Protoplast Culture And Protoplast Symmetric Fusion In Cotton

Cotton is the world's leading natural fiber and the second largest oilseed crop inproduction; cotton genentics and breeding is on the basis of the developmemt of newgermplasm.The abundant cotton wild species has been the valuable genetic germplasm,and which has been significant in the reality and theory, potential in the application. Ithave to establish system for somatic embryogenesis, plant regeneration and protoplastculture if we want to develop protoplast culture and protoplast fusion.Our studies involved factors affecting somatic embryogenesis, protoplast culture in G.arboretum L and protoplast fusion between G. arboretum L and G. davidsonii Kellogg. Itis suitable for somatic embryogenesis to induce calluses with a-Naphthalene acetic acid0.5mg/L+kinetin 0.1mg/L as hormone combination in G. arboretum L, but embryogeniccalluses were not obtained. The factors affecting isolation of G. arboretum L mesophyllprotoplasts were compared. The results showed that the following parameters could gethigher protoplast yield and viability: pretreatment time 12-24 hour, osmoticum with 11%Mannitol ,concentration of enzyme solution (Cellulase 1.5%, Macerozyme 1%,Hemicellulase 2%) , plasmolysis time 3 hour, incubation time 14 hour. This researchwould be of some referential value for establishing high effective regeneration system ofprotoplasts and the related genetic manipulations based on protoplasts in G. arboretum L.This study attempted to find the condition suitable for the electrofuion between G.arboretum L and G davidsonii Kellogg. The results indicated that the conditions couldlead to the best effect of cell fusion were setting the alternating current field strength at100 V/cm, the direct current strength at 1.25 kV/cm, the direct current pulse width at50μs and 3 direct pulse times.