For: Pesticide biochemistry and physiology recG is involved with the resistance of Bt to UV

Xu, Jin; Wu, Chenxu; Yang, Zemin; Liu, Wen‐Cheng; Chen, Hong; Batool, Khadija; Yao, Junmin; Fan, Xuetong; Wu, Juan; Rao, Wenhua; Huang, Tianpei; Xu, Lei; Guan, Xiong; Zhang, Lingling

doi:10.1016/j.pestbp.2020.104599

Cited by 8 publications

(14 citation statements)

References 42 publications

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“…The helicase regulates the microbial response to stress, and the same family helicases can perform different functions [ 31 , 32 ]. In the UV treatment test, the survival rate of tested Bt strains decreased after UV stress.…”

Section: Discussionmentioning

confidence: 99%

“…Knockout gene homologous recombination and identification of knockout strains were performed as previously described [ 31 ]. The mutant strain was obtained by high temperature homologous recombination reaction.…”

Section: Methodsmentioning

confidence: 99%

“…DNA helicase had been found to participate in the stress response of Bt. The DNA helicase recG in B. thuringiensis is associated with UV resistance, and recG knockout strains are more sensitive to UV light [ 31 ]. A variety of RNA helicases have been identified in B. cereus , which can participate in stress reactions under heat, oxidation, and pH stress [ 32 , 33 ].…”

Section: Introductionmentioning

confidence: 99%

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Whole Genome Resequencing Revealed the Effect of Helicase yqhH Gene on Regulating Bacillus thuringiensis LLP29 against Ultraviolet Radiation Stress

Guan

Miao

et al. 2023

IJMS

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Bacillus thuringiensis (Bt) is a widely used microbial pesticide. However, its duration of effectiveness is greatly shortened due to the irradiation of ultraviolet rays, which seriously hinders the application of Bt preparations. Therefore, it is of great importance to study the resistance mechanism of Bt to UV at the molecular level to improve the UV-resistance of Bt strains. In order to know the functional genes in the UV resistance, the genome of UV-induced mutant Bt LLP29-M19 was re-sequenced and compared with the original strain Bt LLP29. It was shown that there were 1318 SNPs, 31 InDels, and 206 SV between the mutant strain and the original strain Bt LLP29 after UV irradiation, which were then analyzed for gene annotation. Additionally, a mutated gene named yqhH, a member of helicase superfamily II, was detected as an important candidate. Then, yqhH was expressed and purified successfully. Through the result of the enzymatic activity in vitro, yqhH was found to have ATP hydrolase and helicase activities. In order to further verify its function, the yqhH gene was knocked out and complemented by homologous recombinant gene knockout technology. The survival rate of the knockout mutant strain Bt LLP29-ΔyqhH was significantly lower than that of the original strain Bt LLP29 and the back-complemented strain Bt LLP29-ΔyqhH-R after treated with UV. Meanwhile, the total helicase activity was not significantly different on whether Bt carried yqhH or not. All of these greatly enrich important molecular mechanisms of Bt when it is in UV stress.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Whole Genome Resequencing Revealed the Effect of Helicase yqhH Gene on Regulating Bacillus thuringiensis LLP29 against Ultraviolet Radiation Stress

Guan

Miao

et al. 2023

IJMS

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“…According to the previously reported method (Zhang et al 2016), the anti-UV-B ability determination of Bt LLP29 and Bt LLP29-M19 were compared as follows. After cultured, collected, and washed twice with 0.01 M PBS, the same volumes of cells (OD 600 = 1.0) were resuspended and added to a 6-deep well plate and placed in an ultraviolet cross-linker (SCIENTZ 03-II, Ningbo Scientz Biotechnology Co., Ltd., China) (Xu et al 2020;Zhang et al 2018). Samples were taken at 0, 60, and 120 min with 50 W UVB lamps and then diluted 10 − 4 times with LB (gradually diluted, 10 − 2 times each time).…”

Section: Anti-uv-b Ability Determination Of Btmentioning

confidence: 99%

“…Because it is environmentally friendly, Bt has gradually become the most widely used microbial pesticide in the eld against a variety of insect species, such as Lepidoptera, Hemiptera, Diptera, and Nematodes (Alam et The persistence of Bt is an important factor in determining the success of this product as a pest control agent. However, the ultraviolet radiation in the environment causes Bt spores and insecticidal crystal proteins (ICPs) to be easily degraded, which greatly reduces the application durability and e ciency of Bt, and then increases the cost of the practical application of Bt, limiting its use on a global scale ; Xu et al 2020; Zhang et al 2016). Therefore, the mechanism and application of Bt anti-ultraviolet rays have been investigated for a long time, including the identi cation and screening of Bt antiultraviolet genes (Xu et al 2020; Zhang et al 2008), mutagenesis of UV radiation resistant strains (Hoti and Balaraman 1993;Patel et al 1996;Saxena et al 2002;Zhang et al 2016), researches on improving melanin production (Hoti and Balaraman 1993;Ruan et al 2004), formulating a chemical that can absorb UV rays or biological control agents and so on (Cohen et al 1991;Kaiser et al 2019;Pan et al 2017).…”

Section: Introductionmentioning

confidence: 99%

TCA Cycle Difference is Greatly Related to the Resistance of Bacillus thuringiensis against UV Stress

Zhang

et al. 2023

Preprint

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Bacillus thuringiensis (Bt) is a popular and environment-friendly biopesticide. However, similar to other microbial pesticides, Bt is limited by ultraviolet (UV) radiation during its application, which greatly reduces its toxicity and persistence. To further know the mechanism of Bt against UV radiation, metabolomic profiles between Bt LLP29 and its UV-resistant mutant LLP29-M19 were compared, analyzed, and annotated in this study, and then a total of 61 metabolites with different abundances were detected. With P < 0.05 as the standard, a total of 12 metabolic pathways were enriched, including the TCA cycle. According to the result of RT-qPCR, the expression levels of the TCA cycle key genes in Bt LL29-M19, such as icd1 citZ, citB, sdhA, sdhB, sdhC, fumA and mdh, were found down-regulated for 85.58%, 37.02%, 70.87%, 85.97%, 76.33%, 83.15%, 87.28%, and 35.77% than those in Bt LLP29. It was consistent with the down-regulation trend of the TCA cycle key enzymes activity in Bt LLP29-M19. Consistently, the enzyme activities of ICDH, SDH, and PDH in LLP29-M19 were detected 86.28%, 43.93%, and 83.03% lower than those in Bt LLP29. It was revealed that the reduced TCA cycle was required for Bt UV radiation resistance, which was also demonstrated by the addition of inhibitors furfural and malonic acid, respectively. Based on the result of RT-qPCR, the gene transcription levels of the main reactive oxygen species (ROS) generation pathways were down-regulated, such as EMP, however, the activity of the main degrading enzymes was up-regulated, which showed the reduction of ROS generation rate was a way for the TCA cycle to regulate the anti-ultraviolet resistance of Bt. All of these provide solid evidence for reprogramming metabolomics to strengthen Bt UV radiation resistance.

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Effect of yhfS gene on Bt LLP29 antioxidant and UV ray resistance

Liu

Zhang

et al. 2023

Pest Management Science

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BACKGROUND: Bacillus thuringiensis (Bt) is a widely used microbial insecticide. However, its persistence is limited because of ultraviolet (UV) rays or other environmental factors. The yhfS gene, which encodes acetyl-CoA acyltransferase, plays an important role in lipid transport and metabolism in many organisms. To explore whether it is related to the stress resistance of Bt LLP29, the yhfS gene knockout strain LLP29 Δ-yhfS and the complementary strain LLP29 R-yhfS were generated successfully by homologous recombination technology, and the related phenotypic changes were compared in this study. RESULTS: Gene yhfS was found to be functional in response to UV radiation in Bt by comparing the survival rates of Bt LLP29 harboring yhfS or not under UV light. Enzyme activity assays of key enzymes showed the the Embden-Meyerhof-Parnas pathway was enhanced yet the tricarboxylic acid cycle as well as butanoate synthesis were repressed when the gene was deleted. At the same time, the amino acid content was decreased, but reduced nicotinamide adenine dinucleotide (NADH) and reactive oxygen species (ROS) content were increased. Most noteworthy, antioxidase (such as superoxide dismutase and peroxidase) activities and contents of some potent antioxidants (such as pyruvate, carotenoids and NADPH) were lower in LLP29 Δ-yhfS than in LLP29.CONCLUSION: These tests revealed that the loss of the yhfS gene led to metabolic disorders and reduction of the antioxidant ability of Bt. Higher ROS level and lower anti-oxidative capacity might be responsible for the reduced UV resistance when the gene was deleted. These results not only greatly enrich understanding of the mechanism of Bt UV resistance, but also provide an important theoretical basis for Bt application.

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For: Pesticide biochemistry and physiology recG is involved with the resistance of Bt to UV

Cited by 8 publications

References 42 publications

Whole Genome Resequencing Revealed the Effect of Helicase yqhH Gene on Regulating Bacillus thuringiensis LLP29 against Ultraviolet Radiation Stress

Whole Genome Resequencing Revealed the Effect of Helicase yqhH Gene on Regulating Bacillus thuringiensis LLP29 against Ultraviolet Radiation Stress

TCA Cycle Difference is Greatly Related to the Resistance of Bacillus thuringiensis against UV Stress

Effect of yhfS gene on Bt LLP29 antioxidant and UV ray resistance

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