INITIATION AND DEVELOPMENT OF ROOT NODULES OF CASUARINA (CASUARINACEAE)

Roots of seedlings of the “beefwood” tree, Casuarina cunninghamiana Miq. grown in nitrogen-free nutrient solution were inoculated with a suspension prepared from crashed root nodules taken from mature plants. Marked deformation of root hairs was evident but no infection threads were observed in root hairs. The mode of infection remains undetermined. Root nodules were initiated within three weeks and thereafter numerous upward-growing nodule roots developed from each nodule. Nodules in this symbiotic nitrogen-fixing plant resulted from an infection caused by an unidentified actinomycete-like soil microorganism. Anatomical analysis of nodule formation showed that nodules are the result of repeated endogenous lateral root initiations, one placed upon another in a complexly branched and truncated root system. The endophyte-infected cortical tissues derived from successive root primordia form the swollen nodular mass. Nodule roots develop from nodule lobes after escaping from the initial inhibitory effects of the endophyte. Included is a discussion of the anatomical similarities between nodules of Casuarina which produce nodule roots and those of Alnus which form coralloid nodules usually lacking nodule roots.     

INTERACTION BETWEEN THE ECTOMYCORRHIZAL FUNGUS PISOLITHUS TINCTORIUS AND ROOT HAIRS OF PICEA MARIANA (PINACEAE)

The ectomycorrhizal fungus Pisolithus tinctorius interacts with roots of Picea mariana to form a typical mantle and Hartig net. Hyphae alter their growth pattern when in contact with susceptible root hairs in the mycorrhizal infection zone and grow acropetally, gradually covering the length of the hair to form a mantlelike structure. Initial contact with the hair may be influenced by a fibrillar material on the root hair surface. Although many root hairs become surrounded by fungal hyphae, they are not penetrated, and therefore are not entry points for this symbiotic fungus.     

SEEDLING DEVELOPMENT OF OPUNTIA BRADTIANA (CACTACEAE)

Seedling development in Opuntia bradtiana, a north-central Mexican endemic, is similar to that of other opuntias, except for the absence of glochids and the fact that germination is extremely slow and germination percentage low. Hypocotyl and root elongation and epicotyl development are rapid for two weeks after rupture of the seed coat. However at this point hypocotyl elongation nearly ceases, while stem and root development continue at a reduced rate. An eight-month seedling is usually not more than 25 mm tall but has numerous areoles with spines and occasionally one subtending leaf. At this time tubercles have begun to coalesce into the vertical rows of ribs characteristic of the section Grusonia.     

STRUCTURAL INVESTIGATIONS OF ASEXUAL REPRODUCTION IN NEPHROLEPIS EXALTATA AND PLATYCERIUM BIFURCATUM

Nephrolepis exaltata cv. Bostoniensis, the Boston fern, exhibits extreme stem dimorphism. The plant has orthotropic, dictyostelic shoots which bear pinnatifid leaves and plagiotropic, protostelic stolons which are aphyllous. Vegetative reproduction occurs by budding from primary and secondary stolons. Secondary stolons arise exogenously from derivatives of the apical cell of the primary stolon, whereas root primordia develop endogenously. Shoots develop in vivo when a creeping stolon makes contact with the substrate via extensive root proliferation. When stolon segments are excised and grown in vitro, secondary stolon primordia expand and initiate leaf primordia, forming new leafy shoots. In Platycerium bifurcatum, the staghorn fern, asexual propagation occurs on ageotropic roots ramifying among the basal nest fronds. Root bud initiation is marked by root tip hypertrophy following cortical parenchyma expansion. Root apical cell derivatives produce the bud apex; the root apical cell remains separate from the developing root bud. Superficially, vegetative reproduction in Nephrolepis and Platycerium appears to involve unusual organs. However, both ferns exhibit leafy bud development from distinct sites of origin, not from undetermined primordia or from direct transformation of root to shoot. Thus, distinctness of organ types is maintained in these two ferns and no evidence for interconvertibility of organ types has been found.     

Scanning Electron Microscopy of Plant Roots

A glycol methacrylate infiltration and polymerization techniquewas used to prepare clover roots inoculated with Rhizobium forscanning reflection electron microscopy. Root hairs and epidermalcells were coated with many bacteria; some bacteria seemed tobe embedded in the wall surface. Root hair tips were often smoothbut some older root hair surfaces showed a fibrillar meshworkpattern. Small granules c. 0.18 µm diameter were presenton the root hair and epidermal cell walls. The root cap, someroot hairs, and some epidermal cells were covered by an amorphousfilm thought to be the mucigel.     

A root hairless barley mutant for elucidating genetic of root hairs and phosphorus uptake

This paper reports a new barley mutant missing root hairs. The mutant was spontaneously discovered among the population of wild type (Pallas, a spring barley cultivar), producing normal, 0.8 mm long root hairs. We have called the mutant bald root barley (brb). Root anatomical studies confirmed the lack of root hairs on mutant roots. Amplified Fragment Length Polymorphism (AFLP) analyses of the genomes of the mutant and Pallas supported that the brb mutant has its genetic background in Pallas. The segregation ratio of selfed F₂ plants, resulting from mutant and Pallas outcross, was 1:3 (–root hairs:+root hairs), suggesting a monogenic recessive mode of inheritance.In rhizosphere studies, Pallas absorbed nearly two times more phosphorus (P) than the mutant. Most of available inorganic P in the root hair zone (0.8 mm) of Pallas was depleted, as indicated by the uniform P depletion profile near its roots. The acid phosphatase (Apase) activity near the roots of Pallas was higher and Pallas mobilised more organic P in the rhizosphere than the mutant. The higher Apase activity near Pallas roots also suggests a link between root hair formation and rhizosphere Apase activity. Hence, root hairs are important for increasing plant P uptake of inorganic as well as mobilisation of organic P in soils.Laboratory, pot and field studies showed that barley cultivars with longer root hairs (1.10 mm), extracted more P from rhizosphere soil, absorbed more P in low-P field (Olsen P=14 mg P kg^–1 soil), and produced more shoot biomass than shorter root hair cultivars (0.63 mm). Especially in low-P soil, the differences in root hair length and P uptake among the cultivars were significantly larger. Based on the results, the perspectives of genetic analysis of root hairs and their importance in P uptake and field performance of cereals are discussed.     

亚热带典型树种根毛特征及其与共生真菌的关系

根毛和共生真菌增加了吸收面积,提高了植物获取磷等土壤资源的能力。由于野外原位观测根表微观结构较为困难,吸收细根、根毛、共生真菌如何相互作用并适应土壤资源供应,缺乏相应的数据和理论。该研究以受磷限制的亚热带森林为对象,选取了21种典型树种,定量了根毛存在情况、属性变异,分析了根毛形态特征与共生真菌侵染率、吸收细根功能属性之间的关系,探讨了根表结构对低磷土壤的响应和适应格局。结果表明:1)在亚热带森林根毛不是普遍存在的, 21个树种中仅发现7个树种存有根毛, 4个为丛枝菌根(AM)树种, 3个为外生菌根(ECM)树种。其中,马尾松(Pinus massoniana)根毛出现率最高,为86%;2)菌根类型是理解根-根毛-共生真菌关系的关键,AM树种根毛密度与共生真菌侵染率正相关,但ECM树种根毛直径与共生真菌侵染率负相关; 3) AM树种根毛长度和根毛直径、ECM树种根毛出现率与土壤有效磷含量呈负相关关系。该研究揭示了不同菌根类型树种根毛-共生真菌-根属性的格局及相互作用,为精细理解养分获取策略奠定了基础。     

<Emphasis Type="Italic">Agrobacterium rhizogenes</Emphasis>: recent developments and promising applications

Agrobacterium rhizogenes is the etiological agent for hairy-root disease (also known as root-mat disease). This bacterium induces the neoplastic growth of plant cells that differentiate to form “hairy roots.” Morphologically, A. rhizogenes-induced hairy roots are very similar in structure to wild-type roots with a few notable exceptions: Root hairs are longer, more numerous, and root systems are more branched and exhibit an agravitropic phenotype. Hairy roots are induced by the incorporation of a bacterial-derived segment of DNA transferred (T-DNA) into the chromosome of the plant cell. The expression of genes encoded within the T-DNA promotes the development and production of roots at the site of infection on most dicotyledonous plants. A key characteristic of hairy roots is their ability to grow quickly in the absence of exogenous plant growth regulators. As a result, hairy roots are widely used as a transgenic tool for the production of metabolites and for the study of gene function in plants. Researchers have utilized this tool to study root development and root–biotic interactions, to overexpress proteins and secondary metabolites, to detoxify environmental pollutants, and to increase drought tolerance. In this review, we provide an up-to-date overview of the current knowledge of how A. rhizogenes induces root formation, on the new uses for A. rhizogenes in tissue culture and composite plant production (wild-type shoots with transgenic roots), and the recent development of a disarmed version of A. rhizogenes for stable transgenic plant production.     

PARASITISM IN PHOLISMA (LENNOACEAE). I. EXTERNAL MORPHOLOGY OF SUBTERRANEAN ORGANS

The external features of the subterranean organs of coastal Californian Pholisma depressum Greene are described. A dimorphic root system exists. Long, stout roots (pilot roots) serve for vegetative propagation in reaching other host roots. Short, unbranching, more slender roots originating from the pilot roots are haustorial in function. Shoots seem to arise only on pilot roots. Host attachment is achieved through the direct transformation of the apical meristem of haustorial roots into the haustorial organ. The basal part of the shoot is also able to produce roots when near host roots, and may form lateral branch shoots either directly or after new haustorial connections are formed. Both types of roots arise endogenously. The invaded host root undergoes considerable hypertrophy. The intrusive organ itself forms a massive but irregular body. More than one haustorial root may enter even a small host root. Self parasitism may occur but is not common.     

SEQUENCE AND PATTERN OF LATERAL ROOT FORMATION IN FIVE SELECTED SPECIES

The proximal-distal distribution of the lateral roots of five species was studied. A detailed investigation was carried out on two of the five species, Ceratopteris thalictroides and Cucurbita maxima. A definite pattern of lateral root arrangement, with a degree of variability related to the number of protoxylem poles, was found in all of the species studied. In the fern Ceratopteris, lateral root initiation was found to be related to the segmentation of the apical cell, which in turn determines the distribution of the laterals. In this species the lateral roots occur in a predictable sequence and they are grouped in pairs. In the angiosperms studied, the pattern of lateral root distribution seemed to depend primarily upon a rather strict longitudinal relationship between the lateral root primordia formed opposite any one protoxylem pole. In Cucurbita maxima, 93.7 ± 5.02% of the lateral root primordia observed were in a specific sequence. The laterals of this species are also arranged in groups. In the other plants studied, Arachis hypogaea, Victoria trickeri, and Eichhornia crassipes, the laterals were not as regularly arranged, but nevertheless they were found to be arranged in groups along the main root axis and not randomly dispersed. Factors controlling the spacing of lateral root primordia include their relationship with the developing vascular system, a direct effect of the parent root apex, and an effect of older lateral root primordia in the same sector of the root.     

Regulation of root hair density by phosphorus availability in Arabidopsis thaliana

We characterized the response of root hair density to phosphorus (P) availability in Arabidopsis thaliana. Arabidopsis plants were grown aseptically in growth media with varied phosphorus concentrations, ranging from 1 mmol m⁻³ to 2000 mmol m⁻³ phosphorus. Root hair density (number of root hairs per mm of root length) was analysed starting at 7 d of growth. Root hair density was highly regulated by phosphorus availability, increasing significantly in roots exposed to low-phosphorus availability. The initial root hairs produced by the radicle were not sensitive to phosphorus availability, but began to respond after 9 d of growth. Root hair density was about five times greater in low phosphorus (1 mmol m⁻³) than in high phosphorus (1000 mmol m⁻³) media. Root hair density decreased logarithmically in response to increasing phosphorus concentrations within that range. Root hair density also increased in response to deficiencies of several other nutrients, but not as strongly as to low phosphorus. Indoleacetic acid (IAA), the auxin transport inhibitor 2-(p-chlorophenoxy)-2-methylpropionic acid (CMPA), the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), and the ethylene synthesis inhibitor amino-oxyacetic acid (AOA) all increased root hair density under high phosphorus but had very little effect under low phosphorus. Low phosphorus significantly changed root anatomy, causing a 9% increase in root diameter, a 31% decrease in the cross-sectional area of individual trichoblasts, a 40% decrease in the cross-sectional area of individual atrichoblasts, and 45% more cortical cells in cross-section. The larger number of cortical cells and smaller epidermal cell size in low phosphorus roots increased the number of trichoblast files from eight to 12. Two-thirds of increased root hair density in low phosphorus roots was caused by increased likelihood of trichoblasts to form hairs, and 33% of the increase was accounted for by changes in low phosphorus root anatomy resulting in an increased number of trichoblast files. These results show that phosphorus availability can fundamentally alter root anatomy, leading to changes in root hair density, which are presumably important for phosphorus acquisition.     

Salt-induced plasticity of root hair development is caused by ion disequilibrium in Arabidopsis thaliana

Root hair development is controlled by environmental signals. Studies on root hair plasticity in Arabidopsis thaliana have mainly focused on phosphate and iron deficiency. Root hair growth and development and their physiological role in response to salt stress are largely unknown. Here, we show that root epidermal cell types and root hair development are highly regulated by salt stress. Root hair length and density decreased significantly in a dose-dependent manner on both primary roots and junction sites between roots and shoots. The root hair growth and development were sensitive to inhibition by ions but not to osmotic stress. High salinity also alters anatomical structure of roots, leading to a decrease in cell number in N positions and enlargement of the cells. Moreover, analysis of the salt overly sensitive mutants indicated that salt-induced root hair response is caused by ion disequilibrium and appears to be an adaptive mechanism that reduces excessive ion uptake. Finally, we show that genes WER, GL3, EGL3, CPC, and GL2 might be involved in cell specification of root epidermis in stressed plants. Taken together, data suggests that salt-induced root hair plasticity represents a coordinated strategy for early stress avoidance and tolerance as well as a morphological sign of stress adaptation.     

Development and function ofAzospirillum-inoculated roots

Summary The surface distribution ofAzospirillum on inoculated roots of maize and wheat is generally similar to that of other members of the rhizoplane microflora. During the first three days, colonization takes place mainly on the root elongation zone, on the base of root hairs and, to a lesser extent, on the surface of young root hairs.Azospirillum has been found in cortical tissues, in regions of lateral root emergence, along the inner cortex, inside xylem vessels and between pith cells. Inoculation of several cultivars of wheat, corn, sorghum and setaria with several strains ofAzospirillum caused morphological changes in root starting immediately after germination. Root length and surface area were differentially affected according to bacterial age and inoculum level. During the first three weeks after germination, the number of root hairs, root hair branches and lateral roots was increased by inoculation, but there was no change in root weight. Root biomass increased at later stages. Cross-sections of inoculated corn and wheat root showed an irregular arrangement of cells in the outer layers of the cortex. These effects on plant morphology may be due to the production of plant growth-promoting substances by the colonizing bacteria or by the plant as a reaction to colonization. Pectic enzymes may also be involved. Morphological changes had a physiological effect on inoculated roots. Specific activities of oxidative enzymes, and lipid and suberin content, were lower in extracts of inoculated roots than in uninoculated controls. This suggests that inoculated roots have a larger proportion of younger roots. The rate of NO^– ₃, K⁺ and H₂PO^– ₄ uptake was greater in inoculated seedlinds. In the field, dry matter, N, P and K accumulated at faster rates, and water content was higher inAzospirillum-inoculated corn, sorghum, wheat and setaria. The above improvements in root development and function lead in many cases to higher crop yield.     

Effects of temperature on parameters of root growth relevant to nutrient uptake: Measurements on oilseed rape and barley grown in flowing nutrient solution

Summary Effects of root temperature on the growth and morphology of roots were measured in oilseed rape (Brassica napus L.) and barley (Hordeum vulgare L.). Plants were grown in flowing solution culture and acclimatized over several weeks to a root temperature of 5°C prior to treatment at a range of root temperatures between 3 and 25°C, with common shoot temperature. Root temperature affected root extension, mean radius, root surface area, numbers and lengths of root hairs. Total root length of rape plants increased with temperature over the range 3–9°C, but was constant at higher temperatures. Root length of barley increased with temperature in the range 3–25°C, by a factor of 27 after 20 days. Root radii had a lognormal distribution and their means decreased with increasing temperature from 0.14 mm at 3°C to 0.08 mm at 25°C. The density of root hairs on the root surface increased by a factor of 4 in rape between 3 and 25°C, but in barley the highest density was at 9°C. The contribution of root hairs to total root surface area was relatively greater in rape than in barley. The changes in root system morphology may be interpreted as adaptive responses to temperature stress on nutrient uptake, providing greater surface area for absorption per unit root weight or length.     

The COW1 locus of arabidopsis acts after RHD2, and in parallel with RHD3 and TIP1, to determine the shape, rate of elongation, and number of root hairs produced from each site of hair formation.

Two recessive mutant alleles at CAN OF WORMS1 (COW1), a new locus involved in root hair morphogenesis, have been identified in Arabidopsis thaliana L. Heynh. Root hairs on Cow1- mutants are short and wide and occasionally formed as pairs at a single site of hair formation. The COW1 locus maps to chromosome 4. Root hairs on Cow1- plants form in the usual positions, suggesting that the phenotype is not the result of abnormal positional signals. Root hairs on Cow1- roots begin hair formation normally, forming a small bulge, or root hair initiation site, of normal size and shape and in the usual position on the hair-forming cell. However, when Cow1- root hairs start to elongate by tip growth, abnormalities in the shape and elongation rate of the hairs become apparent. Genetic evidence from double-mutant analysis of cow1-1 and other loci involved in root hair development supports our conclusion that COW1 is required during root hair elongation.     

Variation in phosphorus uptake efficiency by genotypes of cowpea (Vigna unguiculata) due to differences in root and root hair length and induced rhizosphere processes

The lengths of roots and root hairs and the extent of root-induced processes affect phosphorus (P) uptake efficiency by plants. To assess the influence of variation in the lengths of roots and root hairs and rhizosphere processes on the efficiency of soil phosphorus (P) uptake, a pot experiment with a low-P soil and eight selected genotypes of cowpea (Vigna unguiculata (L) WALP) was conducted. Root length, root diameter and root hair length were measured to estimate the soil volume exploited by roots and root hairs. The total soil P was considered as a pool of Olsen-P, extractable with 0.5 M NaHCO₃ at pH 8.5, and a pool of non-Olsen-P. Model calculations were made to estimate P uptake originated from Olsen-P in the root hair zone and the Olsen-P moving by diffusion into the root hair cylinder and non-Olsen-P uptake. The mean uptake rate of P and the mean rate of non-Olsen-P depletion were also estimated. The genotypes differed significantly in lengths of roots and root hairs, and in P uptake, P uptake rates and growth. From 6 to 85% of total P uptake in the soil volume exploited by roots and root hairs was absorbed from the pool of non-Olsen-P. This indicates a considerable activity of root-induced rhizosphere processes. Hence the large differences show that traits for more P uptake-efficient plants exist in the tested cowpea genotypes. This opens the possibility to breed for more P uptake-efficient varieties as a way to bring more sparingly soluble soil P into cycling in crop production and obtain capitalisation of soil P reserves.     

A Cotyledonary Inhibitor of Root Nodulation in Pisum sativum

Root nodule formation was studied in 11-day-old seedlings of Pisum sativum L. cv. Alaska, which were inoculated with Rhizobium leguminosarum at the time of planting. On intact plants an identical nodulation pattern was observed in the lateral roots attached to the two xylem strands connected to the cotyledons. When one cotyledon was removed before germination, however, a highly significant increase in nodultion was observed on the lateral roots attached to the xylem strand which no longer was connected to a cotyledon. Excision of one cotyledon also caused an alteration in the radial location of root nodules on the primary root. Under these conditions there was a distinct promotion of nodular proliferations in the root cortex opposite the primary phloem poles. The fact that removal of one cotyledon increased nodulation on the lateral roots but had no effect on the rhizobial infection of lateral root hairs suggested that a cotyledonary inhibitor acts at a step between the infection process and the appearance of a macroscopic nodule. The data were interpreted in terms of an inhibitor of cortical cell division which is translocated from the cotyledons to the developing root via the phloem.     

ROOT AERENCHYMA DEVELOPMENT IN SPARTINA PATENS IN RESPONSE TO FLOODING

Root aerenchyma, developed in response to flooding, was measured as specific gravity in previously existing (old) and newly developed (new) roots of Spartina patens in a 25-day greenhouse experiment. Root specific gravity was related to porosity (fractional volume of aerenchyma): porosity = 1.026 – 0.969 × specific gravity, over a wide range of specific gravities (0.5 to 0.9). The specific gravity of flooded roots (new and old) decreased from 0.78 to 0.58 between day 5 and 25, while the specific gravity of old roots that remained drained did not change. After 5 days growth, newly produced roots were found to have less aerenchyma than their older counterparts, but after 25 days the specific gravity of new roots equaled that of old roots. In addition, flooding increased mortality of preexisting roots and inhibited growth of new roots.     

Development of the adhesive pad on climbing fig (Ficus pumila) stems from clusters of adventitious roots

Vines have different climbing strategies; one type, called clinging vines, has developed a specialized structure, the adhesive pad that secretes a sticky substance that adheres to almost any substrate. In this study on the climbing fig, Ficus pumila L., we report on the developmental anatomy of clusters of adventitious roots that become transformed into this unique structure. Clustered adventitious roots in F. pumila are initiated in pairs on either side of a vascular bundle at the 2nd to 3rd internodes of young stems. After emergence through the cortex and epidermis, root hairs form, which secrete a substance that stains positively for polysaccharide and protein. The adventitious roots and root hairs stick together forming the adhesive pad. Pads stick to almost any substrate. If the adventitious roots do not touch a substrate they usually dry up, if they touch moist soil they tend to branch and change to a terrestrial form. We discuss experiments on the shoot developmental behavior leading to adhesive pad formation and the possible roles of auxin.     

Number,position, diameter and initial direction of growth of primary roots in Musa

To understand soil colonization by a root system, information is needed on the architecture of the root system. In monocotyledons, soil exploration is mainly due to the growth of adventitious primary roots. Primary root emergence in banana was quantified in relation to shoot and corm development. Root emergence kinetics were closely related to the development of aerial organs. Root position at emergence on the corm followed an asymptotic function of corm dry weight, so that the age of each root at a given time could be deduced from its position. Root diameter at emergence was related to the position of the roots on the corm, with younger roots being thicker than older ones. However, root diameters were not constant along a given root, but instead decreased with the distance to the base; roots appear to be conical in their basal and apical parts. Root growth directions at emergence were variable, but a high proportion of the primary roots emerged with a low angle to the horizontal. Further research is needed to evaluate whether these initial trajectories are conserved during root development. Results presented in this study are in good agreement with those reported for other monocotyledons such as maize and rice. They give quantitative information that will facilitate the development of models of root system architecture in banana.