Isolation of Mesophyll Protoplasts from Mature Leaves of Soybeans 1

A procedure based on a combined cellulase-Pectolyase Y-23 enzyme digestion and metrizamide-sorbitol gradient purification protocol was developed for isolating mesophyll protoplasts from mature leaves of soybean ( Glycine max L. Merr.). Based on chlorophyll content, this procedure results in a 10 to...

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Bibliographic Details
Published in:	Plant physiology (Bethesda) Vol. 73; no. 4; pp. 1067 - 1069
Main Author:	Lin, Willy
Format:	Journal Article
Language:	English
Published:	01-12-1983
Online Access:	Get full text
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Summary:	A procedure based on a combined cellulase-Pectolyase Y-23 enzyme digestion and metrizamide-sorbitol gradient purification protocol was developed for isolating mesophyll protoplasts from mature leaves of soybean ( Glycine max L. Merr.). Based on chlorophyll content, this procedure results in a 10 to 15% protoplast yield from fully expanded mature leaves and a 20 to 30% yield from young (expanding) leaves within 3 hours. Isolated protoplasts displayed high rates of HCO 3 − -dependent photosynthesis; greater than 75 micromoles O 2 evolved per milligram chlorophyll per hour at 25°C. This photosynthetic rate is comparable to that of mesophyll cells isolated mechanically from the same leaves.
Bibliography:	Contribution No. 3357 from Central Research and Development Department, Experimental Station, E. I. du Pont de Nemours & Company, Wilmington, DE 19898.
ISSN:	0032-0889 1532-2548