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Karbala International Journal of Modern Science

Agrobacterium rhizogenes

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A High Yield Method For Protoplast Isolation And Ease Detection Of Rol B And C Genes In The Hairy Roots Of Cauliflower (Brassica Oleracea L.) Inoculated With Agrobacterium Rhizogenes, Qutaiba Shuaib Al-Nema, Ghazwan Qasim Hasan, Omar Abdulazeez Alhamd Aug 2022

A High Yield Method For Protoplast Isolation And Ease Detection Of Rol B And C Genes In The Hairy Roots Of Cauliflower (Brassica Oleracea L.) Inoculated With Agrobacterium Rhizogenes, Qutaiba Shuaib Al-Nema, Ghazwan Qasim Hasan, Omar Abdulazeez Alhamd

Karbala International Journal of Modern Science

Protoplasts represent a unique experimental system for the circulation and formation of genetically modified plants. Here, protoplasts were isolated from genetically modified hairy root tissues of Brassica oleracea L. induced by the Agrobacterium rhizogenes strain (ATCC13332). The concentration of enzyme solutions utilized for protoplast isolation was 1.5 % Cellulase YC and 0.1 % Pectolyase Y23 in 13% mannitol solution, which resulted in high efficiency of isolation within 8 hours, in which the protoplast yield was 2 × 104 cells ml-1 and the percentage of viability was 72%. Each protoplast has one nucleus with a nucleation of 48%. A polymerase chain …