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N 6 -(2-Hydroxyethyl)-Adenosine Exhibits Insecticidal Activity against Plutella xylostella via Adenosine Receptors

View Article: PubMed Central - PubMed

ABSTRACT

The diamondback moth, Plutella xylostella, is one of the most important pests of cruciferous crops. We have earlier shown that N6-(2-hydroxyethyl)-adenosine (HEA) exhibits insecticidal activity against P. xylostella. In the present study we investigated the possible mechanism of insecticidal action of HEA on P. xylostella. HEA is a derivative of adenosine, therefore, we speculated whether it acts via P. xylostella adenosine receptor (PxAdoR). We used RNAi approach to silence PxAdoR gene and used antagonist of denosine receptor (AdoR) to study the insecticidal effect of HEA. We cloned the whole sequence of PxAdoR gene. A BLAST search using NCBI protein database showed a 61% identity with the Drosophila adenosine receptor (DmAdoR) and a 32–35% identity with human AdoR. Though the amino acids sequence of PxAdoR was different compared to other adenosine receptors, most of the amino acids that are known to be important for adenosine receptor ligand binding and signaling were present. However, only 30% binding sites key residues was similar between PxAdoR and A1R. HEA, at a dose of 1 mg/mL, was found to be lethal to the second-instar larvae of P. xylostella, and a significant reduction of mortality and growth inhibition ratio were obtained when HEA was administered to the larvae along with PxAdoR-dsRNA or antagonist of AdoR (SCH58261) for 36, 48, or 60 h. Especially at 48 h, the rate of growth inhibition of the PxAdoR knockdown group was 3.5-fold less than that of the HEA group, and the corrected mortality of SCH58261 group was reduced almost 2-fold compared with the HEA group. Our findings show that HEA may exert its insecticidal activity against P. xylostella larvae via acting on PxAdoR.

No MeSH data available.


cDNA sequence of Plutella xylostella Adenosine Receptor.The line box area indicates the open reading frame (ORF), which encodes protein of 440 amino acids. The start codon (ATG) and the stop codon (TGA) are highlighted in black. The sequence was deposited in the GenBank (Assession No.KR258794).
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pone.0162859.g001: cDNA sequence of Plutella xylostella Adenosine Receptor.The line box area indicates the open reading frame (ORF), which encodes protein of 440 amino acids. The start codon (ATG) and the stop codon (TGA) are highlighted in black. The sequence was deposited in the GenBank (Assession No.KR258794).

Mentions: Based on the sequence information, specific primers were designed for 5'-and 3'- RACE of the related gene. 5'-RACE generated a 784 bp fragment, and 3'-RACE produced a 960 bp fragment. Splicing the sequences generated a 1828 bp fragment, which was identified as the full-length PxAdoR gene (GenBank accession NO.KR258794). The ORF of PxAdoR cDNA encoded a protein of 440 amino acids residues with a calculated molecular mass of 64.74243 kDa and an isoelectric point of 11.495 (Fig 1).


N 6 -(2-Hydroxyethyl)-Adenosine Exhibits Insecticidal Activity against Plutella xylostella via Adenosine Receptors
cDNA sequence of Plutella xylostella Adenosine Receptor.The line box area indicates the open reading frame (ORF), which encodes protein of 440 amino acids. The start codon (ATG) and the stop codon (TGA) are highlighted in black. The sequence was deposited in the GenBank (Assession No.KR258794).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036850&req=5

pone.0162859.g001: cDNA sequence of Plutella xylostella Adenosine Receptor.The line box area indicates the open reading frame (ORF), which encodes protein of 440 amino acids. The start codon (ATG) and the stop codon (TGA) are highlighted in black. The sequence was deposited in the GenBank (Assession No.KR258794).
Mentions: Based on the sequence information, specific primers were designed for 5'-and 3'- RACE of the related gene. 5'-RACE generated a 784 bp fragment, and 3'-RACE produced a 960 bp fragment. Splicing the sequences generated a 1828 bp fragment, which was identified as the full-length PxAdoR gene (GenBank accession NO.KR258794). The ORF of PxAdoR cDNA encoded a protein of 440 amino acids residues with a calculated molecular mass of 64.74243 kDa and an isoelectric point of 11.495 (Fig 1).

View Article: PubMed Central - PubMed

ABSTRACT

The diamondback moth, Plutella xylostella, is one of the most important pests of cruciferous crops. We have earlier shown that N6-(2-hydroxyethyl)-adenosine (HEA) exhibits insecticidal activity against P. xylostella. In the present study we investigated the possible mechanism of insecticidal action of HEA on P. xylostella. HEA is a derivative of adenosine, therefore, we speculated whether it acts via P. xylostella adenosine receptor (PxAdoR). We used RNAi approach to silence PxAdoR gene and used antagonist of denosine receptor (AdoR) to study the insecticidal effect of HEA. We cloned the whole sequence of PxAdoR gene. A BLAST search using NCBI protein database showed a 61% identity with the Drosophila adenosine receptor (DmAdoR) and a 32–35% identity with human AdoR. Though the amino acids sequence of PxAdoR was different compared to other adenosine receptors, most of the amino acids that are known to be important for adenosine receptor ligand binding and signaling were present. However, only 30% binding sites key residues was similar between PxAdoR and A1R. HEA, at a dose of 1 mg/mL, was found to be lethal to the second-instar larvae of P. xylostella, and a significant reduction of mortality and growth inhibition ratio were obtained when HEA was administered to the larvae along with PxAdoR-dsRNA or antagonist of AdoR (SCH58261) for 36, 48, or 60 h. Especially at 48 h, the rate of growth inhibition of the PxAdoR knockdown group was 3.5-fold less than that of the HEA group, and the corrected mortality of SCH58261 group was reduced almost 2-fold compared with the HEA group. Our findings show that HEA may exert its insecticidal activity against P. xylostella larvae via acting on PxAdoR.

No MeSH data available.