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- Title
Regeneration of sugarcane elite breeding lines and engineering of stem borer resistance.
- Authors
Li-Xing Weng; Haihua Deng; Jin-Ling Xu; Qi Li; Lian-Hui Wang; Zide Jiang; Hai Bao Zhang; Qiwei Li; Lian-Hui Zhang
- Abstract
Five elite sugarcane breeding lines were tested for efficiency in embryogenesis and plant regeneration. All of them produced regenerative embryogenic calli but with varied efficiencies. To engineer strongly insect-resistant sugarcanes, the GC content of a truncated crylAc gene, which encodes the active region of CrylAc insecticidal δ-endotoxin, was increased from the original 37.4 to 47.5% following the sugarcane codon usage pattern. The synthetic cry1Ac gene (s-crylAc) was placed under the control of maize ubiquitin promoter and introduced by microprojectile bombardment into the embryogenic calli of sugarcane lines YT79-177 and ROC16. Southern blotting analysis showed that multicopies of s-cry 1Ac were integrated into the genomes of transgenic sugarcane lines. Immunoblotting analysis identified 18 transgenic lines expressing detectable levels of s-CrylAc, which were estimated in the range of 1.8-10.0 ng mg-1 total soluble proteins. Four transgenic and two parental lines were assayed for sugarcane stem borer resistance in leaf tissue feeding trials and greenhouse plant assays. The results showed that, while the untransformed control lines were severely damaged in both leaves and stems, the transgenic sugarcane lines expressing high levels of s-CrylAc proteins were highly resistant to sugarcane stem borer attack, resulting in complete mortality of the inoculated insects within 1 week after inoculation.
- Subjects
SUGARCANE; REGENERATION (Botany); PLANT growth; GENES; GENOMES; IMMUNOBLOTTING; PLANT inoculation; GREENHOUSE gardening; ANTIGEN analysis
- Publication
Pest Management Science, 2006, Vol 62, Issue 2, p178
- ISSN
1526-498X
- Publication type
Article
- DOI
10.1002/ps.1144