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Detection of Cardamom mosaic virus and Banana bract mosaic virus in cardamom using SYBR Green based reverse transcription-quantitative PCR

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Abstract

Cardamom being perennial, propagated vegetatively, detecting viruses in planting material is important to check the spread of viruses through infected material. Thus development of effective and sensitive assay for detection of viruses is need of the time. In this view, assay for the detection of Cardamom mosaic virus (CdMV) and Banana bract mosaic virus (BBrMV), infecting cardamom was developed using SYBR Green one step reverse transcription-quantitative PCR (RT-qPCR). The RT-qPCR assay amplified all isolates of CdMV and BBrMV tested but no amplification was obtained with RNA of healthy plants. Recombinant plasmids carrying target virus regions corresponding to both viruses were quantified, serially diluted and used as standards in qPCR to develop standard curve to enable quantification. When tenfold serial dilutions of the total RNAs from infected plants were tested through RT-qPCR, the detection limit of the assay was estimated to be 16 copies for CdMV and 10 copies for BBrMV, which was approximately 1,000-fold higher than the conventional RT-PCR. The RT-qPCR assay was validated by testing field samples collected from different cardamom growing regions of India. This is the first report of RT-qPCR assay for the detection of CdMV and BBrMV in cardamom.

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Acknowledgments

We thank Department of Biotechnology, Government of India for financial support; National Research Centre for Banana, Trichy for providing BBrMV antiserum; Distributed Information Sub Centre and Director, Indian Institute of Spices Research, Kozhikode for providing facilities.

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Authors and Affiliations

  1. Division of Crop Protection, Indian Institute of Spices Research, Marikunnu, Kozhikode, 673012, Kerala, India

    A. Siljo & A. I. Bhat

  2. Indian Institute of Spices Research, Cardamom Research Centre, Appangala, Heravanadu Post, Madikeri, 571 201, Karnataka, India

    C. N. Biju

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  1. A. Siljo
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  2. A. I. Bhat
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  3. C. N. Biju
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Correspondence to A. I. Bhat.

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Siljo, A., Bhat, A.I. & Biju, C.N. Detection of Cardamom mosaic virus and Banana bract mosaic virus in cardamom using SYBR Green based reverse transcription-quantitative PCR. VirusDis. 25, 137–141 (2014). https://doi.org/10.1007/s13337-013-0170-z

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  • DOI: https://doi.org/10.1007/s13337-013-0170-z

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