Effect of pectin methylesterase gene expression on pea root development.

Expression of an inducible gene with sequences common to genes encoding pectin methylesterase (PME) was found to be tightly correlated, both spatially and temporally, with border cell separation in pea root caps. Partial inhibition of the gene's expression by antisense mRNA in transgenic pea hairy roots prevented the normal separation of root border cells from the root tip into the external environment. This phenotype was correlated with an increase in extracellular pH, reduced root elongation, and altered cellular morphology. The translation product of the gene exhibited PME activity in vitro. These results are consistent with the long-standing hypothesis that the demethylation of pectin by PME plays a key role in cell wall metabolism.     

Effects of Al<Superscript>3+</Superscript> on the biological characteristics of cowpea root border cells

Root border cells (RBC) are cells surrounding the root apex. They are functionally different from the apex and are considered to play a role in the protection of the root tip from biotic and abiotic stresses. We investigated RBC viability, formation, and pectin methylesterase (PME) activity of the root caps during RBC development in cowpea (Vigna ungniculata ssp. sesquipedalis) under aeroponic culture. The results showed that the border cells formed almost synchronously with the emergence of the root tip. The number of border cells reached the maximum when roots were approximately 15 mm long. Pectin methylesterase (PME) activity of the root cap peaked at a root length of 1 mm. Root border cells separated from the root cap died within 24 h under Al³⁺ stress while those still attached to the root cap maintained 85% viability at 48 h after treatment. The PME activity did not differ significantly under different Al³⁺ treatments.     

Root border cell development is a temperature-insensitive and Al-sensitive process in barley

In vivo and in vitro experiments showed that border cell (BC) survival was dependent on root tip mucigel in barley (Hordeum vulgare L. cv. Hang 981). In aeroponic culture, BC development was an induced process in barley, whereas in hydroponic culture, it was a kinetic equilibrium process during which 300-400 BCs were released into water daily. The response of root elongation to temperatures (10-35 degrees C) was very sensitive but temperature changes had no great effect on barley BC development. At 35 degrees C, the root elongation ceased whereas BC production still continued, indicating that the two processes might be regulated independently under high temperature (35 degrees C) stress. Fifty microM Al could inhibit significantly BC development by inhibiting pectin methylesterase activity in the root cap of cv. 2000-2 (Al-sensitive) and cv. Humai 16 (Al-tolerant), but 20 microM Al could not block BC development in cv. Humai 16. BCs and their mucigel of barley had a limited role in the protection of Al-induced inhibition of root elongation, but played a significant role in the prevention of Al from diffusing into the meristems of the root tip and the root cap. Together, these results suggested that BC development was a temperature-insensitive but Al-sensitive process, and that BCs and their mucigel played an important role in the protection of root tip and root cap meristems from Al toxicity.     

高等植物根边缘细胞的发育调控及其生物学功能

植物根边缘细胞是从根冠表皮游离出来并聚集在根尖周围的一群特殊细胞,以前曾称为根冠脱落细胞.最近的证据表明,绝大多数物种边缘细胞具有生物学活性,其发育是受内外信号调控.边缘细胞一旦从根表皮游离后,其代谢活性大大上升、基因表达明显不同于根冠细胞.最近,与边缘细胞发育早期和晚期相关的两个基因PsUGT1和RCPME1分别被克隆和鉴定.边缘细胞能特异性地合成、分泌一系列的化学物质,包括花色素苷、抗生素、特异性酶类及其他化学物质能抑制或促进根际周围的细菌、真菌、病毒、线虫等的生长以及中和根际周围一些有毒化学物质如铝毒.因此,边缘细胞在植物生长发育过程中起着多种生物学功能.     

高等植物根边缘细胞的发育调控及其生物学功能

植物根边缘细胞是从根冠表皮游离出来并聚集在根尖周围的一群特殊细胞 ,以前曾称为根冠脱落细胞。最近的证据表明 ,绝大多数物种边缘细胞具有生物学活性 ,其发育是受内外信号调控。边缘细胞一旦从根表皮游离后 ,其代谢活性大大上升、基因表达明显不同于根冠细胞。最近 ,与边缘细胞发育早期和晚期相关的两个基因PsUGT1和RCPME1分别被克隆和鉴定。边缘细胞能特异性地合成、分泌一系列的化学物质 ,包括花色素苷、抗生素、特异性酶类及其他化学物质能抑制或促进根际周围的细菌、真菌、病毒、线虫等的生长以及中和根际周围一些有毒化学物质如铝毒。因此 ,边缘细胞在植物生长发育过程中起着多种生物学功能     

反枝苋水浸提液与挥发油对黄瓜根尖的影响

采用悬空气法研究了在入侵植物反枝苋(Amaranthus retroflexus L.)水浸提液和挥发油作用下,黄瓜根缘细胞活性、根冠果胶甲基酯酶(PME)、根尖过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)以及丙二醛(MDA)含量的变化规律.结果表明:反枝苋水浸提液对黄瓜根的生长无显著性影响而挥发油显著抑制黄瓜根的生长,且随浓度增大抑制作用显著增强.PME活性随着水浸提液浓度的增大呈先上升后下降趋势,而随着挥发油浓度的升高呈现逐渐上升的趋势;水浸提液和挥发油均降低了对根缘细胞的存活率,这种抑制作用随浓度的增加而增大;随着处理液浓度增大,黄瓜根尖中MDA含量、CAT活性整体表现为增加,SOD活性先升高后降低,POD活性与对照差异不显著.反枝苋挥发油的化感效应大于水浸提液的化感效应.     

Differential Expression of Proteins and mRNAs from Border Cells and Root Tips of Pea

Many plants release large numbers of metabolically active root border cells into the rhizosphere. We have proposed that border cells, cells produced by the root cap meristem that separate from the rest of the root upon reaching the periphery of the cap, are a singularly differentiated part of the root system that modulates the environment of the plant root by producing specific substances to be released into the rhizosphere. Proteins synthesized in border cells exhibit profiles that are very distinct from those of the root tip (root cap, root meristem, and adjacent cells). In vivo-labeling experiments demonstrate that 13% of the proteins that are abundant in preparations from border cells are undetectable in root tip preparations. Twenty-five percent of the proteins synthesized by border cells in a 1-h period are rapidly excreted into the incubation medium. Quantitative variation in levels of specific marker proteins, including glutamine synthetase, heat-shock protein 70, and isoflavone reductase, also occurs between border cells and cells in the root tip. mRNA differential-display assays demonstrate that these large qualitative and quantitative differences in protein expression are correlated with similarly distinct patterns of gene expression. These observations are consistent with the hypothesis that a major switch in gene expression accompanies differentiation into root border cells, as expected for cells with specialized functions in plant development.     

Meristem-Specific Suppression of Mitosis and a Global Switch in Gene Expression in the Root Cap of Pea by Endogenous Signals

Two functionally distinct sets of meristematic cells exist within root tips of pea (Pisum sativum): the root apical meristem, which gives rise to the body of the root; and the root cap meristem, which gives rise to cells that differentiate progressively through the cap and separate ultimately from its periphery as border cells. When a specific number of border cells has accumulated on the root cap periphery, mitosis within the root cap meristem, but not the apical meristem, is suppressed. When border cells are removed by immersion of the root tip in water, a transient induction of mitosis in the root cap meristem can be detected starting within 5 min. A corresponding switch in gene expression throughout the root cap occurs in parallel with the increase in mitosis, and new border cells begin to separate from the root cap periphery within 1 h. The induction of renewed border cell production is inhibited by incubating root tips in extracellular material released from border cells. The results are consistent with the hypothesis that operation of the root cap meristem and consequent turnover of the root cap is self-regulated by a signal from border cells.     

Correlation of Pectolytic Enzyme Activity with the Programmed Release of Cells from Root Caps of Pea (Pisum sativum)

In many plant species, the daily release of hundreds to thousands of healthy cells from the root cap into the soil is a normal process, whose function is unknown. We studied the separation of the cells in pea (Pisum sativum) using an aeroponic system in which separated cells were retained on the root until they were washed off for counting. We found that cell separation is a developmentally regulated, temperature-sensitive process that appears to be regulated independently of root growth. No cells were released from very young roots. When plants were grown aeroponically, cell numbers increased with increasing root length to a mean of 3400 cells per root, at which point the release of new cells ceased. The process could be reset and synchronized by washing the root in water to remove shed cells. Cell separation from the root cap was correlated with pectolytic enzyme activity in root cap tissue. Because these cells that separate from the root cap ensheath the root as it grows and thus provide a cellular interface between the root surface and the soil, we propose to call the cells “root border cells.”     

真盐生植物盐地碱蓬根系边缘细胞在耐盐中的作用初探

以黄河三角洲潮间带盐地碱蓬种子生成的幼苗为材料,研究了NaCl胁迫对盐地碱蓬生长与根系边缘细胞的影响。盐地碱蓬的第一个边缘细胞几乎与根尖同步产生,当根长达到13mm时,边缘细胞数目达到最大值。NaCl胁迫抑制边缘细胞的活性,但低浓度的NaCl处理增加边缘细胞的数目。低浓度NaCl处理时果胶甲基酯酶（PME）的活性比对照有明显增加,超氧化物歧化酶（SOD）活性随着NaCl浓度的增加呈现先上升后下降的趋势,低浓度NaCl可以增加盐地碱蓬根内过氧化氢酶（CAT）的活性,NaCl处理时间和处理浓度都对过氧化物酶（POD）活性的影响不明显。这些结果表明,盐地碱蓬至少部分通过增加调控活性氧（ROS）水平增加PME活性及根系边缘细胞数目来抵抗NaCl胁迫。     

Polymorphism and modulation of cell wall esterase enzyme activities in the chicory root during the growing season

Pectins are major components of the primary plant cell wall. They can be both methylesterified and acetylesterified and de-esterification occurs by specific esterases. Proteins extracted by NaCl treatment from root cell walls of two chicory varieties (Cichorium intybus L. cv. Nausica and Arancha) sampled in an experimental field every 2 weeks between July 2002 and January 2003 were analysed by isoelectrofocalization, semi-denaturing SDS-PAGE, and quantitative assays for their esterase activity. Zymograms showed that chicory root pectin methylesterases belong to a multigene family. The isoelectric points of the pectin methylesterase isoforms ranged from pI 3.8 to pI 9.0. Concerning acetylesterases, only acidic isoforms between pI 4.1 and pI 5.2 were observed, but a large polymorphism of this class of enzymes could be identified in one variety. The results indicate that the root pectin methylesterase activity of the Nausica variety was correlated with ambient temperature, while no significant effect of temperature could be detected on any acetylesterase isoform.     

绿豆根边缘细胞发育特性

以东北绿豆为试验材料,采用琼脂悬空培养法,研究了绿豆边缘细胞的发育特性。结果表明:绿豆根尖发育初期根边缘细胞呈球形,随着根尖伸长逐渐发育形成椭圆形、长椭圆形和长条形;发育过程中,根边缘细胞具有较高的存活率,在根长大于10mm后根边缘细胞的存活率均在70%～80%之间并趋于稳定;在根长为25～30mm时根边缘细胞数目达到最大值(约13 000个);根冠果胶甲基酯酶(PME)活性在根长5mm时达到最高值(1.486H+μmol.root cap-1.h-1),此后随着根的伸长,根冠PME活性在1.107～1.256H+μmol.root cap-1.h-1间变化并趋于稳定。     

Extracellular proteins in pea root tip and border cell exudates

Newly generated plant tissue is inherently sensitive to infection. Yet, when pea (Pisum sativum) roots are inoculated with the pea pathogen, Nectria haematococca, most newly generated root tips remain uninfected even though most roots develop lesions just behind the tip in the region of elongation. The resistance mechanism is unknown but is correlated spatially with the presence of border cells on the cap periphery. Previously, an array of >100 extracellular proteins was found to be released while border cell separation proceeds. Here we report that protein secretion from pea root caps is induced in correlation with border cell separation. When this root cap secretome was proteolytically degraded during inoculation of pea roots with N. haematococca, the percentage of infected root tips increased from 4% +/- 3% to 100%. In control experiments, protease treatment of conidia or roots had no effect on growth and development of the fungus or the plant. A complex of >100 extracellular proteins was confirmed, by multidimensional protein identification technology, to comprise the root cap secretome. In addition to defense-related and signaling enzymes known to be present in the plant apoplast were ribosomal proteins, 14-3-3 proteins, and others typically associated with intracellular localization but recently shown to be extracellular components of microbial biofilms. We conclude that the root cap, long known to release a high molecular weight polysaccharide mucilage and thousands of living cells into the incipient rhizosphere, also secretes a complex mixture of proteins that appear to function in protection of the root tip from infection.     

Root Caps and Rhizosphere

In this paper we discuss recent work on the physiological, molecular, and mechanical mechanisms that underlie the capacity of root caps to modulate the properties of the rhizosphere and thereby foster plant growth and development. The root cap initially defines the rhizosphere by its direction of growth, which in turn occurs in response to gradients in soil conditions and gravity. The ability of the root cap to modulate its environment is largely a result of the release of exudates and border cells, and so provides a potential method to engineer the rhizosphere. Factors affecting the release of border cells from the outer surface of the root cap, and function of these cells and their exudates in the rhizosphere, are considered in detail. Release of border cells into the rhizosphere depends on soil matric potential and mechanical impedance, in addition to a host of other environmental conditions. There is good evidence of unidentified feedback signals between border cells and the root cap meristem, and some potential mechanisms are discussed. Root border cells play a significant mechanical role in decreasing frictional resistance to root penetration, and a conceptual model for this function is discussed. Root and border cell exudates influence specific interactions between plant hosts and soil organisms, including pathogenic fungi. The area of exudates and border cell function in soil is an exciting and developing one that awaits the production of appropriate mutant and transgenic lines for further study in the soil environment.     

Root Caps and Rhizosphere

In this paper we discuss recent work on the physiological, molecular, and mechanical mechanisms that underlie the capacity of root caps to modulate the properties of the rhizosphere and thereby foster plant growth and development. The root cap initially defines the rhizosphere by its direction of growth, which in turn occurs in response to gradients in soil conditions and gravity. The ability of the root cap to modulate its environment is largely a result of the release of exudates and border cells, and so provides a potential method to engineer the rhizosphere. Factors affecting the release of border cells from the outer surface of the root cap, and function of these cells and their exudates in the rhizosphere, are considered in detail. Release of border cells into the rhizosphere depends on soil matric potential and mechanical impedance, in addition to a host of other environmental conditions. There is good evidence of unidentified feedback signals between border cells and the root cap meristem, and some potential mechanisms are discussed. Root border cells play a significant mechanical role in decreasing frictional resistance to root penetration, and a conceptual model for this function is discussed. Root and border cell exudates influence specific interactions between plant hosts and soil organisms, including pathogenic fungi. The area of exudates and border cell function in soil is an exciting and developing one that awaits the production of appropriate mutant and transgenic lines for further study in the soil environment.     

Root apical organization inArabidopsis thaliana 1. Root cap and protoderm

Summary We investigated the development of the root cap and protoderm inArabidopsis thaliana root tips.A. Thaliana roots have closed apical organization with the peripheral root cap, columella root cap and protoderm developing from the dermatogen/calyptrogen histogen. The columella root cap arises from columella initials. The initials for the peripheral root cap and protoderm are arranged in a collar and the initiation event for these cells occurs in a sequential pattern that is coordinated with the columella initials. The resulting root cap appears as a series of interconnected spiraling cones. The protoderm, in three-dimensions, is a cylinder composed of cell files made up of packets of cells. The number of cell files within the protoderm cylinder increases as the root ages from one to two weeks. The coordinated division sequence of the dermatogen/calyptrogen and the increase in the number of protoderm cell files are both features of post-embryonic development within the primary root meristem.Abbreviations RCP root cap/protoderm - CI columella initial - PI protoderm initial     

A quantitative cytochemical study of UDP-D-glucose: NAD-oxidoreductase (E.C. 1.1.1.22) activity during stelar differentiation in Pisum sativum L. cv Meteor

A quantitative cytochemical assay for UDP-D-glucose dehydrogenase (UDPGD) activity employing scanning and integrating microdensitometry has been revised and applied to a study of this enzyme during the initiation of secondary cell wall biosynthesis during the formation of primary vascular tissues in roots of Pisum sativum L. cv Meteor. The reaction involves the use of NBT as final electron acceptor and is inhibited 10-fold by either 10 mM UDP-D-xylose or 25 mM UDP-D-glucuronic acid, two molecules involved in feed-back inhibition of UDPGD activity in vivo. UDPGD is a far-from equilibrium enzyme representing a flux-generating step in the biosynthesis of precursors for hemicelluloses involved in secondary cell wall construction, and can be demonstrated to increase sharply in activity in cells of the developing primary vascular elements. This changed activity occurs 18-20 cells back from the root cap junction and coincides with the first cells containing the activated programme for secondary cell wall synthesis.     

Root gravitropism

When a plant root is reoriented within the gravity field, it responds by initiating a curvature which eventually results in vertical growth. Gravity sensing occurs primarily in the root tip. It may involve amyloplast sedimentation in the columella cells of the root cap, or the detection of forces exerted by the mass of the protoplast on opposite sides of its cell wall. Gravisensing activates a signal transduction cascade which results in the asymmetric redistribution of auxin and apoplastic Ca²⁺ across the root tip, with accumulation at the bottom side. The resulting lateral asymmetry in Ca²⁺ and auxin concentration is probably transmitted to the elongation zone where differential cellular elongation occurs until the tip resumes vertical growth. The Cholodny-Went theory proposes that gravity-induced auxin redistribution across a gravistimulated plant organ is responsible for the gravitropic response. However, recent data indicate that the gravity-induced reorientation is more complex, involving both auxin gradient-dependent and auxin gradient-independent events.