Cultivation Effect of Chitinase-Transgenic Cotton on Functional Bacteria and Fungi in Rhizosphere and Bulk Soil

Document Type: Research Paper


1 Department of Biotechnology and Plant Breeding, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Plant Biotechnology, Faculty of Bioscience and Biotechnology, Shahid Beheshti University, Tehran, Iran

3 Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran


Background: In consideration for the increasing widespread use of genetically modified (GM) crops, one of the important issues for assessment is the effect of GM crops on soil microbial communities
Objectives: In this study, T2 chitinase-transgenic cotton (line #57) and its non-transgenic line were investigated for bacterial and fungal dynamics during its development stages.
Material and Methods: The assessments were performed by viable plate count and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assays.
Results: Viable plate count analysis showed an increase in community structures and the number of culturable bacteria in rhizosphere of both transgenic and non-transgenic cultivars as compared to bulk soil. PCR-DGGE confirmed results of viable plate count assays of the changes in bacterial and fungal communities for all cotton development stages in rhizosphere and bulk zones. No significant differences in number of functional bacteria were observed between rhizosphere soil of chitinase transgenic and non-chitinase transgenic cotton at one particular stage.
Conclusions: The results indicated that T2 chitinase-transgenic cotton (line #57) might have no adverse effects on community structures and total number of culturable bacteria and fungi in the rhizosphere.


Main Subjects

1.           Bruinsma M, Kowalchuk G, Van Veen J. Effects of genetically modified plants on microbial communities and processes in soil. Biol Fertil oils. 2003;37(6):329-337.
2.           Liu B, Zeng Q, Yan F, Xu H, Xu C. Effects of transgenic plants on soil microorganisms. Plant Soil. 2005;271(1):1-13. doi: 10.1007/s11104-004-1610-8
3.           Icoz I, Stotzky G. Fate and effects of insect-resistant Bt crops in soil ecosystems. Soil Biol Biochem. 2008;40(3):559-586. doi: 10.1016/j.soilbio.2007.11.002
4.           Wei M, Tan F, Zhu H, Cheng K, Wu X, Wang J, et al. Impact of Bt-transgenic rice (SHK601) on soil ecosystems in the rhizosphere during crop development. Plant Soil Environ. 2012;58(5):217-223. doi: 10.17221/627/2011-PSE
5.           Giovannetti M, Sbrana C, Turrini A, editors. The impact of genetically modified crops on soil microbial communities. Rivista di Biologia/Biology Forum; 2005.
6.           Berg G, Smalla K. Plant species and soil type cooperatively shape the structure and function of microbial communities in the rhizosphere. FEMS Microbiol Ecol. 2009;68(1):1-13. doi: 10.1111/j.1574-6941.2009.00654.x pmid: 19243436
7.           Hinsinger P, Bengough AG, Vetterlein D, Young IM. Rhizosphere: biophysics, biogeochemistry and ecological relevance. Plant Soil. 2009;321(1-2):117-152. doi: 10.1007/s11104-008-9885-9
8.           Zaman M, Mirza MS, Irem S, Zafar Y. A Temporal Expression of Cry1Ac Protein in Cotton Plant and its Impact on Soil Health. Int J Agricult Biol. 2015;17(2).
9.           Brusetti L, Francia P, Bertolini C, Pagliuca A, Borin S, Sorlini C, et al. Bacterial communities associated with the rhizosphere of transgenic Bt 176 maize (Zea mays) and its non transgenic counterpart. Plant Soil. 2005;266(1-2):11-21. doi: 10.1007/s11104-005-5399-x
10.        Rui Y-K, Yi G-X, Zhao J, Wang B-M, Li Z-H, Zhai Z-X, et al. Changes of Bt toxin in the rhizosphere of transgenic Bt cotton and its influence on soil functional bacteria. World J Microbiol Biotechnol. 2005;21(6):1279-1284. doi: 10.1007/s11274-005-2303-z
11.        Xue K, Luo HF, Qi HY, Zhang HX. Changes in soil microbial community structure associated with two types of genetically engineered plants analyzing by PLFA. J Environ Sci (China). 2005;17(1):130-134. pmid: 15900775
12.        Filion M. Do transgenic plants affect rhizobacteria populations? Microb Biotechnol. 2008;1(6):463-475. doi: 10.1111/j.1751-7915.2008.00047.x pmid: 21261867
13.        Chen ZH, Chen LJ, Wu ZJ. Relationships among persistence of Bacillus thuringiensis and Cowpea trypsin inhibitor proteins, microbial properties and enzymatic activities in rhizosphere soil after repeated cultivation with transgenic cotton. Appl Soil Ecol. 2012;53:23-30. doi: 10.1016/j.apsoil.2011.10.019
14.        Schmidt TM, Waldron C. Microbial diversity in soils of agricultural landscapes and its relation to ecosystem function. The ecology of agricultural landscapes: long-term research on the path to sustainability. New York: Oxford University Press; 2015. 135-157 p.
15.        Torsvik V, Ovreas L. Microbial diversity and function in soil: from genes to ecosystems. Curr Opin Microbiol. 2002;5(3):240-245. pmid: 12057676
16.        Wang B, Shen H, Yang X, Guo T, Zhang B, Yan W. Effects of chitinase-transgenic (McChit1) tobacco on the rhizospheric microflora and enzyme activities of the purple soil. Plant Soil Environ. 2013;59:241-246. doi: 10.17221/704/2012-PSE
17.        Hirsch PR, Mauchline TH, Clark IM. Culture-independent molecular techniques for soil microbial ecology. Soil Biol Biochem. 2010;42(6):878-887. doi: 10.1016/j.soilbio.2010.02.019
18.        Tohidfar M, Mohammadi M, Ghareyazie B. Agrobacterium-mediated transformation of cotton (Gossypium hirsutum) using a heterologous bean chitinase gene. Plant Cell Tissue Organ Cult. 2005;83(1):83-96. doi: 10.1007/s11240-004-6155-2
19.        Tohidfar M, Hossaini R, Bashir NS, Meisam T. Enhanced Resistance to Verticillium dahliae in Transgenic Cotton Expressing an Endochitinase. Czech J Genet Plant Breed. 2012;48(1):33-41. doi: 10.17221/89/2011-CJGPB
20.        Ben-David A, Davidson CE. Estimation method for serial dilution experiments. J Microbiol Methods. 2014;107:214-221. doi: 10.1016/j.mimet.2014.08.023 pmid: 25205541
21.        Karimi E, Sadeghi A, Abbaszade Dehaji P, Dalvand Y,
Omidvari M, Kakuei Nezhad M. Biocontrol activity of salt tolerant Streptomyces isolates against phytopathogens causing root rot of sugar beet. Biocont Sci Technol. 2012;22(3):333-349. doi: 10.1080/09583157.2012.658552
22.        Hu H-Y, Liu X-X, Zhao Z-W, Sun J-G, Zhang Q-W, Liu X-Z, et al. Effects of repeated cultivation of transgenic Bt cotton on functional bacterial populations in rhizosphere soil. World J Microbiol Biotechnol 2009;25(3):357-366. doi: 10.1007/s11274-008-9899-8
23.        Yuan X, Xu J, Chai H, Lin H, Yang Y, Wo X, et al. Differences of rhizo-bacterial diversity and the content of peimine and peiminine of Fritillaria thunbergii among different habits. J Med Plants Res. 2010;4(6):465-470.
24.        Das M, Royer TV, Leff LG. Diversity of fungi, bacteria, and actinomycetes on leaves decomposing in a stream. Appl Environ Microbiol. 2007;73(3):756-767. doi: 10.1128/AEM.01170-06 pmid: 17142366
25.        Liu Y, Li J, Neal Stewart C, Jr., Luo Z, Xiao N. The effects of the presence of Bt-transgenic oilseed rape in wild mustard populations on the rhizosphere nematode and microbial communities. Sci Total Environ. 2015;530-531:263-270. doi: 10.1016/j.scitotenv.2015.05.073 pmid: 26047860
26.        Zhang Y-J, Xie M, Wu G, Peng D-L, Yu W-B. A 3-year field investigation of impacts of Monsanto’s transgenic Bt-cotton NC 33B on rhizosphere microbial communities in northern China. Appl Soil Ecol. 2015;89:18-24. doi: 10.1016/j.apsoil.2015.01.003
27.        Saxena D, Stotzky G. Bacillus thuringiensis (Bt) toxin released from root exudates and biomass of Bt corn has no apparent effect on earthworms, nematodes, protozoa, bacteria, and fungi in soil. Soil Biol Biochem. 2001;33(9):1225-1230. doi: 10.1016/S0038-0717(01)00027-X
28.        Castaldini M, Turrini A, Sbrana C, Benedetti A, Marchionni M, Mocali S, et al. Impact of Bt corn on rhizospheric and soil eubacterial communities and on beneficial mycorrhizal symbiosis in experimental microcosms. Appl Environ Microbiol. 2005;71(11):6719-6729. doi: 10.1128/AEM.71.11.6719-6729.2005 pmid: 16269702
29.        Turrini A, Sbrana C, Nuti MP, Pietrangeli BM, Giovannetti M. Development of a model system to assess the impact of genetically modified corn and aubergine plants on arbuscular mycorrhizal fungi. Plant Soil. 2005;266(1-2):69-75. doi: 10.1007/s11104-005-4892-6
30.        Liu W, Lu HH, Wu W, Wei QK, Chen YX, Thies JE. Transgenic Bt rice does not affect enzyme activities and microbial composition in the rhizosphere during crop development. Soil Biol Biochem. 2008;40(2):475-486. doi: 10.1016/j.soilbio.2007.09.017