Construction and analysis of cDNA libraries from the antennae of Batocera horsfieldi and expression pattern of putative odorant binding proteins.
A high-quality cDNA library was constructed from female and male antenna of the longhorned beetle, Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae), a serious pest of Populus (Salicales: Salicaceae). The titer was approximately 2.37 × 106 pfu/mL, and this complies with the test requirement. From the libraries, 692 clones were selected randomly, sequenced, and further analyzed, and the recombinational efficiency reached 93.85%. By alignment and cluster analysis, we identified four odorant binding proteins, two pheromone-binding proteins (have the characteristic six conserved cysteine residues), four Minus-C odorant binding proteins (lost two conserved cysteines), and three chemosensory proteins. In this study, we describe the identification and characterization of four new cDNAs that encode Minus-C odorant binding proteins (Minus-C OBPs) from B. horsfieldi antennal cDNA libraries. Our investigation focused on the expression pattern of the Minus-C OBP genes in various tissues in both sexes at different developmental stages, using reverse transcription PCR (RT-PCR) and realtime PCR (qPCR) strategies. Minus-C OBP1, 2, and 3 were expressed in all tested tissues, with the exception of the head (without antenna, labial palps, and maxillary palps). Minus-C OBP4 was expressed in the antenna, legs, and abdomen, but not in the labial palps, maxillary palps, or head. The qPCR results revealed MinusC OBPs were expressed in the antenna throughout the adult life, and that the transcript levels of these genes depended on the sex, age, and mating status of adults.
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, P. R. China Henan University of Technology, Zhengzhou 450001, P. R. China.
Hui
H
Li
Invasive Insect Biocontrol and Behavior Laboratory, USDA-ARS-Plant Sciences Institute, Beltsville, MD 20705- 2350, USA.
Aijun
A
Zhang
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, P. R. China.
Li-Zhen
LZ
Chen
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, P. R. China.
Guoan
G
Zhang
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, P. R. China mqwang@mail.hzau.edu.cn.
Man-Qun
MQ
Wang
D000595:Amino Acid Sequence; D000818:Animals; D057506:Arthropod Antennae / Q000378:metabolism*; D001517:Coleoptera / Q000033:anatomy & histology / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics*; D020224:Expressed Sequence Tags; D005260:Female; D005786:Gene Expression Regulation; D015723:Gene Library; D008297:Male; D008969:Molecular Sequence Data; D011485:Protein Binding; D060888:Real-Time Polymerase Chain Reaction; D018035:Receptors, Odorant / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome
D018076:DNA, Complementary; D018035:Receptors, Odorant
Antennal cDNA library; Minus-C OBPs; RT-PCR; qPCR