ORIGINAL ARTICLE
The toxicity of Cry1Ia5 transgenic soybean plants against Spodoptera littoralis
Reda E.A. Moghaieb 1, A-F
,  
Etr Hussein Khashaba 2, A-F  
,  
 
 
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1
Department of Genetics, Faculty of Agriculture, Cairo University, Giza, Egypt
2
Department of Field Crop Pests, Plant Protection Research Institute, Agriculture Research Center, Dokki, Egypt
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article
CORRESPONDING AUTHOR
Etr Hussein Khashaba   

Department of Field Crop Pests, Plant Protection Research Institute, Agriculture Research Center, Dokki, Egypt
Online publish date: 2019-06-27
Submission date: 2019-02-10
Acceptance date: 2019-05-31
 
Journal of Plant Protection Research 2019;59(2):185–191
KEYWORDS
TOPICS
ABSTRACT
Three transgenic soybean lines expressing the Cry1Ia5 gene were developed using the Agrobacterium transformation system. The integration of the Cry1Ia5 gene in the genome of the transgenic plants was approved using specific primers for PCR and real time PCR analysis, respectively. The insecticidal activity of three transgenic lines (L1, L2 and L3) against 2nd larval instars Spodoptera littoralis was tested. The data indicate that L2 exhibited the highest mortality percentage 9 days post feeding (60%) followed by L3 (40%) then L1 (20%) while the control showed 0% mortality. The larvae fed transgenic material appeared smaller in size than compared to the control larvae. The reduction in insect size and weight was due to the accumulation of higher phenoloxidase activity in insect tissues. The higher mortality observed in L2 was due to a significant decrease in the acetylcholine esterase activity that leads to accumulation of acetylcholin at higher levels which causes paralysis and death. The developed transgenic line 2 could be used to construct an insect resistant soybean cultivar.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
REFERENCES (34)
1.
Ahmad A., Maqbool B.S., Hashsham A.S., Sticklen M. 2005. Determination of cryIAb and cryIAc copy number in transgenic basmati 370 rice (Oryza sativa L.) plants using realtime PCR and its comparison with Southern Blot. Journal of Biological Sciences 5 (3): 283−288.
 
2.
Al-shannaf H.M., Mead H.M., Al-Kazafy H.S. 2012. Toxic and biochemical effects of some bioinsecticides and igrs on american bollworm, Helicoverpa armigera (Hüb.) (Noctuidae: Lepidoptera) in cotton fields. Journal of Biofertilizers and Biopesticides 3: 2. DOI: http://dx.doi.org/10.4172/2155....
 
3.
Bernardi O., Malvestiti G.S., Dourado P.M., Oliveira W.S., Martinelli S., Berger G.U., Head G.P., Omoto C. 2012. Assessment of the high-dose concept and level of control provided by MON 87701 × MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera:.Noctuidae) in Brazil. Pest Management Science 68 (7): 1083–1091. DOI: 10.1002/ps.3271.
 
4.
Bernardi O., Sorgatto R.J., Barbosa A.D., Domingues F.A., Dourado P.M., Carvalho R.A., Martinelli S., Head G.P., Omoto C. 2014. Low susceptibility of Spodoptera cosmioides, Spodoptera eridania and Spodoptera frugiperda (Lepidoptera: Noctuidae) to genetically-modified soybean expressing Cry1Ac protein. Crop Protection 58: 33–40. DOI: https://doi.org/10.1016/j.crop....
 
5.
Bradford M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248−254. DOI: https://doi.org/10.1016/0003-2....
 
6.
Brown E.S., Dewhurst C.F. 1975. The genus Spodoptera (Lepidoptera, Noctuidae) in Africa and the Near East. Bulletin of Entomological Research 65 (2): 221–262. DOI: https://doi.org/10.1017/S00074....
 
7.
Castillo J.C., Reynolds S.E., Eleftherianos I. 2011. Insect immune responses to nematode parasites. Trends in Parasitology 27 (12): 537–547. DOI: https://doi.org/10.1016/j.pt.2....
 
8.
Chen M., Liu Z., Ye G., Shen Z., Hu C., Peng Y. 2007. Impacts of transgenic Cry1Ab rice on non-target planthoppers and their main predator Cyrtorhinus lividipennis (Hemiptera: Miridae) a case study of the compatibility of bt rice with biological control. Biological Control 42 (2): 242–250. DOI: 1016/j.biocontrol.2007.05.005.
 
9.
Ding X., Gopalakrishnan B., Lowell B.J., White F.F., Wang X., Morgan T.D., Kramer K.J., Muthukrishnan S. 1998. Insect resistance of transgenic tobacco expressing an insect chitinase gene. Transgenic Research 7 (2): 77–84. DOI: 10.1023/A:1008820507262.
 
10.
Ebrahimi L., Gholamreza N., Dunphy G.B., Toorchi M. 2014. Effect of an entomopathogenic nematode, Steinernema carpocapsae on haemocyte profile and phenoloxidase activity of the Colorado potato beetle, (Leptinotarsa decemlineata). Journal Biocontrol Science and Technology 24 (12): 1383-1393. DOI: https://doi.org/10.1080/095831....
 
11.
Ericsson J.D., Janmaatb A.F., Lowenbergera C., Myers J.H. 2009. Is decreased generalized immunity a cost of Bt resistance in cabbage loopers Trichoplusia ni? Journal of Invertebrate Pathology 100 (2): 61−67. DOI: https://doi.org/10.1016/j.jip.....
 
12.
Gabriela V.S., Buenob A.F., Bortolottoa O.C., Santosc A.C., Fernandes A.P. 2016. Biological characteristics of black armyworm Spodoptera cosmioides on genetically modified soybean and corn crops that express insecticide Cry proteins. Revista Brasileira de Entomologia 60 (3): 255–259. DOI: http://dx.doi.org/10.1016/j.rb....
 
13.
Gomez I., Sánchez J., Munoz-Garay C., Matus V., Gill S.S., Soberón M., Bravo A. Bacillus thuringiensis Cry1A toxins are versatile proteins with multiple modes of action: two distinct pre-pores are involved in toxicity. Biochemical Journal 459 (2): 383–396. DOI: 10.1042/BJ20131408.
 
14.
Guo J., Wu G., Wan F. 2011. Temporal allocation of metabolic tolerance to transgenic Bt cotton in beet armyworm, Spodoptera exigua (Hübner). Science China Life Sciences 54 (2): 152–158. DOI: 10.1007/s11427-010-4133-y.
 
15.
Ishaaya I., Casida J.E. 1974. Dietary TH6040 alters composition and enzyme activity of housefly larval cuticle. Pesticide Biochemistry and Physiology 4 (4): 484–490. DOI: https://doi.org/10.1016/0048-3....
 
16.
Leonard C., Kenneth S., Ratcliffe N.A. 1985. Studies on prophenoloxidase and protease activity of Blaberus craniifer hemocytes. Insect Biochemistry 15 (6): 803–810. DOI: https://doi.org/10.1016/0020-1....
 
17.
Lokstan M., Li S.W. 1988. Insect Biochemistry. Science Press, Beijing, China, pp. 151–153. Martins-Sallesa S., Machadob V., Massochin-Pintoc L., Fiuzad L.M. 2017. Genetically modified soybean expressing insecticidal protein (Cry1Ac): Management risk and perspectives. Facets 2 (1): 496–512. DOI: 10.1139/facets-2017-0006.
 
18.
Moghaieb R.E.A. 2010. Transgenic rice plants expressing Cry1Ia5 gene are resistant to stem borer (Chilo agamemnon). GM Crops 1 (5): 288–293. DOI: https://doi.org/10.4161/gmcr.1....
 
19.
Murashige T., Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum 15: 473–379. DOI: https://doi.org/10.1111/j.1399....
 
20.
Pham L.N., Schneider D.S. 2008. Evidence for specificity and memory in the insect innate immune response. p. 97−127. In: “Insect Immunology” (N. Beckage, ed.). Academic Press, San Diego, CA, USA.
 
21.
Rahman M., Rashid H., Shahid A.A., Bashir K., Husnain T., Riazuddin S. 2007. Insect resistance and risk assessment studies of advanced generations of basmati rice expressing two genes of Bacillus thuringiensis. Electronic Journal of Biotechnology 10 (2): 240–251. DOI: 10.2225/vol10-issue2-fulltext-3.
 
22.
Roushr T., Mckenzie J.A. 1987. Ecological genetics of insecticide and acaricide resistance. Annual Review of Entomology 32: 361−380. DOI: https://doi.org/10.1146/annure....
 
23.
Schmutz J., Cannon S.B., Schlueter J., Ma J., Therese M., Nelson W., Hyten D.L., Song Q., Thelen J.J., Cheng J., Xu D., Hellsten U., May G.D., Yu Y., Sakurai T., Umezawa T., Bhattacharyya M., Sandhu D., Cregan P. 2010. Genome sequence of the palaeopolyploid soybean. Nature 463: 178–183. DOI: 10.1038/nature08670.
 
24.
Schünemann R., Knaak N., Fiuza L.M. 2014. Mode of action and specificity of Bacillus thuringiensis toxins in the control of caterpillars and stink bugs in soybean culture. International Scholarly Research Notices Microbiology 2014: 12 pp. DOI: http://dx.doi.org/10.1155/2014....
 
25.
Selvapandiyan A., Reddy V.S., Kumar P.A., Tewari K.K., Bhatnagar R.K. 1998. Transformation of Nicotiana tabacum with a native Cry1Ia5 gene confers complete protection against Heliothis armigera. Molecular Breeding 4: 473–478. DOI: 10.1023/A:1009696027993.
 
26.
Shin E.C., Lee J.H., Hwang C.E., Lee B.W., Kim H.T., Ko J.M., Baek I.Y., Shin J.H., Nam S.H., Seo W.T., Cho K.M. 2014. Enhancement of total phenolic and isoflavone-aglycone contents and antioxidant activities during Cheonggukjang fermentation of brown soybeans by the potential probiotic Bacillus subtilis CSY191. Food Science and Biotechnology 23: 531–538. DOI 10.1007/s10068-014-0073-9.
 
27.
Simpson D.R., Bull D.L., Linquist D.A. 1964. A semi micro technique for the estimation of cholinesterase activity in Boll weevils. Annals of the Entomological Society of America 57 (3): 367–371. DOI: https://doi.org/10.1093/aesa/5....
 
28.
Soreq H., Seidman S. 2001. Acetylcholinesterase – new roles for an old actor. Nature Reviews Neuroscience 2 (4): 294–302. DOI: 10.1038/35067589.
 
29.
Spector E. 1978. Organizational frustration: A model and review of the literature. Personnel Psychology 31 (4): 815–829. DOI: https://doi.org/10.1111/j.1744....
 
30.
Stein A.J., Rodríguez-Cerezo E. 2009. A review of the world’s commercial pipeline of GM crops and implications for asynchronous approval and trade. In: Proceedings of the 13th ICABR Conference on “The Emerging Bio-Economy”, 18−20 June 2009, Ravello, Italy.
 
31.
Strizhov N., Keller M., Mathur J., Koncz-Kalman Z., Bosch D., Prudovsky E., Schell J., Sneh B., Koncz C., Zilberstein A. 1996. A synthetic cry1C gene, encoding a Bacillus thuringiensis endotoxin, confers Spodoptera resistance in alfalfa and tobacco. Proceedings of the National Academy of Sciences of the United States of America 93: 15012–15017. DOI: 10.1073/pnas.93.26.15012.
 
32.
Taleh M., Saadati M., Farshbaf R., Khakvar R. 2014. Partial characterization of phenoloxidase enzyme in the hemocytes of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae). Journal of King Saud – University Science 26 (4): 285−289. DOI: https://doi.org/10.1016/j.jksu....
 
33.
Wang G.L., Fang H.J. 2002. Plant Genetic Engineering. Science Press, Beijing, China. (in Chinese) Zakir A., Khallaf M.A., Hansson B.S., Witzgall P., Anderson P. 2017. Herbivore-induced changes in cotton modulates reproductive behavior in the moth Spodoptera littoralis. Frontiers in Ecology and Evolution 5: 49. DOI: https://doi.org/10.3389/fevo.2....
 
34.
Zia M., Rizvi Z.F., Rehman R., Chaudhary M.F. 2010. Agrobacterium mediated transformation of soybean (Glycine max L.): some conditions standardization. Pakistan Journal of Botany 42 (4): 2269–2279.
 
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