Abstract
Newcastle disease virus (NDV) is a lethal virus in avian species with a disastrous effect on the poultry industry. NDV is enveloped by a host-derived membrane with two glycosylated haemagglutinin-neuraminidase (HN) and Fusion (F) proteins. NDV infection usually leads to death within 2–6 days, so the preexisting antibodies provide the most critical protection for this infection. The HN and F glycoproteins are considered the main targets of the immune system. In the present study, two constructs harboring the HN or F epitopes are sub-cloned separately under the control of a root-specific promoter NtREL1 or CaMV35S (35S Cauliflower Mosaic Virus promoter) as a constitutive promoter. The recombinant vectors were transformed into the Agrobacterium tumefaciens strain LBA4404 and then introduced to tobacco (Nicotiana tabacum L.) leaf disk explants. PCR with specific primers was performed to confirm the presence of the hn and f genes in the genome of the regenerated plants. Then, the positive lines were transformed via non-recombinant A. rhizogenes (strain ATCC15834) to develop hairy roots.HN and F were expressed at 0.37% and 0.33% of TSP using the CaMV35S promoter and at 0.75% and 0.54% of TSP using the NtREL1 promoter, respectively. Furthermore, the mice fed transgenic hairy roots showed a high level of antibody responses (IgG and IgA) against rHN and rF proteins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdisa T, Tagesu T (2017) Review on Newcastle disease of poultry and its public health importance. J Vet Sci Technol 8:441. https://doi.org/10.4172/2157-7579.1000441
Amani J, Kazemi R, Abbasi AR, Salmanian AH (2011) A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis. Iranian J Biotechnol 9:69–71
Armstrong EP (2007) Economic benefits and costs associated with target vaccinations. J Manage Care Pharm 13:12–15. https://doi.org/10.18553/jmcp.2007.13.s7-b.12
Berinstein A, Vazquez-Rovere C, Asurmendi S, Gómez E, Zanetti F, Zabal O, Tozzini A, Grand DC, Taboga O, Calamante G (2005) Mucosal and systemic immunization elicited by Newcastle disease virus (NDV) transgenic plants as antigens. Vaccine 23:5583–5589. https://doi.org/10.1016/j.vaccine.2005.06.033
Blokhina EA, Mardanova ES, Stepanova LA, Tsybalova LM, Ravin NV (2020) Plant-produced recombinant influenza A virus candidate vaccine based on flagellin linked to conservative fragments of M2 protein and hemagglutintin. Plants 9:162. https://doi.org/10.3390/plants9020162
Cattoli G, Susta L, Terregino C, Brown C (2011) Newcastle disease: a review of field recognition and current methods of laboratory detection. J Vet Diagn Invest 23:637–656. https://doi.org/10.1177/1040638711407887
Chumbe A, Izquierdo-Lara R, Calderón K, Fernández-Díaz M, Vakharia VN (2017) Development of a novel Newcastle disease virus (NDV) neutralization test based on recombinant NDV expressing enhanced green fluorescent protein. Virol J 14:1–11. https://doi.org/10.1186/s12985-017-0900-8
Das P (2004) Revolutionary vaccine technology breaks the cold chain. Lancet Infect Dis 4:719. https://doi.org/10.1016/s1473-3099(04)01222-8
De Guzman G, Walmsley AM, Webster DE, Hamill JD (2011) Hairy roots cultures from different Solanaceous species have varying capacities to produce E. coli B-subunit heat-labile toxin antigen. Biotechnol Lett 33:2495–2502. https://doi.org/10.1007/s10529-011-0710-9
Dimitrov KM, Afonso CL, Yu Q, Miller PJ (2017) Newcastle disease vaccines—a solved problem or a continuous challenge? Vet Microbiol 206:126–136. https://doi.org/10.1016/j.vetmic.2016.12.019
Elmayan T, Tepfer M (1995) Evaluation in tobacco of the organ specificity and strength of therold promoter, domain a of the 35S promoter and the 35S 2 promoter. Transgenic Res 4:388–396. https://doi.org/10.1007/BF01973757
Fallah A, Akhavian A, Kazemi R, Jafari M, Salmanian A-H (2018) Production of recombinant LSC protein in Nicotiana tabacum hairy root: direct vs indirect, a comparison. J Appl Biotechnol Rep 5:1–7. https://doi.org/10.29252/JABR.01.01.01
Ganapathy M (2016) Plants as bioreactors-a review. Adv Tech Biol Med 4(2379–1764):1000161. https://doi.org/10.4172/2379-1764.1000161
Ganar K, Das M, Sinha S, Kumar S (2014) Newcastle disease virus: current status and our understanding. Virus Res 184:71–81. https://doi.org/10.1016/j.virusres.2014.02.016
Ghaffar Shahriari A, Bagheri A, Bassami MR, Malekzadeh-Shafaroudi S, Afsharifar A, Niazi A (2016) Expression of hemagglutinin-neuraminidase and fusion epitopes of Newcastle disease virus in transgenic tobacco. Electron J Biotechnol 19:38–43. https://doi.org/10.1016/j.ejbt.2016.05.003
Guerrero-Andrade O, Loza-Rubio E, Olivera-Flores T, Fehérvári-Bone T, Gómez-Lim MA (2006) Expression of the Newcastle disease virus fusion protein in transgenic maize and immunological studies. Transgenic Res 15:455–463. https://doi.org/10.1007/s11248-006-0017-0
Haji-Abdolvahab H, Ghalyanchilangeroudi A, Bahonar A, Ghafouri SA, Marandi MV, Mehrabadi MHF, Tehrani F (2019) Prevalence of avian influenza, Newcastle disease, and infectious bronchitis viruses in broiler flocks infected with multifactorial respiratory diseases in Iran, 2015–2016. Trop Anim Health pro 51:689–695. https://doi.org/10.1007/s11250-018-1743-z
Hirlekar R, Bhairy S (2017) Edible vaccines: An advancement in oral immunization. Facilities 16:20. https://doi.org/10.22159/ajpcr.2017.v10i2.15825
Howard JA, Hood EE (2016) Commercial plant-produced recombinant protein products. Springer. https://doi.org/10.1007/978-3-662-43836-7
Kapczynski DR, Afonso CL, Miller PJ (2013) Immune responses of poultry to Newcastle disease virus. Dev Comp Immunol 41:447–453. https://doi.org/10.1016/j.dci.2013.04.012
Kapczynski DR, King DJ (2005) Protection of chickens against overt clinical disease and determination of viral shedding following vaccination with commercially available Newcastle disease virus vaccines upon challenge with highly virulent virus from the California 2002 exotic Newcastle disease outbreak. Vaccine 23:3424–3433. https://doi.org/10.1016/j.vaccine.2005.01.140
Karsunke J, Heiden S, Murr M, Karger A, Franzke K, Mettenleiter TC, Römer-Oberdörfer A (2019) W protein expression by Newcastle disease virus. Virus Res 263:207–216. https://doi.org/10.1016/j.virusres.2019.02.003
Kim S-H, Xiao S, Paldurai A, Collins PL, Samal SK (2014) Role of C596 in the C-terminal extension of the haemagglutinin–neuraminidase protein in replication and pathogenicity of a highly virulent Indonesian strain of Newcastle disease virus. J Gen Virol 95:331. https://doi.org/10.1099/vir.0.055285-0
Koyama T, Ono T, Shimizu M, Jinbo T, Mizuno R, Tomita K, Mitsukawa N, Kawazu T, Kimura T, Ohmiya K (2005) Promoter of Arabidopsis thaliana phosphate transporter gene drives root-specific expression of transgene in rice. J Biosci Bioeng 99:38–42. https://doi.org/10.1263/jbb.99.38
Kumar GS, Ganapathi T, Srinivas L, Revathi C, Bapat V (2006) Expression of hepatitis B surface antigen in potato hairy roots. Plant Sci 170:918–925. https://doi.org/10.1016/j.plantsci.2005.12.015
Lai KS, Yusoff K, Mahmood M (2013) Functional ectodomain of the hemagglutinin-neuraminidase protein is expressed in transgenic tobacco cells as a candidate vaccine against Newcastle disease virus. Plant Cell Tiss Org Cult 112:117–121. https://doi.org/10.1007/s11240-012-0214-x
Lau OS, Sun SS (2009) Plant seeds as bioreactors for recombinant protein production. Biotechnol Adv 27:1015–1022. https://doi.org/10.1016/j.biotechadv.2009.05.005
Leach F, Aoyagi K (1991) Promoter analysis of the highly expressed rolC and rolD root-inducing genes of Agrobacterium rhizogenes: enhancer and tissue-specific DNA determinants are dissociated. Plant Sci 79:69–76. https://doi.org/10.1016/0168-9452(91)90071-F
Liu J-J, Ekramoddoullah AK (2003) Root-specific expression of a western white pine PR10 gene is mediated by different promoter regions in transgenic tobacco. Plant Mol Biol 52:103–120. https://doi.org/10.1023/a:1023930326839
Makhzoum A, Benyammi R, Moustafa K, Trémouillaux-Guiller J (2014) Recent advances on host plants and expression cassettes’ structure and function in plant molecular pharming. BioDrugs 28:145–159. https://doi.org/10.1007/s40259-013-0062-1
Mason HS, Arntzen CJ (1995) Transgenic plants as vaccine production systems. Trends Biotechnol 13:388–392. https://doi.org/10.1016/S0167-7799(00)88986-6
Medina-Bolívar F, Cramer C (2004) Production of recombinant proteins by hairy roots cultured in plastic sleeve bioreactors. Methods Mol Biol 267:351–363. https://doi.org/10.1385/1-59259-774-2:351
Miller PJ, Afonso CL, El Attrache J, Dorsey KM, Courtney SC, Guo Z, Kapczynski DR (2013) Effects of Newcastle disease virus vaccine antibodies on the shedding and transmission of challenge viruses. Dev Comp Immunol 41:505–513. https://doi.org/10.1016/j.dci.2013.06.007
Motamedi MJ, Amani J, Shahsavandi S, Salmanian AH (2014) In silico design of multimeric HN-F antigen as a highly immunogenic peptide vaccine against Newcastle disease virus. Int J Pept Res Ther 20:179–194. https://doi.org/10.1007/s10989-013-9380-x
Motamedi MJ, Ebrahimi MM, Shahsavandi S, Amani J, Kazemi R, Jafari M, Salmanian AH (2020) The immunogenicity of a novel chimeric hemagglutinin-neuraminidase-fusion antigen from Newcastle disease virus by oral delivery of transgenic canola seeds to chickens. Mol Biotechnol 62:344–354. https://doi.org/10.1007/s12033-020-00254-y
Motamedi MJ, Shahsavandi S, Amani J, Kazemi R, Takrim S, Jafari M, Salmanian A-H (2018) Immunogenicity of the multi-epitopic recombinant glycoproteins of Newcastle disease virus: implications for the serodiagnosis applications. Iranian J Biotechnol 16:248–257. https://doi.org/10.21859/IJB.1749
Ozyigit II, Dogan I, Tarhan EA (2013) Agrobacterium rhizogenes-mediated transformation and its biotechnological applications in crops. In: Crop improvement. Springer, pp 1–48. https://doi.org/10.1007/978-1-4614-7028-1_1
Perozo F, Marcano R, Afonso CL (2012) Biological and phylogenetic characterization of a genotype VII Newcastle disease virus from Venezuela: efficacy of field vaccination. J Clin Microbiol 50:1204–1208. https://doi.org/10.1128/JCM.06506-11
Potenza C, Aleman L, Sengupta-Gopalan C (2004) Targeting transgene expression in research, agricultural, and environmental applications: promoters used in plant transformation. In Vitro Cell Dev Biol Plant 40:1–22. https://doi.org/10.1079/IVP2003477
Rehmani SF, Wajid A, Bibi T, Nazir B, Mukhtar N, Hussain A, Lone NA, Yaqub T, Afonso CL (2015) Presence of virulent Newcastle disease virus in vaccinated chickens in farms in Pakistan. J Clin Microbiol 53:1715–1718. https://doi.org/10.1128/JCM.02818-14
Russell DW, Sambrook J (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Cold Spring Harbor, NY
Shahar E, Haddas R, Goldenberg D, Lublin A, Bloch I, Bachner Hinenzon N, Pitcovski J (2018) Newcastle disease virus: is an updated attenuated vaccine needed? Avian Pathol 47:467–478. https://doi.org/10.1080/03079457.2018.1488240
Shahid N, Samiullah TR, Shakoor S, Latif A, Yasmeen A, Azam S, Shahid AA, Husnain T, Rao AQ (2020) Early stage development of a Newcastle disease vaccine candidate in corn. Front Vet Sci 7:499. https://doi.org/10.3389/fvets.2020.00499
Shahriari AG, Bagheri A, Bassami MR, Malekzadeh Shafaroudi S, Afsharifar AR (2015) Cloning and expression of fusion (F) and haemagglutinin-neuraminidase (HN) epitopes in hairy roots of tobacco (Nicotiana tabaccum) as a step toward developing a candidate recombinant vaccine against Newcastle disease. J Cell Mol Res 7:11–18. https://doi.org/10.22067/JCMR.V7I1.41621
Skarjinskaia M, Ruby K, Araujo A, Taylor K, Gopalasamy-Raju V, Musiychuk K, Chichester JA, Palmer GA, de la Rosa P, Mett V (2013) Hairy roots as a vaccine production and delivery system. In: biotechnology of hairy root systems. Springer, pp 115–134. https://doi.org/10.1007/10_2013_184
Thomas DR, Walmsley AM (2018) Plant-made veterinary vaccines for Newcastle disease virus. In: prospects of plant-based vaccines in veterinary medicine. Springer, pp 149–167. https://doi.org/10.1007/978-3-319-90137-4_6
Varasteh-Shams M, Nazarian-Firouzabadi F, Ismaili A (2020) The direct and indirect transformation methods on expressing a recombinant Dermaseptin peptide in tobacco transgenic hairy root clones. Curr Plant Biol 24:100177. https://doi.org/10.1016/j.cpb.2020.100177
Walmsley AM, Alvarez ML, Jin Y, Kirk DD, Lee S, Pinkhasov J, Rigano MM, Arntzen C, Mason HS (2003) Expression of the B subunit of Escherichia coli heat-labile enterotoxin as a fusion protein in transgenic tomato. Plant Cell Rep 21:1020–1026. https://doi.org/10.1007/s00299-003-0619-4
Wang Y, Yu W, Huo N, Wang W, Guo Y, Wei Q, Wang X, Zhang S, Yang Z, Xiao S (2017) Comprehensive analysis of amino acid sequence diversity at the F protein cleavage site of Newcastle disease virus in fusogenic activity. PLoS ONE 12:e0183923. https://doi.org/10.1371/journal.pone.0183923
Wen G, Hu X, Zhao K, Wang H, Zhang Z, Zhang T, Yang J, Luo Q, Zhang R, Pan Z (2016) Molecular basis for the thermostability of Newcastle disease virus. Sci Rep 6:1–9. https://doi.org/10.1038/srep22492
Wongsamuth R, Doran PM (1997) Production of monoclonal antibodies by tobacco hairy roots. Biotechnol Bioeng 54:401–415. https://doi.org/10.1002/(SICI)1097-0290(19970605)54:5%3c401::AID-BIT1%3e3.0.CO;2-I
Xu J, Towler M, Weathers P (2016) Platforms for plant-based protein production. Bioprocess Plant Vitro Sys 1:1–40. https://doi.org/10.1007/978-3-319-32004-5_14-1
Zhang C, Pan S, Chen H, Cai T, Zhuang C, Deng Y, Zhuang Y, Zeng Y, Chen S, Zhuang W (2016) Characterization of NtREL1, a novel root-specific gene from tobacco, and upstream promoter activity analysis in homologous and heterologous hosts. Plant Cell Rep 35:757–769. https://doi.org/10.1007/s00299-015-1918-2
Acknowledgements
We gratefully acknowledge the Iran National Science Foundation (INSF) (Grant No.: 95844522) and the National Institute of Genetic Engineering and Biotechnology (NIGEB) (Grant number: 960827-IV-701) for financially supporting this work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare they have no financial or competing conflict of interest.
Ethical statement
The animals were kept in suitable and pathogen-free conditions in the Animal House of the National Institute of Genetic Engineering and Biotechnology (Tehran, Iran). All experimental procedures were approved by the ethics committee of the institute (IR.NIGEB.EC.1397.8.1.D and IR.NIGEB.EC.1397.8.1.C). At the end of the experiments, the animals were sacrificed according to ethical standards.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Boroujeni, N.A., Khatouni, S.B., Motamedi, M.J. et al. Root-preferential expression of Newcastle virus glycoproteins driven by NtREL1 promoter in tobacco hairy roots and evaluation of oral delivery in mice. Transgenic Res 31, 201–213 (2022). https://doi.org/10.1007/s11248-021-00295-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-021-00295-2