Transformation of Agrobacterium Tumefactions by Conjugation Technique

Raghad N G Al-zaıdy; Najwa I Albarhawı; Abdullah N Al-nıemı

Research Article

Transformation of Agrobacterium Tumefactions by Conjugation Technique

Year 2018, Issue: 3, 28 - 39, 01.12.2018

Raghad N G Al-zaıdy Najwa I Albarhawı Abdullah N Al-nıemı

Abstract

Two genera of Rhizobiaceae ,wild
strain Agrobacterium tumefaciens (AtMcol)
and standerd strain Agrobacterium tumefaciens (C58C1), in addition to
Sinrhizobium melilti are used in this study. The results showed, these two
strains ofA. tumifaciens bacteria were resistant to Trimetheprim and sensitive
to chloromphinicol while the S.melilti bacteria gave the opposite of these
results,and used as genetic markers for conjugation as one of the methods used
in the technique of Horizontal gene transfer, with the use of acardine orange
to cure the bacteria from plasmid.Two conjugation procces were performed,
between Agrobacterium tumefaciens (AtMcol) and S. melilti, and between
Agrobacterium tumefaciens (C58C1)and S.melilti, result showed succecful of
conjugation procces with conjugation frequency 1.8x10-2 and 2.1x10-2
respectively, and the transconjugant bacteria result from procces succeded in
infecting the root tip of alfalfa seedlings to form root nodules.
Characterization of plasmid DNA content for bacterial isolates belonging to conjugated cells, were
carried out by using agarose gel electrophorsis and the results showed isolate
the presence of discrete clear and isolated band of similar size present in
agaros gel.

Keywords

Conjugation technique, Biochemical, DNA

References

Amabile-Cuevas,.F. and Chicurel,M.E.(1992) Bacterial plasmids and gene flux.Cell.70:189-199. Al-Kanaany ,C. F; Hisham, F; Al-Yassri. and Al-Mousawi,A. (2012). Plasmid profile of two isolates of Sinorhizobium meliloti isolated from different soil areas in Basrah Iraq. Bas. J. Agr. Sci., 25: 13-18. Ann,M.Hirsch; Daniela drake.; Thomas W.Jacobs; and Sharon R.Long (1985). Nodules are induced on alfalfa Roots by Agrobacterium tumefacienes and Rhizobium trifolii containin small segments of the Rhizobium meliloti nodulation region.Vo: 161 No(1):223-230. Bono,J.J.; Gressent,F.; Niebel, A.; Cllimore, J.V.; and Rnajeva,R.(1996). Biochemical characterization of nod factor binding sites in Medicago roots cell suspension cultures. In: Biology of Plant Microbe Interaction. (eds.) Stacey, G.; Mullin, B.and Gresshoff, P.M.) International Society for, Molecular Plants Microbe Interactions,St.Paul,M.N.,P. 90-104. Belachew,T. (2010). Intrinsic antibiotic resistance, survival of Rhizobium leguminosarum strains and fixation potential of pea varieties (Pisum sativum L.) In southeast Ethiopia. Int. J. Mic. Res., 1: 75-79. Brody,T.M; Larner,J.; Minneman,L.P. and Nen, H.C.(1994). Human Pharmacology. 2nd ed., Mosby- year book,Inc., St. Louis, Missouri. Bradley,D.E.(1981)conjugative pili plasmids in Eschericah coli K-12 and Pseudomonag species,P.217-226. In S.B Levy,R.C.Clones and E.L.Koenig (ed).Molecular biology,pathogenicity gand ecology of Bacteria plasmids.plenum press, NewYork. Cho,H.;Pinto, U. M. and Winanse,S.C.(2009). Transsexuality in the rhizosphere: quorum sensing reversibly converts Agrobacterium tumefaciens from phenotypically female to male.J.bacteriol.,191, 3375-3383. Dominguez- Ferreras, A.;Perez-Arnedo, R.;Becher ,A.;Olivares ,J.;Soto, M .J.and Sanjuan ,J.(2006). Transcriptome profiling revents the importance of plasmid sym B for osmoadaptation of Sinorhizobium meliloti,J.Bact.,188: 7617-7625. Dale,J.M. and Park, S.F.(2004). Molecular Genetics of Bacteria. 4th ed,John Wiley and Sons,Ltd. Chichster, England. Fullner, K.J.; Cano,L.J.and Nester,E.W. (1996). Pilus assembly by Agrobacterium T- DNA transfer gene. Science, 273: 1107- 1109. Guo,X.;Flores,M;Mavinguip,F.; Hernandez, G;Davila, G.and Palacios, R. (2003). Natural genomic design in Sinorhizobiummeliloti novel genomic architecture.Genome Res .,13:1810-1817. Hartwell,L.; Hood,L.; Glodberg,M.L; Reynold,A.E; Silver,L.M and Vers,R. C.(2000). Genetics from genes to genemes. The McGraw – Hill companies, Inc,Boston,U.S.A. Holt, J.G.; Krieg ,N.R.; Sneath ,P.H. ;Slaley, J.T.and William, S.T. (1994). Bergey's manual of determinative bacteriology 9th edu. Williams and Wilkins Baltimors, Netherlands. html, cited 22 Jan 2009. Hellens,R.; PhilifMulliaeux. and Harry, K.(2000). A guide to Agrobacterium binary Ti Vectors trends in plant. Science, 5(10): 446-451. Hohn,B.and Korn,D.(1969).Co-segregation of sex factor with the Escherichia coli. Chromosome during curing by acridine orange. J. of Molecular biology, 45:385-395. Hahn,F.E. (1976). Elimination of plasmids from bacteria. Antibiotics and Chemotherapy,20, 196-201. Hellens,R.; PhilifMulliaeux. and Harry, K.(2000). A guide to Agrobacterium binary Ti Vectors trends in plant. Science, 5(10): 446-451. Herrera-Cervera,J.A.; Olivares,J.; and Sanjuan,J. (1996). Ammonia inhibition of plamid PRme GR4a Conjugal transfer between Rhiobium meliloti stains. Appl.Environ.Microbiol.,62:1145-1150. Hooykaas,P.J.; Van Brussel,A.A.; Dulk-Ras,H.; Van- Slogteren,G.M. and Schilperroot, R.A. (1981). Sym plasmid of Rhizobium trifolii expression in different rhizobial species and Agrobacterium tumefaciences. Nature, 91:351-353. Hynes, M. F.; Simon, R.; Muller, P.; Niehaus, K.; Labes, M and Puhler, A. (1986) .The two megaplasmids of Sinorhizobium meliloti are involved in the effective nodlation of alfalfa. Mol .gen. Genet., 202; 356-362. Hiroki, N.; Toshiki, C.;Kenichi ,K.;Akihiro, S; Shiro, H and Nikiko, A.(2008). Transposon mediation allows symbiotic plasmid of Rhizobium leguminosarum bv.trifolii to become a symbiosis island Agrobacterium and Rhizobium.. The Journal of General and Applied Microbiology, 54: 107-118. Hirsch,A.M .(1992). Development biology of legume nodulation . New Phytol., 122: 211-237. Hirsch,A.M.H.; Daniela,A; Thomas, W;Jacobs and Sharon, R.L. (1985). Nodules are induced on alfalfa roots by Agrobacterium tumefacienes and Rhizobium trifolii containin small segments of the Rhizobium meliloti nodulation region.J. Bacteriol., 161:223-230. Kondorosi, A.; Kondorgi, E.; Pankhurst, C. E.; Broughton,W. J. and Bauflav, Z.(1982). Mobilization of Rhizobium meliloti megaplamsmid curring nodulation and nitrogen fixation genes into other Rhizobium and Agrobacterium. Mol. Gen.,188:433-439. Lee,L.; Huang,Y. and Chen, C.W.(1996). Two classes of ethidium bromide resistant mutants of streptomycess lividans 66. Microbiology, 142: 1041-1047. Lowry,O.H.;Roisebrough,N.J.;Farr,A.L.and Radad,R.J.(1951). Protein measurment with folin phenol reagent. J. Bio.Chem.,193: 257-265. Lichtenstein,C.and Draper,J.(1986). "In DNA cloning: Apractical Approach",(ed.)Glover,D.M. (IRL,Oxford,Vol.2). Lujan, S. A.; Guagas, M.; Ragonese, H; Matson, S.Wand Redingbo, M. R. (2007). Disrupting antibiotic resistance propagation by inhibiting the conjugative DNA relaxase. P.N.A.S., 104:1228-12287. Lorernz,M.G.and Wackernagel,W .(1994). Bacterial gene transfer. Environment Microbiol.Rev., 58:563-602. Moore, L.W.; Kado, C.I. and Bouzar, H.( 1988). Agrobacterium. In Laboratory guide for identification of plant pathogenic bacteria. 2nd ed. by Schaad, N.W. American Phytopathological Society Press, St. Paul, Minn. pp. 16–36. Marja , K. C., William, R., Horwath, B. D.and Hammock,W.( 2004). \Isolation of a strain of Agrobacterium tumefaciens (Rhizobium radiobacter) utilizing methylene urea (ureaformaldehyde) as nitrogene source .Can. J. Microbiol., 50: 167-174. Mickelsen, P. A.; Plorde, J. J; Gordon, K. P.; Hargisis,C.and Maclure, J. (1985). Intability of antibiotics resistance in an strain.of Staphylococcus epidermidis isolated from on outbreak of prosthetic value endocarditis.J.Infect.Dis.,.152:50-58. Mysore, K.S.;Nam, J and Gelvin, S.B .(2000).An Arbidopsis histone H2 Amutantis deficient in Agrobacterium T-DNA- integration.Proc.Natl.Acad. Sci., 97:748-953. Naher, M.; Zinat, M.; Hasan, M.Z.; Khadiza, Z.and Fahmida , J. (2012). Effects of plasmid curing on Rhziboium spp. J. of Mic., 2: 2074-5346. Prescotta,L.M.; Harley,J.P. and Kelin,D.A (2007). Microbiology Prokaryotes: Bacterial Genetic System Taxonomy Microbiology. bytes.22, No .5. Rasool, S. A.; Ahmad,A.; Khan,S. and Wahab,A. (2003). Plasmid bone antibiotic resistance factors among indigenous Klebsiella pneumonia .J.Bot.,35(2) 243-248. Strohmaier,H.;Noiges,R.;Kotschan,S.;Sawers,G.; Hogenauerg G.; Zechner, E. L.; Koraimann, G.; Zechner, E. L.; Koraimann, G. (1998). Single transduction and bacterial conjugation characterziation of the role of Arc A in regulating conjugation transfer of the resistance plasmid Ri. J.Mol.Biol.,277:309-316. Schacterle,G.R.,and Pollack,R.L.,(1973). A simplified method for the quantitative assay of small amounts of protein in biological material.Anal.Biochem.sl:654-655. Sambrook,J.;Fritsch,T.F. and Maniatis,T. (1989). Molecular cloning a laboratory.New York,USA. Shishido, M. and Pepper,I.L. (1990). Identification of dominant indigenous Rhizobium melilolti by plasmid profiles and intrinsic antibiotic resistance. Soil Bio. Bioch., 22: 11-16. Talaro, K. and Talaro, A.(1996). Foundation in microbiology. 2nd ed., W.M.C. Brown Publishers,PP :281-282 . Tomoeda, M.; Inuzuka, M.; Anto, S. and Konishi, M. (1974). Curing action of sodium dodecyle sulfate on a Proteus mirabilis R+ strain. J. Bacteriol., 120: 1158-1163. Valerie, O. and Sharon, R. L. (1999) .Bacterial genes induced the nodule during the Rhizobium legume symbiosis. Molecular Microbiology, 32(4):837-849. Vincent,J.M.(1970). A manual for the Practical Study of Root Nodule Bactria.IBP Handbook No.15. Oxford: Blackwell Scientific Publication,Oxford,PP.113-131. Williams,H.G.; Day, M. J.; Fry, J. C .and Stewart, G. J.(1996). Natural transformation in River epilithon. Appl.Environ. Microiobl., 62:2994-2998. Willetts, N. and Wilkins, B. (1984) .Processing of plasmid DNA during bacterial conjugation. Microbiologicla Reviews, 48(1):24-41. Wessells,N.K. and Hopson,J.(1988). Biology Rnadom House,INC.,New York.PP.303-320. Wong,C.H.; Pankhurst, C. E.; Kondorosi,A. and N. D Broughton, W. J. (1983). Morphology of root nodules and Nodule- like structures formodby Rhizobium and Agrobacterium strains containing Rhizobium meliloti megaplasmid . J of Cell Biology, 97 :787-794. Xi- Wan; Jan, H.; Allessandra ,L.; Chiara ,G.; Diederikvande ,P.; Elena, F.; Ton, B. and Henk,F. (2007) Medicago trancatula EWoD40-1 and ENOD 40-2 are both involved in nodule initiation and bacteroid development .J. of Experimental Botany., 72: 1-9. معارج، محمد عبد المحسن.(1999). مقدمة في الهندسة الوراثية، كلية الطب البشري، جامعة الموصل. الزبيدي، اريان محمد حامد اسماعيل (2005). نقل بلازميدات بكتريا S.meliloti الى بكتريا E.coli بطريقتي الاقتران البكتري والتحول الوراثي ودورها في تكوين العقد الجذرية. رسالة ماجستير، كلية التربية، جامعة الموصل,العراق.

Year 2018, Issue: 3, 28 - 39, 01.12.2018

Raghad N G Al-zaıdy Najwa I Albarhawı Abdullah N Al-nıemı

Abstract

References

Amabile-Cuevas,.F. and Chicurel,M.E.(1992) Bacterial plasmids and gene flux.Cell.70:189-199. Al-Kanaany ,C. F; Hisham, F; Al-Yassri. and Al-Mousawi,A. (2012). Plasmid profile of two isolates of Sinorhizobium meliloti isolated from different soil areas in Basrah Iraq. Bas. J. Agr. Sci., 25: 13-18. Ann,M.Hirsch; Daniela drake.; Thomas W.Jacobs; and Sharon R.Long (1985). Nodules are induced on alfalfa Roots by Agrobacterium tumefacienes and Rhizobium trifolii containin small segments of the Rhizobium meliloti nodulation region.Vo: 161 No(1):223-230. Bono,J.J.; Gressent,F.; Niebel, A.; Cllimore, J.V.; and Rnajeva,R.(1996). Biochemical characterization of nod factor binding sites in Medicago roots cell suspension cultures. In: Biology of Plant Microbe Interaction. (eds.) Stacey, G.; Mullin, B.and Gresshoff, P.M.) International Society for, Molecular Plants Microbe Interactions,St.Paul,M.N.,P. 90-104. Belachew,T. (2010). Intrinsic antibiotic resistance, survival of Rhizobium leguminosarum strains and fixation potential of pea varieties (Pisum sativum L.) In southeast Ethiopia. Int. J. Mic. Res., 1: 75-79. Brody,T.M; Larner,J.; Minneman,L.P. and Nen, H.C.(1994). Human Pharmacology. 2nd ed., Mosby- year book,Inc., St. Louis, Missouri. Bradley,D.E.(1981)conjugative pili plasmids in Eschericah coli K-12 and Pseudomonag species,P.217-226. In S.B Levy,R.C.Clones and E.L.Koenig (ed).Molecular biology,pathogenicity gand ecology of Bacteria plasmids.plenum press, NewYork. Cho,H.;Pinto, U. M. and Winanse,S.C.(2009). Transsexuality in the rhizosphere: quorum sensing reversibly converts Agrobacterium tumefaciens from phenotypically female to male.J.bacteriol.,191, 3375-3383. Dominguez- Ferreras, A.;Perez-Arnedo, R.;Becher ,A.;Olivares ,J.;Soto, M .J.and Sanjuan ,J.(2006). Transcriptome profiling revents the importance of plasmid sym B for osmoadaptation of Sinorhizobium meliloti,J.Bact.,188: 7617-7625. Dale,J.M. and Park, S.F.(2004). Molecular Genetics of Bacteria. 4th ed,John Wiley and Sons,Ltd. Chichster, England. Fullner, K.J.; Cano,L.J.and Nester,E.W. (1996). Pilus assembly by Agrobacterium T- DNA transfer gene. Science, 273: 1107- 1109. Guo,X.;Flores,M;Mavinguip,F.; Hernandez, G;Davila, G.and Palacios, R. (2003). Natural genomic design in Sinorhizobiummeliloti novel genomic architecture.Genome Res .,13:1810-1817. Hartwell,L.; Hood,L.; Glodberg,M.L; Reynold,A.E; Silver,L.M and Vers,R. C.(2000). Genetics from genes to genemes. The McGraw – Hill companies, Inc,Boston,U.S.A. Holt, J.G.; Krieg ,N.R.; Sneath ,P.H. ;Slaley, J.T.and William, S.T. (1994). Bergey's manual of determinative bacteriology 9th edu. Williams and Wilkins Baltimors, Netherlands. html, cited 22 Jan 2009. Hellens,R.; PhilifMulliaeux. and Harry, K.(2000). A guide to Agrobacterium binary Ti Vectors trends in plant. Science, 5(10): 446-451. Hohn,B.and Korn,D.(1969).Co-segregation of sex factor with the Escherichia coli. Chromosome during curing by acridine orange. J. of Molecular biology, 45:385-395. Hahn,F.E. (1976). Elimination of plasmids from bacteria. Antibiotics and Chemotherapy,20, 196-201. Hellens,R.; PhilifMulliaeux. and Harry, K.(2000). A guide to Agrobacterium binary Ti Vectors trends in plant. Science, 5(10): 446-451. Herrera-Cervera,J.A.; Olivares,J.; and Sanjuan,J. (1996). Ammonia inhibition of plamid PRme GR4a Conjugal transfer between Rhiobium meliloti stains. Appl.Environ.Microbiol.,62:1145-1150. Hooykaas,P.J.; Van Brussel,A.A.; Dulk-Ras,H.; Van- Slogteren,G.M. and Schilperroot, R.A. (1981). Sym plasmid of Rhizobium trifolii expression in different rhizobial species and Agrobacterium tumefaciences. Nature, 91:351-353. Hynes, M. F.; Simon, R.; Muller, P.; Niehaus, K.; Labes, M and Puhler, A. (1986) .The two megaplasmids of Sinorhizobium meliloti are involved in the effective nodlation of alfalfa. Mol .gen. Genet., 202; 356-362. Hiroki, N.; Toshiki, C.;Kenichi ,K.;Akihiro, S; Shiro, H and Nikiko, A.(2008). Transposon mediation allows symbiotic plasmid of Rhizobium leguminosarum bv.trifolii to become a symbiosis island Agrobacterium and Rhizobium.. The Journal of General and Applied Microbiology, 54: 107-118. Hirsch,A.M .(1992). Development biology of legume nodulation . New Phytol., 122: 211-237. Hirsch,A.M.H.; Daniela,A; Thomas, W;Jacobs and Sharon, R.L. (1985). Nodules are induced on alfalfa roots by Agrobacterium tumefacienes and Rhizobium trifolii containin small segments of the Rhizobium meliloti nodulation region.J. Bacteriol., 161:223-230. Kondorosi, A.; Kondorgi, E.; Pankhurst, C. E.; Broughton,W. J. and Bauflav, Z.(1982). Mobilization of Rhizobium meliloti megaplamsmid curring nodulation and nitrogen fixation genes into other Rhizobium and Agrobacterium. Mol. Gen.,188:433-439. Lee,L.; Huang,Y. and Chen, C.W.(1996). Two classes of ethidium bromide resistant mutants of streptomycess lividans 66. Microbiology, 142: 1041-1047. Lowry,O.H.;Roisebrough,N.J.;Farr,A.L.and Radad,R.J.(1951). Protein measurment with folin phenol reagent. J. Bio.Chem.,193: 257-265. Lichtenstein,C.and Draper,J.(1986). "In DNA cloning: Apractical Approach",(ed.)Glover,D.M. (IRL,Oxford,Vol.2). Lujan, S. A.; Guagas, M.; Ragonese, H; Matson, S.Wand Redingbo, M. R. (2007). Disrupting antibiotic resistance propagation by inhibiting the conjugative DNA relaxase. P.N.A.S., 104:1228-12287. Lorernz,M.G.and Wackernagel,W .(1994). Bacterial gene transfer. Environment Microbiol.Rev., 58:563-602. Moore, L.W.; Kado, C.I. and Bouzar, H.( 1988). Agrobacterium. In Laboratory guide for identification of plant pathogenic bacteria. 2nd ed. by Schaad, N.W. American Phytopathological Society Press, St. Paul, Minn. pp. 16–36. Marja , K. C., William, R., Horwath, B. D.and Hammock,W.( 2004). \Isolation of a strain of Agrobacterium tumefaciens (Rhizobium radiobacter) utilizing methylene urea (ureaformaldehyde) as nitrogene source .Can. J. Microbiol., 50: 167-174. Mickelsen, P. A.; Plorde, J. J; Gordon, K. P.; Hargisis,C.and Maclure, J. (1985). Intability of antibiotics resistance in an strain.of Staphylococcus epidermidis isolated from on outbreak of prosthetic value endocarditis.J.Infect.Dis.,.152:50-58. Mysore, K.S.;Nam, J and Gelvin, S.B .(2000).An Arbidopsis histone H2 Amutantis deficient in Agrobacterium T-DNA- integration.Proc.Natl.Acad. Sci., 97:748-953. Naher, M.; Zinat, M.; Hasan, M.Z.; Khadiza, Z.and Fahmida , J. (2012). Effects of plasmid curing on Rhziboium spp. J. of Mic., 2: 2074-5346. Prescotta,L.M.; Harley,J.P. and Kelin,D.A (2007). Microbiology Prokaryotes: Bacterial Genetic System Taxonomy Microbiology. bytes.22, No .5. Rasool, S. A.; Ahmad,A.; Khan,S. and Wahab,A. (2003). Plasmid bone antibiotic resistance factors among indigenous Klebsiella pneumonia .J.Bot.,35(2) 243-248. Strohmaier,H.;Noiges,R.;Kotschan,S.;Sawers,G.; Hogenauerg G.; Zechner, E. L.; Koraimann, G.; Zechner, E. L.; Koraimann, G. (1998). Single transduction and bacterial conjugation characterziation of the role of Arc A in regulating conjugation transfer of the resistance plasmid Ri. J.Mol.Biol.,277:309-316. Schacterle,G.R.,and Pollack,R.L.,(1973). A simplified method for the quantitative assay of small amounts of protein in biological material.Anal.Biochem.sl:654-655. Sambrook,J.;Fritsch,T.F. and Maniatis,T. (1989). Molecular cloning a laboratory.New York,USA. Shishido, M. and Pepper,I.L. (1990). Identification of dominant indigenous Rhizobium melilolti by plasmid profiles and intrinsic antibiotic resistance. Soil Bio. Bioch., 22: 11-16. Talaro, K. and Talaro, A.(1996). Foundation in microbiology. 2nd ed., W.M.C. Brown Publishers,PP :281-282 . Tomoeda, M.; Inuzuka, M.; Anto, S. and Konishi, M. (1974). Curing action of sodium dodecyle sulfate on a Proteus mirabilis R+ strain. J. Bacteriol., 120: 1158-1163. Valerie, O. and Sharon, R. L. (1999) .Bacterial genes induced the nodule during the Rhizobium legume symbiosis. Molecular Microbiology, 32(4):837-849. Vincent,J.M.(1970). A manual for the Practical Study of Root Nodule Bactria.IBP Handbook No.15. Oxford: Blackwell Scientific Publication,Oxford,PP.113-131. Williams,H.G.; Day, M. J.; Fry, J. C .and Stewart, G. J.(1996). Natural transformation in River epilithon. Appl.Environ. Microiobl., 62:2994-2998. Willetts, N. and Wilkins, B. (1984) .Processing of plasmid DNA during bacterial conjugation. Microbiologicla Reviews, 48(1):24-41. Wessells,N.K. and Hopson,J.(1988). Biology Rnadom House,INC.,New York.PP.303-320. Wong,C.H.; Pankhurst, C. E.; Kondorosi,A. and N. D Broughton, W. J. (1983). Morphology of root nodules and Nodule- like structures formodby Rhizobium and Agrobacterium strains containing Rhizobium meliloti megaplasmid . J of Cell Biology, 97 :787-794. Xi- Wan; Jan, H.; Allessandra ,L.; Chiara ,G.; Diederikvande ,P.; Elena, F.; Ton, B. and Henk,F. (2007) Medicago trancatula EWoD40-1 and ENOD 40-2 are both involved in nodule initiation and bacteroid development .J. of Experimental Botany., 72: 1-9. معارج، محمد عبد المحسن.(1999). مقدمة في الهندسة الوراثية، كلية الطب البشري، جامعة الموصل. الزبيدي، اريان محمد حامد اسماعيل (2005). نقل بلازميدات بكتريا S.meliloti الى بكتريا E.coli بطريقتي الاقتران البكتري والتحول الوراثي ودورها في تكوين العقد الجذرية. رسالة ماجستير، كلية التربية، جامعة الموصل,العراق.

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Details

Primary Language	English
Subjects	Engineering
Journal Section	Articles
Authors	Raghad N G Al-zaıdy Najwa I Albarhawı Abdullah N Al-nıemı
Publication Date	December 1, 2018
Published in Issue	Year 2018Issue: 3

Cite

APA	Al-zaıdy, R. N. G., Albarhawı, N. I., & Al-nıemı, A. N. (2018). Transformation of Agrobacterium Tumefactions by Conjugation Technique. The Eurasia Proceedings of Science Technology Engineering and Mathematics(3), 28-39.