A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process
<i>Bacillus thuringiensis</i> (Bt) is used for insect pest control, and its larvicidal activity is primarily attributed to Cry toxins. Other factors participate in infection, and limited information is available regarding factors acting on the peritrophic matrix (PM). This study aimed to...
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MDPI AG
2020-04-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/4/252 |
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| author | Jiaxin Qin Zongxing Tong Yiling Zhan Christophe Buisson Fuping Song Kanglai He Christina Nielsen-LeRoux Shuyuan Guo |
| author_facet | Jiaxin Qin Zongxing Tong Yiling Zhan Christophe Buisson Fuping Song Kanglai He Christina Nielsen-LeRoux Shuyuan Guo |
| author_sort | Jiaxin Qin |
| collection | DOAJ |
| description | <i>Bacillus thuringiensis</i> (Bt) is used for insect pest control, and its larvicidal activity is primarily attributed to Cry toxins. Other factors participate in infection, and limited information is available regarding factors acting on the peritrophic matrix (PM). This study aimed to investigate the role of a Bt chitin-binding protein (CBPA) that had been previously shown to be expressed at pH 9 <i>in vitro</i> and could therefore be expressed in the alkaline gut of lepidopteron larvae. A ∆cbpA mutant was generated that was 10-fold less virulent than wild-type Bt HD73 towards <i>Ostrinia furnacalis</i> neonate larvae, indicating its important role in infection. Purified recombinant <i>Escherichia coli</i> CBPA was shown to have a chitin affinity, thus indicating a possible interaction with the chitin-rich PM. A translational GFP–CBPA fusion elucidated the localization of CBPA on the bacterial surface, and the transcriptional activity of the promoter P<i>cbpA</i> was immediately induced and confirmed at pH 9. Next, in order to connect surface expression and possible <i>in vivo</i> gut activity<i>,</i> last instar <i>Galleria</i> <i>mellonella</i> (Gm) larvae (not susceptible to Bt HD-73) were used as a model to follow CBPA in gut expression, bacterial transit, and PM adhesion. CBPA-GFP was quickly expressed in the Gm gut lumen, and more Bt HD73 strain bacteria adhered to the PM than those of the ∆cbpA mutant strain. Therefore, CBPA may help to retain the bacteria, via the PM binding, close to the gut surface and thus takes part in the early steps of Bt gut interactions. |
| first_indexed | 2024-03-10T20:30:23Z |
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| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-10T20:30:23Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-c94d13d02f934a299fb0b826deb5b24a2023-11-19T21:26:59ZengMDPI AGToxins2072-66512020-04-0112425210.3390/toxins12040252A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection ProcessJiaxin Qin0Zongxing Tong1Yiling Zhan2Christophe Buisson3Fuping Song4Kanglai He5Christina Nielsen-LeRoux6Shuyuan Guo7School of Life Science, Beijing Institute of Technology, Beijing 100081, ChinaSchool of Life Science, Beijing Institute of Technology, Beijing 100081, ChinaSchool of Life Science, Beijing Institute of Technology, Beijing 100081, ChinaMicalis Institute, INRAE, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, FranceState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaMicalis Institute, INRAE, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, FranceSchool of Life Science, Beijing Institute of Technology, Beijing 100081, China<i>Bacillus thuringiensis</i> (Bt) is used for insect pest control, and its larvicidal activity is primarily attributed to Cry toxins. Other factors participate in infection, and limited information is available regarding factors acting on the peritrophic matrix (PM). This study aimed to investigate the role of a Bt chitin-binding protein (CBPA) that had been previously shown to be expressed at pH 9 <i>in vitro</i> and could therefore be expressed in the alkaline gut of lepidopteron larvae. A ∆cbpA mutant was generated that was 10-fold less virulent than wild-type Bt HD73 towards <i>Ostrinia furnacalis</i> neonate larvae, indicating its important role in infection. Purified recombinant <i>Escherichia coli</i> CBPA was shown to have a chitin affinity, thus indicating a possible interaction with the chitin-rich PM. A translational GFP–CBPA fusion elucidated the localization of CBPA on the bacterial surface, and the transcriptional activity of the promoter P<i>cbpA</i> was immediately induced and confirmed at pH 9. Next, in order to connect surface expression and possible <i>in vivo</i> gut activity<i>,</i> last instar <i>Galleria</i> <i>mellonella</i> (Gm) larvae (not susceptible to Bt HD-73) were used as a model to follow CBPA in gut expression, bacterial transit, and PM adhesion. CBPA-GFP was quickly expressed in the Gm gut lumen, and more Bt HD73 strain bacteria adhered to the PM than those of the ∆cbpA mutant strain. Therefore, CBPA may help to retain the bacteria, via the PM binding, close to the gut surface and thus takes part in the early steps of Bt gut interactions.https://www.mdpi.com/2072-6651/12/4/252<i>Bacillus thuringiensis</i>chitin-binding proteinadhesionperitrophic matrix |
| spellingShingle | Jiaxin Qin Zongxing Tong Yiling Zhan Christophe Buisson Fuping Song Kanglai He Christina Nielsen-LeRoux Shuyuan Guo A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process Toxins <i>Bacillus thuringiensis</i> chitin-binding protein adhesion peritrophic matrix |
| title | A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process |
| title_full | A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process |
| title_fullStr | A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process |
| title_full_unstemmed | A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process |
| title_short | A <i>Bacillus thuringiensis</i> Chitin-Binding Protein is Involved in Insect Peritrophic Matrix Adhesion and Takes Part in the Infection Process |
| title_sort | i bacillus thuringiensis i chitin binding protein is involved in insect peritrophic matrix adhesion and takes part in the infection process |
| topic | <i>Bacillus thuringiensis</i> chitin-binding protein adhesion peritrophic matrix |
| url | https://www.mdpi.com/2072-6651/12/4/252 |
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