title: Enhanced whitefly resistance in transgenic tobacco plants expressing double stranded RNA of v-ATPase A gene.
abstract: BACKGROUND
  Expression of double strand RNA (dsRNA) designed against important insect genes in transgenic plants have been shown to give protection against pests through RNA interference (RNAi), thus opening the way for a new generation of insect-resistant crops. We have earlier compared the efficacy of dsRNAs/siRNAs, against a number of target genes, for interference in growth of whitefly (Bemisia tabaci) upon oral feeding. The v-ATPase subunit A (v-ATPaseA) coding gene was identified as a crucial target. We now report the effectiveness of transgenic tobacco plants expressing siRNA to silence v-ATPaseA gene expression for the control of whitefly infestation.
  
  
  RESULTS
  Transgenic tobacco lines were developed for the expression of long dsRNA precursor to make siRNA and knock down the v-ATPaseA mRNA in whitefly. Molecular analysis and insecticidal properties of the transgenic plants established the formation of siRNA targeting the whitefly v-ATPaseA, in the leaves. The transcript level of v-ATPaseA in whiteflies was reduced up to 62% after feeding on the transgenic plants. Heavy infestation of whiteflies on the control plants caused significant loss of sugar content which led to the drooping of leaves. The transgenic plants did not show drooping effect.
  
  
  CONCLUSIONS
  Host plant derived pest resistance was achieved against whiteflies by genetic transformation of tobacco which generated siRNA against the whitefly v-ATPaseA gene. Transgenic tobacco lines expressing dsRNA of v-ATPaseA, delivered sufficient siRNA to whiteflies feeding on them, mounting a significant silencing response, leading to their mortality. The transcript level of the target gene was reduced in whiteflies feeding on transgenic plants. The strategy can be taken up for genetic engineering of plants to control whiteflies in field crops.
authors.affiliation: Plant Molecular Biology Lab, National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Academy of Scientific and Innovative Research, Anusandhan Bhawan, NewDelhi, India.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Thakur
authors.affiliation: Plant Molecular Biology Lab, National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Department of Biotechnology, National Agri-Food Biotechnology Institute, Mohali, Punjab, India.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Upadhyay
authors.affiliation: Plant Molecular Biology Lab, National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Academy of Scientific and Innovative Research, Anusandhan Bhawan, NewDelhi, India.
authors.forename: Praveen C
authors.initials: PC
authors.lastname: Verma
authors.affiliation: Plant Molecular Biology Lab, National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Indian Agricultural Research Institute-Regional Station, Agricultural College Estate, Pune, Maharashtra, India.
authors.forename: Krishnappa
authors.initials: K
authors.lastname: Chandrashekar
authors.affiliation: Department of Biotechnology, National Agri-Food Biotechnology Institute, Mohali, Punjab, India.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Tuli
authors.affiliation: Plant Molecular Biology Lab, National Botanical Research Institute, Lucknow, Uttar Pradesh, India; Academy of Scientific and Innovative Research, Anusandhan Bhawan, NewDelhi, India.
authors.forename: Pradhyumna K
authors.initials: PK
authors.lastname: Singh
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D017931:DNA Primers; D006430:Hemiptera / Q000235:genetics*; D006790:Host-Parasite Interactions; D030821:Plants, Genetically Modified; D012330:RNA, Double-Stranded / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics* / Q000469:parasitology; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics*
chemical_list: D017931:DNA Primers; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D025262:Vacuolar Proton-Translocating ATPases