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Functional analysis of general odorant binding protein 2 from the meadow moth, Loxostege sticticalis L. (Lepidoptera: Pyralidae).

Yin J, Feng H, Sun H, Xi J, Cao Y, Li K - PLoS ONE (2012)

Bottom Line: In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends.Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants.Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China. jyin@ippcaas.cn

ABSTRACT
Odorant binding proteins play a crucial role in transporting semiochemicals across the sensillum lymph to olfactory receptors within the insect antennal sensilla. In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends. Recombinant LstiGOBP2 was expressed in Escherichia coli and purified by Ni ion affinity chromatography. Real-time PCR assays indicated that LstiGOBP2 mRNA is expressed mainly in adult antennae, with expression levels differing with developmental age. Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants. Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.

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Related in: MedlinePlus

Expression pattern analysis of LstiGOBP2.(A) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different tissues. cDNAs were amplified with specific primers from antennae, thoraxes, abdomens, wings, legs, tarsite and heads(without antennae). (B) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different-day old antennae.
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pone-0033589-g003: Expression pattern analysis of LstiGOBP2.(A) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different tissues. cDNAs were amplified with specific primers from antennae, thoraxes, abdomens, wings, legs, tarsite and heads(without antennae). (B) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different-day old antennae.

Mentions: We examined the expression pattern of LstiGOBP2 mRNA in different tissues by qPCR. The desired product was largely amplified from cDNA templates that were reverse-transcribed from total RNA in male and female antennae, with only a few derived from other tissues, suggesting that GOBP2 is mainly expressed in antennae (Fig. 3A). In general, the levels of transcripts were very low in all tissues except the antennae, where LstiGOBP2 was highly expressed. However, the expressed quantity of LstiGOBP2 was different in antennae of different ages and there was a notable difference between males and females, with the quantity in male antennae being significantly higher than in female antennae. The quantity of LstiGOBP2 was the highest for both males and females in four-day-old antennae, which is consistent with the age that adults find host plants (Fig. 3B).


Functional analysis of general odorant binding protein 2 from the meadow moth, Loxostege sticticalis L. (Lepidoptera: Pyralidae).

Yin J, Feng H, Sun H, Xi J, Cao Y, Li K - PLoS ONE (2012)

Expression pattern analysis of LstiGOBP2.(A) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different tissues. cDNAs were amplified with specific primers from antennae, thoraxes, abdomens, wings, legs, tarsite and heads(without antennae). (B) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different-day old antennae.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316592&req=5

pone-0033589-g003: Expression pattern analysis of LstiGOBP2.(A) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different tissues. cDNAs were amplified with specific primers from antennae, thoraxes, abdomens, wings, legs, tarsite and heads(without antennae). (B) qPCR analysis of Loxostege sticticalis general odorant binding protein 2 (LstiGOBP2) expressed in different-day old antennae.
Mentions: We examined the expression pattern of LstiGOBP2 mRNA in different tissues by qPCR. The desired product was largely amplified from cDNA templates that were reverse-transcribed from total RNA in male and female antennae, with only a few derived from other tissues, suggesting that GOBP2 is mainly expressed in antennae (Fig. 3A). In general, the levels of transcripts were very low in all tissues except the antennae, where LstiGOBP2 was highly expressed. However, the expressed quantity of LstiGOBP2 was different in antennae of different ages and there was a notable difference between males and females, with the quantity in male antennae being significantly higher than in female antennae. The quantity of LstiGOBP2 was the highest for both males and females in four-day-old antennae, which is consistent with the age that adults find host plants (Fig. 3B).

Bottom Line: In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends.Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants.Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China. jyin@ippcaas.cn

ABSTRACT
Odorant binding proteins play a crucial role in transporting semiochemicals across the sensillum lymph to olfactory receptors within the insect antennal sensilla. In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends. Recombinant LstiGOBP2 was expressed in Escherichia coli and purified by Ni ion affinity chromatography. Real-time PCR assays indicated that LstiGOBP2 mRNA is expressed mainly in adult antennae, with expression levels differing with developmental age. Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants. Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.

Show MeSH
Related in: MedlinePlus