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Cell wall extension results in the coordinate separation of parallel microfibrils: evidence from scanning electron microscopy and atomic force microscopy

Journal Article · · The Plant Journal
 [1];  [2];  [3];  [4]
  1. University of Missouri, Columbia, MO (United States)
  2. University of Missouri, Columbia, MO (United States); Sherbrooke University, Quebec (Canada)
  3. Pennsylvania State University, University Park, PA (United States)
  4. University of Missouri, Columbia, MO (United States); University of Massachusetts, Amherst, MA (United States)
+ Show Author Affiliations
Enlargement of the cell wall requires separation of cellulose microfibrils, mediated by proteins such as expansin; according to the multi-net growth hypothesis, enlargement passively reorients microfibrils. However, at the molecular scale, little is known about the specific movement of microfibrils. To find out, we examined directly changes in microfibril orientation when walls were extended slowly in vitro under constant load (creep). Frozen-thawed cucumber hypocotyl segments were strained by 20–30% by incubation in pH 4.5 buffer or by incubation of heat-inactivated segments in α-expansin or a fungal endoglucanase (Cel12A). Subsequently, the innermost layer of the cell wall was imaged, with neither extraction nor homogenization, by field-emission scanning electron microscopy (FESEM) and atomic force microscopy (AFM). AFM images revealed that sample preparation for FESEM did not appreciably alter cell wall ultrastructure. In both FESEM and AFM, images from extended and non-extended samples appeared indistinguishable. To quantify orientational order, we used a novel algorithm to characterize the fast Fourier transform of the image as a function of spatial frequency. For both FESEM and AFM images, the transforms of non-extended samples were indistinguishable from those of samples extended by α-expansin or Cel12A, as were AFM images of samples extended by acidic buffer. Here, we conclude that cell walls in vitro can extend slowly by a creep mechanism without passive reorientation of innermost microfibrils, implying that wall loosening agents act selectively on the cross-linking polymers between parallel microfibrils, rather than more generally on the wall matrix.
Research Organization:
University of Massachusetts, Amherst, MA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division (CSGB)
Grant/Contract Number:
FG02-03ER15421
OSTI ID:
2483282
Journal Information:
The Plant Journal, Journal Name: The Plant Journal Journal Issue: 2 Vol. 43; ISSN 0960-7412
Publisher:
Society for Experimental BiologyCopyright Statement
Country of Publication:
United States
Language:
English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Cucumis sativus
cel12A endoglucanase
cell wall creep
elongation
expansin
fast Fourier transform