qPCR analysis of Ty-2 and Ty-3 gene pyramided lines of tomato for resistance to tomato leaf curl New Delhi virus (ToLCNDV)

SUMAN LATA; ZAKIR HUSSAIN; MANISHA MANGAL; R K YADAV; VINUTHA T; GOGRAJ SINGH JAT; GOKUL GOSAVI; PAWAN KUMAR; SHELLY PERVEEN; B S TOMAR

doi:10.56093/ijas.v89i10.94636

Authors

SUMAN LATA Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
ZAKIR HUSSAIN Principal Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
MANISHA MANGAL Principal Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
R K YADAV Principal Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
VINUTHA T Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
GOGRAJ SINGH JAT Scientist, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
GOKUL GOSAVI Senior Research Fellow, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
PAWAN KUMAR Senior Research Fellow, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India
SHELLY PERVEEN Head, Division of Biochemistry, ICAR-IARI, New Delhi
B S TOMAR Head, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India

https://doi.org/10.56093/ijas.v89i10.94636

Keywords:

AC4, Tomato, Tomato leaf curl disease (ToLCD), Tomato leaf curl New Delhi virus (ToLCNDV), qPCR

Abstract

The present study was carried out at the ICAR-IARI, New-Delhi, India during 2017-18 to analyze the tomato genotypes having Ty-2 and Ty-3 genes incorporated through molecular breeding for resistance against the tomato leaf curl New Delhi virus (ToLCNDV).ToLCNDV is most predominant virus and causes huge economic loss in tomato, chilli, many cucurbits and cotton. Presently six genes Ty-1, Ty-2, Ty-3, Ty-4, ty-5 and Ty-6 are being utilized to address the tomato leaf curl disease (ToLCD). The tomato genotypes under study were grown in the field, along with susceptible and resistant checks. Genotyping was done to know the status of Ty-2 and Ty-3 genes in all the tomato samples.The tomato lines showing phenotypic resistance against ToLCNDV in the field were grown under controlled conditions in phytotron and agroinoculated with ToLCNDV genome. ToLCNDV agroinoculated tomato lines were checked for ToLCNDV specific AC4 transcript by qPCR assay.ToLCNDV specific AC4 transcript level was 4 log fold high in the susceptible check (Pusa Ruby) as compared to donor line (EC814916). However, AC4 transcript level was 2.1-2.8 log fold lower in the tomato line 218 (P. Ruby Ã—EC814916) as compared to susceptible var. Pusa Ruby. The study shows the importance of Ty-3 gene for imparting resistance against ToLCD disease in tomato.

Downloads

Download data is not yet available.

References

Chackraborty S. 2009. Tomato leaf curl viruses from India, pp. 339–47. Desk encyclopedia of plant and fungal virology.(Eds) Mahy B and Regenmortel M V.Oxford press, London.

Dong P, Han K, Siddique M I, Kwon J K, Zhao M, Wang F, and Kang B C. 2016. Gene-Based Markers for the Tomato Yellow Leaf Curl Virus Resistance Gene Ty-3. Plant Breeding and Biotechnology 4(1): 79–86. DOI: https://doi.org/10.9787/PBB.2016.4.1.79

Hutton S F, Scott J W and Schuster D J. 2012. Recessive resistance to Tomato yellow leaf curl virus from the tomato cultivar Tyking is located in the same region as Ty-5 on chromosome 4. HortScience 47(3): 324–327. DOI: https://doi.org/10.21273/HORTSCI.47.3.324

Hutton S F and Scott J W. 2013. Fine-mapping and cloning of Ty-1 and Ty-3; and mapping of a new TYLCV resistance locus,“Ty-6”. Tomato Breeders Round Table.

Ito T, Sharma P, Kittipakorn K and Ikegami M. 2008. Complete nucleotide sequence of a new isolate of tomato leaf curl New Delhi virus infecting cucumber, bottle gourd and muskmelon in Thailand. Archives of Virology 153(3): 611–3. DOI: https://doi.org/10.1007/s00705-007-0029-y

Ji Y, Scott J W, Schuster D J and Maxwell D P. 2009. Molecular mapping of Ty-4, a new Tomato yellow leaf curl virus resistance locus on chromosome 3 of tomato. Journal of the American Society for Horticultural Science 134(2): 281–8. DOI: https://doi.org/10.21273/JASHS.134.2.281

Murray M G and Thompson W F. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic acids research 8(19): 4321–6. DOI: https://doi.org/10.1093/nar/8.19.4321

Padidam M, Beachy RN and Fauquet CM. 1995. Tomato leaf curl gemini virus from India has a bipartite genome and coat protein is not essential for infectivity. Journal of General Virology 76(1): 25–35. DOI: https://doi.org/10.1099/0022-1317-76-1-25

Prasanna H C, Sinha D P, Rai G K, Krishna R, Kashyap S P, Singh N K, Singh M and Malathi V G. 2015. Pyramiding Ty-2 and Ty-3 genes for resistance to monopartite and bipartite tomato leaf curl viruses of India. Plant pathology 64(2): 256–64. DOI: https://doi.org/10.1111/ppa.12267

Pratap D, Kashikar A R and Mukherjee S K. 2011. Molecular characterization and infectivity of a Tomato leaf curl New Delhi virus variant associated with newly emerging yellow mosaic disease of eggplant in India. Virology journal 8(1): 305. DOI: https://doi.org/10.1186/1743-422X-8-305

Sahu P P, Rai N K, Chakraborty S, Singh M, Chandrappa P H, Ramesh B, Chattopadhyay D and Prasad M. 2010. Tomato cultivar tolerant to Tomato leaf curl New Delhi virus infection induces virus-specific short interfering RNA accumulation and defence-associated host gene expression. Molecular plant pathology 11(4): 531–44. DOI: https://doi.org/10.1111/j.1364-3703.2010.00630.x

Sohrab S S, Mandal B, Pant R P and Varma A. 2003. First report of association of Tomato leaf curl virus-New Delhi with yellow mosaic disease of Luffa cylindrica in India. Plant disease 87(9): 1148. DOI: https://doi.org/10.1094/PDIS.2003.87.9.1148A

Usharani K S, Surendranath B, Paul-Khurana S M, Garg I D and Malathi V G. 2004. Potato leaf curl-a new disease of potato in northern India caused by a strain of tomato leaf curl New Delhi virus. Plant pathology 53(2): 235. DOI: https://doi.org/10.1111/j.0032-0862.2004.00959.x

Verlaan M G, Hutton S F, Ibrahem R M, Kormelink R, Visser R G, Scott J W, Edwards J D and Bai Y. 2013. The tomato yellow leaf curl virus resistance genes Ty-1 and Ty-3 are allelic and code for DFDGD-class RNA–dependent RNA polymerases. PLoS genetics 9(3): p.e1003399. DOI: https://doi.org/10.1371/journal.pgen.1003399

Vidavski F S. 2007. Exploitation of resistance genes found in wild tomato species to produce resistant cultivars; pile up of resistant genes, pp 363–72. (In) Tomato yellow leaf curl virus disease.Springer, Dordrecht. DOI: https://doi.org/10.1007/978-1-4020-4769-5_21

Yang X, Caro M, Hutton S F, Scott J W, Guo Y, Wang X, Rashid M H, Szinay D, de Jong H, Visser R G and Bai Y. 2014. Fine mapping of the tomato yellow leaf curl virus resistance gene Ty-2 on chromosome 11 of tomato. Molecular Breeding 34(2): 749–60. DOI: https://doi.org/10.1007/s11032-014-0072-9

Livak K J and Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25(4): 402–8. DOI: https://doi.org/10.1006/meth.2001.1262