Protoplasma

, Volume 197, Issue 1, pp 64–75

Cortical microtubule involvement in bordered pit formation in secondary xylem vessel elements ofAesculus hippocastanum L. (Hippocastanaceae): A correlative study using electron microscopy and indirect immunofluorescence microscopy

  • N. J. Chaffey
  • J. R. Barnett
  • P. W. Barlow
Original Papers

DOI: 10.1007/BF01279885

Cite this article as:
Chaffey, N.J., Barnett, J.R. & Barlow, P.W. Protoplasma (1997) 197: 64. doi:10.1007/BF01279885

Summary

A correlative study, using indirect immunofluorescence microscopy (IIF) of anti-α-tubulin stained sections and transmission electron microscopy (TEM), gave details of the involvement of cortical microtubules (CMTs) in the development of bordered pits in secondary xylem vessel elements ofAesculus hippocastanum L. In addition, aspects of wall cytochemistry were studied during this process using the Thiéry (PATAg) test, immunolocalization with the monoclonal antibodies JIM5 and JIM7, and a range of other cytochemical procedures. IIF showed that the alternately-arranged pits are pre-figured as perforations within a reticulum of randomly-oriented CMTs before any secondary wall thickening is obvious. Each incipient pit border is subsequently delimited by a circle of CMTs whose diameter decreases as deposition of secondary wall takes place around the perforation. These IIF observations are corroborated by a parallel TEM study. During the period of bordered pit formation, the secondary walls of the cell are lignifying. At maturity, however, the pit membrane is unlignified and continues to stain strongly with the monoclonal antibody JIM5, a marker of primary, “juvenile” wall. The results are discussed in terms of the relationship of the CMT cytoskeleton with development of bordered pits.

Keywords

Aesculus hippocastanum LMicrotubulesIndirect immunofluorescence microscopyVascular cambiumSecondary xylem vessel elementsBordered pits

Abbrevations

BSA

bovine serum albumin

CMT(s)

cortical micro-tubule(s)

EGTA

ethylene glycol-bis-(ß-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

FITC

fluorescein isothiocyanate

IIP

indirect immunofluorescence

MAP

microtubule-associated protein

mf(s)

wall microfibril(s)

MTSB

microtubule-stabilising buffer

PATAg

periodic acid-thiocarbohydrazide-silver proteinate

PBS

Phosphate-buffered saline

PIPES

piperazine-N,N′-bis-[2-ethylsulphonic acid]

SVS

secondary vascular system

TEM

transmission electron microscopy

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • N. J. Chaffey
    • 1
    • 2
  • J. R. Barnett
    • 1
  • P. W. Barlow
    • 2
  1. 1.Department of Botany, Plant Sciences LaboratoriesThe University of ReadingReading
  2. 2.Institute of Arable Crops Research, Long Ashton Research Station, Department of Agricultural SciencesUniversity of BristolLong AshtonUK