PJB-2019-751
ESTABLISHMENT OF PROTOCOL FOR PROTOPLAST ISOLATION AND FUSION OF CITRUS CULTIVARS/LINES
Saima Mumtaz
Abstract
Abstract This research study provides an operative and useful protocol for protoplast isolation and fusion of citrus cultivars/ lines. For this an effort was done for protoplast isolation and fusion of various citrus cultivars and lines. Protoplasts were successfully isolated from both callus and mesophyll cells by using enzymatic method (2% cellulase and 2% macerozyme). Murcott, HAM-N, Nova, Volk, AMB and SF-146-zw were used as callus donor parents while Rough lemon, Kinnow, Musambi, Troyer citrange and Feutrel's early were acted as mesophyll donor parents. Chemo fusion (PEG mediated) was effectively done to fuse the isolated protoplasts of mesophyll cells and callus cells. Confirmation of fused product (hybrid) was done by cytological analysis using microscopic examination. The consequence of this study provides the way forward for genetic improvement of commercial cultivars by undertaking further investigation like plant regeneration from the fused protoplasts and molecular analysis.
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