Abstract
A procedure has been devised for isolation and recovery of exine that is generally applicable to pollens and spores from a range of common species. Particles are suspended in an aqueous solution of 4- O-methylmorphine N-oxide and cyclohexylamine which swells them and loosens the exine layer. Gentle pressure in a teflon/glass tissue grinder releases protoplasts from exine and subsequent treatment with a mixture of cellulase and pectinase destroys attachments between intine and exine layers. The suspension is placed on a NaCl/Percoll step gradient to remove protoplasts and then the exine is cleanly separated from other cellular fragments on a step gradient of CsCl. The entire procedure can be accomplished at room temperature which greatly reduces possibility of chemical or physical modification of the exine. Moreover, the method is readily scaled up to production of exine in gram amounts which allows future study of the structural properties of its major biopolymer, sporopollenin.
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