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Research Article

volume 40 issue 5 (october 2017) : 830-835,   Doi: 10.18805/LR-317

Caucasian clover (Trifolium ambiguum Bieb.) × white clover (T. repens L.) – interspecific hybrids developed through tissue culture

F
Fan Huang
M
Mingjiu Wang
Z
Zhiyong Li
1Grassland Research Institute, Chinese Academy of Agricultural Sciences, Hohhot-010 010, China.

  • Submitted13-09-2016|

  • Accepted17-02-2017|

  • First Online 18-09-2017|

  • doi 10.18805/LR-317

Cite article:- Huang Fan, Wang Mingjiu, Li Zhiyong (2017). Caucasian clover (Trifolium ambiguum Bieb.) × white clover (T. repens L.) – interspecific hybrids developed through tissue culture. Legume Research. 40(5): 830-835. doi: 10.18805/LR-317.

ABSTRACT

In order to create new forage variety, caucasian clover (Trifolium ambiguum Bieb.) with strong tolerance to drought and cold introduced from New Zealand as the female parent was hybridized with a white clover (T. repens L.) collected from northeast China, which has strong nitrogen fixation capacity. Hybrid embryos of caucasian clover×white clover were successfully raised using embryo rescue. 13.8% of these embryos grew to maturity. The successfully grown plants were transferred to sterilized vermiculite after 5-6 weeks and then to field after 3-4 months. The hybrid plant showed intermediate morphological features and a chromosomal number of 2n=5X=40. In order to obtain more plants for backcross, a tissue culture system was established for the hybrids, using MS+0.1mg l-1 2, 4-D+2mg l-1 6-BA, MS +0.5mg l-1 NAA+1mg l-1 6-BA+1mg l-1 KT and 1/2MS as the medium for bud-induction, differentiation and root-induction respectively. 80% of the cultured plantlets survived these growing conditions. Hybridity of the plant was also confirmed by ISSR markers using 16 primers.

REFERENCES

  1. Abberton, M.T., Michaelson-Yeates, T.P.T, Marshall, A.H, Holdbrook-Smith, K. and Rhodes, I. (1998). Morphological characteristics of hybrids between white clover, Trifolium repens L., and Caucasian clover, Trifolium ambiguum M. Bieb. Plant Breeding 117: 494-496.
  2. Anderson, J.A., Taylor, N.L. and Williams, E.G. (1991). Cytology and fertility of the interspecific hybrid Trifolium ambiguum × T. repens L. and backcross populations. Crop Sci. 31: 683-687.
  3. Barikissou, E. and Baudoin, J. (2012). A review of embryos rescue in food legumes in general and in the genus Phaseolus in particular. Legume Research 16(2):203-215.
  4. Barpete, S., Sharma, N.C. and Kumar, S. (2014). Assessment of somaclonal variation and stability in in vitro regenerated grasspea plants using. Legume Research 37(4):345-352.
  5. Cherney, J.H. 2000. Kura clover for New York pastures. Kura Clover Update, August, (2000). Cornell University
  6. Coolbear, P., Hill, M.J. and Efendi, F. (1994). Relationships between vegetative and reproductive growth in a four year old stand of Caucasian clover (Trifolium. ambiguum M Bieb.) cv. Monaro. Proc. Agron. Soc. N. Z. 24, 77–82.
  7. Ferguson, N.H., Rupert, E.A. and Evans, P.T. (1990). Interspecific Trifolium hybrids produced by embryo and ovule culture. Crop Sci. 30: 1145-1149.
  8. Frame, J. and Newbould, P. (1986). Agronomy of white clover. Adv. Agron. 40: 1-88.
  9. Hely, F.W. (1963). Relationship between effective nodulation and time to initial nodulation in a diploid line of Trifolium ambiguum M. Bieb. Australian Journal of Biological Sciences 10: 1-16.
  10. Hill, M.J. and Mulcahy, C. (1995). Seedling vigour and rhizome development in Trifolium ambiguum M. Bieb. (Caucasian clover) as affected by density of companion grasses, fertility, drought, and defoliation in the first year. Aust. J. Agric. Res. 46: 807-819.
  11. Kaushal, P., Malaviya, D.R., Roy, A.K., Kumar, B. and Tiwari, A. (2005). Trifolium alexandrinum × T. resupinatum –interspecific hybrids developed through embryo rescue. Plant Cell, Tissue and Organ Culture 83: 137-144.
  12. Malaviya, D.R., Roy, A.K., Kaushal, P., Kumar, B. and Tiwari, A. (2004a). Development and characterization of interspecific hybrids of Trifolium alexandrinum × T. apertum using embryo rescue. Plant Breeding 123: 536-542.
  13. Mathiyazhagan, S., Pahuja, S.K. and Ahlawat, A. (2013). Regeneration in cultivated (Cyamopsistetragonoloba L.) and wild species (C. serrata) of guar. Legume research 36(2):180-187.
  14. Meredith, M.R., Michaelson-Yeates, T.P.T., Ougham, H.J. and Thomas, H. (1995). Trifolium ambiguum as a source of variation in the breeding of white clover. Euphytica 82: 185-191
  15. Owczarzak, Alfred. (1952). A rapid method for mounting pollen grains with special regard to sterility studies. Stain Technology 27: 249-251.
  16. Patrick, H.N. and W.L Lowther. (1995). Inoculation of the rhizobia on the nodulation and establishment of Trifolium ambiguum. Soil Biol. Biochem. 27(4/5): 717-720.
  17. Phillips, G.C., Collins, G.B. and Taylor, N.L. (1982). Interspecific hybridization of red clover (Trifolium pretense L.) with T. sarosiense Hazsl. using in vitro embryo rescue. Theor. Appl. Genet. 62: 17-24.
  18. Phillips, G.C., Grosser, J.W., Berger, S., Taylor, N.L. and Collins, G.B. (1992). Interspecific hybridization between red clover and Trifolium alpestre using invitro embryo rescue. Crop Sci. 32: 1113-1115.
  19. Roy, A.K., Malaviya, D.R., Kaushal, P., Kumar, B. and Tiwari, A. (2004). Interspecific hybridization of Trifolium alexandrinum with T. constantinopolitanum using embryo rescue. Plant Cell Rep. 22: 705-710.
  20. Taylor, N.L. and Smith, R.R. 1998. Kura clover (Trifolium ambiguum M. B.) breeding, culture, and utilisation. Adv. Agron. 63: 154-179.    Widdup, K.H., Hussain, S.W., Williams, W.M., Lowther, W.L., Pryor H.N. and Sutherland B.L. (2003|). The development and plant characteristics of interspecific hybrids between white and Caucasian clover. ‘In Legumes for Dryland Pastures. Proceeding New Zealand Grassland Association Symposium’. Grassland Research and Practice Series NO.11: 143-148.
  21. Williams, E. (1978). A hybrid between Trifolium repens and T. ambiguum obtained with the aid of embryo culture. New Zealand J. Bot. 16: 499-506.
  22. Williams, E. and Verry, I.M. (1981).A partially fertile hybrid between Trifolium repens and T. ambiguum. New Zealand J. Bot. 19: 1-7.
  23. Williams, W.M., Verry, I.M., Ansari, H.A. and Hussain, S.W. (2006 b.) First backcrosses of caucasian x white clover hybrids to caucasian clover. In ‘Breeding for success: diversity in action. Proceedings of the 13th Australasian Plant Breeding Conference’. NewZealand977–983.
  24. Williams, W.M., Verry, I.M., Hussain, S.W., Ansari, H.A., Widdup, K.H., Ellison, N.W. and Nichols, S.N. (2013). Widening the adaptation of white clover by incorporation of valuable new traits from wild clover species. In ‘Proceedings of the 22nd International Grasslands Congress’. Sydney. 287–289.
  25. Williams, W.M. (2014). Trifolium interspecific hybridization: widening the white clover gene pool. Crop & pasture science 65: 1091-1106.
  26. Yamada, T. and Fukuoka, H.(1985). Application of ovule culture to interspecific hybridisation between Trifolium repens and T. ambiguum. In Proceedings of the XV Int Grassl Conf:241-243.
  27. Yamada, Y., Fukuoka, H. and Higuchi, S. (1989). Interspecific hybridisation of tetraploid Kura clover (Trifolium ambiguum M. Bieb.) and white clover (T. repens L.) using ovule culture. J. Jap. Soc. Grassl. Sci. 35: 180-185.
  28. Zorin, M., Brockwell, J. and Muller, W.J. 1976. Symbiotic characteristics of Trifolium ambiguum seedlings grown in tube culture as affecting subsequent symbiotic vigour. Australian Journal of Experimental Agriculture and Animal Husbandry 16: 854-862. 
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    Research Article

    volume 40 issue 5 (october 2017) : 830-835,   Doi: 10.18805/LR-317

    Caucasian clover (Trifolium ambiguum Bieb.) × white clover (T. repens L.) – interspecific hybrids developed through tissue culture

    F
    Fan Huang
    M
    Mingjiu Wang
    Z
    Zhiyong Li
    1Grassland Research Institute, Chinese Academy of Agricultural Sciences, Hohhot-010 010, China.

    • Submitted13-09-2016|

    • Accepted17-02-2017|

    • First Online 18-09-2017|

    • doi 10.18805/LR-317

    Cite article:- Huang Fan, Wang Mingjiu, Li Zhiyong (2017). Caucasian clover (Trifolium ambiguum Bieb.) × white clover (T. repens L.) – interspecific hybrids developed through tissue culture. Legume Research. 40(5): 830-835. doi: 10.18805/LR-317.

    ABSTRACT

    In order to create new forage variety, caucasian clover (Trifolium ambiguum Bieb.) with strong tolerance to drought and cold introduced from New Zealand as the female parent was hybridized with a white clover (T. repens L.) collected from northeast China, which has strong nitrogen fixation capacity. Hybrid embryos of caucasian clover×white clover were successfully raised using embryo rescue. 13.8% of these embryos grew to maturity. The successfully grown plants were transferred to sterilized vermiculite after 5-6 weeks and then to field after 3-4 months. The hybrid plant showed intermediate morphological features and a chromosomal number of 2n=5X=40. In order to obtain more plants for backcross, a tissue culture system was established for the hybrids, using MS+0.1mg l-1 2, 4-D+2mg l-1 6-BA, MS +0.5mg l-1 NAA+1mg l-1 6-BA+1mg l-1 KT and 1/2MS as the medium for bud-induction, differentiation and root-induction respectively. 80% of the cultured plantlets survived these growing conditions. Hybridity of the plant was also confirmed by ISSR markers using 16 primers.

    REFERENCES

    1. Abberton, M.T., Michaelson-Yeates, T.P.T, Marshall, A.H, Holdbrook-Smith, K. and Rhodes, I. (1998). Morphological characteristics of hybrids between white clover, Trifolium repens L., and Caucasian clover, Trifolium ambiguum M. Bieb. Plant Breeding 117: 494-496.
    2. Anderson, J.A., Taylor, N.L. and Williams, E.G. (1991). Cytology and fertility of the interspecific hybrid Trifolium ambiguum × T. repens L. and backcross populations. Crop Sci. 31: 683-687.
    3. Barikissou, E. and Baudoin, J. (2012). A review of embryos rescue in food legumes in general and in the genus Phaseolus in particular. Legume Research 16(2):203-215.
    4. Barpete, S., Sharma, N.C. and Kumar, S. (2014). Assessment of somaclonal variation and stability in in vitro regenerated grasspea plants using. Legume Research 37(4):345-352.
    5. Cherney, J.H. 2000. Kura clover for New York pastures. Kura Clover Update, August, (2000). Cornell University
    6. Coolbear, P., Hill, M.J. and Efendi, F. (1994). Relationships between vegetative and reproductive growth in a four year old stand of Caucasian clover (Trifolium. ambiguum M Bieb.) cv. Monaro. Proc. Agron. Soc. N. Z. 24, 77–82.
    7. Ferguson, N.H., Rupert, E.A. and Evans, P.T. (1990). Interspecific Trifolium hybrids produced by embryo and ovule culture. Crop Sci. 30: 1145-1149.
    8. Frame, J. and Newbould, P. (1986). Agronomy of white clover. Adv. Agron. 40: 1-88.
    9. Hely, F.W. (1963). Relationship between effective nodulation and time to initial nodulation in a diploid line of Trifolium ambiguum M. Bieb. Australian Journal of Biological Sciences 10: 1-16.
    10. Hill, M.J. and Mulcahy, C. (1995). Seedling vigour and rhizome development in Trifolium ambiguum M. Bieb. (Caucasian clover) as affected by density of companion grasses, fertility, drought, and defoliation in the first year. Aust. J. Agric. Res. 46: 807-819.
    11. Kaushal, P., Malaviya, D.R., Roy, A.K., Kumar, B. and Tiwari, A. (2005). Trifolium alexandrinum × T. resupinatum –interspecific hybrids developed through embryo rescue. Plant Cell, Tissue and Organ Culture 83: 137-144.
    12. Malaviya, D.R., Roy, A.K., Kaushal, P., Kumar, B. and Tiwari, A. (2004a). Development and characterization of interspecific hybrids of Trifolium alexandrinum × T. apertum using embryo rescue. Plant Breeding 123: 536-542.
    13. Mathiyazhagan, S., Pahuja, S.K. and Ahlawat, A. (2013). Regeneration in cultivated (Cyamopsistetragonoloba L.) and wild species (C. serrata) of guar. Legume research 36(2):180-187.
    14. Meredith, M.R., Michaelson-Yeates, T.P.T., Ougham, H.J. and Thomas, H. (1995). Trifolium ambiguum as a source of variation in the breeding of white clover. Euphytica 82: 185-191
    15. Owczarzak, Alfred. (1952). A rapid method for mounting pollen grains with special regard to sterility studies. Stain Technology 27: 249-251.
    16. Patrick, H.N. and W.L Lowther. (1995). Inoculation of the rhizobia on the nodulation and establishment of Trifolium ambiguum. Soil Biol. Biochem. 27(4/5): 717-720.
    17. Phillips, G.C., Collins, G.B. and Taylor, N.L. (1982). Interspecific hybridization of red clover (Trifolium pretense L.) with T. sarosiense Hazsl. using in vitro embryo rescue. Theor. Appl. Genet. 62: 17-24.
    18. Phillips, G.C., Grosser, J.W., Berger, S., Taylor, N.L. and Collins, G.B. (1992). Interspecific hybridization between red clover and Trifolium alpestre using invitro embryo rescue. Crop Sci. 32: 1113-1115.
    19. Roy, A.K., Malaviya, D.R., Kaushal, P., Kumar, B. and Tiwari, A. (2004). Interspecific hybridization of Trifolium alexandrinum with T. constantinopolitanum using embryo rescue. Plant Cell Rep. 22: 705-710.
    20. Taylor, N.L. and Smith, R.R. 1998. Kura clover (Trifolium ambiguum M. B.) breeding, culture, and utilisation. Adv. Agron. 63: 154-179.    Widdup, K.H., Hussain, S.W., Williams, W.M., Lowther, W.L., Pryor H.N. and Sutherland B.L. (2003|). The development and plant characteristics of interspecific hybrids between white and Caucasian clover. ‘In Legumes for Dryland Pastures. Proceeding New Zealand Grassland Association Symposium’. Grassland Research and Practice Series NO.11: 143-148.
    21. Williams, E. (1978). A hybrid between Trifolium repens and T. ambiguum obtained with the aid of embryo culture. New Zealand J. Bot. 16: 499-506.
    22. Williams, E. and Verry, I.M. (1981).A partially fertile hybrid between Trifolium repens and T. ambiguum. New Zealand J. Bot. 19: 1-7.
    23. Williams, W.M., Verry, I.M., Ansari, H.A. and Hussain, S.W. (2006 b.) First backcrosses of caucasian x white clover hybrids to caucasian clover. In ‘Breeding for success: diversity in action. Proceedings of the 13th Australasian Plant Breeding Conference’. NewZealand977–983.
    24. Williams, W.M., Verry, I.M., Hussain, S.W., Ansari, H.A., Widdup, K.H., Ellison, N.W. and Nichols, S.N. (2013). Widening the adaptation of white clover by incorporation of valuable new traits from wild clover species. In ‘Proceedings of the 22nd International Grasslands Congress’. Sydney. 287–289.
    25. Williams, W.M. (2014). Trifolium interspecific hybridization: widening the white clover gene pool. Crop & pasture science 65: 1091-1106.
    26. Yamada, T. and Fukuoka, H.(1985). Application of ovule culture to interspecific hybridisation between Trifolium repens and T. ambiguum. In Proceedings of the XV Int Grassl Conf:241-243.
    27. Yamada, Y., Fukuoka, H. and Higuchi, S. (1989). Interspecific hybridisation of tetraploid Kura clover (Trifolium ambiguum M. Bieb.) and white clover (T. repens L.) using ovule culture. J. Jap. Soc. Grassl. Sci. 35: 180-185.
    28. Zorin, M., Brockwell, J. and Muller, W.J. 1976. Symbiotic characteristics of Trifolium ambiguum seedlings grown in tube culture as affecting subsequent symbiotic vigour. Australian Journal of Experimental Agriculture and Animal Husbandry 16: 854-862. 
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