Neutral red as a probe for confocal laser scanning microscopy studies of plant roots

被引:55
作者
Dubrovsky, Joseph G. [1 ]
Guttenberger, Martin
Saralegui, Andres
Napsucialy-Mendivil, Selene
Voigt, Boris
Baluska, Frantisek
Menzel, Diedrik
机构
[1] Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Biol Mol Plantas, Cuernavaca 62191, Morelos, Mexico
[2] Univ Nacl Autonoma Mexico, Inst Biotecnol, Unidad Microscopia, Cuernavaca 62191, Morelos, Mexico
[3] Univ Tubingen, Entwicklungsgenet, Tubingen, Germany
[4] Univ Bonn, Inst Mol & Cellular Biol, D-5300 Bonn, Germany
关键词
CLSM; casparian band; xylem; vacuole biogenesis; lateral root development; root development; neutral red;
D O I
10.1093/aob/mcl045
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
center dot Background and Aims Neutral red (NR), a lipophilic phenazine dye, has been widely used in various biological systems as a vital stain for bright-field microscopy. In its unprotonated form it penetrates the plasma membrane and tonoplast of viable plant cells, then due to protonation it becomes trapped in acidic compartments. The possible applications of NR for confocal laser scanning microscopy (CLSM) studies were examined in various aspects of plant root biology. center dot Methods NR was used as a fluorochrome for living roots of Phaseolus vulgaris, Allium cepa, A. porrum and Arabidopsis thaliana (wild-type and transgenic GFP-carrying lines). The tissues were visualized using CLSM. The effect of NR on the integrity of the cytoskeleton and the growth rate of arabidopsis primary roots was analysed to judge potential toxic effects of the dye. center dot Key Results The main advantages of the use of NR are related to the fact that NR rapidly penetrates root tissues, has affinity to suberin and lignin, and accumulates in the vacuoles. It is shown that NR is a suitable probe for visualization of proto- and metaxylem elements, Caspafian bands in the endodermis, and vacuoles in cells of living roots. The actin cytoskeleton and the microtubule system of the cells, as well as the dynamics of root growth, remain unchanged after short-term application of NR, indicating a relatively low toxicity of this chemical. It was also found that NR is a useful probe for the observation of the internal structures of root nodules and of fungal hyphae in vesicular-arbuscular mycorrhizas. center dot Conclusions Ease, low cost and absence of tissue processing make NR a useful probe for structural, developmental and vacuole-biogenetic studies of plant roots with CLSM.
引用
收藏
页码:1127 / 1138
页数:12
相关论文
共 78 条
[1]   The Arabidopsis TDS4 gene encodes leucoanthocyanidin dioxygenase (LDOX) and is essential for proanthocyanidin synthesis and vacuole development [J].
Abrahams, S ;
Lee, E ;
Walker, AR ;
Tanner, GJ ;
Larkin, PJ ;
Ashton, AR .
PLANT JOURNAL, 2003, 35 (05) :624-636
[2]   Complete disintegration of the microtubular cytoskeleton precedes its auxin-mediated reconstruction in postmitotic maize root cells [J].
Baluska, F ;
Barlow, PW ;
Volkmann, D .
PLANT AND CELL PHYSIOLOGY, 1996, 37 (07) :1013-1021
[3]   THE MICROTUBULAR CYTOSKELETON IN CELLS OF COLD-TREATED ROOTS OF MAIZE (ZEA-MAYS L) SHOWS TISSUE-SPECIFIC RESPONSES [J].
BALUSKA, F ;
PARKER, JS ;
BARLOW, PW .
PROTOPLASMA, 1993, 172 (2-4) :84-96
[4]  
Baluska F, 1997, EUR J CELL BIOL, V72, P113
[5]   A polarity crossroad in the transition growth zone of maize root apices: Cytoskeletal and developmental implications [J].
Baluska, F ;
Volkmann, D ;
Barlow, PW .
JOURNAL OF PLANT GROWTH REGULATION, 2001, 20 (02) :170-181
[6]   Latrunculin B-induced plant dwarfism:: Plant cell elongation is F-actin-dependent [J].
Baluska, F ;
Jasik, J ;
Edelmann, HG ;
Salajová, T ;
Volkmann, D .
DEVELOPMENTAL BIOLOGY, 2001, 231 (01) :113-124
[7]   Cytoskeletal, perspectives on root growth and morphogenesis [J].
Barlow, PW ;
Baluska, F .
ANNUAL REVIEW OF PLANT PHYSIOLOGY AND PLANT MOLECULAR BIOLOGY, 2000, 51 :289-322
[8]   Plant growth and phosphorus accumulation of wild type and two root hair mutants of Arabidopsis thaliana (Brassicaceae) [J].
Bates, TR ;
Lynch, JP .
AMERICAN JOURNAL OF BOTANY, 2000, 87 (07) :958-963
[9]   Local, efflux-dependent auxin gradients as a common module for plant organ formation [J].
Benková, E ;
Michniewicz, M ;
Sauer, M ;
Teichmann, T ;
Seifertová, D ;
Jürgens, G ;
Friml, J .
CELL, 2003, 115 (05) :591-602
[10]   Soil CO2 concentration does not affect growth or root respiration in bean or citrus [J].
Bouma, TJ ;
Nielsen, KL ;
Eissenstat, DM ;
Lynch, JP .
PLANT CELL AND ENVIRONMENT, 1997, 20 (12) :1495-1505