Plant tissue culture studies in cajanus cajan by vijay chourey

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  • Callus induction
  • Plant tissue culture studies in cajanus cajan by vijay chourey

    1. 1. VIJAY KUMAR CHOUREY DEPARTMENT OF BIOCHEMISTRY & GENETICS BARKATULLAH UNIVERSITY BHOPAL (M.P.) MULITIPLESHOOT INDUCTION AND PLANT REGENERATION IN PIGEONPEA (CAJANUS CAJAN L.)
    2. 2. Contents Introduction Materials & methods Results Conclusion References
    3. 3. INTRODUCTION
    4. 4. Introduction Cajanus cajan L. belong to family fabaceae also known as red gram, is an important world crop species widely cultivated. Cajanus cajan L. is an excellent source of protein, seeds are using as vegetable flour additive or other food. Its flower are self compatible and usually self pollinated. Its forms root nodules associated with Rhizobium sp. bacteria that function nitrogen fixation. 66 chromosome are present in Cajanus cajan L.
    5. 5. It is usually single stemmed, freely branching, and become woody after a few months. India is the world’s largest pigeonpea producer and grows over 77% of the total world production. Typical nutritional values for seeds are: moisture, 10.1 percent, protein 19.2 percent, fat, 1.5 percent, carbohydrates, 57.3 percent, fiber 8.1 percent, and ash, 3.8 percent.
    6. 6. Present study was carried out with the following objectives: 1. To study of different explant for to callus induction. 2. To study redifferentiation in Cajanus cajan L.
    7. 7. METERIALS AND METHODS
    8. 8. Material & Methods MATERIAL  Healthy seeds of Cajanus cajan L. variety Balwan (NRS) used in the present study were obtained from Mukesh seeds, Bhopal.  All the chemicals used in the present study were analytical grade and were obtained from CDH, Himedia and Qualigens India. Plant growth regulators, gelling agent, sucrose, vitamins, colchicine etc, were obtained from “CDH fine chemicals” and “Himedia” Mumbai.
    9. 9. Method  The tissue culture laboratory was washed with effective disinfectant (Teepol) followed by wiping with 2% sodium hypochlorite solution or 95% ethyl alcohol, extran, Lysol, zephiran or roccal.  MS medium used for present study:  30 g (3% w/v) sucrose.  Required quantities of plant growth regulators .  Micronutrient and Macronutrient were added.  8 g agar.  Make up final volume (i.e. 1000ml),  pH was adjusted between 5.4- 5.6.  Finally, the culture vessels were plugged, labeled and autoclaved at 1210 C and 1.06 kg/cm2 for 20 min.
    10. 10. Explants Selection  The present study, apical, axilliary buds, cotyledonary leaves and hypocotyl parts were used as explants.
    11. 11. RESULTS
    12. 12. Table-1 Callus induction and redifferentiation response with different Axillary Shoot explants of Cajanus cajan L. in MS medium supplemented with variable concentrations of growth adjuvants. Explant MS + Growth adjutants (mg/l) 14 days 21 days NAA BAP Callus induction (%) Redifferentiatio n Response (%) Callus formation in Explants* Axillary Shoot 0.25 0.25 0.00 0.00 0.00 0.50 0.25 40.00 16.00 + 0.75 0.25 60.00 44.00 ++ 1.0 0.25 66.00 50.00 ++ 1.5 0.25 46.00 37.00 + 2.0 0.25 20.00 31.00 + Note:* Relative value of Callus formation on Explants: -, Nil; +, Slight; ++, Moderate; +++, High.
    13. 13. Hypocotyl, 0.25 0.25 40.00 40.00 + 0.50 0.25 53.00 43.00 ++ 0.75 0.25 58.00 53.00 +++ 1.0 0.25 86.00 68.00 ++ 1.5 0.25 60.00 57.00 ++ 2.0 0.25 43.00 34.00 + Stem 0.25 0.25 20.00 23.00 + 0.50 0.25 33.00 26.00 + 0.75 0.25 40.00 32.00 ++ 1.0 0.25 60.00 44.00 + 1.5 0.25 41.00 33.00 + 2.0 0.25 26.00 25.00 +
    14. 14. Cotyledonary leaves. 0.25 0.25 0.00 0.00 - 0.50 0.25 48.00 25.00 + 0.75 0.25 46.00 28.00 + 1.0 0.25 52.00 36.00 ++ 1.5 0.25 38.00 32.00 + 2.0 0.25 33.00 26.00 + Note:* Relative value of Callus formation on Explants: -, Nil; +, Slight; ++, Moderate; +++, High.
    15. 15. Fig. 1: Callus induction and redifferentiation response with different explants of Cajanus cajan L. in MS medium supplemented with variable concentrations of growth adjuvants.
    16. 16. Fig. 1a: Callus induction from Hypocotyl parts
    17. 17. Table 2:Optimum concentration of cytokinin alone in subculture for multiplication and elongation of shoots raised intervening by callus in Cajanus cajan L. Shoots (in 21 days) Explant MS+ BAP (mg/l) Shoot multiplication response (%) Average no. of* shoots (Mean ± S.E.) Average length* of the multiplied (Mean ± S.E.) Callus raised shoot from hypocotyl 0.25 40 3.91 ± 0.39 2.03 ± 0.21 0.50 52 4.33 ± 0.50 2.66 ± 0.17 0.75 55 6.43 ± 0.34 3.08 ± 0.18 1.0 75 13.33 ± 0.41 5.33 ± 0.21 1.25 64 8.58 ± 0.53 4.33 ± 0.23 1.50 44 4.8 ± 0.67 3.00 ± 0.24 Note: * Based on six replicates of five explants each.
    18. 18. Fig 2: Shoot multiplication from callus obtained from hypocotyls explant. Shoot multiplication
    19. 19. Table 3: Root induction with auxin in elongated adventitious raised from hypocotyl in Cajanus cajan L. Explant ½ MS media + Growth regulator (mg/l) Roots (after 21 days) Root Induction (%) Mean no. of* root ± S.E. Mean root* length ± S.E. Elongated shoot through callus derived shoots MS plain 38 2.30 ± 0.20 0.65 ± 0.02 IBA (0.25) 62 2.50 ± 0.24 0.80 ± 0.04 IBA (0.50) 66 3.66 ± 0.30 1.22 ± 0.15 IBA (0.75) 70 5.50 ± 0.24 1.30 ± 0.08 IBA (1.00) 72 5.70 ± 0.32 1.50 ± 0.06 IBA (1.25) 85 6.50 ± 0.34 2.85 ± 0.06 IBA (1.50) 34 3.75 ± 0.31 1.25 ± 0.05 Note: * Based on five replicates of three explants each.
    20. 20. Fig.3: Root induction in callus raised shoots obtained from hypocotyls explant. Root induction
    21. 21. CONCLUSION
    22. 22. Conclusion  The present project report entitled “Tissue Culture Studies in Cajanus cajan L.” presents findings and inferences of tissue culture experiments carried out through different organ explants in order to demonstrate a reproducible protocol for in vitro regeneration in Cajanus cajan L.  Present study principally focuses on regeneration through organogenesis and demonstrates organogenesis competence of hypcotyl portions in Cajanus cajan L., chosen for the study.
    23. 23. References Blümmel, M and Saxena, KB (2005): Genetic variation for forage quality parameters in pigeonpea. Fourth International Food Legumes Research Conference, New Delhi, October 18–22, 2005 Butler, JE (1910): The wilt disease of pigeonpea and the parasitism of Neocosmospora vasinfecta Smith., India. Dept. Agr. Memoir. Bot. Ser 11(9):1–64. George, L; Eapen, SL (1994): Organogenesis and embryogenesis from diverse explants in pigeonpea (Cajanus cajan L.). Plant Cell Rep., 13: 417-420.
    24. 24. Thank

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