Comparative analysis of antennal olfactory gene expression between Culex pipiens pallens and Culex pipiens molestus

doi:10.12140/j.issn.1000-7423.2019.04.015

Abstract

Abstract:

Objective To analyze the difference in antennal olfactory gene expression between Culex pipiens pallens and Cx. pipiens molistus to provide a reference for revealing the characteristics of their genetic differentiation. Methods Total RNAs were extracted from antennas of Cx. pipiens pallens and Cx. pipiens molistus before and after blood feeding, and the RNA libraries were constructed using NEBNext Ultra^TM RNA ^® RNA Library Prep Kit. The transcriptome were obtained by RNAseq and their difference in different species of Cx. was analyzed by FPKM. Those olfactory genes with significant difference before and after blood feeding were identified by qRT-PCR. Results RNA-Seq results identified 50 odorant binding protein (OBP) genes and 31 odor receptor(OR) genes expressed in Cx. pipiens pallens, 36 OBPs and 12 ORs in Cx. pipiens molestus. The regulation trend of OBP44 was, in consistence, the same with the RNAseq results while OBP31 was contrary to that. The expression of OBP17 was significantly higher in Cx. pipiens molestus than in Cx. pipiens pallens before blood-sucking, and the expressions of OBP31 and OBP44 displayed the same trend after blood-sucking while the expression of OBP99a was significantly lower than Cx. pipiens pallens(P < 0.05). Expression of OR31, OR65 and CPIJ001069 were significantly up-regulated in Cx. pipiens molestus post blood feeding, but not in Cx. pipiens pallens, consistent with RNAseq results. Expression of OR65 and CPIJ001069 was significantly higher in Cx. pipiens molestus than that in Cx. pipiens pallens post blood feeding(P < 0.05). There was no significant change of OR gene expression in Cx. pipens pallents post blood feeding. Conclusion OBPs and ORs of Cx. pipiens pallens and Cx. pipiens molestus showed genetic differentiation. OBP17 and OBP44 is possibly involved in the regulation of behavior post blood feeding of Cx. pipiens molestus. OR31, OR65, and CPIJ001069 could be involved in regulating the behavior of Cx. pipiens molestus post blood feeding.

Key words: Culex pipiens pallens, Culex pipiens molestus, High-throughput analysis, Olfactory gene

CLC Number:

R384.1

Jie ZHOU, Chun-xiao LI, Ce-jie LAN, Jian GAO, Qin-mei LIU, Ai-juan SUN, Tong-yan ZHAO. Comparative analysis of antennal olfactory gene expression between Culex pipiens pallens and Culex pipiens molestus[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2019, 37(4): 453-457.

Figures/Tables 3

Table 1

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基因名称 Gene name	引物（5′→3′） Primer sequences( 5′→3′)	产物长度/bp Product length/bp
RPL8	R: GTATCGACAAGCCCATCCTGA F: ACCTGGCGGCGTTCCT	176
OR58	R: ACTGTTCTGCTCGGAGTCTCTGTC F: TTCGGCGTCGTTGAAGTTAGCG	160
OR65	R: ACGCTCTCATGGATCGCAGGATA F: CCAGAACTGTTGGTACGAGGCAA	85
OR31	R: GTCCTGTTCTGGCTGTCGTTGAT F: GAATGTTCGTACCACGGCATATCG	119
CPIJ001069	R: AGGATTCACGCAGGTCGTCACA F: ATCGGAGGCATCTTCGTCTTGGA	108
CPIJ019566	R: AACGCTTGCTGCCGAACTTCC F: TGCTGCTGCTTCTCGGTCCAA	110
CPIJ013183	R: TCGCTCTCGTCGCTGGTGTATT F: GCTCGTGTTGATGTGCTTGATTCG	204
OBP23	R: CAGGAGAAGGCGGAACGGTT F: CAGAAACAGCGAACATCCAGCAAA	157
OBP31	R: AGTTGCGTGATGCAGAAGATTGGA F: TCGTTAGTCGTGCCTTCACATTGG	143
OBP44	R: GAGTTCACATGGACGCCACCTG F: ACAATACGCATCATCCGCCAAGG	131
OBP99a	R: GGACAAGAACACCGACGACAACT F: TGTTAGAGCCAGCGTTTCCTTCAA	184
OBP17	R: GGTGCTTCGTGCAGTGCTTCT F: GGTCAAACTCGGCGGACATCTTT	102
CPIJ016952	R: CGTATGCTCAGGTTCAGCTTCTCC F: CGTAGGCAACTTCGCACTTGTCTT	114