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KMID : 0578319950050010065
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Molecules and Cells
1995 Volume.5 No. 1 p.65 ~ p.71
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In Vitro Translation Inhibition and in Vivo Viral sympton Development Attenuation by Cucumber Mosaic Virus RNA3 cDNA Fragments
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Kim, shin Je
Kim, Byung-Dong/Paek, Kyung-Hee
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Abstract
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Two kinds of cDNA clones of cucumber mosaic virus (CMV) strain Kor were constructed in both sense and antisense orientations in pBluescript sk^(+) or plant expression vector, PMBP 1. The clones cover the movement protein gene and part of the coat protein gene of RNA3. Invitro CMV RNA translation inhibition assay using CMV-Kor or CMV-Y as a substrate was carried out by measuring the incorporation rate of [^(35)S]methionine. CMV RNA3 translation inhibition ranged from 46 to 74% depending on the constructions. An in vivo analysis of these CMV-Kor RNA3 cDNA clones was carried out in transgenic tobacco plants. Each construction in pMBPl was transformed into Agrobacterium tumefaciens and used for developing transgenic tobacco plants. From polymerase chain reaction and Northern blot analyses, it was confirmed that the cDNA clones were stably expressed in the transgenic plants. Trnnsgenic plants expressing CMV-Kor RNA were challenged with CMV-Kor or CMV-Y. Local lesion assays using Chenopodium amaranticolor were carried out for a quantitative estimation of symptom attenuation observed in these transgenic plants. In the case of transgenic plants producing sense transcripts, there was almost no development of disease symptoms upon CMV-Kor or CMV-Y inoculation. On the other hand, in transgenic plants expressing antisense transcripts, the development of disease symptoms was only slightly attenuated.
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