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Genetic transformation with untranslatable coat protein gene of sugarcane yellow leaf virus reduces virus titers in sugarcane

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Abstract

Sugarcane yellow leaf syndrome, characterized by a yellowing of the leaf midrib followed by leaf necrosis and growth suppression, is caused by sugarcane yellow leaf virus (SCYLV). We produced SCYLV-resistant transgenic sugarcane from a susceptible cultivar (H62-4671) and determined the amount of virus present following inoculation. The transgenic plants were produced through biolistic bombardment of cell cultures with an untranslatable coat protein gene. Presence of the transgene in regenerated plants was confirmed using PCR and Southern blot analysis. The transgenic lines were inoculated by viruliferous aphids and the level of SCYLV in the plants was determined. Six out of nine transgenic lines had at least 103-fold lower virus titer than the non-transformed, susceptible parent line. This resistance level, as measured by virus titer and symptom development, was similar to that of a resistant cultivar (H78-4153). The selected SCYLV-resistant transgenic sugarcane lines will be available for integration of the resistance gene into other commercial cultivars and for quantification of viral effects on yield.

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Abbreviations

BA:

Benzyladenine

IBA:

Indolebutyric acid

MS:

Murashige and Skoog plant culture medium

NAA:

Naphthaleneacetic acid, 2, 4-D: 2, 4-dichlorophenoxyacetic acid

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription polymerase chain reaction

SCYLV:

Sugarcane yellow leaf virus

YLS:

Yellow leaf syndrome

Ubi:

Ubiquitin

nptII:

Neomycin phosphotransferase

SCYLVcp:

Sugarcane yellow leaf virus coat protein

TPI:

Triosephosphate isomerase

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Acknowledgments

We thank Dr. E. Mirkov, Texas A&M Agriculture Experiment Station, Weslaco, TX for providing pFM395 construct; Dr. Henrik Albert at USDA, ARS for providing the pHA9 construct containing selection marker; Dr. M. Fitch, USDA, ARS, for advice on sugarcane transformation and clonal propagation and Dr. M. Irey, US Sugar, for providing primer sequences for SCYLV. This work was partially supported by a cooperative agreement (No. CA 58-5320-3-460) between the U.S. Department of Agriculture, Agricultural Research Service, and the Hawaii Agriculture Research Center.

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Authors and Affiliations

  1. Hawaii Agriculture Research Center, 94-340 Kunia Road, Waipahu, HI, 96797, USA

    Yun J. Zhu, Heather McCafferty, Greg Osterman, Steven Lim, Ricelle Agbayani, Axel Lehrer & Susan Schenck

  2. Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, 96822, USA

    Greg Osterman & Ricelle Agbayani

  3. Currently with Hawaii Biotech Inc, 99-193 Aiea Heights Drive, Aiea, HI, 96701, USA

    Axel Lehrer

  4. University of Bayreuth, 95440, Bayreuth, Germany

    Ewald Komor

Authors
  1. Yun J. Zhu
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  2. Heather McCafferty
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  3. Greg Osterman
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  4. Steven Lim
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  5. Ricelle Agbayani
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  6. Axel Lehrer
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  7. Susan Schenck
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  8. Ewald Komor
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Correspondence to Yun J. Zhu.

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Zhu, Y.J., McCafferty, H., Osterman, G. et al. Genetic transformation with untranslatable coat protein gene of sugarcane yellow leaf virus reduces virus titers in sugarcane. Transgenic Res 20, 503–512 (2011). https://doi.org/10.1007/s11248-010-9432-3

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  • DOI: https://doi.org/10.1007/s11248-010-9432-3

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