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Balasubramani, V.
- Transformation of Tobacco (Nicotiana tabaccum) with cry2AX1 Gene and Analysis of Transgenic Plants
Authors
1 Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore (T.N.), IN
Source
International Journal of Forestry and Crop Improvement, Vol 7, No 1 (2016), Pagination: 79-85Abstract
A novel synthetic cry2AX1 gene was codon optimized and a sequence encoding cotton rbcS1b transit peptide was fused upstream of coding sequence. The fusion cry2AX1 gene, driven by maize ubiquitin1 promoter was cloned in a pUH plant transformation vector. Agrobacterium mediated transformation was carried out with pUH-ctp-2AX1 construct using leaf discs of tobacco as model plant. Screening by PCR revealed presence of cry2AX1 gene in all nine putative transformants and expression of cry2AX1 protein in PCR positive T0 tobacco plants ranged from 1.5 to 10.0 ng/g. The detached leaf bit bioassay of tobacco transformants with Helicoverpa armigera showed 30 per cent mortality even at lower level of cry2AX1 expression. The results indicated a newly developed construct was functionally expressed in tobacco plant.Keywords
Tobacco, Transformation, Insect Resistance, Helicoverpa armigera.- Characterisation of native Bacillus thuringiensis isolates toxicity to fall armyworm, Spodoptera frugiperda (J.E. Smith)
Authors
1 Department of Agricultural Entomology, Tamil Nadu Agricultural University, Coimbatore – 641003, Tamil Nadu, IN
2 Department of Plant Biotechnology, Tamil Nadu Agricultural University, Coimbatore – 641003,, IN
3 Department of Plant Biotechnology, Tamil Nadu Agricultural University, Coimbatore – 641003, Tamil Nadu, IN
Source
Journal of Biological Control, Vol 35, No 3 (2021), Pagination: 171-180Abstract
Toxicity of nine indigenous Bacillus thuringiensis (Bt) isolates collected from Tamil Nadu, India were tested against fall armyworm, Spodoptera frugiperda. At 30 ?g/ml concentration, two Bt isolates viz., T350 and T532 recorded 100 per cent mortality whereas isolates T527 and T532 registered 96 per cent mortality against neonate larvae of S. frugiperda in leaf dip bioassay. SDS PAGE analysis of spore crystal mixture revealed the presence of Cry1 and Cry2 proteins with visible bands at 130 kDa and 65 kDa. PCR screening results showed the presence of cry1 (cry1A, cry1Aa, cry1Ab), cry2 (cry2Aa, cry2Ab) in four isolates and vip3A genes in three isolates but cry9 gene was not present in any of the isolates tested.