Indian Phytopathology

, Volume 71, Issue 3, pp 435–443 | Cite as

Prevalence of severe Citrus tristeza virus infection in Khasi mandarin throughout the north-eastern states of India: an evidence study based on coat protein gene analysis

  • Amit Kumar Singh
  • Ng. Tombisana Meetei
  • Monoj Sutradhar
  • Brijesh Kumar Singh
  • Nirmal MandalEmail author
Research Article


Citrus tristeza virus (CTV) infected plant samples from the North Eastern states of India namely Arunachal Pradesh (AR1), Assam (AS1), Nagaland (NL1), Meghalaya (M1), Sikkim (SK1) and Tripura (TR1) were examined through double antibody sandwich enzyme linked immunosorbent assay and reverse transcriptase polymerase chain reaction using coat protein (CP) gene specific primers, yielding amplicon of 672 base pair for all the isolates. The sequence analysis revealed nucleotide (nt) and amino acid (aa) sequence similarity with other exotic CTV isolates. AR1 and AS1 shared similarity with T3 isolate (Florida) at 98–99% and 95–99% for nt-aa respectively, whereas, M1 showed 99% similarity with stem pitting isolate B165 (Bangalore). SK1 and NL1 were 95–97% and 97–99% similar with Hawaiian isolate HA at nt-aa level. TR1 showed 97% homology with seedling yellowing Israeli isolate VT. The CP gene sequence based phylogenetic analysis grouped present isolates into four clades with reference severe isolates (i.e. VT, B165, HA and T3), indicating their probable evolution. TR1 and NL1 are the putative recombinant isolates and ASI and SK1 were the major and minor parents respectively. This might be due to the conserved nature of CP gene. Moreover, the CP gene sequences of present isolates showing nucleotide identity to the VT, T3 and B165 isolates also suggest the prevalence of severe CTV isolates in Khasi mandarin growing areas comes under North East (NE) region of India.


Citrus tristeza virus Coat protein gene DAS-ELISA Evolution North Eastern isolates Phylogenetic analysis 



Authors acknowledge the laboratory as well as library assistance from BCKV, West Bengal. We further are thankful to the DBT for providing grant (BT/04/NE/2009) under the project Advanced level Institutional Biotech Hub, CHF, CAU, Pasighat.

Supplementary material

42360_2018_56_MOESM1_ESM.jpg (94 kb)
Eco RI digested CP gene insert cloned in pGEM-T Easy Vector. Lane M represents 100 bp DNA ladder (Xcelris genomics, Ahmedabad, India) (JPEG 94 kb)
42360_2018_56_MOESM2_ESM.docx (19 kb)
Supplementary material 2 (DOCX 18 kb)
42360_2018_56_MOESM3_ESM.docx (19 kb)
Supplementary material 3 (DOCX 19 kb)


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Copyright information

© Indian Phytopathological Society 2018

Authors and Affiliations

  • Amit Kumar Singh
    • 1
  • Ng. Tombisana Meetei
    • 2
  • Monoj Sutradhar
    • 3
  • Brijesh Kumar Singh
    • 1
  • Nirmal Mandal
    • 3
    Email author
  1. 1.College of Horticulture and ForestryCentral Agriculture UniversityPasighatIndia
  2. 2.College of Post Graduate StudiesCentral Agriculture UniversityUmiamIndia
  3. 3.Department of Agricultural Biotechnology, Faculty of AgricultureBidhan Chandra Krishi ViswavidyalayaNadiaIndia

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