3 Biotech

, 9:177 | Cite as

Development and utilization of gyrA and gyrB gene-based diagnostics for the phytoplasma classified under 16Sr I group in plants and insects

  • Madhupriya
  • Aundy Kumar
  • G. P. RaoEmail author
  • S. M. P. Khurana
Short Reports


In the present study, a new set of primers of gyrA and gyrB genes of the phytoplasma genome were designed and validated for the successful detection and taxonomic classification of the previously identified phytoplasma strains of ‘Candidatus P. asteris’ (16SrI-B subgroup) associated with Catharanthus leaf yellows, sesame phyllody and the leafhopper (Hishimonus phycitis). Our results suggested the ability and sensitivity of gyrA and gyrB genes as an alternative molecular marker to identify the Ca. P. asteris strain up to subgroup level associated both with plants and insects.


Multilocus genes Validation Phytoplasma detection Leafhopper 



The authors wish to express sincere thanks to the Head, Division of Plant Pathology and the Director, Indian Agricultural Research Institute for providing laboratory facilities. Funding was provided by Department of Biotechnology, Ministry of Science and Technology (Grant no: 24/468).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ahrens U, Seemüller E (1992) Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerasechain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828–832CrossRefGoogle Scholar
  2. Bertaccini A, Duduk B, Paltrinieri S, Contaldo N (2014) Phytoplasmas and phytoplasma diseases: a severe threat to agriculture. Am J Plant Sci 5:46–62CrossRefGoogle Scholar
  3. Hodgetts J, Dickinson M (2010) Phytoplasma phylogeny and detection based on genes other than 16S rRNA. In: Weintraub PG, Jones P (eds) Phytoplasmas genomes, plant hosts and vectors. CAB International, Wallingford, pp 93–113Google Scholar
  4. Hodgetts J, Boonham N, Mumford R, Harrison N, Dickinson M (2008) Phytoplasma phylogenetics based on analysis of secA and 23S rRNA gene sequences for improved resolution of a candidate species of ‘Candidatus Phytoplasma’. Int J Syst Evol Microbiol 58:1826–1837CrossRefGoogle Scholar
  5. Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33(7):1870–1874CrossRefGoogle Scholar
  6. Langer M, Maixner M (2004) Molecular characterization of grapevine yellows associated phytoplasmas of the stolbur group based on RFLP-analysis of non-ribosomal DNA. Vitis 43:191–200Google Scholar
  7. Lee IM, Zhao Y, Davis RE, Wei W, Martini M (2007) Prospects of DNA-based systems for differentiation and classification of phytoplasmas. Bull Insectol 60(2):239–244Google Scholar
  8. Madhupriya (2016) Molecular characterization of phytoplasmas associated with important ornamental plant species in Northern India. Ph.D. Thesis, Amity University, Haryana, p 293Google Scholar
  9. Madhupriya, Tiwari AK, Rao GP (2015) Utilization of dnaB gene for characterization of phytoplasmas associated with toria, brinjal and Phlox in India. Phytopathogenic Mollicutes 5:11–12Google Scholar
  10. Martini M, Lee IM, Bottner KD, Zhao Y, Botti S, Bertaccini A, Harrison NA, Carraro L, Marcone C, Khan AJ, Osler R (2007) Ribosomal protein gene-based phylogeny for finer differentiation and classification of phytoplasmas. Int J Syst Evol Microbiol 57(9):2037–2051CrossRefGoogle Scholar
  11. Marzachi C, Verati F, Bosco D (1998) Direct PCR detection of phytoplasmas in experimentally infected insects. Ann Appl Biol 133:45–54CrossRefGoogle Scholar
  12. Schneider B, Gibb KS, Seeümller E (1997) Sequence and RFLP analysis of the elongation factor Tuf gene used in differentiation and classification of phytoplasmas. Microbiology 143:3381–3389CrossRefGoogle Scholar
  13. Yu S, Li Y, Ren Z, Song C, Lin C, Piao C, Tian G (2017) Multilocus sequence analysis for revealing finer genetic variation and phylogenetic interrelatedness of phytoplasma strains in 16SrI group in China. Sci Silv Sin 53(3):105–118Google Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.Division of Plant PathologyICAR-Indian Agricultural Research InstituteNew DelhiIndia
  2. 2.Amity Institute of BiotechnologyAmity UniversityManesarIndia

Personalised recommendations