Publication 163 163. Kudlicka, K., Brown, Jr. R. M., Li, L., Lee, J. H., and S. Kuga. 1995. ß-glucan synthesis in the cotton fiber: IV. In vitro assembly of the cellulose I allomorph. Plant Physiology 107:111-123.

163. Abstract

In vitro assembly of cellulose from plasma membrane extracts of the cotton (Gossypium hirsutum) fiber was enriched by a combination of 3-(N-morpholino) propanesulfonic acid extraction buffer and two independent digitonin solubilization steps consisting of 0.05% digitonin (SE,) followed by 1% digitonin (SE,). Glucan synthase activity assays revealed that, although the SE, fraction possessed higher activity, only 8.6% of the in vitro product survived acetic/nitric acid treatment. On the other hand, the SE, fraction was less active, but 32.1% of the total glucan in vitro product was resistant to acetic/nitric acid. In vitro products synthesized from the SE, fraction contained ß-1,3-glucan and fibrillar cellulose I, whereas the SE, fraction produced ß-1,3-glucan and cellulose II. Both celluloses assembled in vitro were labeled with cellobiohydrolase I-gold complex, and the electron diffraction patterns of both products from SE, and SE, revealed cellulose I and cellulose II, respectively. Contamination of native cellulose was ruled out by extensive evidence from autoradiography of the ethanol-insoluble and acetic/nitric acid-insoluble materials, including three different controls.

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