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Aims: Most plants produce a root tip extracellular matrix that includes viable border cell populations programmed to disperse into soil. Like neutrophils, border cells export structures that trap pathogens and prevent root tip infection. Border cells also trap metals. The goal of this study was to determine if border cells trap Pb.Methods: Border cell responses to Pb were observed microscopically. Border cell impact on Pb-induced injury to roots was assessed using root growth assays. Pb removal from solution was measured using inductively coupled plasma mass spectrometry (ICP-MS). Speciation of Pb associated with border cells was evaluated by synchrotron X-ray absorption spectroscopy (XAS).Results: Increased border cell trap size and number occurred within minutes in response to Pb but not silicon (Si). Transient immersion of root tips into Pb after border cells were removed resulted in growth inhibition. Immersion of root tips and border cells into Pb solution resulted in significant removal of Pb. Si levels in the presence of root tips remained unchanged. The Pb speciation, measured with Pb LIII XAS, altered when reacted with border cells, indicating that direct binding by extracellular traps occurred.Conclusions: Border cells can trap Pb and prevent damage to the root tip. [ABSTRACT FROM AUTHOR]

Background and aims: In a previous study, a compost water extract (CWE) applied to pea seedlings resulted in >95 % protection against root infection. The protection was correlated with retention of a sheath of root border cells surrounding each root tip. A transient exposure to CWE was correlated with 80 % reduction in infection, and with retention of border cell sheaths. Early effects of CWE on border cell dispersal therefore were examined. Materials and methods: Temporal and spatial dynamics of pea, maize, cotton, and cucumber border cell dispersal into water or CWE were measured. Results: Border cells formed a mass surrounding root tips within seconds after exposure to water, and most cells dispersed into suspension spontaneously. In CWE, >90 % of the border cell population instead remained appressed to the root surface, even after vigorous agitation. In cotton, for example, >25,000 border cells dispersed within seconds of immersion in water, but <100 border cells dispersed after >24 h in CWE. Conclusions: Border cells can contribute >90 % of carbon released from young roots, and a single border cell can trap hundreds of bacteria within minutes. The impact of altered border cell dispersal on soil properties, plant nutrition, and root disease development warrants further study. [ABSTRACT FROM AUTHOR]

Background and aims: Recent studies suggest that root border cells function in defense of the root by an extracellular DNA-based trapping mechanism similar to that described in mammalian white blood cells. Genes controlling the specialized properties of border cells as they detach from the root tip therefore are of interest. Methods: mRNA differential display was used to identify Brd13, a sequence expressed in border cells but not other root tissues. RNase protection and mRNA Northern blot analyses, and reporter gene expression under the control of the Brd13 promoter in transgenic hairy roots were used to confirm localized expression. Phenotype analysis of transgenic hairy roots expressing Brd13 antisense mRNA was carried out. Results: Brd13 was expressed constitutively in border cells but not in leaves, stems, or roots without border cells. The predicted protein shares sequence similarity with flavin-binding proteins. Transgenic hairy roots expressing Brd13 antisense mRNA exhibited abnormal growth and morphology. Conclusions: We report here that altered expression of a putative flavin-binding protein in border cells resulted in altered root development. Flavin-binding proteins play key roles in development, defense, and local auxin biosynthesis. The Brd13 gene and its promoter may be useful in creating defined changes in root development and defense. [ABSTRACT FROM AUTHOR]

Background: As roots penetrate soil, specialized cells called 'border cells' separate from root caps and contribute a large proportion of exudates forming the rhizosphere. Their function has been unclear. Recent findings suggest that border cells act in a manner similar to that of white blood cells functioning in defense. Histone-linked extracellular DNA (exDNA) and proteins operate as 'neutrophil extracellular traps' to attract and immobilize animal pathogens. DNase treatment reverses trapping and impairs defense, and mutation of pathogen DNase results in loss of virulence. Scope: Histones are among a group of proteins secreted from living border cells. This observation led to the discovery that exDNA also functions in defense of root caps. Experiments revealed that exDNA is synthesized and exported into the surrounding mucilage which attracts, traps and immobilizes pathogens in a host-microbe specific manner. When this plant exDNA is degraded, the normal resistance of the root cap to infection is abolished. Conclusions: Research to define how exDNA may operate in plant immunity is needed. In the meantime, the specificity and stability of exDNA and its association with distinct microbial species may provide an important new tool to monitor when, where, and how soil microbial populations become established as rhizosphere communities. [ABSTRACT FROM AUTHOR]

Here we describe the use of Pisum sativum L. as a model system to measure how short-term treatment of root tips with soluble metabolites can influence root growth and release of root exudates. The results revealed that even a 3-minute exposure of root tips to metabolites normally released from roots into the rhizosphere (e.g. rhamnose, ferulic acid, salicylic acid) can significantly influence root growth without affecting production of border cells and associated exudates. Conversely, products including caffeine, saccharide lactone, and pisatin alter production of border cells, without affecting root growth. Understanding how root-derived and exogenous metabolites can selectively impact root function may yield benefits in crop production, especially in greenhouse agriculture systems where growing roots can be exposed to a significant accumulation of plant exudates. [ABSTRACT FROM AUTHOR]

Root cap development in cereals and legumes is self-regulated by a repressor that accumulates in the extracellular environment, and immersing the root tip into water results in renewed cap development. By exploiting this phenomenon, root cap mitosis and differentiation can be synchronously induced among populations. In Pisum sativum L., messenger RNA (mRNA) differential display revealed changes in expression of approximately 1% of the sample mRNA population within minutes of induced cap turnover. This profile changes sequentially over a period of 30 min, then stabilizes. Microarray analysis of Medicago truncatula root caps confirmed changes in expression of approximately 1% of the target population, within minutes. A cell specific marker for cap turnover exhibited the same temporal and spatial expression profile in the gymnosperm species Norway spruce ( Picea abies) as in pea. Induced cap development provides a means to profile cell-specific gene expression among phylogenetically diverse species from the early moments of mitosis and cellular differentiation. [ABSTRACT FROM AUTHOR]

Flavonoids have been proposed to play diverse roles in plant growth and development, including defense, symbiosis, pollen development and male fertility, polar auxin transport, and protection against ultraviolet radiation. Recently, a new role in cell cycle regulation has emerged. Genetic alteration of glucuronide metabolism by altered expression of aPisum sativumUDP-glucuronosyltransferase (PsUGT1) results in an altered cell cycle in pea, alfalfa, andArabidopsis. In alfalfa, altered expression ofPsUGT1results in accumulation of a flavonoid-like compound that suppresses growth of cultured cells. The results are consistent with the hypothesis thatPsUGT1functions by controlling cellular levels of a factor controlling cell cycle (FCC). [ABSTRACT FROM AUTHOR]

GUSA from Escherichia coli, encoded by the uidA gene, has been successfully used as a plant reporter system for more than a decade with no reported deleterious effects. However, when expressed in coordination with a UDP-glucuronosyltransferase isolated from the root cap meristem of Pisum sativum (PsUGT1) at the onset of mitosis, GUSA expression was lethal in pea, alfalfa, and Arabidopsis thaliana. These unexpected results indicate that, under some circumstances, using GUSA in plants is incompatible with life and suggest that the cell-specific lethal phenotype might be useful in selecting for genes specifically involved in regulating the G2-M phase of the cell cycle. [ABSTRACT FROM AUTHOR]

Discusses 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. Factors affecting the release of border cells from the outer surface of the root cap; Development and structure of root caps; Model for root cap-rhizosphere interactions.

Two cDNA clones representing mRNAs, highly expressed in pea root tips, were isolated by mRNA differential display. Ribonuclease protection analyses showed different patterns of expression of these two messages in several pea tissues. Sequence analysis showed that the first clone, PsH1b-40, has 100% homology with a previously isolated H1 histone cDNA, PsH1b. However, it has an additional 30 nt at the 3′ end which is absent in PsH1b, suggesting possible multiple polyadenylation sites in the same mRNA. The second clone, PsH1b-41, encodes a deduced 19.5 kDa protein of 185 amino acids with an isoelectric point of 11.5. The putative globular domain of the encoded protein showed 67-71% residue identity with globular domains of 28 kDa pea PsH1b H1 histone and Arabidopsis thaliana H1-1 H1 histone. It has 9 repeating motifs of (T/S)XXK. In the C-terminal domain, there are four lysine-rich repeating motifs of SXK(T/S)PXKKXK which may be involved in chromatin condensation and decondensation. Southern blot analysis of nuclear DNA shows that PsH1-41 belongs to a multigene family. [ABSTRACT FROM AUTHOR]

Removal of border cells from pea roots synchronizes and induces root cap cell division, wall biogenesis and differentiation. Three messages which are expressed differentially in such induced root caps have been cloned. Sequence analyses showed that the PsHRGP1-encoded protein has high homology with a hydroxyproline-rich glycoprotein. The PsCaP23-encoded protein has high homology with an alfalfa callus protein or translationally controlled human or mouse tumor protein P23. The PsRbL41-encoded protein has high homology with a highly basic 60S ribosomal protein L41. In situ hybridization showed that PsHRGP1, PsCaP23 and PsRbL41 messages are localized within dividing cells of the root cap. PsHRGP1 is highly expressed in uninduced root caps, but its message is repressed by 10–11 times as soon as cell division and differentiation begin. Expression of PsHRGP1 recovers to higher than (180%) its initial level in 30 min. PsHRGP1 is root-specific. PsCaP23 and PsRbL41 messages increase ca. 3-fold within 15 min after root cap induction. All three genes represent small families of 3–5 closely related genes in the pea genome. [ABSTRACT FROM AUTHOR]

A procedure is presented for the detection of mRNA in a single root border cell of pea seedlings using RT-PCR (reverse transcription-polymerase chain reaction) and direct incorporation of digoxigenin-dUTP. It includes cell preservation with aldehyde. Signal is detected using a conjugate of anti-DIG and alkaline phosphatase. [ABSTRACT FROM AUTHOR]

In a world where there is growing pressure to grow more with a smaller environmental footprint, alternative forms of plant protection are needed. The root tips of most plants produce border cells in a mucilage that also contains extracellular DNA (exDNA), which is known to be involved in plant defence. A decade after we first demonstrated that there was varietal difference in the number of border cells in Australian cotton cultivars, we enumerated current commercial cultivars and assessed the level of exDNA produced by individual root tips. The results exposed that there has been a change in the number of border cells per root tip, that cultivar variation still exists and that the recovered levels of exDNA also differs. However, there was no correlation between border cell number and disease resistance to two of the major wilt pathogens and the levels of exDNA did not change when a root tip suspension was incubated with spores of fungal pathogens. The results imply that, while there is potential for border cells and root tip properties to be incorporated into breeding programmes, we still need to develop a better understanding of how root tips are able to influence disease epidemiology if we are to capitalise on this phenotypic property. [ABSTRACT FROM AUTHOR]