Gene expression studies and molecular characterization of a cathepsin L-like from the Asian citrus psyllid Diaphorina citri, vector of Huanglongbing
Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri Kuwayama. Diaphorina citri belongs to Hemiptera order, which has cysteine peptidases as the most abundant proteolytic enzymes present in dig...
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Published in | International journal of biological macromolecules Vol. 158; pp. 375 - 383 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Elsevier B.V
01.09.2020
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Abstract | Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri Kuwayama. Diaphorina citri belongs to Hemiptera order, which has cysteine peptidases as the most abundant proteolytic enzymes present in digestive tract. As cysteine peptidases are involved in different insect development processes, this class of enzymes has acquired biotechnological importance. In this context, we identified a cathepsin L-like (DCcathL1) from the Diaphorina citri transcriptome database and expressed the enzyme in E. coli. Quantitative real-time RT-PCR was conducted to determine DCcathL1 gene expression in different parts and developmental phases of the insect. We observed that DCcathL1 expression in the gut was 2.59 and 2.87-fold higher than in the head and carcass, respectively. Furthermore, DCcathL1 expression was greater in eggs than in nymphs and adults, suggesting a putative role of the enzyme in the embryonic development. In addition, enzymatic inhibitory activity using four recombinant Citrus cystatins were performed. Among them, CsinCPI-2 was the strongest DCcathL1 inhibitor with a Ki value of 0.005 nM. Our results may contribute in the development of strategies for D. citri control, such as silencing the DCcathL1 gene and the use of transgenic plants that overexpress peptidase inhibitors. |
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AbstractList | Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri Kuwayama. Diaphorina citri belongs to Hemiptera order, which has cysteine peptidases as the most abundant proteolytic enzymes present in digestive tract. As cysteine peptidases are involved in different insect development processes, this class of enzymes has acquired biotechnological importance. In this context, we identified a cathepsin L-like (DCcathL1) from the Diaphorina citri transcriptome database and expressed the enzyme in E. coli. Quantitative real-time RT-PCR was conducted to determine DCcathL1 gene expression in different parts and developmental phases of the insect. We observed that DCcathL1 expression in the gut was 2.59 and 2.87-fold higher than in the head and carcass, respectively. Furthermore, DCcathL1 expression was greater in eggs than in nymphs and adults, suggesting a putative role of the enzyme in the embryonic development. In addition, enzymatic inhibitory activity using four recombinant Citrus cystatins were performed. Among them, CsinCPI-2 was the strongest DCcathL1 inhibitor with a Ki value of 0.005 nM. Our results may contribute in the development of strategies for D. citri control, such as silencing the DCcathL1 gene and the use of transgenic plants that overexpress peptidase inhibitors. Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri Kuwayama. Diaphorina citri belongs to Hemiptera order, which has cysteine peptidases as the most abundant proteolytic enzymes present in digestive tract. As cysteine peptidases are involved in different insect development processes, this class of enzymes has acquired biotechnological importance. In this context, we identified a cathepsin L-like (DCcathL1) from the Diaphorina citri transcriptome database and expressed the enzyme in E. coli. Quantitative real-time RT-PCR was conducted to determine DCcathL1 gene expression in different parts and developmental phases of the insect. We observed that DCcathL1 expression in the gut was 2.59 and 2.87-fold higher than in the head and carcass, respectively. Furthermore, DCcathL1 expression was greater in eggs than in nymphs and adults, suggesting a putative role of the enzyme in the embryonic development. In addition, enzymatic inhibitory activity using four recombinant Citrus cystatins were performed. Among them, CsinCPI-2 was the strongest DCcathL1 inhibitor with a K value of 0.005 nM. Our results may contribute in the development of strategies for D. citri control, such as silencing the DCcathL1 gene and the use of transgenic plants that overexpress peptidase inhibitors. Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri Kuwayama. Diaphorina citri belongs to Hemiptera order, which has cysteine peptidases as the most abundant proteolytic enzymes present in digestive tract. As cysteine peptidases are involved in different insect development processes, this class of enzymes has acquired biotechnological importance. In this context, we identified a cathepsin L-like (DCcathL1) from the Diaphorina citri transcriptome database and expressed the enzyme in E. coli. Quantitative real-time RT-PCR was conducted to determine DCcathL1 gene expression in different parts and developmental phases of the insect. We observed that DCcathL1 expression in the gut was 2.59 and 2.87-fold higher than in the head and carcass, respectively. Furthermore, DCcathL1 expression was greater in eggs than in nymphs and adults, suggesting a putative role of the enzyme in the embryonic development. In addition, enzymatic inhibitory activity using four recombinant Citrus cystatins were performed. Among them, CsinCPI-2 was the strongest DCcathL1 inhibitor with a Ki value of 0.005 nM. Our results may contribute in the development of strategies for D. citri control, such as silencing the DCcathL1 gene and the use of transgenic plants that overexpress peptidase inhibitors. |
Author | Machado, Marcelo F.M. Henrique-Silva, Flavio Schneider, Vanessa K. Lima, Patricia Silva Soares-Costa, Andrea Ferrara, Taise F.S. Bronze, Fellipe S. Carmona, Adriana K. |
Author_xml | – sequence: 1 givenname: Taise F.S. surname: Ferrara fullname: Ferrara, Taise F.S. organization: Departamento de Biofísica, Universidade Federal de São Paulo, 04039-032 São Paulo, Brazil – sequence: 2 givenname: Vanessa K. surname: Schneider fullname: Schneider, Vanessa K. organization: Departamento de Genética e Evolução, Universidade Federal de São Carlos, 13565-905 São Paulo, Brazil – sequence: 3 givenname: Patricia Silva surname: Lima fullname: Lima, Patricia Silva organization: Departamento de Biofísica, Universidade Federal de São Paulo, 04039-032 São Paulo, Brazil – sequence: 4 givenname: Fellipe S. surname: Bronze fullname: Bronze, Fellipe S. organization: Departamento de Biofísica, Universidade Federal de São Paulo, 04039-032 São Paulo, Brazil – sequence: 5 givenname: Marcelo F.M. surname: Machado fullname: Machado, Marcelo F.M. organization: Departamento de Biofísica, Universidade Federal de São Paulo, 04039-032 São Paulo, Brazil – sequence: 6 givenname: Flavio surname: Henrique-Silva fullname: Henrique-Silva, Flavio organization: Departamento de Genética e Evolução, Universidade Federal de São Carlos, 13565-905 São Paulo, Brazil – sequence: 7 givenname: Andrea surname: Soares-Costa fullname: Soares-Costa, Andrea email: andreasc@ufscar.br organization: Departamento de Genética e Evolução, Universidade Federal de São Carlos, 13565-905 São Paulo, Brazil – sequence: 8 givenname: Adriana K. surname: Carmona fullname: Carmona, Adriana K. email: ak.carmona@unifesp.br organization: Departamento de Biofísica, Universidade Federal de São Paulo, 04039-032 São Paulo, Brazil |
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CitedBy_id | crossref_primary_10_3389_fpls_2023_1219319 crossref_primary_10_1016_j_pep_2022_106174 crossref_primary_10_1016_j_jip_2024_108122 |
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Keywords | Cysteine peptidases Huanglongbing Diaphorina citri Citrus cystatins Cathepsin L-like |
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Snippet | Huanglongbing (HLB) is a devastating citrus disease associated with Candidatus Liberibacter asiaticus (CLas) and is transmitted by the psyllid Diaphorina citri... |
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SubjectTerms | Cathepsin L-like Citrus cystatins Cysteine peptidases Diaphorina citri Huanglongbing |
Title | Gene expression studies and molecular characterization of a cathepsin L-like from the Asian citrus psyllid Diaphorina citri, vector of Huanglongbing |
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