Lupin'den Lupinus spp Protoplast İzolasyonu

Abstract

Bu araşt ı rman ı n amac ı değişik lüpin tür'erinden Lupinus spp protoplast izole etmek için uygun yöntem geliştirmektir. Al 10 ml r , CaCl2. 2H20 0.103 g I , Cellulase Onozuka R-10 1.5 %, Macerozyme R-10 0.1%, mannitol 8.96 % ve BSA 0.1% kar ışımı serada yetiştirilerblüpin türleri yapraklar ı ndan protoplast elde etmeye uygun bir enzim kar ışımı olarak belirlendi. Karanl ı kta, buzdolabı nda +4 C 14-b5 saat hücre duvar ı n ı enzimle uzakla ştı rma bunu takiben de oda s ı caklığı nda 30-60 dakika inkube etme ve daha sonra 25 C de 70 devir! dk. 30 dakika sallama ile ara ştı rmada kullan ı lan Bütün lüpin türlerinden yüksek kalite ve miktarda protoplast elde edildi.

Keywords

• Lupin
• Lupinus
• protoplast
• izolasyon

Protoplast Isolation in Lupin Lupinus spp

Abstract

The aim of this wprk was to find out a method for isolation of+ protoplasts from various lupin Lupinus spp species. Al 10 ml I , CaCl2. 2H20 0.103 g I , Cellulase Onozuka R-10 1.5 %, Macerozyme R-10 0.1%, mannitol 8.96 % and BSA 0.1% have been found to be a suitable enzyme mixt yre for isolating protoplasts from the leaves of glasshouse-grown lupin species. Digestion in the dark in the refrigerator C for 14-15 hours followed by incubation at room temperature for 30- 60 minutes and then shaking at 70 rpm at 25 C for 30 minutes released a high quality and number of protoplasts in all lupin species examined.

Keywords

• Lupinus
• lupin
• protoplast
• isolation

References

1. Bhojwani, S.S.,and M.K. Razdan,1983. Plant tissue culture: Theory and Practice Davey, M.R and A. Kumar (1983), Higher plant protoplasts-Retrospect and prospect. Ir, intemational review of cytology, supplement 16. plant protoplasts ed. K.L. Giles pp:1-3
2. Dupuis,J.M; M. Pean and Chagvardieff (1990), Plant donor tissue and isolation procedure efbct on early formation of embryoids from protoplasts of Hellanthus annuus L. Plant Cell, Tissue and Organ Culture 22: 183-189
3. Fellner,M and P. Havranek (1992), Isolation of Album pollen protoplasts. Plant Cell, Tissue and Organ Culture 29: 275-279
4. Fitterr, M.S., and A.D. Krikorian, 1989. Plant protoplasts some guidlines for their preparabon and manipulation in culture. Calbiochem Corporation.
5. Gilbert, J.E.,1991. Interaction between mixing and density in protoplast cultures of Solanum tuberosum L. PhD thesis, University of Reading, Department of Agriculturai Botany.
6. Larkin, P.j., 1976. Purification and viability determinations of plant protoplasts. Planta 128: 213-216
7. Lee,N., H.Y. Wetzstein and C.H. Borman (1989) Cortical microtubule organization in Vibs protoplasts affected by concentration of enzyme isolation medium and duration of incubation. Physioloa plantamm 77: 27-32
8. Sarhan, F., and D. Cesar, 1988. High yield isolation of mesophyll protoplasts from wheat, barley and rye. Physiologia Plantarum, 72: 337-342.

9. Schafer-Menuhr, A., 1987. Isolation und kultur von lupinen protoplasten. I. Protoplasten aus blattern von Lupinus angustifolius sorte kubesa. Landbaufarschung volkenrode, 37(3): 117-120
10. Schafer-Menuhr, A. (1988) Isolation und kultur vonlupinen protoplasten. III. Protoplasten aus zellsuspensionskulturen von Lupinus polyphyllus. Landbaufarschung volkenrode, 38. Heft 2, Seite 99-102
11. Schafer-Menuhr, A.(1989) Isolation und kultur von lupinen protoplasten. IV. Regeneration von pflanzen aus protoplasten. Landbaufarschung volkenrode, 39. Heft 3, Seite 133-136
12. Schafer-Menuhr, A (1990) Electrical fusion of lupin species. In, International lupin associallon 6th intemallonal lupin conference proceedings. Temuco-Pucan Chile. Edit. by D. von Barer pp: 373-375
13. Schafer-Menuhr, A (1991) regeneration of shoots from protoplasts of lupins. Lupin newsletter. 14: 42-44
14. Schafer-Menuhr, A.,and S. Sturmer, 1987. Isolation und kultur von lupinen protoplasten. Il. Modifikation von nahrmedien zur beschleunigten teilung von protoplasten aus blattern von Lupinus angustifolius sorte kubesa. Landbaufarschung volkenrode. 37(4) 231-234
15. Vasil, V and I.K. Vasil (1980) Isolation and culture of cereal protoplats. Il Embryogenesis and plantlet formation from protoplasts of Pennisetum americanum. Theor Apll. Genet. 56: 97