Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (12): 2461-2471.doi: 10.3864/j.issn.0578-1752.2022.12.016

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Molecular Cloning and Expression Pattern Analysis of NPC2 Gene Family of Apis cerana cerana

ZHANG Li(),ZHANG Nan,JIANG HuQiang,WU Fan,LI HongLiang()   

  1. College of Life Sciences, China Jiliang University/Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018
  • Received:2021-12-19 Accepted:2022-01-21 Online:2022-06-16 Published:2022-06-23
  • Contact: HongLiang LI E-mail:18852648434@163.com;hlli@cjlu.edu.cn

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Abstract:

【Background】As an important native resource insect, the Chinese honeybee (Apis cerana cerana) plays important ecological roles in pollinating the plants that bloom at low temperatures in early winter in China, and the pollination behavior of A. c. cerana is closely related to its olfactory system. According to the analysis of antenna transcriptome data collected from foragers treated at high and low temperatures, it was found that the Niemann-Pick type C2 protein (NPC2) gene family related to insect olfaction was up-regulated expression at low temperatures.【Objective】Therefore, this study aims on the A. c. cerana NPC2 family genes, including cloning and analysis of their structural characteristics and expression profiles. Moreover, the NPC2 family gene expression under high and low temperatures was also studied. It will provide an evidence of the AcNPC2 gene family in the low-temperature adaptation involved in the chemosensory and olfactory function of A. c. cerana.【Method】Based on the results of high and low temperatures transcriptome sequencing of A. c. cerana, the ORF sequence of AcNPC2 genes was cloned by RT-PCR, and phylogenetic tree analysis and three-dimensional structure prediction were performed. Then, the spatio-temporal expression profile of AcNPC2 genes in different developmental stages and different tissues, as well as the amount of expression at high and low temperatures of A. c. cerana were analyzed by qRT-PCR.【Result】The full length ORFs of four NPC2 genes of A. c. cerana (AcNPC2a, AcNPC2b, AcNPC2c, and AcNPC2d) were obtained as 447, 480, 459, and 465 bp, respectively, encoding 148, 159, 152, and 154 amino acids. The predicted protein molecular weight is 16.12-18.53 kD, and the isoelectric points are 7.98, 7.57, 6.56, and 6.34, respectively. Phylogenetic tree analysis showed that AcNPC2 sequences were most close to the NPC2 homologous sequence of Apis mellifera ligustica. qRT-PCR results showed that the expression level of AcNPC2a was the highest in the abdomen of the newborns, followed by the abdomen of the nurses and the larval stage. The expression of AcNPC2b was the highest in the thorax of the newborn bees, followed by the head, thorax and metapodium of the foragers. AcNPC2c was notably expressed in high abundance in the antennae of the nurses and the foragers. AcNPC2d had the highest expression in the head of the foragers. After low temperature treatment, the expression levels of all AcNPC2 genes in the forager antennae increased, but there was no significant difference.【Conclusion】AcNPC2 has the conserved structure of NPC2 protein family, and its members show diversity in the spatio-temporal expression profile of A. c. cerana. Among them, AcNPC2c is highly expressed in the antennae, indicating that it is closely related to the olfactory function of A. c. cerana. The expression of the whole AcNPC2 family genes increased in the antennae of the foragers at lower temperature, indicating that these genes might be involved in the low-temperature adaptability of A. c. cerana and pollination behavior in early winter.

Key words: Apis cerana cerana, NPC2, gene cloning, spatio-temporal expression profile

Table 1

List of the primer information of the NPC2 genes"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 产物长度
Product length (bp)		 退火温度
Tm (℃)
用于基因克隆 For gene cloning
NPC2a NPC2e-Fa CCGAATTCATGGCAATCCTCACCTATGTT 463 55
NPC2e-Ra GGCTCGAGTTAGTCGACAACTTTAGCTGG
NPC2b NPC2e-Fb CCGAATTCATGAATCGAAACGGTATGTTC 496 58
NPC2e-Rb GGCTCGAGTTAGTCGACAACTTTAGCTGG
NPC2c NPC2e-Fc CCGAATTCATGTATCGAATGATTGCTGTAA 475 60
NPC2e-Rc GGCTCGAGCTATTTAATTCGAGCCGGAAT
NPC2d NPC2e-Fd CCGAATTCATGAGGGGAACCATTCTCGTT 481 61
NPC2e-Rd GGCTCGAGTCAATTGGTGATCTTCGCTG
用于实时荧光定量PCR For qRT-PCR
NPC2a NPC2q-Fa ACGTTGACTTTAAGGCTGTTGC 129 51
NPC2q-Ra ACTGTCCGCTCACCAGATTCTT
NPC2b NPC2q-Fb TCCATTATACGCCTGCCTTCTCCT 116 53
NPC2q-Rb GGAAGTTGACAAGCAAGCATCGG
NPC2c NPC2q-Fc GATGCTTGCCAAACTCCTGATTCT 116 55
NPC2q-Rc ACAGTAACACTCACCTTAGGATACG
NPC2d NPC2q-Fd TTCGTAGCTCTGCTCGTAGTTGTCT 138 54
NPC2q-Rd CGCCTTCGTCCTTCGTTTCAGTT
内参
Reference		 β-actinq-F TGCCAACACTGTCCTTTCTGGA 96 53
β-actinq-R TTCATGGTGGATGGTGCTAGGG
酶切位点用下划线表示The underlines represent the restriction sites

Table 2

Sequence characteristics and physicochemical properties of AcNPC2 gene family"

基因
Gene		 GenBank登录号
GenBank accession number		 开放阅读框全长
ORF full length (bp)		 编码蛋白质全长
Length of encoding protein (aa)		 分子量
Molecular weight
(kD)		 等电点
Isoelectric point (pI)		 与意蜂核苷酸序列的相似性
Similarity to the nucleotide sequence of A. m. ligustica (%, E value)
AcNPC2a KJ633823.1 447 148 16.12 7.98 94.59%, 1e-98
AcNPC2b OL741688 480 159 18.53 7.57 96.86%, 2e-113
AcNPC2c OL741689 459 152 16.96 6.56 98.03%, 8e-103
AcNPC2d OL741690 465 154 16.78 6.34 99.35%, 2e-108

Fig. 1

Multiple sequences alignment between AcNPC2s and homologous protein sequences The N-terminal box part is the predicted signal peptide sequence; Asterisks mean the conserved cysteine; The different amino acids between AcNPC2s and AmNPC2s were shown by bold fonts and underline; HomNPC2, BovNPC2 and CjapNPC2 belong to Homo sapiens NPC2, bovine NPC2 and Camponotus japonicus NPC2, respectively"

Fig. 2

Phylogenetic tree based on AcNPC2 gene family"

Table 3

Three-dimensional model scoring of AcNPC2 proteins"

蛋白Protein 描述Description Template Seq Identity GMQE QMEAN
AcNPC2a 人Homo sapiens 5kwy.1.C 32.74% 0.68 -1.25
AcNPC2b 牛Bovine 2hka.1.A 28.57% 0.55 -2.20
AcNPC2c 日本弓背蚁C. japonicus 3wea.1.A 39.53% 0.77 -0.15
AcNPC2d 日本弓背蚁C. japonicus 3wea.2.A 33.06% 0.63 -2.96

Fig. 3

Three-dimensional structure prediction of AcNPC2 gene family protein A-H are the best predicted three-dimensional models of AcNPC2 proteins and their corresponding surface models, respectively, red indicates disulfide bonds, the yellow parts are the β-fold structures, and the arrow indicates the amino acid sequence from N-terminal to C-terminal"

Fig. 4

Expression profile of AcNPC2 gene family in tissues of worker, and in eggs, larvae, and pupae"

Fig. 5

Expression of AcNPC2 gene family in the antenna of foragers at 12℃ and 25℃"

[1] 徐祖荫, 龙立炎. 论中蜂在我国养蜂生产中的地位和作用. 中国蜂业, 2019, 70(2): 63-65.
XU Z Y, LONG L Y. On the position and function of Chinese honeybee in our country’s beekeeping production. Apiculture of China, 2019, 70(2): 63-65. (in Chinese)
[2] RADLOFF S E, HEPBURN C, HEPBURN H R, FUCHS S, HADISOESILO S, TAN K, ENGEL M S, KUZNETSOV V. Population structure and classification of Apis cerana. Apidologie, 2010, 41(6): 589-601.
doi: 10.1051/apido/2010008
[3] 谭荣德, 李宏芳, 陆超丽, 吴殿军. 中华蜜蜂对茶花蜜源的利用研究. 中国蜂业, 2013, 64(16): 33-35.
TAN R D, LI H F, LU C L, WU D J. Study on the utilization of tea nectar by Chinese honeybees. Apiculture of China, 2013, 64(16): 33-35. (in Chinese)
[4] 周冰峰, 许正鼎. 蜜蜂低温采集活动的研究. 中国蜂业, 1988(5): 7-9.
ZHOU B F, XU Z D. Study on low temperature collection of bee. Apiculture of China, 1988(5): 7-9. (in Chinese)
[5] 余林生, 邹运鼎, 曹义锋, 毕守东, 巫厚长, 丁建, 解文飞. 意大利蜜蜂(Apis mellifera ligustica)与中华蜜蜂(Apis cerana cerana)的生态位比较. 生态学报, 2008, 28(9): 4575-4581.
YU L S, ZOU Y D, CAO Y F, BI S D, WU H Z, DING J, XIE W F. Comparative study on the niches of Apis mellifera ligustica and Apis cerana cerana. Acta Ecologica Sinica, 2008, 28(9): 4575-4581. (in Chinese)
[6] LEAL W S. Odorant reception in insects: Roles of receptors, binding proteins, and degrading enzymes. Annual Review of Entomology, 2013, 58: 373-391.
doi: 10.1146/annurev-ento-120811-153635
[7] 莫建初, 王成盼, 尉吉乾. 昆虫外周嗅觉系统研究进展. 江西农业大学学报, 2019, 41(1): 50-57.
MO J C, WANG C P, WEI J Q. Advance in the research on insect peripheral olfactory system. Acta Agriculturae Universitatis Jianxienses, 2019, 41(1): 50-57. (in Chinese)
[8] JIAO Z, GUO M, BAN L, SONG L M, LIU Y, PELOSI P, WANG G. Niemann-Pick C 2 proteins: A new function for an old family. Frontiers in Physiology, 2018, 9: 52.
doi: 10.3389/fphys.2018.00052
[9] 李红亮, 张林雅, 庄树林, 倪翠侠, 韩宝瑜, 商晗武. 中华蜜蜂普通气味结合蛋白ASP2的气味结合功能模式分析. 中国农业科学, 2013, 46(1): 154-161.
LI H L, ZHANG L Y, ZHUANG S L, NI C X, HAN B Y, SHANG H W. Interpretation of odorant binding function and mode of general odorant binding protein ASP2 in Chinese honeybee (Apis cerana cerana). Scientia Agricultura Sinica, 2013, 46(1): 154-161. (in Chinese)
[10] LI H L, SONG X M, WU F, QIU Y L, FU X B, ZHANG L Y, TAN J. Chemical structure of semiochemicals and key binding sites together determine the olfactory functional modes of odorant-binding protein 2 in Eastern honey bee, Apis cerana. International Journal of Biological Macromolecules, 2020, 145: 876-884.
doi: 10.1016/j.ijbiomac.2019.11.189
[11] 倪翠侠, 张林雅, 李红亮, 商晗武. 中华蜜蜂化学感受蛋白基因家族克隆及表达特征分析. 中国农业科学, 2013, 46(8): 1706-1715.
NI C X, ZHANG L Y, LI H L, SHANG H W. Molecular cloning and expression profiles analysis of chemosensory protein genes family in the Chinese honeybee (Apis cerana cerana). Scientia Agricultura Sinica, 2013, 46(8): 1706-1715. (in Chinese)
[12] LI H L, NI C X, TAN J, ZHANG L Y, HU F L. Chemosensory proteins of the Eastern honeybee, Apis cerana: Identification, tissue distribution and olfactory related functional characterization. Comparative Biochemistry and Physiology. Part B, Biochemistry and Molecular Biology, 2016, 194/195: 11-19.
doi: 10.1016/j.cbpb.2015.11.014
[13] PELOSI P, IOVINELLA I, FELICIOLI A, DANI F R. Soluble proteins of chemical communication: An overview across arthropods. Frontiers in Physiology, 2014, 5: 320.
[14] ISHIDA Y, TSUCHIYA W, FUJII T, FUJIMOTO Z, MIYAZAWA M, ISHIBASHI J, MATSUYAMA S, ISHIKAWA Y, YAMAZAKI T. Niemann-Pick type C2 protein mediating chemical communication in the worker ant. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(10): 3847-3852.
[15] IOVINELLA I, BAN L, SONG L, PELOSI P, DANI F R. Proteomic analysis of castor bean tick Ixodes ricinus: A focus on chemosensory organs. Insect Biochemistry and Molecular Biology, 2016, 78: 58-68.
doi: 10.1016/j.ibmb.2016.09.004
[16] ZHENG Y, WANG S N, PENG Y, LU Z Y, SHAN S, YANG Y Q, LI R J, ZHANG Y J, GUO Y Y. Functional characterization of a Niemann-Pick type C2 protein in the parasitoid wasp Microplitis mediator. Insect Science, 2018, 25(5): 765-777.
doi: 10.1111/1744-7917.12473
[17] LEE K S, PARK H G, DENG Y J, KIM B Y, KYUNG S S, CHOI Y S, YOON H J, LI M, JIN B R. Molecular characterization of a Niemann-Pick disease type C2 protein from the honeybee Apis cerana. Journal of Asia-Pacific Entomology, 2014, 17(3): 555-560.
doi: 10.1016/j.aspen.2014.05.005
[18] ARNOLD K, BORDOLI L, KOPP J, SCHWEDE T. The SWISS- MODEL workspace: A web-based environment for protein structure homology modelling. Bioinformatics, 2006, 22(2): 195-201.
doi: 10.1093/bioinformatics/bti770
[19] LI X, SAHA P, LI J, BLOBEL G, PFEFFER S R. Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(36): 10079-10084.
[20] XU S, BENOFF B, LIOU H L, LOBEL P, STOCK A M. Structural basis of sterol binding by NPC2, a lysosomal protein deficient in Niemann-Pick type C2 disease. The Journal of Biological Chemistry, 2007, 282(32): 23525-23531.
doi: 10.1074/jbc.M703848200
[21] MC CAULIFF L A, XU Z, LI R, KODUKULA S, KO D C, SCOTT M P, KAHN P C, STORCH J. Multiple surface regions on the Niemann-Pick C2 protein facilitate intracellular cholesterol transport. The Journal of Biological Chemistry, 2015, 290(45): 27321-27331.
doi: 10.1074/jbc.M115.667469
[22] LIOU H L, DIXIT S S, XU S, TINT G S, STOCK A M, LOBEL P. NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. The Journal of Biological Chemistry, 2006, 281(48): 36710-36723.
doi: 10.1074/jbc.M608743200
[23] STORCH J, ZHI X. Niemann-Pick C 2 (NPC2) and intracellular cholesterol trafficking. Biochimica et Biophysica Acta, 2009, 1791(7): 671-678.
[24] 吴帆, 张莉, 邱一蕾, 李红亮. 昆虫嗅觉结合蛋白研究进展. 昆虫学报, 2021, 64(4): 523-535.
WU F, ZHANG L, QIU Y L, LI H L. Research progress of olfactory binding proteins in insects. Acta Entomologica Sinica, 2021, 64(4): 523-535. (in Chinese)
[25] HUANG X, WARREN J T, BUCHANAN J, GILBERT L I, SCOTT M P. Drosophila Niemann-Pick type C-2 genes control sterol homeostasis and steroid biosynthesis: A model of human neurodegenerative disease. Development, 2007, 134(20): 3733-3742.
doi: 10.1242/dev.004572
[26] CAICEDO P A, SERRATO I M, SIM S, DIMOPOULOS G, COATSWORTH H, LOWENBERGER C, OCAMPO C B. Immune response-related genes associated to blocking midgut dengue virus infection in Aedes aegypti strains that differ in susceptibility. Insect Science, 2019, 26(4): 635-648.
doi: 10.1111/1744-7917.12573
[27] SHI X Z, ZHONG X, YU X Q. Drosophila melanogaster NPC2 proteins bind bacterial cell wall components and may function in immune signal pathways. Insect Biochemistry and Molecular Biology, 2012, 42(8): 545-556.
doi: 10.1016/j.ibmb.2012.04.002
[28] 陈剑, 王兆祥, 杨岭, 叶海霞, 毛丽, 桂连友, 张国辉. 柑橘大实蝇NPC2基因的序列分析和组织表达模式. 植物保护, 2020, 46(4): 132-136.
CHEN J, WANG Z X, YANG L, YE H X, MAO L, GUI L Y, ZHANG G H. Sequence analysis and tissue expression pattern of Niemann- Pick type C2 gene in Bactrocera minax. Plant Protection, 2020, 46(4): 132-136. (in Chinese)
[29] CHEN M M, ZHONG L, ZHAO C S, WANG F C, JI W J, ZHANG B, LIU S Y, LIU Y Q, LI X S. Characterization of an ecdysteroid- regulated 16 kDa protein gene in Chinese oak silkworm, Antheraea pernyi (Lepidoptera: Saturniidae). Journal of Insect Science, 2020, 20(3): 4.
[30] RENTHAL R, MANGHNANI L, BERNAL S, QU Y, GRIFFITH W P, LOHMEYER K, GUERRERO F D, BORGES L M F, DE LEÓN A P . The chemosensory appendage proteome of Amblyomma americanum (Acari: Ixodidae) reveals putative odorant-binding and other chemoreception-related proteins. Insect Science, 2017, 24(5): 730-742.
doi: 10.1111/1744-7917.12368
[31] ZHOU J J. Odorant-binding proteins in insects. Vitamins 2010, 83: 241-272.
[32] LARTIGUE A, CAMPANACCI V, ROUSSEL A, LARSSON A, JONES T, TEGONI M, CAMBILLAU C. X-ray structure and ligand binding study of a moth chemosensory protein. The Journal of Biological Chemistry, 2002, 277(35): 32094-32098.
doi: 10.1074/jbc.M204371200
[33] PELOSI P, IOVINELLA I, ZHU J, WANG G, DANI F R. Beyond chemoreception: Diverse tasks of soluble olfactory proteins in insects. Biological Reviews of the Cambridge Philosophical Society, 2018, 93(1): 184-200.
doi: 10.1111/brv.12339
[34] FORÊT S, MALESZKA R. Function and evolution of a gene family encoding odorant binding-like proteins in a social insect, the honey bee (Apis mellifera). Genome Research, 2006, 16(11): 1404-1413.
doi: 10.1101/gr.5075706
[35] FORÊT S, WANNER K W, MALESZKA R. Chemosensory proteins in the honey bee: Insights from the annotated genome, comparative analyses and expressional profiling. Insect Biochemistry and Molecular Biology, 2007, 37(1): 19-28.
doi: 10.1016/j.ibmb.2006.09.009
[36] 秦明, 王红芳, 刘振国, 王颖, 王帅, 郗学鹏, 刘春蕾, 张卫星, 胥保华. 中华蜜蜂和意大利蜜蜂耐寒性能差异比较. 中国农业科学, 2017, 50(12): 2380-2388.
QIN M, WANG H F, LIU Z G, WANG Y, WANG S, CHI X P, LIU C L, ZHANG W X, XU B H. Comparison of different cold resistance between Apis cerana cerana and Apis mellifera ligustica. Scientia Agricultura Sinica, 2017, 50(12): 2380-2388. (in Chinese)
[37] 杨冠煌, 孙东江, 肖京城, 孙庆海, 林桂莲. 中华蜜蜂群体内温度、湿度及CO2浓度的变化及调节研究. 中国农业科学, 1999, 32(3): 96-101.
YANG G H, SUN D J, XIAO J C, SUN Q H, LIN G L. Study on the regulation ability of Apis cerana cerana on temperature, relative humidity and CO2 concentration in its colony. Scientia Agricultura Sinica, 1999, 32(3): 96-101. (in Chinese)
[38] RATNIEKS F L W. Asian honey bees: Biology, conservation, and human interactions. Nature, 2006, 442(7100): 249.
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