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Molecular Characterization and Expression Pattern of Two General Odorant Binding Proteins from the Diamondback Moth, Plutella xylostella

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Abstract

In the Lepidoptera, odorant signals are thought to be mediated by general odorant binding proteins (GOBPs) in the sensillar lymph surrounding the olfactory receptors. We describe the identification and characterization of two new cDNAs encoding GOBPs from the antennae of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), a species for which no GOBPs have been identified to date. We focused our investigation on this olfactory protein family by using reverse transcription–polymerase chain reaction strategies. The deduced amino acid sequences of PxylGOBP1 and PxylGOBP2 revealed open reading frames of 168 and 163 amino acids, respectively, with six cysteine residues in conserved positions relative to other known GOBPs. The alignment of the mature PxylGOBPs with other Lepidoptera GOBPs showed high sequence identity (70–80%) with other full-length sequences from GenBank. Sequence identity between PxylGOBP1and PxylGOBP2 was only 50%, suggesting that the two proteins belong to different classes of lepidopteran GOBPs. The expression patterns of the two PxylGOBP genes, with respect to tissue distribution and sex, were further investigated by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR. Although the two GOBP genes were expressed only in the antennae of both sexes, reflecting the antennal specificity of GOBPs, the transcription levels of these genes depended on the sex, the age, the mating status, and the genes.

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Acknowledgments

This work was funded by a grant from the National Natural Science Foundation of China (No. 30571505).

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Authors and Affiliations

  1. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China

    Zhi-Chun Zhang, Man-Qun Wang & Guoan Zhang

  2. Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China

    Yao-Bin Lu

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  1. Zhi-Chun Zhang
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Correspondence to Man-Qun Wang.

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Supplemental Data Table 1

Relative quantification of expression of PxylGOBP1 and PxylGOBP2 at various times in the life cycle of Plutella xylostella (DOC 72 kb)

Supplemental Data Table 2

Relative quantification of expression of PxylGOBP1 and PxylGOBP2 amplified at various times by real-time PCR (DOC 71 kb)

Supplemental Data Fig. 1

The Rn vs Cycle of PxylGOBP1 real-time PCR. (DOC 138 kb)

Supplemental Data Fig. 2

The Standard Curve for PxylGOBP1 real-time PCR: K = 3.25, B = −28.63,R2 = 1. (DOC 95 kb)

Supplemental Data Fig. 3

The Rn vs Cycle of PxylGOBP2 real-time PCR. (DOC 132 kb)

Supplemental Data Fig. 4

The Standard Curve for PxylGOBP2 real-time PCR: K = −3.05, B = 27.89,R2 = 0.996. (DOC 95 kb)

Supplemental Data Fig. 5

The Rn vs Cycle of PxylActin real-time PCR. (DOC 143 kb)

Supplemental Data Fig. 6

The Standard Curve of PxylActin real-time PCR: K = −3.1, B = 31.94, R2 = 0.99. (DOC 95 kb)

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Zhang, ZC., Wang, MQ., Lu, YB. et al. Molecular Characterization and Expression Pattern of Two General Odorant Binding Proteins from the Diamondback Moth, Plutella xylostella . J Chem Ecol 35, 1188–1196 (2009). https://doi.org/10.1007/s10886-009-9697-2

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