INVESTIGATING RHIZOBIUM BACTERIA AND OBTAINING SALINITY TOLRANCE ISOLATES USING GENETIC TRANSFORMATION METHOD

KH. OTHMAN, SHAIMAA; F. ABD-AL-KADER, DHAFER

doi:10.37652/juaps.2014.122637

Document Type : Research Paper

Authors

University of Anbar ,Collage of Education

10.37652/juaps.2014.122637

Abstract

This study was conducted to isolate Rhizobium from the roots of wild and field cereal plants (Trifollium spp. , Medicago sativa , Ficia vaba , Pisum sativum , Medicago polymorpha L. , Melilotus indica (L.) ALL , Sesbania spp. , (Trigonella coerulescens M.Bieb ) ,30 isolates strains from plants roots were obtained , 15 from wild cereal and 15 from field cereal, The most efficient nitrogen fixing 8 isolates have been selected on the basis of the diameter of the colony and that of the change of the blue color of the medium of the promothymol blue indicator YEMA-BTB .The optimal conditions for the development of the selected isolates have been studied in terms of temperature, pH and their endurance of the salinity of the medium. Consequently, the best wild isolate,PW7 is taken from wild Sesbania the best field isolate designated PF12 isolated from Medicago sativa , the genetic content of these selected strains were studied, the results showed that the two strains contain small plasmide bands, PF12 strains contains an additional large plasmide band, in order to study the role of these plasmids in the genetic experiment was conducted to neutralize these plasmids by using salsalic acid, results showed that the neutralize cells lost their salinity resistance when tested on a YEMA medium with varying concentrations , genetic transformation was conducted by carrying the DNA extracted from strain PW7 into strain PF12.results showed the transference of the plasmids from PW7 to PF12 which transferred the salinity resistance trait . This has been confirmed by re-culturing the transformed isolate onto the YEMA medium with different salinity concentrations. The modified isolate could grow in a salinity concentration of 8% NaCL.

Keywords

Rhizobium

Main Subjects

Biology

References

1-Sessitsch,A.Howieson,J.G.;Perret, X.and Amtoun,H.(2002).advances in Rhizobium research. Crtitical Reviews in plant Scienes. 21(4);323-358.

2-Keneni,A.;Assefa,F. and Prabu,C.P.(2010). Characterization of acid and salt tolerant Rhizobial strains isolated from Faba Bean Fields of wollo. Northren Ethiopia.J. Agr. Sci. Tech.12;365-376.

3-Shameseldin,A.and Werner,D.(2005).high salt and high PH tolerance of new isolated Rhizobium etli strains from egyption soils. Current Microbiol.50(1);11-16.

4-Shameseldin,A.;Nyalwidhe,J.and Werner ,D.(2006).aproteomic approach to wards the analysis of salt tolerance in Rhizobium etil and Sinorhizobium meliloti strains. Current Microbiol.52(5);333-339.

5-Adal,M.(2009).phenotypic and symbiotic characterization of grass pea (Lathyrus sativus) rhizobial isolates from some major growing areas of south wollo and west shoa, Ethiopia.Addis Ababa University.Masters of Science in Biotechnology.

6--Afrasayab , S.; Faisal, M. and Hsamain,S. (2010). comparative study of wild and transformed salt tolerant bacterial strains of Triticum aestivum growth under salt stress.Bra. J. Micro. 41; 946-955.

7-Shamseldin,A.(2008).plasmid content of salt stress-tolerant Rhizobium strains from egyption soils nodulating common bean (Phaseolus vulgaris L.).World J.Microbiol Biotchnol.24;1603-1606.

8-Beck,D.P.;Materon,L.A.; and Afnadi, F. (1993).practical Rhizobium legume technology manual. Technical Manual.19; ICARDA.

9-Domenico,P.;Schwatz,S.;and Cunha, B. (1989).reduction of capsular polysaccharide production in Klebsiella pneumonia by sodium salicylate.J.Infect.Immun.57;3778-3782.

10-Sambrook,J.;Fritgah,E.and Maniatis,T. (1989).molecular colning.alaboratory manual.Cold Spring Harbour Laboratory. Newyork.

11-Rhijn,P.and Vanderleyden, J.(1995).the Rhizobium plant symbiosis. Microbiological Reviews.Mar.59(1);124-142.

12-Mensah,J.K.;Esumeh,F.;Lyamu,M.and Omoifo,C(2006). Effects of different salt on centration and PHon groth of Rhizobium sp. American. Eurasion J.Agric& Environ. Sci. 1(3);198-202.

13-Watkin,E.L.Ohara,G.W.and Glenn,A.R. (2003).physological responses to acid stress of an acid soil tolerant and an acid soil sensitive strain of Rhizobium leguminosarum bv. Trifolii. Soil Biol.Biochem.35;621-624.

14-Kucuk,K.;Kivanc,M.;and Engin,K. (2006).characterization of Rhizobium sp. Isolated from bean.Turk J Biol.30;124-132.

15-Abdel-salam,M.S.;Ibahim,M.M.,Abdel-Halim,F.M.; and Badawy,S.E. (2010). phenotypic characterization of indigenous egyption Rhizobial strains for stresses performance. Journal of American Science.6(9);498-503.

16-Garatezi,S. and Ahmad, D. (2001). biodiversty and performance of Rhizobia in relation to plasmid genes and their transformation products.Journal of Biological Sciences.1(12) ;1134-1137.

-17الحسینی، انوار علی عبدالله،(2005). دراسة بکتریلوجیة ووراثیة على البکتریا المسببة لحب الشباب،رسالة ماجستیر-کلیة العلوم-جامعة بابل.

18- Fox, M.A. (2005). Adaptation of Rhizobium to environmental stress. degree of Doctor of philosophy ,university of Reading.

Journal of University of Anbar for Pure Science

INVESTIGATING RHIZOBIUM BACTERIA AND OBTAINING SALINITY TOLRANCE ISOLATES USING GENETIC TRANSFORMATION METHOD

References

References

Volume 8, Issue 1
April 2014
Page 41-47

INVESTIGATING RHIZOBIUM BACTERIA AND OBTAINING SALINITY TOLRANCE ISOLATES USING GENETIC TRANSFORMATION METHOD

References

References

Volume 8, Issue 1April 2014Page 41-47

Volume 8, Issue 1
April 2014
Page 41-47