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Estimation of Variability of Five Enzyme Systems Among Wild and Cultivated Species of Trifolium


The genus Trifolium comprises of 290 annual and perennial species of which the species such as T. repens, T. hybridum, T. pratense, T. ambiguum, T. resupinatum, T. alexandrinum are economically important. Boundaries between species in many cases have been difficult to define because of wide range of diversity caused by primary polymorphism. Hence, inter- and intraspecies variation in Trifolium, for zymogram pattern of five enzyme system was made to work out estimate of variability for isozymic banding pattern and get an insight into the species relationship. A total of 25 species represented by 134 accessions were compared for 5 enzymes viz. peroxidase, esterase, superoxide dismutase, acid phosphatase, and glutamate oxalo acetate transaminase using starch gel electrophoresis. Forty-six types of zymograms for Est isozyme pattern were observed amounting to 4.38 estimate of variability. The estimates of variability revealed maximum variation in T. resupinatum (4.24) followed with 3.02 in T. nigrescens. Estimate of variability for superoxide dismutase ranged from 0.46 to 2.67 among species amounting to 1.08 total variability across species. A total of 28 types of glutamate oxalo acetate transaminase zymograms were observed accounting for 2.48 estimate of variability. All but one band attributed to an estimate of variability of 1.43 in the genus for ACP and 16 different types of ACP zymograms were noticed. Highest variation for ACP was observed in T. resupinatum (4.53). Estimate of variation for peroxidase was 4.83 and 51 types of zymogram were observed. The species differed markedly for zymograms. The species distributed both in temperate and tropical parts like T. resupinatum had more variability as compared to cultivated species like T. alexandrinum and T. pratense. The rich variability present among these species can provide good source of gene transfer from wild to cultivated species which otherwise have no specific zymogram and exhibit low variability. The species sharing zymogram pattern for one or more enzymes with cultivated species were considered to have affinity and can further be utilized in attempting interspecific hybridization.

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  • F.M. Abdel-Tawab M.A. Rashid A.A. El-Seoudy E.M. Fahmi A.M. Rammah F.M. Ali (1985) ArticleTitlePrediction of yield potentials in berseem clover (Trifolium alexandrinum L.) by biochemical genetic markers Egypt J. Genet. Cytol. 14 283–292

    Google Scholar 

  • S.J. Bennett A. Mathews (2003) ArticleTitleAssessment of genetic diversity in clover species from SardiniaItaly, using AFLP analysis Plant Breed. 122 362–367 Occurrence Handle10.1046/j.1439-0523.2003.00860.x Occurrence Handle1:CAS:528:DC%2BD3sXot1Sjsbg%3D

    Article  CAS  Google Scholar 

  • A.H.D. Brown B.S. Weir (1983) Measuring genetic variability in plant populations S.D. Tanksely T.J. Orton (Eds) Isozymes in Plant Genetics and Breeding Elsevier Science Publishing Co Amsterdam 219–239

    Google Scholar 

  • Z. Bulinska-Radomska (1992) Morphological and enzymatic variation of Polish populations of Trifolium medium L F. Veronesi S. Bullitto S. Caredda (Eds) Ploidy and Chromosome Manipulation in Forage Breeding, Proceedings of 17th meeting of the Fodder Crop Section of Eucarpia14–18 Oct. 1991 Algharo Rome, Italy 153–155

    Google Scholar 

  • Z. Bulinska-Radomska (1994) Patterns of genetic variation in some Trifolium species F. Balfourier M.R. Perretant (Eds) Evolution and Exploitation of Genetic Resources: Pre-breeding Clermont- Ferrand France Versailles, France 43–46

    Google Scholar 

  • W.J. Collins R.C. Rossiter Y. Haines (1984) ArticleTitleIdentification of subterranean clover cultivars and their relationship by isozyme analysis Aust. J. Agric. Res. 35 399–411 Occurrence Handle10.1071/AR9840399

    Article  Google Scholar 

  • L.D. Gottlieb (1981) ArticleTitleElectrophoretic evidence and plant populations Prog. Phytochem. 7 1–46 Occurrence Handle1:CAS:528:DyaL3MXitVGgtLo%3D

    CAS  Google Scholar 

  • I. Hagima F. Moisa (1992) ArticleTitleIsozyme variation in diploid and tetraploid genotypes of red clover (Trifolium pratense L.) Probleme de Genetica Teoritica Si Applicata 24 125–129

    Google Scholar 

  • J.L. Hamrick M.J.W. Godt (1990) Allozyme diversity in plant species A.H.D. Brown M.T. Clegg A.L. Kahler B.S. Weir (Eds) Plant Population Genetics, Breeding and Genetic Resources Sinauer Associates Inc. MA 43–63

    Google Scholar 

  • O. Lange M.T. Schifino-Wittmann (2000) ArticleTitleIsozyme variation in wild and cultivated species of the genus Trifolium L. (Leguminosae) Ann. Bot. 86 339–345 Occurrence Handle10.1006/anbo.2000.1190 Occurrence Handle1:CAS:528:DC%2BD3cXltlOqsL4%3D

    Article  CAS  Google Scholar 

  • C.K. Lee H.A. Eagles N.M. Mc Farlane K.B. Kelly (1993) ArticleTitleGenetical variation within white clover (Trifolium repens L.) population from North Central Victoria Austr. J. Experim. Agric. 33 333–336

    Google Scholar 

  • F. Molina-Freaner S.K. Jain (1992) ArticleTitleIsozyme variation in California and Turkish populations of the colonizing species Trifolium hirtum J. Hered. 83 423–430

    Google Scholar 

  • M.D. Poulik (1957) ArticleTitleStarch gel electrophoresis in a discontinuous system of buffers Nature 180 1477–1479 Occurrence Handle1:CAS:528:DyaG1cXjtlamsQ%3D%3D Occurrence Handle13493570

    CAS  PubMed  Google Scholar 

  • W. Powell M. Morgante C. Andre M. Hanafey J. Vogel S. Tingey A. Rafalski (1996) ArticleTitleThe comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis Mol. Breed. 2 225–238 Occurrence Handle10.1007/BF00564200 Occurrence Handle1:CAS:528:DyaK28XntlGrtLk%3D

    Article  CAS  Google Scholar 

  • J.R. Russell J.D. Fuller M. Macaulay B.G. Hatz A. Jahoor W. Powell R. Waugh (1997) ArticleTitleDirect comparison of levels of genetic variation among barley accessions detected by RFLP's, AFLP's, SSR's and RAPD's Theor. Appl. Genet. 95 714–722 Occurrence Handle10.1007/s001220050617 Occurrence Handle1:CAS:528:DyaK2sXmvFGgt7k%3D

    Article  CAS  Google Scholar 

  • B.B. Shahi H. Morishima H.I. Oka (1969) ArticleTitleA survey of variations in peroxidase and esterase isozymes of wild and cultivated Oryza species Jpn. J. Genet. 44 IssueID5 303–319 Occurrence Handle1:CAS:528:DyaE3MXkvVGhtLo%3D

    CAS  Google Scholar 

  • O. Smithies (1995) ArticleTitleZone electrophoresis in starch gel: group variation in the serum proteins of normal human adults Biochem. J. 61 629–641

    Google Scholar 

  • J.A. Veech (1969) ArticleTitleLocalization of peroxidase in infected tobaccos susceptible and resistant to black shank Phytopathology 59 556–571

    Google Scholar 

  • J.F. Wendel N.F. Weeden (1989) Visualization and interpretation of plant isozymes D.S. Soltis P.S. Soltis (Eds) Isozymes in Plant Biology Chapman and Hall London 5–45

    Google Scholar 

  • M. Zohary (1972) ArticleTitleOrigins and evolution in the genus Trifolium Bot. Notiser 125 501–511

    Google Scholar 

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Correspondence to D. R. Malaviya.

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Malaviya, D.R., Kumar, B., Roy, A.K. et al. Estimation of Variability of Five Enzyme Systems Among Wild and Cultivated Species of Trifolium. Genet Resour Crop Evol 52, 967–976 (2005).

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  • Isozyme
  • Trifolium intraspecies variation
  • Isozymic polymorphism