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Serine proteases-like genes in the asian rice gall midge show differential expression in compatible and incompatible interactions with rice.

Sinha DK, Lakshmi M, Anuradha G, Rahman SJ, Siddiq EA, Bentur JS, Nair S - Int J Mol Sci (2011)

Bottom Line: Quantitative real time PCR analysis revealed that both the genes were significantly upregulated in larvae feeding on resistant cultivar than in those feeding on susceptible cultivar.These results provide an opportunity to understand the gut physiology of the insect under compatible or incompatible interactions with the host.Phylogenetic analysis grouped these genes in the clade containing proteases of phytophagous insects away from hematophagous insects.

View Article: PubMed Central - PubMed

Affiliation: Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India; E-Mail: deepak22sinha@yahoo.co.in.

ABSTRACT
The Asian rice gall midge, Orseolia oryzae (Wood-Mason), is a serious pest of rice. Investigations into the gall midge-rice interaction will unveil the underlying molecular mechanisms which, in turn, can be used as a tool to assist in developing suitable integrated pest management strategies. The insect gut is known to be involved in various physiological and biological processes including digestion, detoxification and interaction with the host. We have cloned and identified two genes, OoprotI and OoprotII, homologous to serine proteases with the conserved His(87), Asp(136) and Ser(241) residues. OoProtI shared 52.26% identity with mosquito-type trypsin from Hessian fly whereas OoProtII showed 52.49% identity to complement component activated C1s from the Hessian fly. Quantitative real time PCR analysis revealed that both the genes were significantly upregulated in larvae feeding on resistant cultivar than in those feeding on susceptible cultivar. These results provide an opportunity to understand the gut physiology of the insect under compatible or incompatible interactions with the host. Phylogenetic analysis grouped these genes in the clade containing proteases of phytophagous insects away from hematophagous insects.

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Multiple sequence alignment of OoprotI (A) and OoprotII (B) of Orseolia oryzae showing homology to different insects. Conserved amino acid residues are highlighted and active sites are boxed. Horizontal arrows show the regions from which the RT-PCR primers were made.
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f1-ijms-12-02842: Multiple sequence alignment of OoprotI (A) and OoprotII (B) of Orseolia oryzae showing homology to different insects. Conserved amino acid residues are highlighted and active sites are boxed. Horizontal arrows show the regions from which the RT-PCR primers were made.

Mentions: Here we report, for the first time, the cloning and transcriptional expression patterns of two serine protease-like genes (designated as OoprotI (Acc. No. HQ587043) and OoprotII (Acc. No. HQ587044)) from the Asian rice gall midge. BLAST searches of the amino acid sequences inferred from RNA sequence data revealed that OoProtI has 52.26% identity with mosquito-type trypsin from M. destructor (Hessian fly). OoProtII showed 52.49% identity to complement component activated C1s from the Hessian fly [calcium-dependent serine proteinase, C1 esterase (MER048620)]. The classification of these genes was primarily based on the identity shared at the amino acid level with those from other dipterans. Further, the conserved amino acid residues were correctly positioned with reference to the other previously identified proteases and, therefore, likely to confer serine proteases-like specificity to the enzymes (Figures 1A and 1B). Within the predicted amino acid sequences of OoProtI and OoProtII, the active site triad (Histidine-87, Aspartic acid-136, Serine-241) was conserved. Phylogenetic analysis (Figure 2A) revealed that OoProtI, coding for a trypsin-like protein, was the closest in terms of homology to that of the Hessian fly. OoProtII (Figure 2B), in terms of phylogeny, falls in a completely different group of its own, while all the proteases of the hematophagous insects are grouped into a single clade. Importantly, in this phylogenetic grouping, other than Apis, all the non-hematophagous insects fall into separate groups i.e., Drosophila, Mayetiola and Orseolia. This type of a grouping is expected as proteases of phytophagous insects and those from the hamatophagous insects differ probably in the substrate amino acid constituents on which respective enzymes have to act.


Serine proteases-like genes in the asian rice gall midge show differential expression in compatible and incompatible interactions with rice.

Sinha DK, Lakshmi M, Anuradha G, Rahman SJ, Siddiq EA, Bentur JS, Nair S - Int J Mol Sci (2011)

Multiple sequence alignment of OoprotI (A) and OoprotII (B) of Orseolia oryzae showing homology to different insects. Conserved amino acid residues are highlighted and active sites are boxed. Horizontal arrows show the regions from which the RT-PCR primers were made.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3116160&req=5

f1-ijms-12-02842: Multiple sequence alignment of OoprotI (A) and OoprotII (B) of Orseolia oryzae showing homology to different insects. Conserved amino acid residues are highlighted and active sites are boxed. Horizontal arrows show the regions from which the RT-PCR primers were made.
Mentions: Here we report, for the first time, the cloning and transcriptional expression patterns of two serine protease-like genes (designated as OoprotI (Acc. No. HQ587043) and OoprotII (Acc. No. HQ587044)) from the Asian rice gall midge. BLAST searches of the amino acid sequences inferred from RNA sequence data revealed that OoProtI has 52.26% identity with mosquito-type trypsin from M. destructor (Hessian fly). OoProtII showed 52.49% identity to complement component activated C1s from the Hessian fly [calcium-dependent serine proteinase, C1 esterase (MER048620)]. The classification of these genes was primarily based on the identity shared at the amino acid level with those from other dipterans. Further, the conserved amino acid residues were correctly positioned with reference to the other previously identified proteases and, therefore, likely to confer serine proteases-like specificity to the enzymes (Figures 1A and 1B). Within the predicted amino acid sequences of OoProtI and OoProtII, the active site triad (Histidine-87, Aspartic acid-136, Serine-241) was conserved. Phylogenetic analysis (Figure 2A) revealed that OoProtI, coding for a trypsin-like protein, was the closest in terms of homology to that of the Hessian fly. OoProtII (Figure 2B), in terms of phylogeny, falls in a completely different group of its own, while all the proteases of the hematophagous insects are grouped into a single clade. Importantly, in this phylogenetic grouping, other than Apis, all the non-hematophagous insects fall into separate groups i.e., Drosophila, Mayetiola and Orseolia. This type of a grouping is expected as proteases of phytophagous insects and those from the hamatophagous insects differ probably in the substrate amino acid constituents on which respective enzymes have to act.

Bottom Line: Quantitative real time PCR analysis revealed that both the genes were significantly upregulated in larvae feeding on resistant cultivar than in those feeding on susceptible cultivar.These results provide an opportunity to understand the gut physiology of the insect under compatible or incompatible interactions with the host.Phylogenetic analysis grouped these genes in the clade containing proteases of phytophagous insects away from hematophagous insects.

View Article: PubMed Central - PubMed

Affiliation: Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India; E-Mail: deepak22sinha@yahoo.co.in.

ABSTRACT
The Asian rice gall midge, Orseolia oryzae (Wood-Mason), is a serious pest of rice. Investigations into the gall midge-rice interaction will unveil the underlying molecular mechanisms which, in turn, can be used as a tool to assist in developing suitable integrated pest management strategies. The insect gut is known to be involved in various physiological and biological processes including digestion, detoxification and interaction with the host. We have cloned and identified two genes, OoprotI and OoprotII, homologous to serine proteases with the conserved His(87), Asp(136) and Ser(241) residues. OoProtI shared 52.26% identity with mosquito-type trypsin from Hessian fly whereas OoProtII showed 52.49% identity to complement component activated C1s from the Hessian fly. Quantitative real time PCR analysis revealed that both the genes were significantly upregulated in larvae feeding on resistant cultivar than in those feeding on susceptible cultivar. These results provide an opportunity to understand the gut physiology of the insect under compatible or incompatible interactions with the host. Phylogenetic analysis grouped these genes in the clade containing proteases of phytophagous insects away from hematophagous insects.

Show MeSH
Related in: MedlinePlus