NDLI: RNA Interference-Based Transgenic Maize Resistant to Maize Dwarf Mosaic Virus

Content Provider	Springer Nature Link
Author	Zhang, Zhi Yong Fu, Feng Ling Gou, Lin Wang, Han Guang Li, Wan Chen
Copyright Year	2010
Abstract	Maize dwarf mosaic virus (MDMV) is a widespread pathogenic virus that causes serious loss of yield in maize (Zea mays). RNA interference (RNAi) triggered by hairpin RNA (hpRNA) transcribed from a transgenic inverted-repeat sequence is an effective way to defend against viruses in plants. In this study, an hpRNA expression vector containing a sense arm and an antisense arm of 150 bp separated by an intron of the maize actin gene was constructed to target the P1 protein (protease) gene of MDMV and used to transform Agrobacterium tumefaciens strain EHA105. The transformed Agrobacterium strain was used to transform maize embryonic calli isolated from immature embryos by an improved culture technique. In all, 46 plants were regenerated after stringent hygromycin B selection, and 18 of them were certified to be positive by PCR amplification. Of these positive plants, 13 were grown to produce offspring, and nine were identified by Southern blotting to have the transgene integrated with one or two copies. The resistance of three T$_{2}$ lines was evaluated in a field trial of dual MDMV inoculation in two environments and was found to be improved compared with the non-transformed control. The disease indexes of the transgenic plant lines h2, 13, and h1 were not significantly different from the highly resistant control line H9-21. The viral titers of the inoculated plants were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), and the result was in accord with the resistance evaluated in the field trial. The addition of uniconazole S3307 (0.25 mg l$^{−1}$) and ABT root-promoting powder (0.5 mg l$^{−1}$) showed a significant improvement of hardening in regenerated plantlets, which were stronger and generated a better fibrous root system than the control. This improvement could facilitate the transgenic operation of maize.
Starting Page	297
Ending Page	305
Page Count	9
File Format	PDF
ISSN	12269239
Journal	Journal of Plant Biology
Volume Number	53
Issue Number	4
e-ISSN	18670725
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-07-07
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Agrobacterium-mediated transformation Embryonic callus Hairpin RNA Maize dwarf mosaic virus Resistance RNA interference Plant Ecology Plant Systematics/Taxonomy/Biogeography Plant Genetics & Genomics Plant Breeding/Biotechnology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Plant Science

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