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Improvement of resistance to maize dwarf mosaic virus mediated by transgenic RNA interference

Paper ID Volume ID Publish Year Pages File Format Full-Text
23836 43476 2011 7 PDF Available
Title
Improvement of resistance to maize dwarf mosaic virus mediated by transgenic RNA interference
Abstract

To overcome the low efficiency of agronomic protection from maize dwarf mosaic disease, susceptible maize inbred line was transformed by Agrobacterium tumefaciens harboring hpRNA expression vectors containing inverted-repeat sequences of different lengths targeting coat protein gene (CP) of maize dwarf mosaic virus (MDMV). After PCR screening and Southern blotting, the flanking sequences of the integration sites were amplified by thermal asymmetric interlaced PCR (TAIL-PCR) and used for analysis of T-DNA integration patterns. The T2 plant lines were evaluated for their MDMV resistance in field inoculation trials under two environments. Of the nineteen T2 plant lines positive in Southern blotting, six were evaluated as resistant to MDMV, and four of them had resistance non-significantly different from the highly resistant control “H9-21”, while the resistance of the other eleven was proved to be significantly improved when compared to their non-transformed parent line. These improvements in MDMV resistance were verified by the relative amount of virus CP gene expression measured by quantitative real time PCR. Comparing the results of Southern blotting and TAIL-PCR analysis, different integration patterns of one or two copies of the inverted-repeat sequences were identified from non-repetitive and repetitive sequences of the maize genome. The MDMV resistance mediated by RNA interference is relative to the length of the inverted-repeat sequence, the copy number of T-DNA integration and the repeatability of integration sites. A longer hpRNA expression construct shows more efficiency than a shorter one.

Keywords
Integration pattern; Inverted-repeat sequence; Maize dwarf mosaic virus; RNA interference; Transgene; Resistance
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Improvement of resistance to maize dwarf mosaic virus mediated by transgenic RNA interference
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 153, Issues 3–4, 20 May 2011, Pages 181–187
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us