PCR-based method for the detection of cry genes in local isolates of Bacillus thuringiensis from Thailand

Thammasittirong, Anon; Attathom, Tipvadee

doi:10.1016/j.jip.2008.03.001

Cited by 29 publications

(19 citation statements)

References 17 publications

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“…As indicated previously (Bravo et al, 1998;Thammasittirong and Attathom, 2008;Nazarian et al, 2009;Vidal-Quest et al, 2009), we also observed that the cry2 gene was present in combination with the cry1 gene and this combination was the most frequent (40%) out of all of the cry gene combinations (Figure 1). Moreover, some of the Bt strains were found to contain other combinations of the cry genes, such as the lepidopteran-active cry1 gene and the coleopteran-active cry3 gene.…”

Section: Discussionsupporting

confidence: 89%

Distribution, occurrence of cry genes, and lepidopteran toxicity of native Bacillus thuringiensis isolated from fig tree environments in Aydın Province

Alper¹,

Güneş

Tatlipinar³

et al. 2014

Turk J Agric For

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Bacillus thuringiensis (Bt) has a significant impact on biological pest control because of the insecticidal activity through its parasporal inclusion bodies (crystal proteins). Fig is an economically important plant in Turkey; agricultural pests result in a considerable economic loss in fig quality and cultivation. The aim of this work was to isolate, characterize, and determine the lepidopteran toxicity of Bt obtained from fig groves in Aydın Province. A total of 606 colonies (out of 1167) obtained from 380 samples were identified as Bt based on parasporal crystal formation. The highest Bt index of 0.60 was observed in the Kuyucak region. A total of 288 Bt isolates were characterized in terms of cry gene content by PCR analysis. It was found that the cry1 plus cry2 genotype was the most abundant (40%) in our collection. Bioactivity tests indicated that 6 isolates exhibited high mortalities against Cadra cautella and 3 isolates were found to exhibit high toxicity against Carpophilus hemipterus. Moreover, 13 Bt isolates exhibiting toxic activity against fig pests were further characterized based on specific cry gene content, protein profiles, and PCR-RFLP analysis. Among cry1 genes, the cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1D, and cry1Ea genes were the most frequent (100%). Protein profiles of isolates toxic to C. cautella were different from those of isolates toxic to C. hemipterus. PCR-RFLP analysis indicated that toxic isolates differed from the reference strain with respect to cry1 type gene. Finally, it was concluded that Bt strains isolated from fig groves showed high level of toxicity against fig pests. These strains can serve as potential biopesticides for the control of C. cautella in the region as well as alternative biopesticides in the case of pesticide resistance in insects.

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Section: Discussionsupporting

confidence: 89%

Distribution, occurrence of cry genes, and lepidopteran toxicity of native Bacillus thuringiensis isolated from fig tree environments in Aydın Province

Alper¹,

Güneş

Tatlipinar³

et al. 2014

Turk J Agric For

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“…The results reported here differ from the results of a recent survey of Bt isolates obtained in Thailand by Thammasittirong and Attathom 12 in which cry1A, found at a relatively low frequency in the current work, was the most prevalent type. Moreover, cry1F was not found in the previous study 12 , but occurred at a relatively high frequency (22.6%) in the present study. Such variability in cry1 gene types and their frequencies is thought to be due to differences in the geography and ecology of the collection sites and/or artefacts arising from the methods used for gene type identification 12,[27][28][29] .…”

Section: Discussioncontrasting

confidence: 83%

“…Moreover, cry1F was not found in the previous study 12 , but occurred at a relatively high frequency (22.6%) in the present study. Such variability in cry1 gene types and their frequencies is thought to be due to differences in the geography and ecology of the collection sites and/or artefacts arising from the methods used for gene type identification 12,[27][28][29] . The presence of more than one cry-type gene in individual isolates was not surprising since, in addition to their own chromosomal cry gene, strains can carry plasmids that encode additional cry genes 31,32 .…”

Section: Discussioncontrasting

confidence: 83%

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Lertwiriyawong¹,

Pinthong²,

Chanpaisaeng³

et al. 2010

ScienceAsia

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ABSTRACT:The novel δ-endotoxin from Bacillus thuringiensis (Bt) is one of the alternative measures for lepidopteran pest control. A survey of 31 Bt isolates, obtained from Thailand and previously screened for their toxicity to lepidopterans, was conducted to determine the presence of cry1-type genes, using polymerase chain reaction -restriction fragment length polymorphism analysis. Seven distinct types of cry1 genes: cry1Aa, cry1B, cry1C, cry1Cb, cry1D, cry1E, and cry1F, were identified. The most common of the cry1-type genes was cry1Cb, followed by cry1C and cry1D, which covered 64.6, 48.4, and 25.8%, respectively. Besides cry1C and cry1Cb, two candidate cry genes, cry1E and cry1D, of isolate JC 190 (harbouring cry1C/1E), with cotton bollworm toxin specificity, showed 99% amino acid sequence identity to Cry1Ea of B. thuringiensis subsp. kenyae, while isolate JC 291 (containing cry1C/1Cb/1D), with Asian corn borer toxin specificity, harboured Cry1Dc, which exhibited only 84% amino acid sequence identity to Cry1Da of B. thuringiensis subsp. aizawai. The C-terminus of the JC291Cry1Dc possessed a unique sequence, 812-NVPIIPIISPW-822. Furthermore, the major differences between the 3D structure of Cry1Dc and Cry1Aa were confined to Domain II as three loop structures.

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“…In addition to 16S rDNA sequence analysis, PCR detection of the cry2 gene specific for most B. thuringiensis was performed. The amplification was achieved using the cry2-F (5ʹ-TACCTTTATTTGCACAGGCA-3ʹ) and cry2-R (5ʹ-CTACCGTTTATAGTAACTCG-3ʹ) primers following the previously described PCR conditions (Thammasittirong and Attathom, 2008). …”

Section: Identification Of the Most Potent Phb-producing Isolatementioning

confidence: 99%

Efficient polyhydroxybutyrate production from Bacillus thuringiensis using sugarcane juice substrate

2017

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The present study focused on the screening and optimization of biopolymer polyhydroxybutyrate (PHB) production by Bacillus spp. using cost-effective substrates. Among 602 local Bacillus isolates, Bacillus thuringiensis B417-5 produced the highest amount of PHB (2.278 g/L, 60.07% of dry cell weight, DCW). 1 H NMR and FTIR analyses of the extracted polymer revealed the characteristic peaks of PHB. The optimization results showed that the highest PHB accumulation (2.768 g/L, 72.08% of DCW) was achieved when culturing B. thuringiensis B417-5 in a nitrogen-deficient medium containing 1% total sugar from sugarcane juice and 0.5% yeast extract, with a pH of 7.0 and an incubation temperature of 37 °C for 48 h. B. thuringiensis B417-5 can thus be considered a good candidate for large-scale production of PHB. We are reporting for the first time that sugarcane juice is a promising carbon source for economical PHB production by B. thuringiensis.

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PCR-based method for the detection of cry genes in local isolates of Bacillus thuringiensis from Thailand

Cited by 29 publications

References 17 publications

Distribution, occurrence of cry genes, and lepidopteran toxicity of native Bacillus thuringiensis isolated from fig tree environments in Aydın Province

Distribution, occurrence of cry genes, and lepidopteran toxicity of native Bacillus thuringiensis isolated from fig tree environments in Aydın Province

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Efficient polyhydroxybutyrate production from Bacillus thuringiensis using sugarcane juice substrate

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