A Comparative Studies of the Activities of Phenoloxidase, Carboxylesterase and Alkaline Phosphatase from Different Geographic Strains of Flacherie-Infected Bombyx mori

Volume 6, Issue 3, June 2021     |     PP. 25-47      |     PDF (607 K)    |     Pub. Date: May 17, 2021
DOI: 10.54647/agriculture21204    105 Downloads     5327 Views  

Author(s)

Wenchu Li, Department of Sericultural Science, College of Animal Science, South China Agricultural University, Guangzhou, China.
Jingyu Zhang, Department of Sericultural Science, College of Animal Science, South China Agricultural University, Guangzhou, China.
Shizhang Feng, Department of Sericultural Science, College of Animal Science, South China Agricultural University, Guangzhou, China.

Abstract
Flacherie diseases of silkworm caused by complex pathogens but mainly Bacillus spp. in springtime. The paper aims to explore interaction between different geographic strains of pathogens and host, and focuses on studying phenoloxidase (PO) and other enzymes activities in hemolymph, midgut and silk glands. PO activity increased in silkworms infected with Bacillus spp. geographic strain of BsE, and BsC. The activities of PO and carboxylesterase (CE) in the midgut of infected silkworms gradually increased in fourth larval instars, while it decreased in fifth larval instars. Alkaline phosphatase (ALKP) activity was present on the inner and outer membranes of the silk gland of diseased silkworms. The outer membrane of the silk gland was dyed with Diaminoben-zidine (DAB), but the inner membrane excluded dye. A mixture of the Nitro-Blue-Tetrazolium/ 5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) and DAB methods was used to treat the fat body, and both reactions could be detected. Current research into the PO cascade, CE, and ALKP is further reviewed and discussed in detail.

Keywords
flacherie; Bombyx mori; enzymes; silk gland; pathology; insect immunity

Cite this paper
Wenchu Li, Jingyu Zhang, Shizhang Feng, A Comparative Studies of the Activities of Phenoloxidase, Carboxylesterase and Alkaline Phosphatase from Different Geographic Strains of Flacherie-Infected Bombyx mori , SCIREA Journal of Agriculture. Volume 6, Issue 3, June 2021 | PP. 25-47. 10.54647/agriculture21204

References

[ 1 ] Albert Š, Gätschenberger H, Azzami K, Gimple O, Grimmer G, Sumner S, Fujiyuki T, Tautz J, Mueller MJ. (2011) Evidence of a novel immune responsive protein in the Hymenoptera. Insect Biochem Mol Biol. 41: 968-981. Doi:10.1016/j.ibmb. 2011.09.006.
[ 2 ] Andersen SO. (2010) Insect cuticular sclerotization: A review. Insect Biochem Mol Biol.,40:166-178. Doi:10.1016/j.ibmb.2009.10.007.
[ 3 ] Aruga Hisao. (1994) Principles of Sericulture (Translated from Japanese). Oxford and IBH Publishing Co.Pvt.Ltd.66 Janpath, New Delhi, 110001.
[ 4 ] Ashida M and Brey P.T.( 1998) Recent advances in research on the insect prophenoloxidase cascade. In “Molecular Mechanisms of Immune Responses in Insects”, 135-172. Edit by Paul T. Brey and Dan Hultmark. Chapman & Hall, 2-6 Boundary Row, London SE1 8HN.
[ 5 ] Ashida M. and Dohke K.(1980) Activation of prophenoloxidase by the activating enzyme of the silkworm, Bombyx mori. Insect Biochem.10: 27-47. Doi:10.1016/0020- 1790 (80)90036-0.
[ 6 ] Azevedo AM, Martins VC, Prazeres DMF, Vojinović V, Cabral JMS, Fonseca LP. (2003) Horseradish peroxidase: a valuable tool in biotechnology. Biotechnol.Annu. Rev. 9: 199–247. Doi:10.1016/S1387-2656(03)09003-3.
[ 7 ] BaoYY, Tang XD, Lv ZY, Wang XY, Tian CH, XuYP, Zhang CX. (2009) Gene expression profiling of resistant and susceptible Bombyx mori strains reveals nucleopolyhedro- virus-associated variations in host gene transcript levels. Genomics. 94: 138–145. Doi:10.1016/j.ygeno.2009.04.003
[ 8 ] Bublitz R., Armesto J., Hoffmann-Blume E., Schulze M., Rhode H., Horn A,, Aulwurm S., Hannappel E. and Fischer W. (1993) Heterogeneity of glycosylphosphatidyl- inositol-anchored alkaline phosphatase of calf intestine. Eur. J. Biochem. 217: 199- 207. Doi:10.1111/j.1432-1033. 1993.tb18234.x
[ 9 ] Casartelli M, Corti P, Cermenati G, Grimaldi A, Fiandra L, Santo N, Pennacchio F, Giordana B. (2007) Absorption of horseradish peroxidase in Bombyx mori larval midgut. J Insect Physiol.53:517–525. Doi:10.1016/j.jinsphys.2007.02.004.
[ 10 ] Chen KK, Liu C, He Y, Jiang HB, Lu ZQ. (2014) A short-type peptidoglycan recognition protein from the silkworm: expression, characterization and involvement in the prophenoloxidase activation pathway. Dev. Comp. Immunol. 4: 1-9. Doi: 10.1016/j.dci.2019.01.015.
[ 11 ] Chen TT, Hu N, Tan LR, Xiao Q, Dong ZQ, Chen P, Xu AY, Pan MH, Lu C. (2019) Resistant silkworm strain block viral infection independent of melanization. Pesti Biochem Physiol.154:88-96. Doi:10.1016/j.pestbp.2018.12.012.
[ 12 ] Choudhury T P, Lalita G, Kumar S. (2019 ) Identification, characterization and expression analysis of Anopheles stephensi double peroxidase. Acta Tropica. 190: 210–219. Doi: 10.1016/j.actatropica.2018.10.008.
[ 13 ] Corona M, Robinson GE. (2006) Genes of the antioxidant system of the honey bee: annotation and phylogeny. Insect Mol. Biol. 15 (5): 687–701. Doi:10.1111 /j.1365-2583.2006.00695.x.
[ 14 ] Cui F, Lin Z, Wang HS, Liu SL, Chang HJ, Reeck G, Qiao CL, Raymond M, Kang L. (2011) Two single mutations commonly cause qualitative change of nonspecific carboxyl-esterases in insects. Insect Biochem Mol Biol. 41: 1-8. Doi:10.1016/ j.ibmb.2010.09.004
[ 15 ] Eguchi M, Azuma M, Yamamoto H, Takeda S. (1990)Genetically defined membrane-bound and soluble alkaline phosphatases of the silkworm: their discrete localization and properties. Prog Clin Biol Res. 344:267-87.PMID:2392471.
[ 16 ] Eguchi M. and Kuriyama K. (1985) Purification and characterization of membrane-bound alkaline proteases from midgut tissue of the silkworm, Bombyx mori. J. Biochem. 97:1437-1445. Doi: 10.1093/oxfordjournals.jbchem.a135198.
[ 17 ] Eguchi M., Sawaki M. and Suzuki Y. (1972) Multiple forms of midgut alkaline phosphatase in the silkworm: separation and comparison of two isoenzymes. Insect Biochem. 2:167-174. Doi:10.1016/0020-1790(72)90050-9.
[ 18 ] Ganga G. and J.Sulochana Chetty. (1991).An Introduction to Sericulture. Oxford and IBH Publishing Co. Pvt.Ltd. 66 Janpath, New Delhi, 110001.
[ 19 ] Grabowski NT, Olivas JF, Lozano DG, Kehrenberg C, Aguilar DG. (2018) Assessment of pasteurisation of edible insects using enzymatic tests (activity of alkaline phosphatase and lactoperoxidase) applied in dairy products. Food Sci Technol Int. 24(8):699-704. Doi: 10.1177/1082013218789815.
[ 20 ] Gray JS, Yun Yang B, Montgomery R. (1998) Heterogeneity of glycans at each N-glycosylation site of horseradish peroxidase. Carbohydr Res. 311:61–9. Doi: 10.1016/s0008-6215(98)00209-2.
[ 21 ] Gu ZY, Li FC, Wang BB, Xu KZ, Ni M, Zhang H, Shen WD, Li B. (2015) Differentially expressed genes in the fat body of Bombyx mori in response to phoxim insecticide. Pesti Biochem Physiol.117: 47–53. Doi: 10.1016/j.pestbp. 2014.10.007.
[ 22 ] Guo HZ, Jiang L, Xia QY. (2016) Selection of reference genes for analysis of stress-responsive genes after challenge with viruses and temperature changes in the silkworm Bombyx mori. Mol. Genet. Genomics..291(2):999-1004. Doi：10.1007/ s00438-015-1125-4.
[ 23 ] Harlow E and Lane D. (2006) Preparing paraffin tissue sections for immunostaining. CSH Protoc. (1) pii: pdb.prot4329. Doi: 10.1101/pdb.prot4329.
[ 24 ] Hiroto O, Hiroshi O, Naotaka H, Tetsuya I, Yoshihisa O, Shigeru M, Kenji K.( 2012) A secreted placental alkaline phosphatase-based reporter assay system for screening of compounds acting at an octopamine receptor stably expressed in a mammalian cell line. Biosci. Biotechnol. Biochem. 76 (1): 209–211. Doi:10.1271/bbb.110690.
[ 25 ] Ishida Y and Leal WS. (2005) Rapid inactivation of a moth pheromone. Proc. Natl. Acad.Sci. U. S. A. 102: 14075–14079. Doi:10.1073pnas.0505340102 PN.
[ 26 ] Kausar S, Abbas MN, Qian C, Zhu BJ, Sun Y, Sun YX, Wang L, Wei GQ, Maqsood I, Liu CL. (2017 ) Serpin-14 negatively regulates prophenoloxidase activation and expression of antimicrobial peptides in Chinese oak silkworm Antheraea pernyi. Dev Com Immunol. 76: 45-55 Doi:10.1016/j.dci.2017.05.017
[ 27 ] Kawamoto M., Jouraku A., Toyodac A., Yokoi K., Minakuchi Y, Katsuma S, Fujiyama A, Kiuchi T, Yamamoto K, Shimada T. (2019) High-quality genome assembly of the silkworm, Bombyx mori. Insect Biochem Mol Biol. 107 :53–62. Doi:10.1016 /j.ibmb.2019.02.002.
[ 28 ] Lee KS, Kim BY, Choo YM, Jin BR. (2018) Dual role of the serine protease homolog BmSPH-1 in the development and immunity of the silkworm Bombyx mori. Dev Com Immunol. 85: 170-176. Doi:10.1016/j.dci. 2018.04.011.
[ 29 ] Leonard C. Ratcliffe N. A. and Rowley A. F. (1985) The role of prophenoloxidase actiation in non-self recognition and phagocytosis by insect blood cells. J. Insect Physiol. 31(10): 789-799. Doi：10.1016/0022-1910(85)90072-1.
[ 30 ] Li B, Yu HZ, Ye CJ, Ma Y, Li X, Fan T, Chen FS, Xu JP.( 2017) Bombyx mori Serpin-6 regulates prophenoloxidase activity and the expression of antimicrobial proteins. Gene. 30;610:64-70. Doi: 10.1016/j.gene.2017.02.011.
[ 31 ] Li JL, Li M, Zhe L, Zhen Z, Lu ZQ. (2016) Serpin-5 regulates prophenoloxidase activation and antimicrobial peptide pathways in the silkworm, Bombyx mori. Insect Biochem Mol Biol. 73: 27-37. Doi: 10.1016/j.ibmb.2016.04.003
[ 32 ] Liu DG, Wang L, Yang L, Qian C, Wei GQ, Dai LS, Li J, Zhu BJ, Liu CL. (2015) Serpin-15 from Bombyx mori inhibits prophenoloxidase activation and expression of antimicrobial peptides. Dev Com Immunol. 51: 22–28 . Doi:10.1016/j.ibmb. 2016.04.003
[ 33 ] Low M. G., Ferguson M. A. J., Futerman A. H. And Silman I. (1986) Covalently attached phosphatidylinositol as a hydrophobic anchor for membrane proteins. Trends Biochem. Sci. 11: 212-215. Doi:10.1016/0968-0004(86)90009-5.
[ 34 ] Ma L, Zhou LZ, Lin JS, Ji JY, Wang Y, Jiang HB, Shen XH, Lu ZQ. (2019) Manipulation of the silkworm immune system by a metalloprotease from the pathogenic bacterium Pseudomonas aeruginosa. Dev Comp Immunol. 90:176-185. Doi: 10.1016/j.dci.2018.09.017.
[ 35 ] Mao TT, Li FC, Fang YL, Wang H, Chen J, Li MX, Lu ZT, Qu JW, Li JX, Hu JH, Cheng XY, Ni M, Li B. (2019) Effects of chlorantraniliprole exposure on detoxification enzyme activities and detoxification-related gene expression in the fat body of the silkworm, Bombyx mori. Ecotoxicol Environ Safety. 176: 58–63. Doi: 10.1016/j.ecoenv.2019.03.074.
[ 36 ] Miao YG. (2002) Studies on the activity of the alkaline phosphatase in the midgut of infected silkworm, Bombyx mori L. J. Appl. Ent. 126:138-142. Doi: 10.1046/j.1439- 0418.2002.00625.x
[ 37 ] Patmawati, Minamihata K, Tatsuke T, Lee J M, Kusakabe T, Kamiya N. (2019) Functional horseradish peroxidase−streptavidin chimeric proteins prepared using a silkworm-baculovirus expression system for diagnostic purposes. J Biotechnol. 297: 28–31. Doi: 10.1016/j.jbiotec.2019.03.007.
[ 38 ] Qu Z, Steiner H, Engström A, Bennich H, Boman HG. (1982) Insect immunity: isolation and structure of cecropins B and D from pupae of the Chinese oak silk moth, Antheraea pernyi. Eur J Biochem. 127(1):219-224. Doi: 10.1111/j.1432-1033. 1982. tb06858.x
[ 39 ] Rao XJ, Yu XQ. (2010) Innate immune responses in the Chinese oak silkworm, Antheraea pernyi. Dev Com Immunol. 34: 1119–1128. Doi:10.1016/j.dci.2010.06.007.
[ 40 ] Ratcliffe NA, Leonard C, Rowley A.F. (1984) Prophenoloxidase activation: nonself recognition and cell cooperation in insect immunity. Sci. 226(4674): 557-559. PMID: 17821514.
[ 41 ] Shao Q, Yang B, Xu Q, Li X, Lu Z,Wang C, Huang Y, Soderhall K, Ling E. (2012) Hindgut innate immunity and regulation of fecal microbiota through melanization in insects. J Biol Chem. 287: 14270-14279. Doi: 10.1074/jbc.M112.354548
[ 42 ] Shiotsuki T, Kato Y. (1999,) Induction of carboxylesterase isozymes in Bombyx mori by E. coli infection. Insect Biochem Mol Biol. 29: 731–736. Doi:10.1016/S0965- 1748(99)00054-5
[ 43 ] Shu M, Mang DZ, Fu GS, Tanaka S, Endo H, Kikuta S, Sato R. (2016) Mechanisms of nodule-specific melanization in the hemocoel of the silkworm, Bombyx mori. Insect Biochem Mol Biol. 70 :10-23, Doi: 10.1016/j.ibmb.2015.12.005
[ 44 ] Son H F, Kim K J. (2018) Structural basis for substrate specificity of meso-diaminopimelic acid decarboxylase from Corynebacterium glutamicum. Biochem Biophysi Res Commun. 495 :1815e1821. Doi:10.1016/j.bbrc.2017.11.097.
[ 45 ] Sumida M, Ichimori H, Johchi S, Takaoka A, Yuhki T, Mori H, Matsubara F. (1992) Antibacterial activity inducible in the haemolymph of the silkworm, Bombyx mori, by injection of formalin-treated Escherichia coli K-12 during the fifth larval instar and pharate adult development. Comp Biochem Physiol B.,101(1-2):165-71. Doi: 10.1016/0305-0491(92)90174-P.
[ 46 ] Takeda S,  Azuma M,  ItohM, Eguchi M. (1993) The strain difference and analysis of polymorphic nature of membrane-bound alkaline phosphatase in the midgut epithelium of the silkworm, Bombyx mori. Com Biochem Physiol. Part B: Com Biochem.  104(1): 81-89. Doi:10.1016/0305-0491(93)90341-2
[ 47 ] Tanaka H., Ishibashi J., Fujita K., Nakajima Y., Sagisaka A., Tomimoto K., Suzuki N., Yoshiyama M., Kaneko Y., Iwasaki T., Sunagawa T., Yamaji K., Asapka A., Mita K., Yamakawa M., (2008) A genome-wide analysis of genes and gene families involved in innate immunity of Bombyx mori. Insect Biochem. Mol. Biol. 38:1087–1110. DoiOI:10.1016/j.ibmb.2008.09.001.
[ 48 ] Tokura A, Fu GS, Sakamoto M, Endo H, Tanaka S, Kikuta S, Tabunoki H, Sato R. (2014) Factors functioning in nodule melanization of insects and their mechanisms of accumulation in nodules. J Insect Physiol. 60: 40-49. Doi:10.1016/j.jinsphys. 2013.11.003.
[ 49 ] Wang L, Yang L, Zhou XS, Li TH, Liu CL. (2018) A clip domain serine protease stimulates melanization activation and expression of antimicrobial peptides in the Chinese oak silkworm, Antheraea pernyi. J Asia-Pacific Entomol. 21: 864–871. Doi: 10.1016/j.aspen.2018.06.008.
[ 50 ] Wang LL, Liu HW, Fu HY, Zhang L, Guo PC, Xia QY, Zhao P. (2019) Silkworm serpin32 functions as a negative-regulator in prophenoloxidase activation. Dev Com Immunol. 91: 123–131. Doi:10.1016/j.dci.2018.10.006.
[ 51 ] Wang Q, Ren MJ, Liu XY, Xia HC, Chen KP. (2019) Peptidoglycan recognition proteins in insect immunity. Mol Immunol. 106: 69–76. Doi:10.1016/j.molimm.2018.12.021.
[ 52 ] Wang XL, Wang KL, He YY, Lu XR, Wen DH, Wu CF, Zhang JH, Zhang R. (2017) The functions of serpin-3, a negative-regulator involved in prophenoloxidase activation and antimicrobial peptides expression of Chinese oak silkworm, Antheraea pernyi. Dev Com Immunol. 69: 1-11. Doi:10.1016/j.dci. 2016.11.022
[ 53 ] Wang Y, Jiang HB, (2017) Prophenoloxidase activation and antimicrobial peptide expression induced by the recombinant microbe binding protein of Manduca sexta. Insect Biochem Mol Biol. 83: 35–43. Doi:10.1016/j.ibmb.2016.10.006.
[ 54 ] Whitten, M.M.A., Coates, C.J. (2017) Re-evaluation of insect melanogenesis research: views from the dark side. Pigment cell Melano Res. 30: 386–401. Doi: 10. 1111 /pcmr.12590.
[ 55 ] Wu K, Yang B, Huang W, Leonard D, Song HS, Ling EJ. (2016) Gut immunity in Lepidop- teran insects. Dev Com Immunol, 64: 65-74, Doi: 10.1016/j.dci.2016. 02.010
[ 56 ] Wu P, Han S, Chen T, Qin G, Li L, Guo X. (2013) Involvement of microRNAs in infection of silkworm with Bombyx mori cytoplasmic polyhedrosis virus (BmCPV). PLoS ONE, 8(7): e68209. Doi:10.1371/journal.pone.0068209.
[ 57 ] Wu, K., Zhang, J., Zhang, Q., Zhu, S., Shao, Q., Clark, K.D., Liu, Y., Ling, E., (2015) Plant phenolics are detoxified by prophenoloxidase in the insect gut. Sci Rep. 5:16823. Doi: 10.1038/srep16823.
[ 58 ] Zhang SD, Shen ZJ, Li Z, Wu FM, Zhang BY, Liu YJ, Zhang QW, Liu XX. ( 2015) Identification of a thioredoxin peroxidase gene involved in resistance to nucleopolyhedrovirus infection in Helicoverpa armigera with RNA interference. J Insect Physiol. 82: 17–27. Doi:10.1016/j.jinsphys.2015.07.017.
[ 59 ] Zhan XM, Liu HJ, Miao YG, Liu WP. (2006) A comparative study of rac- and S-metolachlor on some activities and metabolism of silkworm, Bombyx mori L. Pesti Biochem Physiol. 85: 133–138.Doi:10.1016/j.pestbp.2005.12.003.
[ 60 ] Zhang XL, Guo R, Kumar D, Ma HY, Liu JB, Hu XL, Cao GL, Xue RY, Gong CL. (2015) Identification, gene expression and immune function of the novel Bm-STAT gene in virus-infected Bombyx mori. Gene, 577(1): 82-88. Doi:10.1016/j.gene.2015. 11.027.
[ 61 ] Zhao GD, Huang MX, Zhang YL, Wang XC, Du J, Li B, Chen YH, Xu YY, Shen WD, Wei ZG. (2014 ) Expression analysis and RNA interference of BmCarE-10 gene from Bombyx mori. Mol Biol Rep. 41:1607–1616. Doi: 10.1007/s11033-013-3007-3.
[ 62 ] Zhao P, Wang GH, Dong ZM, Duan J, Xu PZ, Cheng TC, Xiang ZH, Xia QY, (2010) Genome-wide identification and expression analysis of serine proteases and homologs in the silkworm Bombyx mori. BMC Gen. 11:405. Doi:10.1186/1471- 2164-11-405.
[ 63 ] Zhou XS, Chen C, Li TH, Tang JJ, Zhu BJ, Wei GQ, Qian C, Liu CL, Wang L. (2019) A QM protein from Bombyx mori negatively regulates prophenoloxidase activation and melanization by interacting with Jun protein. Insect Mol Biol. 28:1-13. Doi: 10.1111/imb.12573.
[ 64 ] Zou FM, Lee KS, Kim BY, Kim HJ, Gui ZZ, Zhang GZ, Guo XJ, Jin BR. (2015) Differential and spatial regulation of the prophenoloxidase (proPO) and proPO-activating enzyme in cuticular melanization and innate immunity in Bombyx mori pupae. J Asia-Pacific Entomol. 18: 757–764. Doi:10.1016/j.aspen.2015.09.007.