Table Of ContentRECENT ADVANCES OF PLANT ROOT STRUCTURE AND FUNCTION
Developments in Plant and Soil Sciences
VOLUME 90
RECENT ADVANCES OF PLANT ROOT
STRUCTURE AND FUNCTION
PROCEEDINGS OF THE 5TH INTERNATIONAL
SYMPOSIUM ON STRUCTURE AND FUNCTION
OF ROOTS
Stant Lensna, Slovakia, 30 August - 4 September, 1998
Guest Editors:
Otflia GaspanKova,
Milada Ciamporova,
Igor Mistrik and
Frantisek Baluska
....
"
SPRINGER-SCIENCE+BUSINESS MEDIA, B.V.
A C.LP. Catalogue record for this book is available from
the library of Congress.
ISBN 978-90-481-5598-9 ISBN 978-94-017-2858-4 (eBook)
DOI 10.1007/978-94-017-2858-4
Printed an acid-free paper
AII Rights reserved
© 2001 Springer Science+Business Media Dordrecht
Originally published by Kluwer Academic Publishers in 2001
Softcover reprint of tbe hardcover 1s t edition 200 1
N o part of the material protected by this copyright notice
may be reproduced ar utilized in any form or by any means,
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CONTENTS
Preface
I. STRUCTURAL AND MOLECULAR ASPECTS OF ROOT GROWTH AND DEVELOPMENT
• Histochemical analysis of root meristem activity in Arabidopsis thaliana using a cyclin:GUS «(3-
glucuronidase) marker line
M.-T. Hauser and E. Bauer 3
• Ultrastructural effects of the herbicide chlorpropham (CIPC) in root tip cells of wheat
E.P. Eleftheriou and E. Bekiari 13
• Measurement of viscoelastic properties of root cell walls affected by low pH in lateral roots of Pisum
sativum L.
E. Tanimoto, S. Fujii, R. Yamamoto and S. Inanaga 23
Secondary dilatation growth in the root endodermis
A. Lux and M. Luxova 31
Changes of root structure in connection with the development of lateral root primordia in wheat and
pumpkins
K.N. Demchenko and N.P. Demchenko 39
Some aspects of endodermis and cortex structure in Dryas roots
K.M. Barmicheva and M.E Danilova 49
• IAA and ZR content in leek (Allium porrum L.), as influenced by P nutrition and arbuscular mycorrhizae,
in relation to plant development
A. Torelli, A. Trotta, L. Acerbi, G. Arcidiacono, G. Berta and C. Branca 55
• Polyploidy in tomato roots as affected by arbuscular mycorrhizal colonization
G. Berta, A. Fusconi, S. Sampo, G. Lingua, S. Perticone and O. Repetto 63
II. PHYSIOLOGY OF WATER AND ION UPTAKE
• Water uptake by plant roots: an integration of views
E. Steudle 71
• Root-shoot interactions in mineral nutrition
W.D. Jeschke and W. Hartung 83
Calcium channels in the plasma membrane of root cells: Their roles in mineral nutrition and cell signalling
PJ. White and M.S. Ridout 97
• Nitrogen and base cation uptake in seedlings of Acer pseudoplatanus and Calamagrostis villosa exposed
to an acidified environment
V. Gloser and J. Gloser 103
• Nitrate uptake by bean (Phaseolus vulgaris L.) roots under phosphate deficiency
A. Gniazdowska and A.M. Rychter III
Nitrate reductase in roots: Succinate-and NADH-dependent plasma membrane-bound forms
C. StOhr, S. Wienkoop and w.R. Ullrich 119
• Impact of Agrobacterium tumefaciens-induced stem tumors on N03 uptake in Ricinus communis
1. Mistrik, J. Pavlovkin, R. Wachter, K.S. Pradel, K. Schwalm, W. Hartung, U. Mathesius, C. StOhr and
C.L Ullrich 123
Fast shoot responses to root treatment. Are hormones involved?
G. Kudoyarova, D. Veselov, M. Symonyan, A. Mustafina, A. Dedov, L. Vysotzkaya and S. Veselov 135
III. ROOT UNDER STRESS CONDITIONS
• Response of tomato plants to chilling stress in association with nutrient or phosphorus starvation
z. Starck, B. Niemyska, 1. Bogdan and R.N. Akour Tawalbeh 143
Effect of water stress upon root meristems of pea seedlings: The role of quantitative and qualitative changes
in protein patterns
D. Chiatante, L. Maiuro, G.S. Scippa and A. Di Iorio 151
• The distribution of lead in duckweed (Lemna minor L.) root tip
S. Samardakiewicz and A. Wozny 159
Lead effects on cereal roots in terms of cell growth, root architecture and metal accumulation
N.v. Obroucheva, V.B. Ivanov, M. Sobotik, H. Bergmann, O.v. Antipova, E.L Bystrova, LV. Seregin and 165
L.K. Shpigun
• Diverse responses of root cell structure to aluminium stress
M. Ciamporova 171
• Relationship between microtubule orientation and patterns of cell expansion in developing cortical cells
of Lemna minor roots
S. Inada and S. Sato 175
* Reprinted from Plant and Soil Volume 226, No.1 (2000).
Recent Advances of Plant Root Structure and Function, 1-2
Edited by O. Gasparllwva, M. Ciamporova, I. Mistrlk and F. HaluSka
© 2001 Kluwer Academic Publishers.
Preface
It has been claimed that many of the most im supplied by roots comprise roughly half the atoms of
portant chemical and physical processes which drive each photosynthetically assembled glucose molecule
change in the material and biological world take place and thus the many substances derived from it that build
at interfaces. It is here that processes such as adsorp and maintain living organisms of all kinds. If we add to
tion and de-absorption, uptake and efflux, attraction this the importance for respiration of the oxygen gen
and repulsion, and slip and slide re-arrange molecules erated from this same water we are left in little doubt
and elements in ways that alter their properties rad about the key role played by roots and their interface
ically by provoking new interactions. It is difficult with the soil in the original colonisation of the land
to find a functional interface of greater significance and its healthy continuance.
for terrestrial life than the one that exists between Thus placing roots in their true perspective makes a
plant roots and the soil they exploit. Only when this stirring story that contrasts strongly with their modest
complex interface between bio-, geo-and hydrophase outward appearance, at least as individual roots axes.
became established in relatively recent biological and That roots are largely hidden from our view adds a
geological time (400 million years ago) could ex further piquancy since, for the majority of plant sci
tensive land colonization and evolution of terrestrial entists, out of sight has meant out of mind despite the
life begin in earnest. Likely reasons for this depend overwhelming importance of the activities of roots to
ency on roots and their interactions with mineral-and living things. There can be little doubt that the excite
water-bearing substrates are not hard to find. At their ment created by the opening up this hitherto secret but
heart are the gravity-guided penetrative and branch vital world of plant roots through experimental and
ing properties of roots that enable soil and more rocky observational studies inspired Professor Iozef Kolek
substrates to be explored thoroughly and in a steadily and his Slovak colleagues to arrange the first Interna
expanding way that keeps in proportion to the aerial tional Symposium on Structure and Function of Roots
shoot system it supports both physically (for stability) at Tatranska Lomnica in 1971. This sparked a suc
and chemically. This presents an enormous absorptive cessful and influential series of international symposia
interface to the soil (several hundred square meters devoted to the investigation of plant roots and atten
even in a modest herbaceous plant) that selectively ded by root devotees from many countries, almost as a
takes-in the twenty four or so essential elements from necessary pilgrimage. The most recent of these meet
highly diluted solutions (J-LM concentrations) plus a ings (the fifth) held at Stara Lesna, Slovak Republic in
few others, even gold. This impressive permeation el September 1998 was attended by root scientist from 17
evates metabolically driven nanoscale absorption pro countries. The symposium was organized by Dr. Otilia
cesses in to a vast mining operation that sustains the Ga panKova and her colleagues of the Institute of
mineral requirements not only of plants but almost Botany and the Commenius University in Bratislava.
all other forms of terrestrial life. Without the action They have sustained and nourished a fine tradition for
of roots the minerals would remain locked away and which a grateful botanical community benefits much.
mostly unavailable to non-aquatic life. The root/soil Some chauvinists may claim that plant science has
interface is also essential for water uptake in amounts reached a stage where focusing on different organs,
that maintains cell hydration and expansive growth, species, even phyla is a distraction as we increasingly
and creates the medium for cellular biochemistry. Of recognize the extensive homeology between the gen
even more fundamental significance perhaps is that omes of disparate plant species and also their closely
water supplied by roots serves as a co-substrate in similar regulatory mechanisms such as those of signal
photosynthesis along with carbon dioxide. This means sensing and transduction. Gene sequence databases
that hydrogen atoms split-off by photolysis from water contain no taxonomic or morphological boundaries
2
and it can be argued that roots should be studied only to the next international conference about plant root
where they offer advantages for understanding basic systems. This book presents selected contributions to
processes that apply more widely. However, as we the Fifth International Symposium on Structure and
have seen, roots possess unique characteristics that Function of Roots.
played a crucial role in establishing life on the land
and continue to sustain it. The human race depends on Michael B. Jackson
them and this alone is reason enough to study roots in IACR-Long Ashton Research Station
their own right and for us to look forward expectantly
Recent Advances of Plant Root Structure and Function, 3-12 3
Edited by 0, Gasparlkova, M. Ciamporova, I. Mistrfk and F Baluska
© 2001 Kluwer Academic Publishers.
Histochemical analysis of root meristem activity in Arabidopsis thaliana
using a cyclin:GUS (f3 -glucuronidase) marker line
Marie-Theres Hauser and Elisabeth Bauer
Center of Applied Genetics, University of Agricultural Sciences Vienna, Muthgasse 18, A-1190 Vienna, Austria.
E-mail: [email protected]
Key words: Arabidopsis thaliana, cell cycle, root meristem, mitotic cyclin, postembryonic development
Abstract
We used a transgenic Arabidopsis line expressing a translational fusion between a mitotic cyclin and the reporter
gene ,B-glucuronidase (GUS) to investigate cell divisions in postembryonic root meristems. The fusion protein
contains the cyclin destruction box (CDB) and this leads to a rapid degradation of the chimeric GUS-protein after
mitosis. Hence, the staining pattern of the meristem marks dividing cells. We observed that upon germination the
first cell divisions occur in epidermis cells at the junction with the hypocotyl. Moreover, the accelerated root growth
on media supplemented with sucrose correlates with an increased number of dividing cells and an enlargement of
the root meristematic zone. The conditional root expansion mutants porn poml and procusteJ (quill) suppress
this sugar effect leading to a smaller meristematic zone. Simultaneous visualisation of the nucleus revealed that
the CYCATl:CDB:GUS expression is subcellularly localised around the nucleus. This particular staining starts at
prophase and disappears after the completion of the new cell wall. In metaphase the staining invades the cytoplasm
whereas in the telophase it concentrates again around the nucleus. This cell cycle-dependent distribution was
used to characterise the two root specific cytokinesis mutants pleiadel and hyadel. In both mutants, cells which
fail to develop a complete cell wall during cytokinesis divide synchronously in further cell divisions leading to
multinucleate cells. These experiments demonstrate the usefulness of the CYCATl :CDB:GUS marker line for
studying cell division of wild-type and mutants. Furthermore, this line can be used to analyse the influence of
biotic and abiotic signals on the rate and spatial distribution of cell divisions.
Introduction cell length measurements, rates of local cell produc
tion (cells per mm and h) and cell division (cells per
Most of the plant organs are generated by postem cell and h) for the root cortex and the two epidermal
bryonic cell divisions realised by meristems at the end cells types, trichoblasts and atrichoblasts in Arabidop
of the apical-basal axis of the plant. The meristem sis root meristems. All these methods were applied
activity, i.e. the rate at which daughter cells are pro on already established postembryonic root meristems.
duced is developmentally controlled and modified by We wanted to analyse cell division frequencies during
environmental signals such as light or nutrient avail germination and under different growth conditions as
ability. Classical experimental approaches to asses the well as in different root morphogenesis mutants.
meristematic activity are 3H-thymidine and bromod As scoring mitotic frequencies in sections is dif
eoxyuridine feeding which specifically labels cells in ficult, we decided to use a mitosis specific marker
S-phase (Dolan et aI., 1993; Scheres et aI., 1994). line to test their usefulness for studying and measuring
Other studies counted mitotic figures in serial sec meristem activity.
tions of root meristems. An extensive overview of the The cell cycle machinery is regulated by the cyc
microscopic and mathematical methods used to de lins which together with cyclin-dependent kinases
termine the relationship between cell proliferation and (CDKs) are necessary for cell cycle progression
elongation is given by Ivanov (1997). Recently, Beem (Evans et aI., 1983). While plant CDKs are expressed
ster and Baskin (1998) calculated from kinematic and throughout the cell cycle in division competent cells
4
(Hemedy et al., 1993; Martinez et al., 1992), expres Materials and methods
sion of cyclins oscillates in these cells (Doerner et
al., 1996; Fobert et al., 1994; Hemedy et al., 1992). Plant strains and growth conditions
Different classes of cyclins regulate similar cell cycle
controls and checkpoints in animals and plants. For The CYCATl:CDB:GUS line of ecotype Columbia
example, A type cyclins are expressed during S-phase was kindly provided by John Celenza. The mutants
until the G2 phase and are destroyed during M phase poml-5, poml-21, prcl-ll (former name: quil-3),
whereas the B type cyclins accumulate only late dur hyal-l and plel-3 were isolated in the lab during sev
ing G2 phase and start also to be destroyed parallel to eral mutant screens (Hauser and Benfey, 1994; Hauser
the end of mitosis (Ferreira et aI., 1994; Gentschik, et al., 1995; Hauser, unpublished). Genetic crosses
et al., 1998; Hirt et al., 1992; Kouchi et al., 1995; between these mutants and the CYCAT1:CDB:GUS
Meskiene et aI., 1995; Setiady et aI., 1995; Shaul et aI., line were performed in both directions and Fl plants
1996). The A and B type cyclins contain a conserved were allowed to self-fertilise. F2 seeds were plated as
motif of nine amino acids at their N-terminus which described below and wild type as well as mutant plants
resembles the destruction box of mitotic cyclins of an were separately stained for GUS activity (see below).
imals and fungi. Minor structural differences of this To observe the root phenotype and to analyse
motif probably reflect the functional distinctions in the young seedlings, seeds were sterilised with 5% so
mode and timing of cyclin degradation (Renaudin et dium hypochlorite, 0.5% Tween-20 solution for about
al., 1996). 3 min, washed twice with sterile water and plated on
The cyclin Arath:CycBl;1 (former name: At agar-solidified Murashige & Skoog (1962) medium
cyc1; Renaudin et aI., 1996) which was used for the without or with 4.5% (~130 mM) sucrose. After 2
CYCATl :CDB:GUS marker line belongs to the B type days of stratification at 4°C 12x12 cm square tis
cylins and is known to be expressed during mitosis sue culture plates were vertically placed in a growth
in root meristematic cells (Doerner et al., 1996). For chamber under 16 h light (80-90 ILmol m-2 s-l) and
the translational fusion the first 150 amino acids ofthe 8 h dark cycles at 22°C.
cyclin protein were combined with the normally stable
f3-g1ucuronidase gene (John Celenza, personal com Histological staining
munication). The expressed chimeric protein therefore
contains the cyclin B type destruction box (CDB) Seedlings were stained from 30 min to several hours in
which resides between amino acid 40 and 50 of the 250 ILL freshly prepared staining solution by incuba
N-terminus of the Arath:CycBl;1. Hence, the activ tion at 37 °C on microscope slides. A 1-2 mm high
ity of the chimeric GUS protein detected by histo barrier of vaseline on the microscope slide avoided
chemical analysis marks cells in late G2 and mitosis. pressure damage of the plant organs and reduced
Moreover, GUS staining of transgenic Arabidopsis evaporation. The staining solution contained 100 mM
plants expressing this construct have a spotty pattern TrislHCI pH 7.5, 50 mM NaCl, 2mM K3Fe(CN)6,
indicating that the chimeric protein is degraded rap 20% MeOH and 0.5-1 mg/ml X-Gluc (5-bromo-4-
idly. In this paper we describe the difference between chloro-3-indolyl-f3-D-glucuronic acid cyclohexylam
embryonic and postembryonic root meristems. We use monium salt, Biomol). Pictures were taken from
the CYCATl:CDB:GUS marker line to visualise and cleared preparations. Clearing was done in microtiter
quantify the temporal and spatial distribution of divid plates after the following protocol. Mter staining,
ing cells within the root meristem of wild type and root seedlings were treated for 15 min at 57°C in 0.24
morphogensis mutants. Furthermore, the marker line N HCl in 20% MeOH, then transferred to 7% NaOH
facilitates the quantification of the stimulatory effect in 60% EtOH for 15 min at room temperature. Seed
of sucrose on the activity of the root meristem. We re lings were rehydrated in 40%, 20% and 10% EtOH
port some preliminary observations on the subcellular each 5 min and finally 5% EtOH, 25% glycerol for 15
distribution of the cyclin destruction box containing min. For visualising the nuclei, the GUS stained seed
reporter gene during cell division. lings were incubated with the following solutions for
15 min each: 50% EtOH, 75% EtOH, 1 ILg/ml DAPI
(4,6,-diamino-2-phenylindo1e, Sigma) in H20, 30%
glycerol and 50% glycerol and mounted in 70% gly
cerol on microscope slides. All samples were observed
