Characterization and cloning of the cry2A gene from indigenous isolates of Bacillus thuringiensis

Manikandan, R.; Ramalakshmi, A.; Balasubramani, V.; Udayasuriyan, V.

doi:10.1134/s0026893315040111

Cited by 6 publications

(4 citation statements)

References 24 publications

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“…While screening local isolates of Bt, Manikandan et al (2015) observed bipyramidal shape of crystals in T30 and T48 which was reported toxic against Spodoptera litura, in the present study, Bt isolates T350, T352, T527, T532 toxic against S. frugiperda were found to have bipyramidal shape of crystal inclusions. Bipyramidal shape of crystal structure was found to be dominant and more effective against lepidopteran larvae (Bernard et al, 1997;Swamy et al, 2013).…”

Section: Discussionsupporting

confidence: 48%

Characterisation of native Bacillus thuringiensis isolates toxicity to fall armyworm, Spodoptera frugiperda (J.E. Smith)

Maheesha

Balasubramani

Murugan

et al. 2022

JBC

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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.

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Section: Discussionsupporting

confidence: 48%

Characterisation of native Bacillus thuringiensis isolates toxicity to fall armyworm, Spodoptera frugiperda (J.E. Smith)

Maheesha

Balasubramani

Murugan

et al. 2022

JBC

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“…Bright field microscopic observation at 100x magnification showed the presence of bipyramidal shape of crystals produced by Bt isolate T352. Manikandan et al [26] observed the occurrence of bipyramidal and cuboidal shapes of inclusions in Bt isolate T30 whereas isolate T48 contains only bipyramidal shaped inclusions. While, Navya et al [32] observed that Bt isolate RM11 effective against Plutella xylostella was found to possess three different shapes of crystals such as bipyramidal, spherical and cuboidal.…”

Section: Colony and Crystal Morphologymentioning

confidence: 99%

Characterization and Evaluation of Indigenous Bacillus thuringiensis Isolate T352 against Fall Armyworm, Spodoptera frugiperda (J.E. Smith)

Karuppaiyan

Balasubramani²,

Murugan³

et al. 2022

IJPSS

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Bacillus thuringiensis (Bt) Berliner is a ubiquitous soil bacterium with commercial bio pesticidal value and widely used for effective control of various important agricultural insect pests. Invasion of Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) into India caused potential yield loss in maize production, and also it threats the cultivation of other related crops. This study was aimed to characterize and evaluate the pathogenic activity of indigenous Bt isolate T352 against fall armyworm (FAW), S. frugiperda. The Bt isolate T352 was creamy white in colour and had irregular shaped flat colonies with undulated margin. Bipyramidal shape of parasporal inclusions was found in isolate T352. The isolate produced protein bands of ̴ 130 kDa and ̴ 65 kDa size in SDS PAGE analysis. PCR screening also confirmed the presence of cry1Ab, cry1Ac, cry2Aa, cry2Ab and vip3A genes. During probit analysis, isolate T352 exhibited the LC50 of 1.927 µg/ml as against 0.421µg/ml in positive standard strain HD- 1 based on mortality observed at 72 h after treatment in leaf disc bioassay with spore crystal mixtures.

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“…The gene sequence and its deduced amino acid sequence were submitted to Bacillus thuringiensis delta endotoxin nomenclature committee and it was named as cry2Aa23. Manikandan et al (2015) reported the cloning of cry2A gene from an indigenous Bt isolate using T/A cloning vector and the sequencing results revealed 99 per cent homology of the sequence with that of cry2Aa1. Likewise cloning of cry1Ab (Darsi et al, 2010), cry2Ab (Jain et al, 2006 andPan et al, 2014), cry2Ac (Sasaki et al, 1997) and cry9 (Ben-Dov et al, 1999;Wasano et al, 2005) genes were cloned and characterized.…”

Section: Resultsmentioning

confidence: 99%

Molecular characterization of an indigenous lepidopteran toxic Bacillus thuringiensis strain T532

Kondabattula

Reyaz

Balakrishnan

2018

Journal of Biological Control

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Bacillus thuringiensis Berliner (Bt) being an eco-friendly bioinsecticide is effectively used in pest management strategies and, therefore, isolation and identification of new strains effective against a broad range of target pests is important. In the present study, four indigenous Bt isolates (T489, T491, T495 and T532) were taken and investigated for their properties such as crystal morphology, cry gene(s) content and insecticidal activity. Bipyramidal, cuboidal and spherical crystals were observed. Among the four isolates only one isolate was found positive for lepidopteran specific cry genes. The isolate T532 which showed the presence of cry1, cry2Aa and cry2Ab genes was selected for protein profiling and bioassay. The isolate showed the presence of two proteins of molecular weights at ~135 and ~65 kDa sizes. Artificial diet based bioassay resulted in 100% mortality of the neonate larvae of H. armigera and S. litura for the isolate T532 which was comparable to that of the positive control (HD1). The full length cry2A gene was amplified and cloned into E. coli DH5α strain. The recombinant plasmids were sequenced and the sequence was deposited in GenBank nucleotide database. The accession number of the gene is MH475905. The gene was named by the Bacillus thuringiensis delta endotoxin nomenclature committee as cry2Aa23.

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Characterization and cloning of the cry2A gene from indigenous isolates of Bacillus thuringiensis

Cited by 6 publications

References 24 publications

Characterisation of native <i>Bacillus thuringiensis</i> isolates toxicity to fall armyworm, <i>Spodoptera frugiperda</i> (J.E. Smith)

Characterisation of native <i>Bacillus thuringiensis</i> isolates toxicity to fall armyworm, <i>Spodoptera frugiperda</i> (J.E. Smith)

Characterization and Evaluation of Indigenous Bacillus thuringiensis Isolate T352 against Fall Armyworm, Spodoptera frugiperda (J.E. Smith)

Molecular characterization of an indigenous lepidopteran toxic Bacillus thuringiensis strain T532

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