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Multilocus genotyping of ‘Candidatus Phytoplasma aurantifolia’ associated with witches’ broom disease in Citrus spp.

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Abstract

Witches’ broom disease of lime (WBDL), associated with ‘Candidatus Phytoplasma aurantifolia,’ is a serious disease of acid lime in Southern Iran. In order to characterize ‘Ca. P. aurantifolia’ strains, a multilocus sequence analysis protocol was developed based on different genomic loci, including 16S rRNA, 16S-23S rRNA intergenic spacer (IS), ribosomal protein genes rpl22 and rps3, cell division protein, and group II intron reverse transcriptase/mature gene, in 46 phytoplasma strains. Analyses of virtual RFLP, phylogenetics, and DNA sequence identity of 16S rRNA of Iranian phytoplasma strains associated with symptomatic citrus plants indicated that the Iranian strains were classified in ribosomal subgroup 16SrII-B. In total, three genotypes were recognized based on 16Sr virtual RFLP profiles. Phylogenetic analysis based on the rp and ftsH gene sequences showed clustering of all the strains as one group. Comparative sequence and phylogenetic analysis of group II intron genes indicated the highest differentiation of ‘Ca. P. aurantifolia’–related strains in four genotypes. The polymorphism of group II intron reverse transcriptase/mature genes fragmentation provides finer differentiation of closely related ‘Ca P. aurantifolia’ strains and reveals higher genetic diversity than previously reported.

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Acknowledgments

The authors would like to express their gratitude to Tim Dumonceaux (Agriculture and Agri-Food Canada), Xavier Foissac (University of Bordeaux), and Jaime Cubero Dabrio (The National Institute for Agricultural and Food Research and Technology, Spain) for critical commentary and editing prior to submission.

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Authors and Affiliations

  1. Plant Protection Department, Agriculture faculty, Tabriz University, Tabriz, Iran

    Sina Noorizadeh & Reza Khakvar

  2. Horticultural Science Research Institute, Citrus and Subtropical Fruits Research Center, Agricultural Research Education and Extension Organization (AREEO), Ramsar, Iran

    Morteza Golmohammadi & Seyed Mehdi Bani Hashemian

  3. Plant Protection Research Department, Fars Agricultural and Natural Resources Research and Education Center, Agricultural Research Education and Extension Organization (AREEO), Zarghan, Iran

    Mohammad Mehdi Faghihi

Authors
  1. Sina Noorizadeh
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  2. Reza Khakvar
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  3. Morteza Golmohammadi
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  4. Seyed Mehdi Bani Hashemian
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  5. Mohammad Mehdi Faghihi
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Contributions

The first author (Sina Noorizadeh) carried out all the experiments as his PhD thesis. Reza Khakvar and Morteza Golmohammadi were supervisor and co-supervisor for this thesis, who allowed Mr. Noorizadeh to work in their lab and use equipment and chemicals; and Seyed Mehdi Bani Hashemian and Mohammad Mehdi Faghihi were advisors for this thesis, who helped Mr. Noorizadeh during this thesis and in writing the manuscript.

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Correspondence to Reza Khakvar or Morteza Golmohammadi.

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Noorizadeh, S., Khakvar, R., Golmohammadi, M. et al. Multilocus genotyping of ‘Candidatus Phytoplasma aurantifolia’ associated with witches’ broom disease in Citrus spp.. Trop. plant pathol. 46, 218–231 (2021). https://doi.org/10.1007/s40858-020-00404-2

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  • DOI: https://doi.org/10.1007/s40858-020-00404-2

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