“…The highest mortality for E. kuehniella happened by the Tunisia isolate, BLB1, harboring cry1 and cry2 in comparison with Btk HD1 (Saadaoui et al, 2010). In comparison with the current bioassay result, some Turkish Bt strains harboring cry2Ab and cry2Aa1 led to 42% mortality in E. kuehniella (higher than Btk) (Alper et al, 2016).…”
Section: Discussioncontrasting
confidence: 44%
“…A previous investigation revealed that molecular mass of Cry1Aa, Cry1Ac, and Cry1C proteins were 133.2, 133.3, and 134.8 kDa analyzed through the SDS-PAGE, respectively (Kalman et al, 1995). All strains studied by Alper et al (2016), harboring the cry1 and cry2 genes, and the reference strain, Btk, almost revealed similar crystal protein contents. Similar results have been achieved for the most Iranian strains studied in the present research.…”
Background
Insecticidal crystal proteins (encoded by cry genes) produced by Bacillus thuringiensis (Bt) are fatal for insects of different orders such as Lepidoptera. The genes that encoded these crystal proteins can be detected on plasmids and chromosomal DNA and show different types in various strains. Therefore, the objective of this study was to determine molecular characteristics of Iranian Bt strains as well as their toxicity against Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae).
Results
The collection sites included fields, gardens, and desert and semi-desert areas in 8 provinces of Iran. For crystal formation, each isolate was cultured in T3 medium. The results showed that the Bt isolates produced different types of crystals including spherical (73.33%), bipyramidal (53.33%), irregular (40%), cubical (33.33%), and elliptical (13.33%). Plasmid DNA extraction was performed and showed that most of the strains exhibited similar pattern in the number and the size of the plasmid bands to those of Bt kurstaki (Btk). Some specific primers were used for PCR amplification to distinguish different crystal genes including cry1 (A, C, and D) and cry2A (a and b). The primers related to cry1D and cry2Aa1 genes produced no amplicons. The results revealed that the most abundant gene was cry1-type. All strains analyzed for the cry2Ab2 gene presented unexpected bands. Electrophoretic profile of the protein crystals showed bands with different diversity in number, and size ranged from about 16 to 140 KDa. The bioassay result of some more toxic strains exhibited that the pathogenicity of 1019 was higher than the rest, even the reference strain, Btk. However, the toxicity of other strains was the same as Btk. According to the phylogenetic tree, 1019 was located in the same group with Bt subspecies coreanensis, Bt subspecies indiana and Bt subspecies tolworthi.
Conclusions
The investigated Iranian strains had the lepidopteran-active cry genes. The strains with the same toxicity to E. kuehniella had various cry genes, plasmid, and crystal protein profiles and vice versa. Therefore, characterization of cry genes of native strains could lead to access potent isolates as biocontrol agents against native insect pests.
“…The highest mortality for E. kuehniella happened by the Tunisia isolate, BLB1, harboring cry1 and cry2 in comparison with Btk HD1 (Saadaoui et al, 2010). In comparison with the current bioassay result, some Turkish Bt strains harboring cry2Ab and cry2Aa1 led to 42% mortality in E. kuehniella (higher than Btk) (Alper et al, 2016).…”
Section: Discussioncontrasting
confidence: 44%
“…A previous investigation revealed that molecular mass of Cry1Aa, Cry1Ac, and Cry1C proteins were 133.2, 133.3, and 134.8 kDa analyzed through the SDS-PAGE, respectively (Kalman et al, 1995). All strains studied by Alper et al (2016), harboring the cry1 and cry2 genes, and the reference strain, Btk, almost revealed similar crystal protein contents. Similar results have been achieved for the most Iranian strains studied in the present research.…”
Background
Insecticidal crystal proteins (encoded by cry genes) produced by Bacillus thuringiensis (Bt) are fatal for insects of different orders such as Lepidoptera. The genes that encoded these crystal proteins can be detected on plasmids and chromosomal DNA and show different types in various strains. Therefore, the objective of this study was to determine molecular characteristics of Iranian Bt strains as well as their toxicity against Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae).
Results
The collection sites included fields, gardens, and desert and semi-desert areas in 8 provinces of Iran. For crystal formation, each isolate was cultured in T3 medium. The results showed that the Bt isolates produced different types of crystals including spherical (73.33%), bipyramidal (53.33%), irregular (40%), cubical (33.33%), and elliptical (13.33%). Plasmid DNA extraction was performed and showed that most of the strains exhibited similar pattern in the number and the size of the plasmid bands to those of Bt kurstaki (Btk). Some specific primers were used for PCR amplification to distinguish different crystal genes including cry1 (A, C, and D) and cry2A (a and b). The primers related to cry1D and cry2Aa1 genes produced no amplicons. The results revealed that the most abundant gene was cry1-type. All strains analyzed for the cry2Ab2 gene presented unexpected bands. Electrophoretic profile of the protein crystals showed bands with different diversity in number, and size ranged from about 16 to 140 KDa. The bioassay result of some more toxic strains exhibited that the pathogenicity of 1019 was higher than the rest, even the reference strain, Btk. However, the toxicity of other strains was the same as Btk. According to the phylogenetic tree, 1019 was located in the same group with Bt subspecies coreanensis, Bt subspecies indiana and Bt subspecies tolworthi.
Conclusions
The investigated Iranian strains had the lepidopteran-active cry genes. The strains with the same toxicity to E. kuehniella had various cry genes, plasmid, and crystal protein profiles and vice versa. Therefore, characterization of cry genes of native strains could lead to access potent isolates as biocontrol agents against native insect pests.
“…So far a large number of Bt strains have been isolated and characterized for their sequence similarities within these cry genes. A total of 700 cry genes have been isolated till date (Alper et al 2016) and interestingly, single strains of Bt may contain more than one crystal protein genes (Letowski et al 2005;Pedro and Ibarra 2010). Although there are variations within the cry genes, the three domain of Cry proteins have conserved domains in their tertiary structures of the active toxin.…”
We have identified both crystalliferous and acrystalliferous Bt isolates from the Assam soil of North East India for the first time. A total of 301 type colonies were selected based on their appearance and colony morphology. Out of these colonies, 42 isolates had characteristics similar to Bt isolates on MYP (Mannitol Egg Yolk Polymyxin) agar base medium. The ERIC-PCR and 16S rDNA analyses confirmed that 42 isolates are. Phase contrast microscopy showed that 37 isolates produced crystal endospore during the sporulation phase and 5 acrystalliferous isolates were also found. Amplification of gene was carried out using general Cry primers along with one gene specific primer. Out of 42 isolates, 50% of the isolates showed presence of gene followed by (40.47) and (40.47). Moreover, 21.42% of isolates showed the presence of more than one genes. We also screened these isolates for the possibility of having new Bt genes using universal primer and found two strains having a new type of gene with 82 and 85% similarities with the available gene sequences. Thus, these new types of Bt gene could be useful for Bt-based bioformulations and generation of transgenic plants.
“…The researchers concluded that this was due to the decrease in the number of eggs laid by females in plants sprayed with Dipel. In another study, Alper et al (2013), in their study examining the effect of a spore-crystal mixture of 31 natural B. thuringiensis isolates on T. urticae nymphs, noted that some isolates may have a certain potential in the development of biopesticides that especially retard the growth of nymphs. Moreover, Neethu et al (2016), stated that toxin-producing bacteria such as B. thuringiensis have been widely used as bio-acaricide in recent years.…”
The purpose of this study was to assess the usability of two bacterial strains, namely Bacillus subtilis PA1 and Paenibacillus azotofixans PA2, for the biological control of red spider species, and their effects on plant quality and yield in cotton under field conditions. The experiments were conducted at three different locations with multiple replications. As a control, a commercial preparation containing Lambda-Cyhalothrin as the active ingredient was used. The obtained results from the study revealed that the application of the bioagent formulation led to a significant decrease in the density of Tetranychus spp. at different biological stages, ranging from 59.22% to 61.07%, when compared to the control group. Additionally, several important plant growth parameters showed remarkable improvements. The number of fruit branches increased by 130.20%, plant crown diameter by 88.16%, plant height by 40.15%, the number of flowers by 21.25%, the number of wood branches by 18.13%, the average number of cocoons by 126.53%, and cocoon weights by 54.65% significantly across all three trial parcels. The successful implementation of the bacterial application for pest control had a positive impact on cotton yield. Bulk cotton yield increased by 80.03%, and fiber yield increased by 82.17%. Consequently, the bacterial formulation containing these two bacteria demonstrated its potential as a biopesticide in cotton cultivation, effectively controlling pests while also playing a crucial role in enhancing productivity. Overall, the study suggests that using the bioagent formulation consisting of Bacillus subtilis PA1 and Paenibacillus azotofixans PA2 could be an effective and environmentally friendly approach for pest control in cotton farming, leading to increased productivity.
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