date status of alfalfa witches’ broom disease in Iran. Phytopathogenic Mollicutes, 5 (1): 9-18.
FAOSTAT, 2013. Food and Agriculture Organization of the United Nations, http:// FAOSTAT3. fao. org.
Getachew, M. A., Mitchell, A., Gurr, G. M., Fletcher, M. J., Pilkington, L. J., Nikandrow, A. and Elliott, E. 2007. First report of a ‘Candidatus Phytoplasma australiense’-related strain in lucerne (Medicago sativa) in Australasian Plant Disease Notes, 111: 91.
Gundersen, D. E. and Lee, I. M. 1996. Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopath Medit 35: 114-151.
Haskell RJ 1926. Diseases of cereal crops and forage crops in in the United States in 1925. Plant Disease Report Suppl, 48: 367.
Jomantiene, R., Davis, R. E., Antoniuk, L. and Staniulis, J. 2000. First report of a phytoplasma in soybean, alfalfa and Lupinus sp. in Lithuania. Plant Disease, 84: 198-199.
Jones, P., Arocha, Y. and Plata, G. 2005. First report of ‘Candidatus Phytoplasma asteris’ associated isolate with witches’ broom disease of alfalfa in Bolivia. Plant Pathology, 54: 559.
Khadhair, A. H., Hiruki, C. H. and Wang, S. 1997. Molecular detection of alfalfa witches’ broom phytoplasma in four leafhopper species associated with infected alfalfa plants. Microbiological Research, 152: 269-275.
Khan, A. J., Botti, S., Alsubhi, A. M., Gundersen-Rindal, D. E. and Bertaccini, A. 2002. Molecular identification of a new phytoplasma associated with alfalfa witches’ broom in Oman. Phytopathology, 92: 1038-1047.
Lee, I. M., Gundersen, D. E., Hammond, R. W., Davis, R. E. and Bartoszyk, I. M. 1998. Revised classification scheme for phytoplasma based on RFLP analysis of 16S rRNA and ribosomal protein gene sequences. International Journal of Systematic Bacteriology, 48: 1153-1169.
Li, Z. N., Zhang, L., Man, J. Y. and Wu, Y. F. 2012. Detection and identification of elm yellows group phytoplasma (16SrV) associated with alfalfa witches’ broom disease. Journal of Phytopathology, 160: 311-313.
Marzachì, C., Veratti, F., d’Aquilio, M., Vischi, A., Conti, M. and Boccardo, G. 2000. Molecular hybridization and PCR amplification of nonribosomal DNA to detect and differentiate stolbur phytoplasma isolates from Italy. Journal of Plant Pathology, 82: 201-212.
Mirzaie, A., Esmailzadeh Hosseini, S. A., Jafari-Nodooshan, A. and Rahimian, H. 2007. Molecular characterization and potential insect vector of a phytoplasma associated with garden beet witches’ broom in Yazd, Iran. Journal of Phytopathology, 155 (4): 198-203.
Peters, R. D., Lee, I. M., Grau, C. R., Driscoll, S. J., Winberg, R. M. and Kutzweil, N. C. 1999. First report of aster yellows phytoplasma in alfalfa. Plant Disease, 83: 488.
Raoofi, D. and Salehi, M. 2012. Partial characterization of a phytoplasma associated with alfalfa witches’ broom in Bushehr province. Iranian Journal of Plant Pathology, 48 (3): 423-431.
Rashidi, M., Ghosta, Y. and Bahar, M. 2010. Molecular identification of a phytoplasma associated with Russian olive witches’ broom in Iran. European Journal of Plant Pathology, 127: 157-159.
Salehi, M. and Izadpanah, K. 1993a. A comparative study of transmission, host range and symptomatology of sesame phyllody and alfalfa witches’ broom. Iranian Journal of Plant Pathology,29: 157-158.
Salehi, M. and Izadpanah, K. 1993b. Etiology of alfalfa witches’ broom disease in Iran. Iranian Journal of Plant Pathology,29:92-93.
Salehi, M., Esmailzadeh Hosseini, S. A. and Salehi, E. 2015. Characterisation of a phytoplasma associated with sunflower phyllody in Fars, Isfahan and Yazd provinces of Iran. New Disease Reports, 3: 6.
Salehi, M., Izadpanah, K. and Ebrahimnesbat, F. 1995. Etiology, transmission and host range of alfalfa witches’ broom in Iran. Iranian Journal of Plant Pathology, 31 (1-4): 1-9.
Schneider, B., Seemüller, E., Smart, C. D. And Kirkpatrick, B. C. 1995. Phylogenetic classification of plant pathogenic mycoplasmalike organisms or phytoplasmas. In S. Razin & J. G. Tully (Eds.), Molecular and diagnostic procedures in mycoplasmology (pp. 369–380). San Diego, CA: Academic.
Smarz, J., Ulrychova, M. and Jokes, M. 1981. Alfalfa witches' broom in Czechoslovakia and demonstration of mycoplasma-like organism in the affected plants. Biologia Plantarum,23: 381-383.
Starović, M., Kuzmanović, S., Gavrilović, V., Aleksić, G., Popović, T., Stojanović, S. and Jošić, D. 2012. Detection and identification of two phytoplasmas (16SrIII-B and 16SrXII-A) from alfalfa (Medicago sativa) in Serbia. Journal of Phytopathology, 160: 758-760.
Tamura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar, S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30: 2725-2729.
Zhang, Y. P., Uyemoto, J. K. and Kirkpatrick, B. C. 1998. A small-scale procedure for extracting nucleic acids from woody plants infected with various phytoplasmas for PCR assay. Journal of Virological Methodes, 71: 45-50.
Zhao, Y., Wei, W., Lee, I. M., Shao, J., Suo, X. and Davis, R. E. 2009.) Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII). International Journal of Systematic and Evolutionary Microbiology, 59: 2582-2593.
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