Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

Jang, Eun-Kyung; Jung, Byung Kwon; Park, Gun-Seok; Khan, Abdur Rahim; Hong, Sung-Jun; Park, Yeong-Jun; Kim, Wonchan; Shin, Jae‐Ho; Al‐Ghamdi, Khalid A.; Johny, Bassam Oudh Al; Anwar, Yasir; Siddiqui, Muhammad Faisal; Ullah, Ihsan

doi:10.1016/j.aspen.2019.12.013

Cited by 6 publications

(2 citation statements)

References 25 publications

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“…The representative insecticidal proteins of Xenorhabdus include Xpt [ 8 ], Txp40 toxin [ 9 ], XaxAB [ 10 ], XnGroE [ 11 ], and PirAB [ 12 ]. Photorhabdus include a wide range of insecticidal proteins including multiunit toxin complexes (Tc), Photorhabdus insect-related (PirAB) toxins, XaxAB, and Photox binary toxins, Makes caterpillar floppy (Mcf), Photorhabdus virulence cassettes (PVC), Photorhabdus insecticidal toxin (Pit), and a ubiquitous Txp40 toxin [ 13 , 14 , 15 , 16 , 17 ]. In addition to the insecticidal proteins without catalytic activity of symbiotic bacteria, digestive enzymes that degrade an insect body are produced by bacteria to provide food for both bacteria and nematodes [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes

Son,

Kim,

Choo

et al. 2024

Toxins

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Xenorhabdus and Photorhabdus, bacterial symbionts of entomopathogenic nematodes Steinernema and Heterorhabditis, respectively, have several biological activities including insecticidal and antimicrobial activities. Thus, XnChi, XhChi, and PtChi, chitinases of X. nematophila, X. hominickii, and P. temperata isolated from Korean indigenous EPNs S. carpocapsae GJ1-2, S. monticolum GJ11-1, and H. megidis GJ1-2 were cloned and expressed in Escherichia coli BL21 to compare their biological activities. Chitinase proteins of these bacterial symbionts purified using the Ni-NTA system showed different chitobiosidase and endochitinase activities, but N-acetylglucosamidinase activities were not shown in the measuring of chitinolytic activity through N-acetyl-D-glucosarmine oligomers. In addition, the proteins showed different insecticidal and antifungal activities. XnChi showed the highest insecticidal activity against Galleria mellonella, followed by PtChi and XhChi. In antifungal activity, XhChi showed the highest half-maximal inhibitory concentration (IC50) against Fusarium oxysporum with 0.031 mg/mL, followed by PtChi with 0.046 mg/mL, and XnChi with 0.072 mg/mL. XhChi also showed the highest IC50 against F. graminearum with 0.040 mg/mL, but XnChi was more toxic than PtChi with 0.055 mg/mL and 0.133 mg/mL, respectively. This study provides an innovative approach to the biological control of insect pests and fungal diseases of plants with the biological activity of symbiotic bacterial chitinases of entomopathogenic nematodes.

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Section: Introductionmentioning

confidence: 99%

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes

Son,

Kim,

Choo

et al. 2024

Toxins

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“…On the contrary, other Photorhabdus toxins are insecticidal per se by injection [ 13 ]. A number of Tc orthologues has been identified in other bacterial genera including Xenorhabdus, Yersinia, Serratia, Pseudomonas, Burkholderia, Shewanella, Erwinia and Aeromonas , suggesting that Tc gene is highly conserved in the Enterobacteriales order [ 6 , 14 ]. In addition, Tc orthologues were reported from gram-positive genera Bacillus [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

The induced knockdown of GmCAD receptor protein encoding gene in Galleria mellonella decreased the insect susceptibility to a Photorhabdus akhurstii oral toxin

et al. 2021

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Photorhabdus bacteria secrete a repertoire of protein toxins that can kill the host insect. Among them, toxin complex (Tc) proteins have gained significant attention due to their wider conservation across the different bacterial genera. In our laboratory, a C-terminal domain of TcaB protein was characterized from P. akhurstii bacterium that conferred the potent oral insecticidal effect on Galleria mellonella . However, the role of insect gut receptors in the TcaB intoxication process was yet to be investigated. In the current study, we examined the transcription of candidate midgut receptors in TcaB-infected larvae and subsequently cloned a cadherin-like gene, GmCAD , from G. mellonella. GmCAD was highly transcribed in the fourth-instar larval stage and specifically in the midgut tissues. Our ligand blot and binding ELISA assays indicated that TcaB binds to the truncated peptides from the GmCAD transmembrane-proximal region with greater affinity than that from the transmembrane-distal region. Oral administration of bacterially expressed GmCAD dsRNA in G. mellonella severely attenuated the expression of target mRNA, which in turn alleviated the negative effect of TcaB on insect survival (TcaB-induced mortality in CAD dsRNA pretreated larvae reduced by 72–83% compared to control), implying the association of GmCAD in the TcaB intoxication process. Present findings form a basis of future research related to the insect gut receptor interactions with Photorhabdus toxins.

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Larvicidal activity of plant extracts by inhibition of detoxification enzymes in Culex pipiens

Al-Solami

2021

Journal of King Saud University - Science

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Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system

Cited by 6 publications

References 25 publications

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes

The induced knockdown of GmCAD receptor protein encoding gene in Galleria mellonella decreased the insect susceptibility to a Photorhabdus akhurstii oral toxin

Larvicidal activity of plant extracts by inhibition of detoxification enzymes in Culex pipiens

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