Archives of Virology

, Volume 164, Issue 4, pp 1005–1013 | Cite as

Assessment of resistance to rice tungro disease in popular rice varieties in India by introgression of a transgene against Rice tungro bacilliform virus

  • G. Kumar
  • M. Jyothsna
  • P. Valarmathi
  • S. Roy
  • A. Banerjee
  • J. Tarafdar
  • B. K. Senapati
  • S. Robin
  • S. Manonmani
  • R. Rabindran
  • I. DasguptaEmail author
Original Article


Rice crops in South and Southeast Asian countries suffer critical yield losses due to rice tungro disease caused by joint infection with rice tungro bacilliform virus (RTBV) and rice tungro spherical virus (RTSV). Previously, for generating RNA interference-based transgenic resistance against tungro viruses, RTBV ORF IV was used as a transgene to develop RTBV resistance in a popular high-yielding scented rice variety. The transgene from this line was then introgressed into five popular high-yielding but tungro-susceptible rice varieties by marker-assisted backcross breeding with a view to combine the resistant trait with the agronomic traits. The present work includes a resistance assay of the BC3F5 lines of these varieties under glasshouse conditions. Out of a total of 28 lines tested, each consisting of 12 individual plants, eight lines showed significant amelioration in height reduction and 100- to 1000-fold reduction in RTBV titers. The RNAi-mediated resistance was clearly manifested by the presence of virus-derived small RNA (vsRNA) specific for RTBV ORF IV in the transgenic backcrossed lines.



This work was funded by Department of Biotechnology, Government of India (Grant No. BT/PR-15033/AGR/02/773/2011). GK is indebted to University Grants Commission, New Delhi, for research fellowships during this work. Funds received from R&D Grant of University of Delhi, DST-PURSE Grant, and the departmental infrastructure grant from DST-FIST are also gratefully acknowledged.

Compliance with ethical standards

This work was performed keeping in mind all applicable ethical standards.

Conflict of interest

There exists no conflict of interest among the authors pertaining to this work.

Supplementary material

705_2019_4159_MOESM1_ESM.tif (160 kb)
Suppl. Figure S1 Schematic representation of the orientation of forward and reverse primers in the two strategies (a and b) used for validation of pRTBV-Inf infection efficiency
705_2019_4159_MOESM2_ESM.tif (51 kb)
Suppl. Figure S2 Confirmation of RTBV in rice plants upon agroinoculation. M, marker; lane 1, negative control; lanes 2 to 13, 12 plants selected for PCR; lane 14, RTBV-positive genomic DNA (1/10th dilution); lane 15, positive control (pRTBV-Inf plasmid)
705_2019_4159_MOESM3_ESM.tif (101 kb)
Suppl. Figure S3 Validation of infection efficiency of the pRTBV-Inf agroinfectious clone. (a) With primer ISEPCIRCU FP/RP: lanes 1-6 and 9-14, plant samples; lane 7, negative control; lane 8, pRTBV-Inf plasmid. (b) With primer IINSVEC FP/RP: lane 1, negative control; lanes 2-13, plant samples; lane 14, positive control (pRTBV-Inf plasmid)
705_2019_4159_MOESM4_ESM.tif (193 kb)
Suppl. Figure S4 Comparison of virus titer of the test TN1 plants at two time points
705_2019_4159_MOESM5_ESM.tif (131 kb)
Suppl. Figure S5 Confirmation of RTBV (amplified as approximately 1-kb bands) in ASD 16 parent and backcrossed lines along with susceptible TN1 and resistant Utri Merah variety. –ve is the mock-inoculated DNA, while +ve is the 1/10th-diluted plasmid. ‘M’ corresponds to the 1-kb DNA marker
705_2019_4159_MOESM6_ESM.tif (173 kb)
Suppl. Figure S6 Confirmation of RTBV (amplified as approximately 1-kb bands) in BPT 5204 parent and backcrossed lines. –ve is the mock-inoculated DNA, while +ve is the 1/10th-diluted plasmid. ‘M’ corresponds to the 1-kb DNA marker
705_2019_4159_MOESM7_ESM.tif (293 kb)
Suppl. Figure S7 Confirmation of RTBV (amplified as approximately 1-kb bands) in CR 1009 parent and backcrossed lines. –ve is the mock-inoculated DNA, while +ve is the 1/10th-diluted plasmid. ‘M’ corresponds to the 1-kb DNA marker
705_2019_4159_MOESM8_ESM.tif (91 kb)
Suppl. Figure S8 Confirmation of RTBV (amplified as approximately 1-kb bands) in Shatabdi and Khitish parent and backcrossed lines. –ve is the mock-inoculated DNA, while +ve is the 1/10th-diluted plasmid. ‘M’ corresponds to the 1-kb DNA marker
705_2019_4159_MOESM9_ESM.tif (84 kb)
Suppl. Figure S9 Standard curve for estimation of RTBV titers
705_2019_4159_MOESM10_ESM.pdf (8.4 mb)
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Supplementary material 11 (DOCX 13 kb)
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Supplementary material 16 (DOCX 67 kb)
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Supplementary material 17 (DOCX 103 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Plant Molecular BiologyUniversity of Delhi South CampusNew DelhiIndia
  2. 2.Department of RiceTamil Nadu Agricultural UniversityCoimbatoreIndia
  3. 3.Department of Plant PathologyTamil Nadu Agricultural UniversityCoimbatoreIndia
  4. 4.Department of Plant BreedingBidhan Chandra Krishi ViswavidyalayaKalyaniIndia
  5. 5.Department of Plant PathologyBidhan Chandra Krishi ViswavidyalayaKalyaniIndia

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