Sugar Tech

, Volume 17, Issue 2, pp 156–161 | Cite as

Characterization of a 16SrI-B Subgroup Phytoplasma Associated with Sugarcane Leaf Yellows Disease in India

  • Shailender Kumar
  • Ajay K. Tiwari
  • Somnath K. Holkar
  • S. K. Duttamajumder
  • G. P. RaoEmail author
Research Article


Leaf mid-rib yellowing along with yellowing of adjoining lamina and development of irregular yellow patches on leaf lamina was observed on two sugarcane varieties (CoLk 94184, CoSe 92423) during 2013–2014 crop season. Phytoplasma association with leaf yellows and leaf lamina with yellow patches symptomatic samples was confirmed through PCR and nested PCR assays using phytoplasma specific primers P1/P6 followed by R16F2n/R16R2. Sequencing and BLASTn analysis of ~1.2 kb product of 16Sr RNA gene fragments of the two phytoplasmas shared 99 and 100 % sequence identity with strains of ‘Candidatus Phytoplasma asteris’ (16SrI). Further characterization through in silico restriction enzyme digestion of 16Sr DNA using iPhyClassifier online tool assigned both the strains into subgroup 16SrI-B and phylogenetic analyses confirmed that both the phytoplasmas were closely related to 16SrI group. This is the first report of association of 16SrI-B subgroup phytoplasma with sugarcane leaf yellows disease in India.


Sugarcane leaf yellows phytoplasma Candidatus Phytoplasma asteris’ Phylogeny 16SrI-B YLD YLS SCLP 



The authors are thankful to the Department of Science and Technology, New Delhi, India, for providing financial assistance during the course of the study. The authors wish to express sincere thanks to the Head, Division of Plant Pathology, Indian Agricultural Research Institute, for providing laboratory facilities. The help rendered by Director, Indian Institute of Sugarcane Research, Lucknow, India during field visit is sincerely acknowledged.


  1. Abu Ahmad, Y., J.C. Girard, and E. Fernandez. 2007. Variation in virus populations and growth characteristics of two sugarcane cultivars naturally infected by Sugarcane yellow leaf virus in different geographical locations. Plant Pathology 56: 743–754.CrossRefGoogle Scholar
  2. Ahrens, U., and E. Seemüller. 1992. Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82: 828–832.CrossRefGoogle Scholar
  3. Aljanabi, S.M., Y. Parmessur, Y. Moutia, S. Saumtally, and A. Dookun. 2001. Further evidence of the association of a phytoplasma and a virus with yellow leaf syndrome in sugarcane. Plant Pathology 50: 628–636.CrossRefGoogle Scholar
  4. Arocha, Y., L. Gonc, E.L. Peralta, and P. Jones. 1999. First report of virus and phytoplasma pathogens associated with yellow leaf syndrome of sugarcane in Cuba. Plant Disease 83: 1177.CrossRefGoogle Scholar
  5. Arocha, Y., P. Jones, I. Sumac, and E.L. Peralta. 2000. Detection of phytoplasmas associated with yellow leaf syndrome in Cuba. Revista de Protección Vegetal 15: 81–87.Google Scholar
  6. Azadvar, M., V.K. Baranwal, and D.K. Yadav. 2009. First report of a 16SrIX (Pigeon pea witches’ broom) phytoplasma associated with toria (Brassica rapa cv. toria) phyllody disease in India. New Disease Reports 20: 27.Google Scholar
  7. Bailey, R., G. Bechet, and P. Cronjé. 1996. Notes on the occurrence of yellow leaf syndrome of sugarcane in southern Africa. Proceedings of the South African Sugar Technologists Association 70: 3–6.Google Scholar
  8. Comstock, J., M. Irey, B. Lockhart, and Z. Wang. 1998. Incidence of yellow leaf syndrome in CP cultivars based on polymerase chain reaction and serological techniques. Sugar Cane 4: 21–24.Google Scholar
  9. Cronje, C.P.R., and R.A. Bailey. 1999. Association of phytoplasmas with yellow leaf syndrome of sugarcane. Proceedings International Society for Sugarcane Technologists 23: 371–381.Google Scholar
  10. Cronjé, P., A. Tymon, P. Jones, and R. Bailey. 1998. Association of a phytoplasma with a yellow leaf syndrome of sugarcane in Africa. Annals of Applied Biology 133: 177–186.CrossRefGoogle Scholar
  11. Deng, S., and C. Hiruki. 1991. Amplification of 16S rRNA genes from culturable and non-culturable Mollicutes. Journal of Microbiology Methods 14: 53–61.CrossRefGoogle Scholar
  12. Elsayed, A.I., and M. Boulila. 2014. Molecular identification and phylogenetic analysis of sugarcane yellow leaf phytoplasma (SCYLP) in Egypt. Journal of Phytopathology 162: 89–97.CrossRefGoogle Scholar
  13. Gaur, R., R. Raizada, and G.P. Rao. 2008. Sugarcane yellow leaf phytoplasma associated for the first time with sugarcane yellow leaf syndrome in India. New Disease Reports 16: 43.Google Scholar
  14. Grisham, M.P., Y.B. Pan, B.L. Legendre, M.A. Gidshall, and G. Eggleston. 2001. Effect of sugarcane yellow leaf syndrome on sugarcane yield and juice quality. Proceedings International Society for Sugarcane Technologists 24: 434–438.Google Scholar
  15. Gundersen, D.E., and I.-M. Lee. 1996. Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathol Mediterrenea 35: 144–151.Google Scholar
  16. Khan, J.A., S.K. Singh, and J. Ahmad. 2008. Characterization and phylogeny of a phytoplasma inducing sandal spike disease in sandal (Santalum album). Annales of Applied Biology 153: 365–372.CrossRefGoogle Scholar
  17. Kumar, S., and A.S. Byadgi. 2012. Molecular detection and characterization of phytoplasma associated with periwinkle. Vegetos 25(2): 127–135.Google Scholar
  18. Lee, I.-M., D.E. Gundersen, R.E. Davis, and I.M. Bartosyzk. 1998. Revised classification scheme of phytoplasma based on RFLP analysis of 16S rRNA and ribosomal protein gene sequences. International Journal of Systematic Bacteriology 48: 1153–1169.CrossRefGoogle Scholar
  19. Lockhart, B., and C.P.R. Cronje. 2000. Yellow leaf syndrome. In A Guide to sugarcane diseases, ed. P. Root, R.A. Bailey, J.C. Comstock, B.J. Croft, and A.S. Saumtally, 291–295. Montpellier: CIRAD.Google Scholar
  20. Marcone, C. 2002. Phytoplasma disease of sugarcane. Sugar Tech 4: 79–85.CrossRefGoogle Scholar
  21. Rao, G.P., S. Srivastava, P.S. Gupta, A. Singh, M. Singh, and C. Marcone. 2008. Detection of sugarcane grassy shoot phytoplasma infecting sugarcane in India and its phylogenetic relationships to closely related phytoplasmas. Sugar Tech 10: 74–80.CrossRefGoogle Scholar
  22. Rao, G.P., S. Mall, and C. Marcone. 2010. ‘Candidatus Phytoplasma cynodontis’ (16SrXIV group) affecting Oplismenus burmannii (Retz.) P. Beauv. And Digitaria sanguinalis (L.) Scop. in India. Australasian Plant Disease Notes 5: 93–95.CrossRefGoogle Scholar
  23. Rao, G.P., S. Mall, and C. Marcone. 2012. Recent biotechnological approaches in diagnosis and management of sugarcane phytoplasma diseases. In Functional plant science & biotechnology, eds. Rajarshi Kumar Gaur et al. 6, Special issue 2, 19–29. Recent trends in biotechnology and microbiology. USA: Global Science Books.Google Scholar
  24. Rao, G.P., Madhupriya., A.K. Tiwari, S. Kumar, and V.K. Baranwal. 2013. Identification of sugarcane grassy shoot-associated phytoplasma and one of its putative vectors in India. Phytoparasitica (in press). DOI  10.1007/s12600-013-0366-1.
  25. Schenck, S., J. Hu, and B. Lockhart. 1997. Use of a tissue blot immunoassay to determine the distribution of sugarcane yellow leaf virus in Hawaii. Sugar Cane 4: 5–8.Google Scholar
  26. Silva, E.G., I.P. Bedendo, M.V. Casagrande, and V.A. Moraes. 2009. Molecular identification and phylogenetic analysis of a group 16SrI-B phytoplasma associated with sugarcane yellow leaf syndrome in Brazil. Journal of Phytopathology 157: 771–774.CrossRefGoogle Scholar
  27. Soufi Z., S. Sakuanrungsirikul, T. Wongwarat, T. Hamarn, S. Srisink, and E. Komor. 2013. Sugarcane yellow leaf symptomatic plants in Thailand are infected by white leaf phytoplasma, not by leaf yellows phytoplasma. Australasian Plant Pathology 42: 723–729.Google Scholar
  28. Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar. 2011. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28: 2731–2739.CrossRefPubMedCentralPubMedGoogle Scholar
  29. Thompson, J.D., D.G. Higgins, and T.J. Gibson. 1994. CLUSTALW: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties, and weight matrix choice. Nucleic Acids Research 22: 4673–4680.CrossRefPubMedCentralPubMedGoogle Scholar
  30. Tran-Ngyuen, L., K.R. Blanche, and K.S. Gibb. 2000. Diversity of phytoplasma in northern Australian sugarcane and other grasses. Plant Disease 49: 666–679.Google Scholar
  31. Vega, J., S.M. Scagliusi, and E.C. Ulian. 1997. Sugarcane yellow leaf disease in Brazil: evidence of association with a luteovirus. Plant Disease 81: 21–26.CrossRefGoogle Scholar
  32. Viswanathan, R., M. Balamuralikrishnan, and R. Karuppaiah. 2008. Identification of three genotypes of sugarcane yellow leaf virus causing yellow leaf disease from India and their molecular characterization. Virus Genes 37: 368–379.CrossRefPubMedGoogle Scholar
  33. Viswanathan, R., C. Chinnaraja, R. Karuppaiah, K.V. Ganesh, R.J.J. Jenshi, and P. Malathi. 2011. Genetic diversity of sugarcane grassy shoot (SCGS)-phytoplasmas causing grassy shoot disease in India. Sugar Tech 13: 220–228.CrossRefGoogle Scholar
  34. Zhao, Y., W. Wei, I.M. Lee, J. Shao, X. Suo, and R.E. Davis. 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 Systematics and Evolutionary Microbiology 59: 2582–2593.CrossRefGoogle Scholar

Copyright information

© Society for Sugar Research & Promotion 2014

Authors and Affiliations

  • Shailender Kumar
    • 1
  • Ajay K. Tiwari
    • 2
  • Somnath K. Holkar
    • 3
  • S. K. Duttamajumder
    • 3
  • G. P. Rao
    • 1
    Email author
  1. 1.Division of Plant PathologyIndian Institute of Agricultural ResearchNew DelhiIndia
  2. 2.Sugarcane Research InstituteUP Council of Sugarcane ResearchShahjahanpurIndia
  3. 3.Division of Crop ProtectionIndian Institute of Sugarcane ResearchLucknowIndia

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