An integrated method for quantifying root architecture of field-grown maize

Background and Aims

A number of techniques have recently been developed for studying the root system architecture (RSA) of seedlings grown in various media. In contrast, methods for sampling and analysis of the RSA of field-grown plants, particularly for details of the lateral root components, are generally inadequate.

Methods

An integrated methodology was developed that includes a custom-made root-core sampling system for extracting intact root systems of individual maize plants, a combination of proprietary software and a novel program used for collecting individual RSA information, and software for visualizing the measured individual nodal root architecture.

Key Results

Example experiments show that large root cores can be sampled, and topological and geometrical structure of field-grown maize root systems can be quantified and reconstructed using this method. Second- and higher order laterals are found to contribute substantially to total root number and length. The length of laterals of distinct orders varies significantly. Abundant higher order laterals can arise from a single first-order lateral, and they concentrate in the proximal axile branching zone.

Conclusions

The new method allows more meaningful sampling than conventional methods because of its easily opened, wide corer and sampling machinery, and effective analysis of RSA using the software. This provides a novel technique for quantifying RSA of field-grown maize and also provides a unique evaluation of the contribution of lateral roots. The method also offers valuable potential for parameterization of root architectural models.     

Root pruning reduces root competition and increases crop growth in a living mulch cropping system

Abstract

In two field experiments, growth of white cabbage in pure stands was compared with that of cabbage grown in living mulch systems to reduce pest attacks. The roots of the living mulch were pruned early in the season, with the aim of reducing competition and increasing growth of the white cabbage. Root pruning was shown to increase the above-ground biomass of white cabbage, with two prunings giving higher cabbage yields than one, but there were clear differences between the living mulch species tested (red clover, birdsfoot trefoil, salad burnet, winter rye). Below-ground growth and competition were examined by measuring root distribution in minirhizotrons and uptake of ¹⁵N placed at different soil depths. These studies showed that the ability of mulch species to compete for resources at depth was restricted by pruning, and that this was crucial for the development of the white cabbage crop.     

The longevity and activity of the primary root of maize

The longevity of the main root cylinder and the laterals of the primary root of maize plants was determined under controlled greenhouse conditions by means of nuclear staining with acridine orange. The cortex of the main root was found to be alive for the whole life-span of the plant, whereas the life-span of the root hairs was only 2 to 3 days as evidenced by electronmicroscopical examination of cell integrity. The onset of senescence of laterals was observed at the older part of the main root at the 6-leaf stage of the plant. Senescence of 1st and 2nd order laterals commenced near the root tip a few days after their protrusion and advanced towards the basal region of the root. In any root segment death of the cortex cells preceeded that of the stele. At the late grain filling stage all laterals along the main root exhibited advanced senescence, but stainable nuclei were seen in the root tissues of the basal part of 1^st order laterals (both cortex and stele) as well as of the 2^nd order laterals which emerged from that root segment. The pattern of the dying of the root tissue is discussed with regard to the P-nutrition of the shoot system by the primary root.     

Light-regulated protein and mRNA synthesis in root caps of maize

Illumination of maize roots initiates changes in mRNA levels and in the activities of proteins within the root cap. Using Northern analysis we showed a 5–6-fold increase in the levels of three specific mRNAs and a 14-fold increase in plastid mRNA. This increase is rapid, occurring within 30 minutes of illumination. With prolonged periods of darkness following illumination, messages return to levels observed in dark, control caps. For two species of mRNA illumination results in a reduction in message levels. Light-stimulated increases in the levels of specific mRNAs are proportionally greater than are increases in the activities of corresponding proteins. We suggest that the light-stimulated increase in protein activity in root caps may be preceded by and occur as a consequence of enhanced levels of mRNA. Our work suggests that photomorphogenesis in roots could involve changes in the levels of a wide variety of mRNAs within the root cap.     

Mixed cropping with maize combined with moderate UV-B radiations lead to enhanced flavonoid production and root growth in faba bean

Abstract

Flavonoids have recently been proposed to function as developmental regulators and/or signaling molecules under biotic or abiotic stress. The aim of this study was to determine the composition and concentration of fiavonoid aglycones (kaempferol, luteolin, and quercetin) in faba bean shoots and roots, as affected by interspecific root interactions with maize and moderate UV-B radiation. Independent of the UV-B treatment, interspecific root interactions with maize enhanced the concentration of both quercetin and luteolin in the faba bean shoots and roots by 50 and 97.8%, respectively, and improved the root length by 14.6%. In addition, moderate UV-B radiation facilitated a systematic increase of both aglycones in both shoots and roots without affecting plant growth. To our knowledge, this report is the first work documenting the response of faba bean flavonoids to interspecific root interactions with maize and moderate UV-B radiation, and it provides a new perspective for understanding interspecific interactions.     

Geometrical properties of simulated maize root systems: consequences for length density and intersection density

The spatial distribution of root length density (RLD) is important because it affects water and nutrient uptake. It is difficult to obtain reliable estimates of RLD because root systems are very variable and heterogeneous. We identified systematic trends, clustering, and anisotropy as geometrical properties of root systems, and studied their consequences for the sampling and observation of roots. We determined the degree of clustering by comparing the coefficient of variation of a simulated root system with that of a Boolean model. We also present an alternative theoretical derivation of the relation between RLD and root intersection density (RID) based on the theory of random processes of fibres. We show how systematic trends, clustering and anisotropy affect the theoretical relation between RLD and RID, and the consequences this has for measurement of RID in the field. We simulated the root systems of one hundred maize crops grown for a thermal time of 600 K d, and analysed the distribution of RLD and root intersection density RID on regular grids of locations throughout the simulated root systems. Systematic trends were most important in the surface layers, decreasing with depth. Clustering and anisotropy both increased with depth. Roots at depth had a bimodal distribution of root orientation, causing changes in the ratio of RLD/RID. The close proximity of the emerging lateral roots and the parent axis caused clustering which increased the coefficient of variation.     

The porous-membrane technique for root studies of field-grown crops

A porous-membrane technique has been successfully used in root studies of field-grown soybean,Glycine max L. Merr., for several years. In order to evaluate this technique on other crops, a study was conducted to compare growth parameters of individual soybean, cotton,Gossypium hirsutum L., and corn,Zea mays L., plants grown with and without their root systems confined within porous membranes. Results indicate that plant species respond differently to root confinement.Root confinement reduced all shoot growth and yield parameters of the crop species, however the reduction was generally greatest for cotton. Apparently, cotton is less adaptable to the restricted rooting volume. Root confinement reduced levels of potassium and phosphorus in soybean and nitrogen in corn and cotton. Fertilizer rates higher than soil test recommendations should be applied to membrane-grown plants to prevent reductions in nutrient levels. It may be possible that modifying the size or shape of the membrane may enhance root and shoot growth of specific crop plants like cotton and increase the ability of this technique to be used on a broader spectrum of crop plants.     

Development and replenishment of the P-depletion zone around the primary root of maize during the vegetation period

The dynamics of the development and replenishment of P-depletion zones around the primary root of maize (Zea mays L. cv ‘Garbo’) was studied during a vegetation period (80 days) under greenhouse conditions in a loamy sand of low P-availability. A recently described freeze-cutting technique was used to determine radial diffusion of labelled phosphate to the primary root. The development of the depletion zone was biphasic. In the initial phase after two days of growth of the primary root in a soil layer labelled with³³P a minimum of isotopically exchangeable P (EP) was observed which had decreased to about 30% of its original amount at the root surface. At that time the corresponding P-concentration in the soil solution was calculated to be as low as 5×10⁻⁷ M. The depletion zone had already spread 0.4 mm from the root surface. During the second phase, between the 10th and 20th day of plant growth the concentration of EP at the root surface increased slowly but did not change markedly. However, the depletion zone continued to spread and after the 20th day of growth reached its maximal diameter (1.07 mm from the root surface) but remained completely within the root hair cyclinder; the single root hairs never exceeded 1.14 mm in length. The biphasic growth of the depletion zone was probably caused by proton extrusion of the root tip. Acidification of the soil solution from pH 5.8 to about 3.9 results in an about 3-fold rise of the concentration of desorbed phosphate and might also have activated acidophilic P-translocators of the root during the initial phase. Anion over cation uptake normally prevailing during the later stage of root development might resulted in a rise of the soil pH within the root hair zone. Consequently P-availability, as well as P-uptake capacity declined, but P-uptake by the seminal root still continued until the 20th day. Subsequently, the P-concentration within the depletion zone increased again while simultaneously its extent was reduced until it was almost completely replenished after 60 days indicating a loss of P-uptake capacity of the primary root. Within the root tissue³³P was accumulated to about twice the concentration of that in the undepleted soils. This accumulation corresponded to periods of high uptake due to the development of root laterals. In the root cortex a high P-content was observed during the first 30 days of growth. At the onset of the reproductive stage of the plant the P-content of the shoot and especially in the developing seeds rose considerably at the cost of phosphate stored in the root cortex. The accumulation of³³P in the root tissue indicated that nutrient gain was mainly achieved during the early stages of plant development and that P was temporarily stored to some extent within the root system.     

Different populations of bacteria associated with sheathed and bare regions of roots of field-grown maize

It has been previously shown that soil sheaths cling tightly to some portions of all axile roots and cover all but the growing tips of the young roots of field-grown maize. These sheaths overlie immature regions of the roots which have intact epidermal cells with root hairs, and living, thus non-conducting, late metaxylem elements. Loss of the soil sheath in the proximal region coincides with the opening of these large metaxylem vessels. Now, total, and viable counts have been recorded of bacteria associated with the root surface and adhering soil of sheathed and bare regions. These showed some common features, in that populations of similar size were associated with the two root regions in plants beginning to flower. Each population included about the same numbers of bacteria that were viable on each of three selective media (nitrogen-free, Pseudomonas F or MacConkey). However, more spore-formers capable of growth on nitrogen-free media and more fluorescent bacteria were isolated from the sheathed regions. Actinomycetes were absent from sheathed but plentiful on bare regions.The high numbers of diverse types of bacteria associated with both root surfaces can be related to the previously demonstrated similarity in amounts of organic carbon released from each region. The proliferation of actionomycetes on the bare roots and their exclusion from sheathed roots may in part be due to the lower water status of the bare region, which is related to its greater axial conducting capacity. Thus the distribution of the two types of root surface within an individual root system has important implications for the choice of root and rhizosphere sampling techniques and for root bacterization work.     

Complementarity in root architecture for nutrient uptake in ancient maize/bean and maize/bean/squash polycultures

Background and Aims

During their domestication, maize, bean and squash evolved in polycultures grown by small-scale farmers in the Americas. Polycultures often overyield on low-fertility soils, which are a primary production constraint in low-input agriculture. We hypothesized that root architectural differences among these crops causes niche complementarity and thereby greater nutrient acquisition than corresponding monocultures.

Methods

A functional–structural plant model, SimRoot, was used to simulate the first 40 d of growth of these crops in monoculture and polyculture and to determine the effects of root competition on nutrient uptake and biomass production of each plant on low-nitrogen, -phosphorus and -potassium soils.

Key Results

Squash, the earliest domesticated crop, was most sensitive to low soil fertility, while bean, the most recently domesticated crop, was least sensitive to low soil fertility. Nitrate uptake and biomass production were up to 7 % greater in the polycultures than in the monocultures, but only when root architecture was taken into account. Enhanced nitrogen capture in polycultures was independent of nitrogen fixation by bean. Root competition had negligible effects on phosphorus or potassium uptake or biomass production.

Conclusions

We conclude that spatial niche differentiation caused by differences in root architecture allows polycultures to overyield when plants are competing for mobile soil resources. However, direct competition for immobile resources might be negligible in agricultural systems. Interspecies root spacing may also be too large to allow maize to benefit from root exudates of bean or squash. Above-ground competition for light, however, may have strong feedbacks on root foraging for immobile nutrients, which may increase cereal growth more than it will decrease the growth of the other crops. We note that the order of domestication of crops correlates with increasing nutrient efficiency, rather than production potential.     

Relationship between root elongation rate and diameter and duration of growth of lateral roots of maize

The objective of this work was to describe the relationship between elongation rate and diameter of maize roots and to estimate the length and growth duration of lateral roots of maize. Diameters and elongation rates of roots were measuredin situ on plants grown 5 weeks in small rhizotrons under greenhouse conditions. At the end of the experimental period the roots were harvested and diameters of axile and lateral roots were measured. The frequency distribution of diameters of harvested roots was bimodal with a minimum at 0.6 mm; 97% of axile roots were larger than this value and 98% of the lateral roots were smaller. Root elongation per day increased as diameter increased but the slope of the relationship with lateral roots was about 2.5 times that with axile roots when separate linear regressions were fitted to the two populations. The length of lateral roots found on axillary roots between the base and about 30 cm from the apex was approximately 2.2 cm. All of the data was consistent with the hypothesis that the lateral roots grew for about 2.5 days and then ceased growing. The axillary roots continued to grow throughout the experimental period at a rate of about 3 cm day⁻¹. Contribution from the Department of Agronomy, New York State College of Agriculture and Life Sciences, Cornell University, Ithaca, NY 14853. Agronomy paper No. 1661. This research is part of the program of the Center for Root-Soil Research.     

Phytotropin-induced root phototropism in maize

Phytotropins, even those not absorbing in the visible region of the spectrum, can induce a phototropic response in maize ( Zea mays L. cv. PX-75) roots when illuminated unilaterally with white light. The most active phytotropin, 2-(1-pyrenoyl) benzoic acid (PBA) can elicit a full response at 10 μ M , while the other active molecules, 2-carboxyphenyl-3-phenylpropane-1,3-dione (CPD), 2-carboxyphenyl-3-phenyl-1,2-pyrazole (CPP), 1-N-naphthylphthalamic acid (NPA) and erythrosin elicit a full response at 100 μ M . The less active phytotropins BBA and fluorescein give a reduced response. It is suggested that the observed effect cannot be explained solely on the basis of auxin transport inhibition. There is a photoreceptor in the extension zone of the root, which may be associated in some way with the receptor for NPA. The results are consistent with the proposal that the phototropic process may form part of the root gravitropic response mechanism.     

Influence of maize root mucilage on soil aggregate stability

This study was undertaken to determine the effects of root exudates on soil aggregate stability. Root mucilage was collected from two-month old maize plants (Zea mays L.) Mucilage and glucose solutions were added at a rate of 2.45 g C kg⁻¹ dry soil to silty clay and silt loam soils. Amended soils, placed in serum flasks, were incubated for 42 d with a drying-wetting cycle after 21 d. Evolved CO₂ was measured periodically as well as the water-stable aggregates and soluble sugar and polysaccharide content of the soil. In mucilage-amended soils CO₂ evolution started with a lag phase of 2–3 days, which was not observed in glucose-amended soils. There was then a sharp increase in evolved CO₂ up to day 7. During the second incubation period there were only small differences in evolved C between treatments. Incorporation of mucilage in both soils resulted in a spectacular and immediate increase in soil aggregate stability. Thereafter, the percent of water-stable aggregates quickly decreased parallel to microbial degradation. On completion of the incubation, aggregate stability in the silty clay soil was still significantly higher in the presence of mucilage than in the control. This work supports the assumption that freshly released mucilage is able to stick very rapidly to soil particles and may protect the newly formed aggregates against water destruction. On the silty clay, microbial activity contributes to a stabilization of these established organo-mineral bounds.     

Sieve size effects on root length and biomass measurements of maize (Zea mays) and Grevillea robusta

The purpose of this study was to investigate the effects of different mesh sizes on the recovery of root length and biomass and to determine whether the degree of recovery was influenced by plant species and sample location. Sieves of 2.0, 1.0, 0.5 and 0.25 mm (4.0, 1.0, 0.25 and 0.06 mm2) mesh sizes were used to recover and measure the root length and biomass of Zea mays L. (maize) at 0–15 cm and 30–45 cm depths and of Grevillea robusta A. Cunn. ex R. Br. (grevillea) at the same depths 1.0 m and 4.5 m from a line of grevillea trees. At 0–15 cm, the coarser sieves (sum collected with 2.0 and 1.0 mm sieves) recovered approximately 80% of the total root biomass measured, but only 60% of the root length. The proportion of total maize root length and biomass recovered by the coarser sieves decreased with soil depth. The proportion of total grevillea root length recovered by the coarser sieves was similar at the two soil depths, but increased slightly with distance from the tree line. The ≥ 0.5 mm sieves recovered between 93 and 96% of grevillea and maize root biomass and between 73 and 98% of their root length, depending on the sample location. Roots passing through the 0.5 mm sieve, but recovered by the 0.25 mm sieve were about 20% of total maize root length and grevillea root length at 1.0 m from the tree line but < 5% of the total grevillea root length at 4.5 m from the tree. Roots passing through the 0.5 mm sieve but recovered by the 0.25 mm sieve contributed only slightly to root biomass. Although the ≥ 0.5 mm sieves provided adequate measurements of root biomass, the ≥ 0.25 mm sieves were required for accurate measurement of fine root length. There was no universal correction for root length and biomass underestimation when large sieve sizes were used because the proportions of length and biomass recovered depended on the plant species and on soil depth and distance from the plant. This revised version was published online in August 2006 with corrections to the Cover Date.     

Hairy roots induced by Agrobacterium rhizogenes and production of regenerative plants in hairy root cultures in maize

Before the late 1980s, although the majority of Agrobacterium-mediated gene transfer experiments have been performed with A. tumefaciens[1―3], some work has also been done with its close relative, Agro-bacterium rhizogene. It has been considered that onl…