Complete Genome Sequence of Bacillus thuringiensis Serovar Israelensis Strain HD-789

Doggett, Norman A.; Stubben, Chris; Chertkov, Olga; Bruce, David; Detter, John C.; Johnson, Shannon L.; Han, Cliff

doi:10.1128/genomea.01023-13

Cited by 29 publications

(44 citation statements)

References 11 publications

(10 reference statements)

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“…In addition, the SDS-PAGE profiles obtained from different total protein extractions from crystal/spore mixture, were all strikingly different from those of Bti -H14 despite sharing many protein bands in Cry and Cyt regions [43]. In all cases, Bt 63 showed an extra band of (68–72 to 99 kDa), possibly belonging to the naturally truncated Cry11, truncated fragment of a second cry 4B gene or of cry 60 genes [49]. …”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.)

et al. 2016

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BackgroundWorldwide, mosquito vectors are transmitting several etiological agents of important human diseases, including malaria, causing millions of deaths every year. In Saudi Arabia, as elsewhere, vector-control is based mostly on chemical insecticides which may be toxic and cause environmental deprivation. Here, to support the development of bio-pesticide alternatives, a study was conducted to identify native Bacillus thuringiensis (Bt) isolates with improved toxicity against the malaria vector, Anopheles gambiae (s.l.).MethodsSixty-eight Bt isolates were obtained from 300 soil and other samples collected from 16 sites across Saudi Arabia. Bt identification was based on morphological characteristics of colonies, shape of parasporal crystals and biochemical profiles. After characterization of their mosquitocidal activity, larvicidal strains were described through 16S ribosomal DNA gene sequencing, cry, cyt and chi genes PCR-amplification profiles, and SDS-PAGE protein analyses.ResultsSpherical Bt crystals were predominant amongst the 68 isolates (34%), while irregular, bi-pyramidal and spore-attached crystals were found in 32, 13 and 21% of strains, respectively. LC50 and LC90 bioassays showed that 23/68 isolates were larvicidal, with distinct biochemical activity profiles compared to non-larvicidal Bt strains. Eight larvicidal strains showed larvicidal activity up to 3.4-fold higher (LC50 range: 3.90–7.40 μg/ml) than the reference Bti-H14 strain (LC50 = 13.33 μg/ml). Of these, 6 strains had cry and cyt gene profiles similar to Bti-H14 (cry4Aa, cry4Ba, cry10, cry11, cyt1Aa, cyt1Ab, cyt2Aa). The seventh strain (Bt63) displaying the highest larvicidal activity (LC50 = 3.90 μg/ml) missed the cry4Aa and cyt1Ab genes and had SDS-PAGE protein profiles and spore/crystal sizes distinct from Bti-H14. The eight strain (Bt55) with LC50 of 4.11μg/ml had cry and cyt gene profiles similar to Bti-H14 but gave a chi gene PCR product size of 2027bp. No strains harbouring cry2, cry17 + 27, cry24 + 40, cry25, cry29, cry30, or cyt2Ba were detected.ConclusionThis study represents the first report of several Saudi indigenous Bt strains with significantly higher larvicidal efficacy against An. gambiae than the reference Bti-H14 strain. The very high toxicity of the Bt63 strain, combined with distinct cry and cyt genes and SDS-PAGE-protein profiles makes it a promising candidate for future applications in mosquito bio-control.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1922-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.)

et al. 2016

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“…Interestingly, the flanking sequences matched the recently released plasmid p01 (349,599 bp) from B. thuringiensis sv. israelensis HD-789 (accession number: CP003764) (Doggett et al, 2013). Several PCR amplicons of pXO16 from p01-based primers were also sequenced and always perfectly matched with p01 (data not shown).…”

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confidence: 99%

pXO16 from Bacillus thuringiensis serovar israelensis: Almost 350 kb of terra incognita

Makart

Gillis

Mahillon

2015

Plasmid

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“…Based on whole-genome comparisons, two intergenic regions were selected for designing primer pairs targeting the Bcg and B. thuringiensis serovar israelensis, respectively ( Table 2). The region flanked by the primers Bcg1_for/rev (Table 2) is located across an intergenic region and a gene predicted as a transcriptional regulator (region 5274038 to 5274283 of reference genome Bacillus thuringiensis serovar israelensis HD-789; NCBI accession number CP003763) (35). For the primers Bti1_for/rev, the target is located in an intergenic region (positions 3152129 to 3152331) (35).…”

Section: Resultsmentioning

confidence: 99%

“…The region flanked by the primers Bcg1_for/rev (Table 2) is located across an intergenic region and a gene predicted as a transcriptional regulator (region 5274038 to 5274283 of reference genome Bacillus thuringiensis serovar israelensis HD-789; NCBI accession number CP003763) (35). For the primers Bti1_for/rev, the target is located in an intergenic region (positions 3152129 to 3152331) (35). The BLAST similarity searches with concatenated Bcg-targeting primer sequences on the WGS database of NCBI recovered 253 hits with 100% sequence similarity for the full query length ( 2015).…”

Section: Resultsmentioning

confidence: 99%

Chromosome-Directed PCR-Based Detection and Quantification of Bacillus cereus Group Members with Focus on B. thuringiensis Serovar israelensis Active against Nematoceran Larvae

Schneider

Hendriksen

Melin

et al. 2015

Appl Environ Microbiol

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e Bacillus thuringiensis serovar israelensis is a wide-spread soil bacterium affiliated with the B. cereus group (Bcg) and is widely used in biocontrol products applied against mosquito and black fly larvae. For monitoring and quantification of applied B. thuringiensis serovar israelensis and its effect on indigenous B. thuringiensis serovar israelensis and Bcg assemblages, efficient and reliable tools are essential. The abundance and properties of B. thuringiensis serovar israelensis strains in the environment traditionally have been investigated with cultivation-dependent techniques, which are hampered by low sensitivity and the morphological similarity between B. cereus and B. thuringiensis. Currently available PCR-based detection and quantification tools target markers located on plasmids. In this study, a new cultivation-independent PCR-based method for efficient and specific quantification of B. thuringiensis serovar israelensis and Bcg is presented, utilizing two sets of PCR primers targeting the bacterial chromosome. Sequence database searches and empirical tests performed on target and nontarget species, as well as on bulk soil DNA samples, demonstrated that this diagnostic tool is specific for B. thuringiensis serovar israelensis and Bcg. The method will be useful for comparisons of Bcg and B. thuringiensis serovar israelensis abundances in the same samples. Moreover, the effect of B. thuringiensis serovar israelensis-based insecticide application on the total Bcg assemblages, including indigenous populations, can be investigated. This type of information is valuable in risk assessment and policy making for use of B. thuringiensis serovar israelensis in the environment. The bacterium Bacillus thuringiensis (Bt) is one of the grand successes in microbial pest control (1). After its first discovery in 1901 and description by Berliner in 1911, the first biocontrol product based on Bt, Sporein, for control of the European corn borer (Ostrinia nubilalis), was commercially available already in 1938 (1). The crystalline, proteinaceous ␦-endotoxins of Bt are formed during sporulation, and their lethal effect is manifested only after larval ingestion of spores or free crystals (2, 3). Bt is a member of the Bacillus cereus group (Bcg), which consists of the Gram-positive rod-shaped spore-forming bacterial species B. cereus, Bt, B. anthracis, B. mycoides, B. pseudomycoides, and B. weihenstephanensis (4,5). However, a multidatatype phylogenetic analysis of Bcg strains isolated from various sample types, including environmental, clinical, food, and diary sources, did not reflect current Bcg taxonomy well. This finding indicated that the phylogeny of this bacterial group is more complicated than reflected by the division into the current six species (6).The bacterium B. thuringiensis serovar israelensis produces toxins active against nematoceran larvae and has been widely used for biocontrol of mosquitoes and black flies (3). B. thuringiensis serovar israelensis was described in 1976 (7) and has been shown to have ...

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Complete Genome Sequence of Bacillus thuringiensis Serovar Israelensis Strain HD-789

Cited by 29 publications

References 11 publications

Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.)

Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.)

pXO16 from Bacillus thuringiensis serovar israelensis: Almost 350 kb of terra incognita

Chromosome-Directed PCR-Based Detection and Quantification of Bacillus cereus Group Members with Focus on B. thuringiensis Serovar israelensis Active against Nematoceran Larvae

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