Lupin'den Lupinus spp Protoplast İzolasyonu

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.

Anahtar Kelimeler:

Lupin, Lupinus, protoplast, izolasyon

Protoplast Isolation in Lupin Lupinus spp

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,

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