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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.


Relative PxAdoR mRNA level.(A) Effect of PxAdoR silencing by 500 ng/mL dsRNA feeding. (B) Effect of HEA feeding after silencing PxAdoR. Analyzed sample were cDNAs that were reverse-transcribed from pooled RNA samples of 10 larvae. Bars represent the means ± standard deviations of technical replicates. *P < 0.05; **P < 0.001 (A) compared with the dsgfp group. (B) compared with the control.
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pone.0162859.g003: Relative PxAdoR mRNA level.(A) Effect of PxAdoR silencing by 500 ng/mL dsRNA feeding. (B) Effect of HEA feeding after silencing PxAdoR. Analyzed sample were cDNAs that were reverse-transcribed from pooled RNA samples of 10 larvae. Bars represent the means ± standard deviations of technical replicates. *P < 0.05; **P < 0.001 (A) compared with the dsgfp group. (B) compared with the control.

Mentions: Quantitative RT-PCR analyzes the RNAi experiments showed that PxAdoR gene expression in the second instar larvae after RNAi was reduced by 2.36-fold compared to dsGFP-fed control (Table 3). The findings show that dsRNA feeding resulted in a significantly reduced the expression of PxAdoR gene (Fig 3A).


N 6 -(2-Hydroxyethyl)-Adenosine Exhibits Insecticidal Activity against Plutella xylostella via Adenosine Receptors
Relative PxAdoR mRNA level.(A) Effect of PxAdoR silencing by 500 ng/mL dsRNA feeding. (B) Effect of HEA feeding after silencing PxAdoR. Analyzed sample were cDNAs that were reverse-transcribed from pooled RNA samples of 10 larvae. Bars represent the means ± standard deviations of technical replicates. *P < 0.05; **P < 0.001 (A) compared with the dsgfp group. (B) compared with the control.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5036850&req=5

pone.0162859.g003: Relative PxAdoR mRNA level.(A) Effect of PxAdoR silencing by 500 ng/mL dsRNA feeding. (B) Effect of HEA feeding after silencing PxAdoR. Analyzed sample were cDNAs that were reverse-transcribed from pooled RNA samples of 10 larvae. Bars represent the means ± standard deviations of technical replicates. *P < 0.05; **P < 0.001 (A) compared with the dsgfp group. (B) compared with the control.
Mentions: Quantitative RT-PCR analyzes the RNAi experiments showed that PxAdoR gene expression in the second instar larvae after RNAi was reduced by 2.36-fold compared to dsGFP-fed control (Table 3). The findings show that dsRNA feeding resulted in a significantly reduced the expression of PxAdoR gene (Fig 3A).

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&ndash;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.