An efficient and rapid procedure for plantlet regeneration from chicory mesophyll protoplasts |
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Authors: | Nenz E Varotto S Lucchin M Parrini P |
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Institution: | (1) Department of Agronomy and Crop Production, University of Padova, Agripolis, Via Romea 16, 35020 Legnaro (PD), Italy |
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Abstract: | An efficient procedure for plantlet regeneration from chicory mesophyll protoplasts has been developed in order to perform
protoplast fusion experiments. Protoplasts were isolated from a genotype of Italian red chicory (CH 363) and purified by centrifugation
in a solution containing 13% (w/v) sucrose to collect uniform protoplasts in size. After 2 days culture at a density of 2×104 protoplasts ml−1 of liquid medium, protoplasts were cultured following three different procedures: in liquid medium, stratified in semi-solid
medium, and embedded in Ca-alginate droplets. Four different media were used and culture procedures were evaluated recording
the protoplast viability, protoplast division frequency and plating efficiency for each experiment. The embedding of protoplasts
in Ca-alginate droplets enhanced both division frequency and plating efficiency for chicory mesophyll cells. Furthermore,
this procedure shortened the cycle of plant regeneration from protoplasts, which could be completed in eight weeks.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Ca-alginate droplets chicory mesophyll protoplasts organogenesis |
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