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Research Article

Effect of Gamma Irradiation on Physiological and Biochemical Traits in Cowpea, Vigna unguiculata (L.) Walp Inoculated with New Recombinant Isolates of Bradyrhizobium

Zaied K.A. , F.S. Faris and A.M. Assar
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The symbiotic interaction between rhizobia and legume roots is characterized by a high degree of specificity. Two varieties of cowpea were gamma irradiated as a one method to create genetic variation resulting in new varieties with better characteristics in nodulation and nitrogen fixation processes. Conjugation is the second method used in this study, a cell contact-dependent DNA transfer mechanism, which has served as elegant tool in the development of genetic engineering technology. The possibility of horizontal gene transfer to other rhizobia, revealed that it is necessary, in view of possibility of deliberate release of a variety of recombinant rhizobia into the environment for such agricultural purposes as improving nitrogen fixation. New recombinants revealed higher amounts of indole compounds from tryptophan above the mid-parents in two out of six transconjugants resulted from the cross between P1 x P3. Significant number of nodules were developed on the root system of V2-variety in M4 generation treated with 20 krad in response to inoculation with the parental strains (P2 and P3) and also in response to inoculation with triparental transconjugants (Tr4 and Tr5), above that developed on the plants fertilized with recommended dose of N. The results revealed the success of rhizobial strains and their recombinants to colonize and infect roots of cowpea, because of significant dry weight of nodules per plant which can be obtained in V1-variety treated with 20 krad in M4 generation inoculated with the parental strain (P3), above that on the plants fertilized with recommended dose of N. Total chlorophyll formation in V1-variety inoculated with di-parental transconjugants (DPM-Tr2 and DPM-Tr3) at all doses of gamma irradiation was significantly increase above that in the plants fertilized with recommended dose and the mid-parents, with the exception at 30 krad if compared with the mid-parents. Significant increase was resulted in fresh weight of pods developed per plant above the mid-parents in M3 generation of V1-variety at doses zero and 10 krad, in response to inoculation with di-parental transconjugant, DPM-Tr2. While, the same trend was also achieved above the full dose in M3 generation at 10 krad in response to inoculation with DPM-Tr2, DPM-Tr3, TPM-Tr4 and TPM-Tr5. The highest nitrogen content was appeared in the shoots of V1-variety at all doses of gamma irradiation in response to inoculation with diparental transconjugant (DPM-Tr2). However, V2-variety had the lowest nitrogen content in relation to the plants fertilized with recommended dose of nitrogen and to the mid-parents of rhizobial transconjugants. The genetic variability of grain-protein content appeared that V2-variety treated with 10 krad had significant increase in protein content above that in the plants fertilized with recommended dose of N among M3 and M4 generations, in response to inoculation with parental strains and most of their transconjugants. The same trend was also shown in M4 generation of V1-variety treated with 20 and 30 krad above the plants fertilized with recommended dose of nitrogen, in response to inoculation with di-parental transconjugants. All biochemical traits studied were more affected by biofertilization than the doses of gamma rays and the interaction between biofertilization x doses. This indicated that the significance of treatments was mainly due to inoculation and particularly to gamma irradiation and the interaction between both of them.

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Zaied K.A. , F.S. Faris and A.M. Assar , 2005. Effect of Gamma Irradiation on Physiological and Biochemical Traits in Cowpea, Vigna unguiculata (L.) Walp Inoculated with New Recombinant Isolates of Bradyrhizobium. Pakistan Journal of Biological Sciences, 8: 1173-1191.

DOI: 10.3923/pjbs.2005.1173.1191


1:  Hadri, A.E., H.P. Spaink, T. Bisseling and N.J. Brewin, 1998. Diversity of Root Nodulation and Rhizobial Infection Processes. In: The Rhizobiaceae, Spaink, H.P., A. Kondorosi and P.J.J. Hooykaas (Eds.). Dordrecht, Kluwer, pp: 347-360.

2:  Singh, B.B., O.L. Chambliss and B. Sharma, 1997. Recent Advances in Cowpea Breeding. In: Advances in Cowpea Research, Singh, B.B., D.R. Mohan Raj, K.E. Dashiell and L.E.N. Jackai (Eds.). IITA and JIRCAS, Hong Kong, pp: 30-50.

3:  Nielsen, S., T. Ohler and C. Mitchell, 1997. Cowpea for Human Consumption: Production, Utilization and Nutrient Composition. In: Advances in Cowpea Research, Singh, B., D.M. Raj, K. Dashiell and L. Jackai (Eds.). IITA/JIRCASS, Ibadan, Nigeria, pp: 326-332.

4:  Ehlers, J.D. and A.E. Hall, 1997. Cowpea (Vigna unguiculata, L. Walp.). Field Crops Res., 53: 187-204.

5:  Laity, F., D. Diaga, A. Mame, N. Fall, A.B. Francois and G. Mamadou, 2003. Genetic diversity in cowpea [Vigna unguiculata (L)Walp varieties determined by ARA and RAPO African. J. Biotechnol., 2: 48-50.

6:  Davis, D.W., E.A. Oelke, E.S.Oplinger, J.D. Doll, C.V. Hanson and D.H. Putnam, 1991. Cowpea. University of Winsconsin-Madison, USA.

7:  Assar, A.M., 2001. Effect of radiation on important economical traits in cowpea. M.Sc. Thesis, Mansoura University, Egypt, pp: 19-20.

8:  Collins, C.H. and P.M. Lyne, 1985. Microbiological Methods. 5th Edn., Butterworths, London, pp: 167-181.

9:  Toda, M., S. Okuba, R. Hily and S. Shimamura, 1989. The bacterial activity of tea and coffee. Lett. Applied Microbiol., 8: 123-125.
Direct Link  |  

10:  Lessel, M., D. Blazer, K. Weyrauch and E. Lanka, 1993. The mating pair formation system of plasmid RP4 defined by RSF1010 mobilization and donor-specific phage propagation. J. Bacteriol., 175: 6415-6425.
Direct Link  |  

11:  Pilet, P.E. and R. Chollet, 1970. Sur le dosage colorimtrique de lacide indolylacetique. C.R. Acad. Sci. Ser. D., 271: 1675-1678.

12:  Glickmann, E. and Y. Dessaux, 1995. A critical examination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogenic bacteria. Applied Environ. Microbiol., 61: 793-796.
Direct Link  |  

13:  APHA, 1992. Standard Methods for the Examination of Water and Wastewater. 18th Edn., American Public Health Association, American Water Works Association, Water Environment Federation, Washington, DC., USA.

14:  Brandl, M.T. and S.E. Lindow, 1998. Environmental signals modulate the expression of an indole-3-acetic acid biosynthetic gene in Erwinia herbicola. Mol. Plant Microbe Interact., 10: 499-505.
CrossRef  |  

15:  Kittell, B.L., D.R. Helinski and G.S. Ditta, 1989. Aromatic aminotransferase activity and indoleacetic acid production in Rhizobium meliloti. J. Bacteriol., 171: 5458-5466.
Direct Link  |  

16:  Brandl, M.T. and S.E. Lindow, 1996. Cloning and characterization of a locus encoding an indolepyruvate decarboxylase involved in indole-3-acetic acid synthesis in Erwinia herbicola. Applied Environ. Microbiol., 62: 4121-4128.
Direct Link  |  

17:  Mayak, S., T. Tirosh and B.R. Glick, 1997. The Influence of Plant Growth Promoting Rhizobacterium Pseudomonas putida GR12-2 on the Rooting of Mung Bean Cuttings. In: Plant Growth-Promoting Rhizobacteria: Present Status and Future Prospects, Ogoshi A., K. Kobayashi, Y. Homma, F. Kodama, N. Kondo and S. Akino (Eds.). OECD, Paris, France, pp: 313-315.

18:  Xie, H., J.J. Pasternak and B.R. Glick, 1996. Isolation and characterization of mutants of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2 that overproduce indoleacetic acid. Curr. Microbiol., 32: 67-71.
CrossRef  |  Direct Link  |  

19:  Mandal, J., A. Chattopadhyay, P. Hazra, T. Dasgupta and M.G. Som, 1999. Genetic variability for three biological nitrogen fixation components in cowpea [Vigna unguiculata L. Walp. cultivars. Crop Res., 18: 222-225.

20:  Buttery, B.R., S.J. Park and D.J. Hume, 1992. Potential for increasing nitrogen fixation in grain legumes. Can. J. Plant Sci., 72: 323-349.

21:  Deshwal, V.K., R.C. Dubey and D.K. Maheshwari, 2003. Isolation of plant growth-promoting strains of Bradyrhizobium (Arachis) sp. With biocontrol potential against Macrophomina phaseolina causing charcoal rot of peanut. Curr. Sci., 48: 443-448.
Direct Link  |  

22:  Rubaihayo, P.R., 1976. Utilization of gamma-rays for soybean improvement. Egypt. J. Genet. Cytol., 5: 136-140.

23:  Sharma, S.K. and B. Sharma, 1984. Pattern of induced macro and micro-mutations with gamma-rays in lentil. Environ. Exp. Bot., 24: 343-351.
Direct Link  |  

24:  Kumar, D., 1977. Studies on 60Co gamma-ray induced variability in common wheat cultivar K 68. Egypt. J. Genet. Cytol., 6: 229-243.

25:  Kimani, P.M., 1988. Improvement of food beans (Phaseolus vulgaris L.) through mutation breeding. Acta Hortic., 218: 251-260.

26:  Gunasekaran, M., U. Selvaraj and T.S. Raveemdram, 1998. Induced polygenic mutations in cowpea (Vigna unguiculata L. Walp). South-Indian Hortic., 46: 13-17.

27:  Graham, R.A. and T.W. Scott, 1983. Varietal characteristics and nitrogen fixation in cowpea. Tropical Agric., 60: 269-271.

28:  Sanginga, N., O. Lyasse and B.B. Singh, 2000. Phosphorus use efficiency and nitrogen balance of cowpea breeding lines in a low P soil of the derived savanna zone in West Africa. Plant Soil, 220: 119-128.
CrossRef  |  Direct Link  |  

29:  Dubeikovsky, A.N., E.A. Mordukkova, V.V. Kochethov, F.Y. Polikarpova and A.M. Boronin, 1993. Growth promotion of black currant soft woodcuttings by recombinant strain Pseudomonas fluorescens BSP53a synthesizing an increased amount of indole-3-acetic acid. Soil Biol. Biochem., 25: 1277-1281.

30:  Matiru, V.N. and F.D. Dakora, 2004. Potential use of rhizobial bacteria as promoters of plant growth for increased yield in landraces of African cereal crops. Afr. J. Biotechnol., 3: 1-7.
Direct Link  |  

31:  Dakora, F.D., V. Matiru, M. King and D.A. Phillips, 2002. Plant Growth Promotion in Legumes and Cereals by Lumichrome, a Rhizobial Signal Metabolite. In: Nitrogen Fixation: Global Perspectives, Finan, T.M., M.R. O'Brian, D.B. Layzell, K. Vessey and W.E. Newton (Eds.). CABI Publishing, Wallingford, UK., pp: 321-322.

32:  Mujeeb, K.A. and J.K. Greig, 1973. Gamma irradiation induced variability in Phaseolus vulgaris L. cv. Blue Lake. Radi. Bot., 13: 121-126.

33:  Ramulu, K.S., 1970. Mutation in sorghum. Mut. Res., 10: 197-205.

34:  Vencatasamy, D.R., 1984. The effects of Rhizobium genotype, host genotype and their interactions on nitrogen fixation in Phaseolus vulgaris. Proceedings of the 1st Conference African Assocation Biology Nitrogen Fixation (AABNF), Jul. 1984, Nairobi, Kenya, pp: 23-27.

35:  Johonson, V.A., P.J. Mattern, D.A. Whited and J.W. Schmidt, 1969. Breeding for high protein content and quality in wheat. Proceedings of the New Approaches to Breeding for Improved Plant Protein, (NABIPP'1969), IAEA, Vienna, pp: 29-40.

36:  Singh, B.B., 1999. Breeding for improved quality. IITA Annual Report, 1999, Project No. 11, pp: 31-32.

37:  Fashakin, J.B. and J.I. Fasanya, 1988. Chemical composition and nutritive changes of some improved varieties of cowpea (Vigna unguiculata). 1: Some selected varieties from the International Institute of Tropical Agriculture, Ibadan, Nigeria. Tropical Sci., 28: 111-118.
Direct Link  |  

38:  Roest, H.P., L. Goosen-de Roo, C.A. Wiffelman, R.A. de Maagd and B.J.J. Lugtenberg, 1995. Outer membrane protein changes during bacteroid development are independent of nitrogen fixation and differ between indeterminant and determinant nodulating host plants of Rhizobium leguminosarum. Mol. Plant Microbe Interact., 8: 14-22.

39:  Newcomb, W., 1980. Control of Morphogenesis and Differentiation of Pea Root Nodules. In: Nitrogen Fixation, Newton W.E. and W.H. Orme-Johnson (Eds.). University Park Press, Baltimore, pp: 87-102.

40:  Kessel, C. and C. Hartley, 2000. Agricultural management of grain legumes: Has it led to an increase in nitrogen fixation? Field Crop Res., 65: 165-181.
CrossRef  |  

41:  Antoun, H., C.J. Beauchamp, N. Goussard, R. Chabot and R. Lalande, 1998. Potential of Rhizobium and Bradyrhizobium species as plant growth promoting rhizobacteria on non-legumes: Effects of radishes (Rhaphanus sativus L.). Plant Soil, 204: 57-67.
Direct Link  |  

42:  Subba-Rao, N.S., 1988. Biofertilizers in Agriculture. 1st Edn., Sunil Printers, Naranina, New Delhi, pp: 208.

43:  Caron, M., C.L. Patten, S. Ghosh and R. Glick, 1995. Effects of the plant growth promoting rhizobacterium Pseudomonas putida GR12-2 on the physiology of canola roots. Plant Growth Regul. Soc. Am. Q., 23: 297-302.

44:  Glick, B.R., D.M. Penrose and J. Li, 1998. A model for the lowering of plant ethylene concentrations by plant growth-promoting bacteria. J. Theoret. Biol., 190: 63-68.
CrossRef  |  Direct Link  |  

45:  Davison, J., 1999. Genetic exchange between bacteria in the environment. Plasmid, 42: 73-91.
CrossRef  |  PubMed  |  Direct Link  |  

46:  Van Elsas, J.D., J.T. Trevors and M.E. Stardub, 1989. Bacterial conjugation between Pseudomonas in the rhizosphere of wheat. FEMS Microbiol. Ecol., 53: 299-306.

47:  Patten, C.L. and B.R. Glick, 2002. Role of Pseudomonas putida indoleacetic acid in development of the host plant root system. Applied Environ. Microbiol., 68: 3795-3801.
CrossRef  |  PubMed  |  Direct Link  |  

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