Characteristics and Bacterial-induced Expression Analysis of Two Prophenoloxidase Genes in The Brown Planthopper, Nilaparvata lugens (Hemiptera Delphacidae)

doi:10.16409/j.cnki.2095-039x.2018.01.004

Abstract

Abstract: This study aims to explore the developmental expression pattern and biological function of prophenoloxidase in Nilaparvata lugens. The cDNA sequences encoding NlPPO1 (GenBank accession number:ALE32751) and NlPPO2 (GenBank accession number:ALE32752) were obtained from RNA-seq transcriptome sequencing database and rapid amplification of cDNA ends (RACE). The sequence analysis showed that the full-length cDNA sequence of NlPPO1 is 2316 bp, which contains an open reading frame (ORF) of 2073 bp encoding 690 amino acid-residues. And the full-length cDNA sequence of NlPPO2 is 2738 bp, which contains an ORF of 2100 bp encoding 699 amino acid-residues. The expression patterns and the changes of NlPPO mRNA induced by injecting bacteria were analyzed by real-time quantitative PCR. The developmental expression patterns showed that NlPPO1 and NlPPO2 were highly expressed in the gut, with a low mRNA level in the epidermis cell and fat body, respectively. The NlPPO1 mRNA expression increased with time and the NlPPO2 mRNA expression remained at a relatively high level in 5th instar larvae stage. The NlPPO2 mRNA expression decreased with time and the NlPPO1 mRNA expression remained at a relatively low level in adult stage. The NlPPO1 mRNA expression level significantly increased while the NlPPO2 mRNA expression level did not change significantly at 24 h after injection of Escherichia coli in N. lugens. The NlPPO1 and NlPPO2 mRNA expression levels significantly increased at 24 h after injection of Bacillus subtilis in N. lugens. The results showed that there are some differences between NlPPO1 and NlPPO2 between developmental stages and tissues in N. lugens. And the mRNA expression levels of NlPPO1 and NlPPO2 differ in the immune responses to bacteria, suggesting that the two NlPPO genes have different biological functions. The results provide a good foundation for further research on immune responses of NlPPO in the brown planthopper.

Key words: Nilaparvata lugens, prophenoloxidase, characteristics, expression pattern, inducible expression

CLC Number:

Q966

CHEN Jing, ZHANG Daowei, QIAN Zhengmin. Characteristics and Bacterial-induced Expression Analysis of Two Prophenoloxidase Genes in The Brown Planthopper, Nilaparvata lugens (Hemiptera Delphacidae)[J]. journal1, 2018, 34(1): 44-51.

References

[1] Strand M R. The insect cellular immune response[J]. Insect Science, 2008, 15(1):1-14.
[2] Lavine M D, Strand M R. Insect hemocytes and their role in immunity[J]. Insect Biochemistry and Molecular Biology, 2002, 32(10):1295-1309.
[3] Lemaitre B, Hoffmann J. The host defense of Drosophila melanogaster[J]. Annual Review of Immunology, 2007, 25(1):697-743.
[4] Kanost M R, Gorman M J. Insect Immunology[M]. San Diego, CA:Academic Press, 2008, 69-96.
[5] Nam H J, Jang I H, You H, et al. Genetic evidence of a redox-dependent systemic wound response via Hayan protease-phenoloxidase system in Drosophila[J]. The EMBO Journal, 2012, 31(5):1253-1265.
[6] Lu A, Zhang Q, Zhang J, et al. Insect prophenoloxidase:the view beyond immunity[J]. Frontiers in Physiology, 2014, 5(252):1-15.
[7] Cerenius L, Lee B L, Soderhall K. The proPO-system:pros and cons for its role in invertebrate immunity[J]. Trends Immunology, 2008, 29(6):263-271.
[8] An C, Zhang M, Chu Y, et al. Serine protease MP2 activates prophenoloxidase in the melanization immune response of Drosophila melanogaster[J]. PLoS ONE, 2013, 8(11):e79533.
[9] Bao Y Y, Wang Y, Wu W J, et al. De novo intestine-specific transcriptome of the brown planthopper Nilaparvata lugens revealed potential functions in digestion, detoxification and immune response[J]. Genomics, 2012, 99(4):256-264.
[10] Xue J, Bao Y Y, Li B L, et al. Transcriptome analysis of the brown planthopper Nilaparvata lugens[J]. PLoS ONE, 2010, 5(12):e14233.
[11] Waterhouse R M, Kriventseva E V, Meister S, et al. Evolutionary dynamics of immune-related genes and pathways in disease-vector mosquitoes[J]. Science, 2007, 316(5832):1738-1743.
[12] 张永亮, 朱勇. 家蚕和野桑蚕酚氧化酶原基因的组织表达差异比较[J]. 蚕业科学, 2009, 35(3):630-633.
[13] 邱宗文, 张锡林, 徐文岳. 按蚊中肠前酚氧化酶原与约氏疟原虫卵囊黑化的关系[J]. 中国寄生虫病防治杂志, 2003, 16(6):321-324.
[14] Wang Z, Lu A, Li X, et al, A systematic study on hemocyte identification and plasma prophenoloxidase from Culex pipiens quinquefasciatus at different developmental stages[J]. Experimental Parasitology, 2011, 127(1):135-141.
[15] Shao Q, Yang B, Xu Q, et al. Hindgut innate immunity and regulation of fecal microbiota through melanization in insects[J]. The Journal of Biological Chemistry, 2012, 287(17):14270-14279.
[16] Diao Y, Lu A, Yang B, et al. Existence of prophenoloxidase in wing discs:a source of plasma prophenoloxidase in the silkworm, Bombyx mori[J]. PLoS ONE, 2012, 7(7):e41416.
[17] Dittmer N T, Hiromasa Y, Tomich J M, et al. Proteomic and transcriptomic analyses of rigid and membranous cuticles and epidermis from the elytra and hindwings of the red flour beetle, Tribolium castaneum[J]. Journal of Proteome Research, 2012, 11(1):269-278.
[18] Tsao I Y, Lin U S, Christensen B M, et al. Armigeres subalbatus prophenoloxidase Ⅲ:cloning, characterization and potential role in morphogenesis[J]. Insect Biochemistry and Molecular Biology, 2009, 39(2):96-104.
[19] Tsao I Y, Christensen B, Chen C C. Armigeres subalbatus (Diptera:Culicidae) prophenoloxidase Ⅲ is required for mosquito cuticle formation:ultrastructural study on dsRNA-knockdown mosquitoes[J]. Journal of Medical Entomology, 2010, 47(4):495-503.
[20] Lai S C, Chen C C, Hou R F. Immunolocalization of prophenoloxidase in the process of wound healing in the mosquito Armigeres subalbatus (Diptera:Culicidae)[J]. Journal of Medical Entomology, 2002, 39(2):266-274.
[21] Karlsson C, Korayem A M, Scherfer C, et al. Proteomic analysis of the Drosophila larval hemolymph[J]. The Journal of Biological Chemistry, 2004, 279(50):52033-52041.
[22] Binggeli O, Neyen C, Poidevin M, et al. Prophenoloxidase activation is required for survival to microbial infections in Drosophila[J]. PLoS Pathogens, 2014, 10(5):e1004067.