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A Single Chimeric Transgene Derived from Two Distinct Viruses for Multiple Virus Reslstance

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Abstract

A chimeric transgene composed of two different viral gene segments, was employed to confer multiple virus resistance in transformed tomato through homology-dependent gene silencing. Two important viruses of tomato, Cucumber mosaic virus (CMV) and Tomato leaf curl virus (ToLCV) were targeted. Comparison of 657 nucleotide coat protein (CP) region of CMV isolate (CMV -B) with those of five other CMV-Indian isolates showed more than 95% identity both at the nucleotide as well as amino acid level. Our previous studies showed the presence of conserved domain in the replicase gene of ToLCV comprising of 319 nts at 3’ of the gene. With this in view, gene encoding full-length transcript of CP of CMV-B was fused with the conserved domain of replicase gene of ToLCV. This is a first attempt of the transcriptional fusion of viral gene segments derived from a DNA virus (ToLCV) and a RNA virus (CMV). The chimeric gene construct was transformed in L. esculentum for multiple virus resistance, which is expected to work through homology - dependent gene silencing.

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Abbreviations

ToLCBV II:

Tomato leaf curl Bangalore virus II isolate

ToLCNDV-Svr:

Tomato leaf curl New Delhi virus Severe isolate

ToLCNDV-Mld:

Tomato leaf curl New Delhi virus Mild isolate

ToLCV-Dh:

Tomato leaf curl virus Dharwad isolate

ToLCV-Jb:

Tomato leaf curl virus Jabalpur isolate

ToLCV-IARI:

Tomato leaf curl virus IARI isolate

ToLCNDV-Luc:

Tomato leaf curl New Delhi virus Lucknowisolate

CMV-Li:

Cucumber mosaic virus Lilly isolate

CMV-B:

Cucumbermosaicvirus Bananaisolate

CMVEH:

Cucumber mosaic virus Egyptian Henbane isolate

CMV-Da:

Cucumber mosaic virus Datura isolate

CMV-Am:

Cucumber mosaic virus Amaranthus isolate

CMV-Ni:

Cucumber mosaic virus Nicotiana isolate

CMV:

Cucumber mosaic virus

ToLCV:

Tomato leaf curl virus

CP:

Coat protein gene

Rep:

Replication initiator protein gene

PTGS:

Post- transcriptional gene silencing

ORF:

Open reading frame

NAA:

(X- Naphthalene acetic acid

BAp:

Benzyl amino purine

IBA:

Indol-3- butyric acid

ZYMV:

Zucchini yellow mosaic virus

WMV:

Watermelon mosaic virus-2.

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Correspondence to Shelly Praveen.

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Part of M.Sc. thesis submitted by Ginny to Postgraduate School, IARI, New Delhi 110012

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Antony, G., Mishrab, A.K. & Praveen, S. A Single Chimeric Transgene Derived from Two Distinct Viruses for Multiple Virus Reslstance. J. Plant Biochem. Biotechnol. 14, 101–105 (2005). https://doi.org/10.1007/BF03263236

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