Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex
<i>Xenorhabdus nematophila</i> HB310 secreted the insecticidal protein toxin complex (Tc). The <i>chi60</i> and <i>chi70</i> chitinase genes are located on the gene cluster encoding Tc toxins. To clarify the insecticidal activity of chitinases and their relationsh...
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MDPI AG
2022-09-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/9/646 |
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| author | Jia Liu Hui Bai Ping Song Ziyan Nangong Zhiping Dong Zhiyong Li Qinying Wang |
| author_facet | Jia Liu Hui Bai Ping Song Ziyan Nangong Zhiping Dong Zhiyong Li Qinying Wang |
| author_sort | Jia Liu |
| collection | DOAJ |
| description | <i>Xenorhabdus nematophila</i> HB310 secreted the insecticidal protein toxin complex (Tc). The <i>chi60</i> and <i>chi70</i> chitinase genes are located on the gene cluster encoding Tc toxins. To clarify the insecticidal activity of chitinases and their relationship with Tc toxins, the insecticidal activity of the chitinases was assessed on <i>Helicoverpa armigera</i>. Then, the <i>chi60</i> and <i>chi70</i> genes of <i>X. nematophila</i> HB310 were knocked out by the pJQ200SK suicide plasmid knockout system. The insecticidal activity of Tc toxin from the wild-type strain (WT) and mutant strains was carried out. The results demonstrate that Chi60 and Chi70 had an obvious growth inhibition effect against the second instar larvae of <i>H. armigera</i> with growth-inhibiting rates of 81.99% and 90.51%, respectively. Chi70 had a synergistic effect with the insecticidal toxicity of Tc toxins, but Chi60 had no synergistic effect with Tc toxins. After feeding Chi60 and Chi70, the peritrophic membrane of <i>H. armigera</i> became inelastic, was easily broken and leaked blue dextran. The Δ<i>chi60</i>, Δ<i>chi70</i> and Δ<i>chi60</i>-<i>chi70</i> mutant strains were successfully screened. The toxicity of Tc toxins from the WT, Δ<i>chi60</i>, Δ<i>chi70</i> and Δ<i>chi60</i>-<i>chi70</i> was 196.11 μg/mL, 757.25 μg/mL, 885.74 μg/mL and 20,049.83 μg/mL, respectively. The insecticidal activity of Tc toxins from Δ<i>chi60</i> and Δ<i>chi70</i> was 3.861 and 4.517 times lower than that of Tc toxins from the WT, respectively, while the insecticidal activity of Tc toxins from the Δ<i>chi60</i>-<i>chi70</i> mutant strain almost disappeared. These results indicate that the presence of <i>chi60</i> and <i>chi70</i> is indispensable for the toxicity of Tc toxins. |
| first_indexed | 2024-03-09T22:19:21Z |
| format | Article |
| id | doaj.art-896d48e5bdde46d48424375ba49ecc2e |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T22:19:21Z |
| publishDate | 2022-09-01 |
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| spelling | doaj.art-896d48e5bdde46d48424375ba49ecc2e2023-11-23T19:17:19ZengMDPI AGToxins2072-66512022-09-0114964610.3390/toxins14090646Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin ComplexJia Liu0Hui Bai1Ping Song2Ziyan Nangong3Zhiping Dong4Zhiyong Li5Qinying Wang6Institute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, National Foxtail Millet Improvement Center, Minor Cereal Crops Laboratory of Hebei Province, Shijiazhuang 050035, ChinaInstitute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, National Foxtail Millet Improvement Center, Minor Cereal Crops Laboratory of Hebei Province, Shijiazhuang 050035, ChinaCollege of Plant Protection, Hebei Agricultural University, Baoding 071000, ChinaCollege of Plant Protection, Hebei Agricultural University, Baoding 071000, ChinaInstitute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, National Foxtail Millet Improvement Center, Minor Cereal Crops Laboratory of Hebei Province, Shijiazhuang 050035, ChinaInstitute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, National Foxtail Millet Improvement Center, Minor Cereal Crops Laboratory of Hebei Province, Shijiazhuang 050035, ChinaCollege of Plant Protection, Hebei Agricultural University, Baoding 071000, China<i>Xenorhabdus nematophila</i> HB310 secreted the insecticidal protein toxin complex (Tc). The <i>chi60</i> and <i>chi70</i> chitinase genes are located on the gene cluster encoding Tc toxins. To clarify the insecticidal activity of chitinases and their relationship with Tc toxins, the insecticidal activity of the chitinases was assessed on <i>Helicoverpa armigera</i>. Then, the <i>chi60</i> and <i>chi70</i> genes of <i>X. nematophila</i> HB310 were knocked out by the pJQ200SK suicide plasmid knockout system. The insecticidal activity of Tc toxin from the wild-type strain (WT) and mutant strains was carried out. The results demonstrate that Chi60 and Chi70 had an obvious growth inhibition effect against the second instar larvae of <i>H. armigera</i> with growth-inhibiting rates of 81.99% and 90.51%, respectively. Chi70 had a synergistic effect with the insecticidal toxicity of Tc toxins, but Chi60 had no synergistic effect with Tc toxins. After feeding Chi60 and Chi70, the peritrophic membrane of <i>H. armigera</i> became inelastic, was easily broken and leaked blue dextran. The Δ<i>chi60</i>, Δ<i>chi70</i> and Δ<i>chi60</i>-<i>chi70</i> mutant strains were successfully screened. The toxicity of Tc toxins from the WT, Δ<i>chi60</i>, Δ<i>chi70</i> and Δ<i>chi60</i>-<i>chi70</i> was 196.11 μg/mL, 757.25 μg/mL, 885.74 μg/mL and 20,049.83 μg/mL, respectively. The insecticidal activity of Tc toxins from Δ<i>chi60</i> and Δ<i>chi70</i> was 3.861 and 4.517 times lower than that of Tc toxins from the WT, respectively, while the insecticidal activity of Tc toxins from the Δ<i>chi60</i>-<i>chi70</i> mutant strain almost disappeared. These results indicate that the presence of <i>chi60</i> and <i>chi70</i> is indispensable for the toxicity of Tc toxins.https://www.mdpi.com/2072-6651/14/9/646<i>Xenorhabdus nematophila</i> HB310chitinasesgene knock outtoxin complexinsecticidal activity |
| spellingShingle | Jia Liu Hui Bai Ping Song Ziyan Nangong Zhiping Dong Zhiyong Li Qinying Wang Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex Toxins <i>Xenorhabdus nematophila</i> HB310 chitinases gene knock out toxin complex insecticidal activity |
| title | Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex |
| title_full | Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex |
| title_fullStr | Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex |
| title_full_unstemmed | Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex |
| title_short | Insecticidal Activity of Chitinases from <i>Xenorhabdus nematophila</i> HB310 and Its Relationship with the Toxin Complex |
| title_sort | insecticidal activity of chitinases from i xenorhabdus nematophila i hb310 and its relationship with the toxin complex |
| topic | <i>Xenorhabdus nematophila</i> HB310 chitinases gene knock out toxin complex insecticidal activity |
| url | https://www.mdpi.com/2072-6651/14/9/646 |
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