RNA Silencing-Mediated Control of Odontoglossum ringspot virus (ORSV) Infection ()
ABSTRACT
Odontoglossum ringspot virus (ORSV)
infects perennial orchids (Phalaenopsis amabilis) and
causes a widespread viral disease. RNA-silencing of viral genes is a promising
and effective way of controlling viral infection in plants. An inverted repeat
(IR) fragment of the ORSV coat protein gene, cp, was inserted into the pXGY1 vector to
generate the silencing construct, pXGY1-ORSV, which was introduced into Nicotiana benthamiana via
Agrobacterium-mediated infiltration. A total of 15 homozygous pXGY1-ORSV transgenic N. benthamiana T1
plants were obtained from five transgenic lines, and ORSV cp gene
multiplication was reduced by at least 75% - 95% in 12 T2 plants, demonstrating
their increased resistance to ORSV. An infectious ORSV clone, pCAMBIA2300-ORSV,
was generated to facilitate rigorous analyses of plant viral resistance.
Semi-quantitative RT-PCR (sqRT-PCR) and northern-blot analyses revealed that
levels of ORSV multiplication and ORSV coat protein were significantly reduced
in pXGY1-ORSV transgenic N. benthamiana. Western-blot
from pXGY1-ORSV inoculated leaves of ORSV infected P. amabilis also
revealed the significant decrease and even degradation of ORSV-CP protein. Disease
symptoms were not observed in transgenic plants. These results indicate a high
level of ORSV-resistance in pXGY1-ORSV transgenic N. benthamiana.
Share and Cite:
Zhang, X. , Hu, Y. , Chen, Z. , Zhang, P. , Li, H. and Shi, N. (2019) RNA Silencing-Mediated Control of
Odontoglossum ringspot virus (ORSV) Infection.
American Journal of Plant Sciences,
10, 147-161. doi:
10.4236/ajps.2019.101013.
Cited by
No relevant information.