Field resistance to Tomato spotted wilt virus in transgenic peanut (Arachis hypogaea L.) expressing an antisense nucleocapsid gene sequence

Citation
Zv. Magbanua et al., Field resistance to Tomato spotted wilt virus in transgenic peanut (Arachis hypogaea L.) expressing an antisense nucleocapsid gene sequence, MOL BREED, 6(2), 2000, pp. 227-236
Citations number
38
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
MOLECULAR BREEDING
ISSN journal
1380-3743 → ACNP
Volume
6
Issue
2
Year of publication
2000
Pages
227 - 236
Database
ISI
SICI code
1380-3743(200004)6:2<227:FRTTSW>2.0.ZU;2-1
Abstract
Peanut (Arachis hypogaea L.) lines transgenic for the antisense nucleocapsi d (N) gene of a Tomato spotted wilt virus (TSWV) strain isolated from peanu t were generated by microprojectile-mediated transformation of repetitive s omatic embryos of cultivars VC1 and AT120. The selectable marker (hygromyci n resistance) and the N gene were on separate plasmids. A total of 207 VC1 and 120 AT120 hygromycin-resistant lines were produced. Of all the VC1 plan ts recovered 71% were cotransformed with the N gene (N+), but all plants we re sterile. For AT120, 48 of the transgenic cell lines converted into plant s. Polymerase chain reaction (PCR) screening showed 15 of the lines were tr ansgenic for the N gene (N+), and two of these lines were fertile. A field test was conducted in 1998 at Ashburn, GA, using seeds from each fertile li ne, along with segregated and non-transgenic controls. Plants from four ran domly selected field plots were examined for symptoms and analyzed by doubl e-antibody sandwich enzyme-linked immunoabsorbent assay and PCR at 10 and 1 4 weeks after planting. At 14 weeks, 76% of the N+ plants were symptomless, while 2% were severely symptomatic or dead. In contrast, only 42% of the p lants lacking the N gene were symptomless and 50% were severely symptomatic or dead. Northern blot analysis of selected field-resistant plants detecte d transgene RNA, and the transcript level appeared undiminished after viral exposure.