Cloning and Bioinformatics Analysis of Voltage-gated Sodium Channel Gene of Myzus persicae

doi:10.16409/j.cnki.2095-039x.2020.03.029

Abstract

Abstract: The cDNA sequence of the voltage-gated sodium channel gene of Myzus persicae was cloned and the typical characteristics of sodium channel were determined, which laid a foundation for studying the molecular mechanism of resistance to Aphis gossypii. The experimental techniques were mainly RT-PCR and PCR, and the cDNA sequence of the sodium ion channel gene was cloned, and the sequence bioinformatics analysis was carried out by using relevant software. Cloning two cDNA sequences MpNa_v-1 (NCBI accession number:MN124170) and MpNa_v-2 (NCBI accession number:MN176136). MpNa_v-1 is 2945 bp in length and includes a 2877 bp complete open reading frame encoding a total of 958 amino acids. MpNa_v-2 is 3546 bp in length and includes a 3486 bp complete open reading frame encoding a total of 1161 amino acids. MpNa_v-1 and MpNa_v-2 together form the sodium channel of Myzus persicae, MpNa_v-1 contains a homologous domain I and a homologous domain II MpNa_v-2 contains a homology domain III and a homologous domain IV. The homology alignment found that the similarity between the Acyrthosiphon pisum and the Melanaphis sacchari sorghum sodium channel was as high as 97.67% and 97.65%, respectively. The cloned sequence contains typical features of the insect sodium ion channel and contains DENS and MFM. Successfully cloned the sodium ion channel gene of Myzus persicae, laying the foundation for elucidating its molecular mechanism of target resistance to pyrethroids.

Key words: Myzus persicae, sodium channel, gene cloning, bioinformatics analysis, phylogenetic analysis

CLC Number:

S476

WANG Hao, DUAN Wenbo, LI Fen, LI Weizheng, WU Shaoying. Cloning and Bioinformatics Analysis of Voltage-gated Sodium Channel Gene of Myzus persicae[J]. Chinese Journal Of Biological Control, 2020, 36(3): 443-451.

References

[1] 蒲颇. 基于微卫星标记的烟蚜种群遗传结构分析[D]. 重庆:西南大学, 2016..
[2] 贾京州. 基于混合推理技术的蚜虫危害诊断与防治专家系统关键技术研究[D]. 杨凌:西北农林科技大学, 2009.
[3] Williamson M S, Denholm I, Bell C A, et al. Knockdown resistance (kdr) to DDT and pyrethroid insecticides maps to a sodium channel gene locus in the housefly (Musca domestica)[J]. Molecular and General Genetics, 1993, 240(1):17-22.
[4] 沈荣. 钠钾离子通道蛋白质结构与功能关联的分子物理力学研究[D]. 南京:南京航空航天大学, 2012.
[5] Goldin A L. Evolution of voltage-gated Na⁺ channels[J]. Journal of Experimental Biology 2002, 205(5):575-584.
[6] Catterall W. From ionic currents to molecular mechanisms:the structure and function of voltage-gated sodium channels[J]. Neuron 2000, 26(1):13-25.
[7] Soderlund D M, Knipple D C. The molecular biology of knockdown resistance to pyrethroid insecticides[J]. Insect Biochemistry and Molecular Biology, 2003, 33(6):570-577.
[8] Loughney K, Kreber R, Ganetzky B. Molecular analysis of the para locus, a sodium channel gene in Drosophila[J]. Cell, 1989, 58(6):1143-1154.
[9] Ingles P J, Adams P M, Knipple D C, et al. Characterization of voltage-sensitive sodium channel gene coding sequences from insecticide-susceptible and knockdown-resistant house fly strains[J]. Insect Biochemistry and Molecular Biology, 1996, 26(4):320-326.
[10] Dong K. A single amino acid change in the para sodium channel protein is associated with knockdown-resistance (kdr) to pyrethroid insecticides in German cockroach[J]. Insect Biochemistry & Molecular Biology, 1997, 27(2):93.
[11] Shao Y M, Dong K, Tang Z H, et al. Molecular characterization of a sodium channel gene from the Silkworm Bombyx mori[J]. Insect Biochemistry and Molecular Biology, 2009, 39(2):150-151.
[12] Dong K. Insect sodium channels and insecticide resistance[J]. Invertebrate Neuroscience, 2007, 7(1):17-30.
[13] 张建亮. 桃蚜的农药生态学研究[D]. 福州:福建农林大学, 2001.
[14] Liu N, Yue X. Genetics of pyrethroid resistance in a strain (ALHF) of house flies (Diptera:Muscidae)[J]. Pestic Biochem Physiol 2001, 70(3):151-158.
[15] Littleton J, Ganetzky B. Ion channels and synaptic organization:analysis of the Drosophila `genome[J]. Neuron, 2000, 26(1):35-43.
[16] Field L M, Blackman R L, Tyler-Smith C, et al. Relationship between amount of esterase and gene copy number in insecticide-resistant Myzus persicae (Sulzer)[J]. Biochemical Journal, 1999, 339(3):737-742.
[17] Zhou W, Chung I, Goldin A, et al. A voltage-gated calcium-selective channel encoded by a sodium channel-like gene[J]. Neuron 2004, 42(1):101-112.
[18] Song W, Liu Z, Tan J, et al. RNA editing generates tissue-specific sodium channels with distinct gating properties[J]. The Journal of Biological Chemistry, 2004, 279(31):32554-32561.
[19] Catterall W A. From ionic currents to molecular review mechanisms:The structure and function of voltage-gated sodium channels[J]. Neuron, 2000, 26(1):13-25.
[20] 万敬员, 张力, 叶笃筠. 选择性剪接调控机制的研究进展[J]. 医学分子生物学杂志, 2003, 25(6):342-345.
[21] Tan J, Liu Z, Nomura Y, et al. Alternative splicing of an insect sodium channel gene generates pharmacologically distinct sodium channels[J]. Journal of Neuroscience, 2002, 22(13):5300-5309.
[22] Rachel O'Donnell Olson, Liu, Z, et al. Molecular and functional characterization of voltage-gated sodium channel variants from drosophila melanogaster[J]. Insect Biochemistry & Molecular Biology, 2008, 38(5):0-610.
[23] Song W, Liu Z, Tan J, et al. RNA editing generates tissue-specific sodium channels with distinct gating properties[J]. Journal of Biological Chemistry, 2004, 279(31):32554-32561.
[24] Lee S, Ingles P J, Knipple D C, et al. Developmental regulation of alternative exon usage in the house flyvssc1sodium channel gene[J]. Invertebrate Neuroscience, 2002, 4(3):125-133.
[25] Wang R, Huang Z Y, Dong K. Molecular characterization of an arachnid sodium channel gene from the varroa mite (Varroa destructor)[J]. Insect Biochemistry and Molecular Biology, 2003, 33(7):730-739.
[26] Chang C, Shen W K, Wang T T, et al. A novel amino acid substitution in a voltage-gated sodium channel is associated with knockdown resistance to permethrin in aedes aegypti[J]. Insect Biochemistry and Molecular Biology, 2009, 39(4):270-278.
[27] Field L M, James A A, Park Y, et al. Voltage-gated sodium channel genes hscp and hDSC1 of Heliothis virescens F. genomic organization[J]. Insect Molecular Biology, 1999, 8(2):161-170.
[28] Shao Y M, Dong K, Tang, Z H, et al. Molecular characterization of a sodium channel gene from the silkworm bombyx mori[J]. Insect Biochemistry and Molecular Biology, 2009, 39(2):150-151.
[29] Davies T G E, Field, L M, Usherwood P N R, et al. A comparative study of voltage-gated sodium channels in the insecta:implications for pyrethroid resistance in anopheline and other neopteran species[J]. Insect Molecular Biology, 2010, 16(3):361-375.
[30] Palladino M J, Keegan L P, O'Connell M A. Reenan RNA:A-to-I pre-mRNA editing in Drosophilais primarily involved in adult nervous system function and integrity[J]. Cell, 2000, 102(4):437-449.
[31] Rieder L E, Staber C J, Hoopengardner B, et al. Tertiary structural elements determine extent and specificity of messenger RNA editing[J]. Nature Communications, 2013, 4:2232.
[32] Liu Z, Song W, Dong K. Persistent tetrodotoxin-sensitive sodium current resulting from u-to-c RNA editing of an insect sodium channel[J]. Proceedings of the National Academy of Sciences, 2004, 101(32):11862-11867.
[33] Lindsay H A, Baines R, Ffrench-Constant R, et al. The dominant cold-sensitive out-cold mutants of drosophila melanogaster have novel missense mutations in the voltage-gated sodium channel gene paralytic[J]. Genetics, 2008, 180(2):873-884.
[34] Song W, Du Y, Liu Z, et al. Substitutions in the domain III voltage sensing module enhance the sensitivity of an insect sodium channel to a scorpion beta-toxin[J]. Journal of Biological Chemistry, 2011, 18(286):15781-15783.