Engineering sugarcane cultivars with bovine pancreatic trypsin inhibitor (aprotinin) gene for protection against top borer (Scirpophaga excerptalis Walker) |
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Authors: | Leela Amala Christy S Arvinth M Saravanakumar M Kanchana N Mukunthan J Srikanth George Thomas N Subramonian |
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Institution: | (1) Sugarcane Breeding Institute, Coimbatore, 641007, India;(2) Interfield Laboratories, Cochin, 682005, India |
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Abstract: | The inhibitory activity of bovine pancreatic trypsin inhibitor (aprotinin), a natural polypeptide and a proteinase inhibitor,
was demonstrated on gut proteinases of three lepidopteran borers of sugarcane using commercially available aprotinin. A synthetic
gene coding for aprotinin, designed and codon optimized for better expression in plant system (Shantaram 1999), was transferred to two sugarcane cultivars namely CoC 92061 and Co 86032 through particle bombardment. Aprotinin gene expression
was driven by maize ubiquitin promoter and the plant selection marker used was hygromycin resistance. The integration, expression
and functionality of the transgene was confirmed by Southern, Western and insect bioassay, respectively. Southern analysis
showed two to four integration sites of the transgene in the transformed plants. Independent transgenic events showed varied
levels of transgene expression resulting in different levels (0.16–0.50%) of aprotinin. In in vivo bioassay studies, larvae
of top borer Scirpophaga excerptalis Walker (Lepidoptera: Pyralidae) fed on transgenics showed significant reduction in larval weight which indicated impairment
of their development. Results of this study show the possibility of deploying aprotinin gene for the development of transgenic
sugarcane cultivars resistant to top borer. |
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Keywords: | Transgenics Sugarcane Aprotinin Bovine pancreatic trypsin inhibitor Borers Scirpophaga excerptalis In vitro bioassay In vivo bioassay |
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