Produce Cluster Roots Research Articles

Phosphorus absorption kinetics and exudation strategies of roots developed by three lupin species to tackle P deficiency.

Different lupin species exhibited varied biomass, P allocation, and physiological responses to P-deprivation. White and yellow lupins had higher carboxylate exudation rates, while blue lupin showed the highest phosphatase activity. White lupin (Lupinus albus) can produce specialized root structures, called cluster roots, which are adapted to low-phosphorus (P) soil. Blue lupin (L. angustifolius) and yellow lupin (L. luteus), which are two close relatives of white lupin, do not produce cluster roots. This study characterized plant responses to nutrient limitation by analyzing biomass accumulation and P distribution, absorption kinetics and root exudation in white, blue, and yellow lupins. Plants were grown in hydroponic culture with (64µM NaH2PO4) or without P for 31days. Under P limitation, more biomass was allocated to roots to improve P absorption. Furthermore, the relative growth rate of blue lupin showed the strongest inhibition. Under + P conditions, the plant total-P contents of blue lupin and yellow lupin were higher than that of white lupin. To elucidate the responses of lupins via the perspective of absorption kinetics and secretion analysis, blue and yellow lupins were confirmed to have stronger affinity and absorption capacity for orthophosphate after P-deprivation cultivation, whereas white lupin and yellow lupin had greater ability to secrete organic acids. The exudation of blue lupin had higher acid phosphatase activity. This study elucidated that blue lupin was more sensitive to P-scarcity stress and yellow had the greater tolerance of P-deficient condition than either of the other two lupin species. The three lupin species have evolved different adaptation strategies to cope with P deficiency.

Dynamic Development of White Lupin Rootlets Along a Cluster Root.

White lupin produces cluster roots in response to phosphorus deficiency. Along the cluster root, numerous short rootlets successively appear, creating a spatial and temporal gradient of developmental stages that constitutes a powerful biological model to study the dynamics of the structural and functional evolution of these organs. The present study proposes a fine histochemical, transcriptomic and functional analysis of the rootlet development from its emergence to its final length. Between these two stages, the tissue structures of the rootlets were observed, the course of transcript expressions for the genes differentially expressed was monitored and some physiological events linked to Pi nutrition were followed. A switch between (i) a growing phase, in which a normal apical meristem is present and (ii) a specialized phase for nutrition, in which the rootlet is completely differentiated, was highlighted. In the final stage of its determinate growth, the rootlet is an organ with a very active metabolism, especially for the solubilization and absorption of several nutrients. This work discusses how the transition between a growing to a determinate state in response to nutritional stresses is found in other species and underlines the fundamental dilemma of roots between soil exploration and soil exploitation.

Cluster roots of Embothrium coccineum modify their metabolism and show differential gene expression in response to phosphorus supply

Embothrium coccineum produces cluster roots (CR) to acquire sparingly soluble phosphorus (P) from the soil through the exudation of organic compounds. However, the physiological mechanisms involved in carbon drainage through its roots, as well as the gene expression involved in the biosynthesis of carboxylates and P uptake, have not been explored. In this work, we evaluated the relationship between carboxylate exudation rate and phosphoenolpyruvate carboxylase (PEPC) activity in roots of E. coccineum seedlings grown in a nutrient-poor volcanic substrate. Second, we evaluated CR formation and the expression of genes involved in the production of carboxylates (PEPC) and P uptake (PHT1) in E. coccineum seedlings grown under three different P supplies in hydroponic conditions. Our results showed that the carboxylate exudation rate was higher in CR than in non-CR, which was consistent with the higher PEPC activity in CR. We found higher CR formation in seedlings grown at 5 μM of P supply, concomitant with a higher expression of EcPEPC and EcPHT1 in CR than in non-CR. Overall, mature CR of E. coccineum seedlings growing on volcanic substrates poor in nutrients modify their metabolism compared to non-CR, enhancing carboxylate biosynthesis and subsequent carboxylate exudation. Additionally, transcriptional responses of EcPEPC and EcPHT1 were induced simultaneously when E. coccineum seedlings were grown in P-limited conditions that favored CR formation. Our results showed, for the first time, changes at the molecular level in CR of a species of the Proteaceae family, demonstrating that these root structures are highly specialized in P mobilization and uptake.

Root positioning and trait shifts in Hibbertia racemosa as dependent on its neighbour's nutrient-acquisition strategy.

Nutrient-poor ecosystems globally exhibit high plant diversity. One mechanism enabling the co-existence of species in such ecosystems is facilitation among plants with contrasting nutrient-acquisition strategies. The ecophysiological processes underlying these interactions remain poorly understood. We hypothesized that root positioning plays a role between sympatric species in nutrient-poor vegetation. We investigated how the growth traits of the focal mycorrhizal non-cluster-rooted Hibbertia racemosa change when grown in proximity of non-mycorrhizal Banksia attenuata, which produces cluster roots that increase nutrient availability, compared with growth with conspecifics. Focal plants were placed in the centre of rhizoboxes, and biomass allocation, root system architecture, specific root length (SRL), and leaf nutrient concentration were assessed. When grown with B. attenuata, focal plants decreased root investment, increased root growth towards B. attenuata, and positioned their roots near B. attenuata cluster roots. SRL was greater, and the degree of localized root investment correlated positively with B. attenuata cluster-root biomass. Total nutrient contents in the focal individuals were greater when grown with B. attenuata. Focal plants directed their root growth towards the putatively facilitating neighbour's cluster roots, modifying root traits and investment. Preferential root positioning and root morphological traits play important roles in positive plant-plant interactions.

Contrasting responses of cluster roots formation induced by phosphorus and nitrogen supply in Embothrium coccineum populations from different geographical origin

Embothrium coccineum is a pioneer tree that produces cluster roots (CR) induced by phosphorus (P) or nitrogen (N) deficiency, but the role which both N and P play in CR formation for different E. coccineum populations is still unknown. We hypothesized that in E. coccineum, N influences the CR formation response, primarily induced by P limitation. However, this response depends on local adaptations to their specific conditions of geographic origin. Seedlings from three contrasting edaphoclimatic conditions (Northern, Central and Southern populations) were grown in hydroponic culture under high and low N supply and at five different P supply. Morphophysiological responses, including CR number, growth, biomass, P and N plant concentration, were studied. Our results showed that at high N supply, CR formation was stimulated with increased P supply, but just until it reaches a maximum of 5 μM, above this concentration CR formation decreases. However, at low N supply, a constitutive response was observed regardless of P supply. Moreover, each of these responses depends on the populations. Our study suggests that N drives CR formation induced by P supply and that a differential response exists among E. coccineum populations, that could be related to their origin edaphoclimatic conditions.

Root dynamics and survival in a nutrient-poor and species-rich woodland under a drying climate

In Australia’s Mediterranean hyperdiverse vegetation, species that produce cluster roots to mobilise poorly-available nutrients (e.g. Banksia spp.) are an important functional and structural component. Cluster roots are only active during the wet season, indicating a strong dependence on suitable surface soil moisture conditions. Winter rainfall in this region is declining due to global climate change, with a delayed commencement of rains and a decline in precipitation. It is unknown how lower soil moisture levels will affect the root dynamics of these globally-significant plant communities. We determined the root dynamics and root lifespan with minirhizotrons with or without irrigation to simulate reduced rainfall scenarios. We found a major effect of irrigation on the early production (0.24 m m−2 d−1 increase), occurrence (97% increase) of cluster roots and only slight effects on lifespan (~10 days less) of all root types. With irrigation, the resultant greater soil moisture levels increased the deployment of cluster roots. Apart from cluster roots, the dynamics of other roots did not decline at lower soil moisture levels, suggesting that this system shows some resilience to decreased rainfall. Future research should focus on assessing if climate-altered cluster-root activity may be promoting compositional shifts in plant communities with additional restraining effects on root trait diversity.

Comparison of the response to phosphorus deficiency in two lupin species, Lupinus albus and L. angustifolius, with contrasting root morphology

White lupin (Lupinus albus) produces cluster roots, an adaptation to low soil phosphorus (P). Cluster roots exude large levels of P-solubilizing compounds such as citrate and malate. In contrast, narrow leaf lupin (L. angustifolius) is closely related to L. albus, but does not produce cluster roots. To examine the different strategies for P acquisition, we compared the growth, biomass allocation, respiratory properties and construction cost between L. albus and L. angustifolius under P-deficient conditions. Both Lupinus species were grown in hydroponic culture with 1 or 100 μM P. Under the P-deficient regime, L. albus produced cluster roots with little change in biomass allocation, while L. angustifolius significantly increased biomass allocation to roots. The rate of cyanide-resistant SHAM (salicylhydroxamic acid)-sensitive respiration was high in cluster roots and very low in roots of L. angustifolius. These results suggest a low alternative oxidase (AOX) activity in L. angustifolius roots, and thus, ATP would be produced efficiently in L. angustifolius roots. The construction cost was highest in cluster roots and lowest in L. angustifolius roots. This study shows that under P deficiency, L. albus produces high-cost cluster roots to increase the P availability, while L. angustifolius produces large quantities of low-cost roots to enhance P uptake.

The alternative respiratory pathway mediates carboxylate synthesis in white lupin cluster roots under phosphorus deprivation

Plant adaptations associated with a high efficiency of phosphorus (P) acquisition can be used to increase productivity and sustainability in a world with a growing population and decreasing rock phosphate reserves. White lupin (Lupinus albus) produces cluster roots that release carboxylates to efficiently mobilize P from P-sorbing soils. It has been hypothesized that an increase in the activity of the alternative oxidase (AOX) would allow for the mitochondrial oxidation of NAD(P)H produced during citrate synthesis in cluster roots at a developmental stage when there is a low demand for ATP. We used the oxygen-isotope fractionation technique to study the in vivo respiratory activities of the cytochrome oxidase pathway (COP) and the alternative oxidase pathway (AOP) in different root sections of white lupins grown hydroponically with and without P. In parallel, AOX protein levels and internal carboxylate concentrations were determined in cluster and non-cluster roots. Higher in vivo AOP activity was measured in cluster roots when malate and citrate concentrations were also high, thus confirming our hypothesis. AOX protein levels were not always correlated with in vivo AOP activity, suggesting post-translational regulation of AOX.

Cluster-root formation and carboxylate release in three Lupinus species as dependent on phosphorus supply, internal phosphorus concentration and relative growth rate

Some Lupinus species produce cluster roots in response to low plant phosphorus (P) status. The cause of variation in cluster-root formation among cluster-root-forming Lupinus species is unknown. The aim of this study was to investigate if cluster-root formation is, in part, dependent on different relative growth rates (RGRs) among Lupinus species when they show similar shoot P status. Three cluster-root-forming Lupinus species, L. albus, L. pilosus and L. atlanticus, were grown in washed river sand at 0, 7·5, 15 or 40 mg P kg(-1) dry sand. Plants were harvested at 34, 42 or 62 d after sowing, and fresh and dry weight of leaves, stems, cluster roots and non-cluster roots of different ages were measured. The percentage of cluster roots, tissue P concentrations, root exudates and plant RGR were determined. Phosphorus treatments had major effects on cluster-root allocation, with a significant but incomplete suppression in L. albus and L. pilosus when P supply exceeded 15 mg P kg(-1) sand. Complete suppression was found in L. atlanticus at the highest P supply; this species never invested more than 20 % of its root weight in cluster roots. For L. pilosus and L. atlanticus, cluster-root formation was decreased at high internal P concentration, irrespective of RGR. For L. albus, there was a trend in the same direction, but this was not significant. Cluster-root formation in all three Lupinus species was suppressed at high leaf P concentration, irrespective of RGR. Variation in cluster-root formation among the three species cannot be explained by species-specific variation in RGR or leaf P concentration.

How a phosphorus‐acquisition strategy based on carboxylate exudation powers the success and agronomic potential of lupines (Lupinus, Fabaceae)

Lupines (Lupinus species; Fabaceae) are an ancient crop with great potential to be developed further for high-protein feed and food, cover crops, and phytoremediation. Being legumes, they are capable of symbiotically fixing atmospheric nitrogen. However, Lupinus species appear to be nonmycorrhizal or weakly mycorrhizal at most; instead some produce cluster roots, which release vast amounts of phosphate-mobilizing carboxylates (inorganic anions). Other lupines produce cluster-like roots, which function in a similar manner, and some release large amounts of carboxylates without specialized roots. These traits associated with nutrient acquisition make lupines ideally suited for either impoverished soils or soils with large amounts of phosphorus that is poorly available for most plants, e.g., acidic or alkaline soils. Here we explore how common the nonmycorrhizal phosphorus-acquisition strategy based on exudation of carboxylates is in the genus Lupinus, concluding it is very likely more widespread than generally acknowledged. This trait may partly account for the role of lupines as pioneers or invasive species, but also makes them suitable crop plants while we reach "peak phosphorus".

The effect of phosphorus on growth and cluster-root formation in the Chilean Proteaceae: Embothrium coccineum (R. et J. Forst.)

One of the main factors that favours the formation of cluster roots is a low supply of phosphorus (P). The soils of southern Chile are mainly formed from volcanic ash, characterized by low levels of available P. Embothrium coccineum, a Chilean Proteaceae species produces cluster roots (CR). The factors that control CR formation in Chilean Proteaceae have not been extensively studied. The objective of this work was to assess the effects of P on the growth and cluster-root formation of E. coccineum. Plants were produced from seeds collected at two different locations: Valdivia and Pichicolo both at 39oS. They were cultured under similar greenhouse conditions, from June to September, watered twice a week using: distilled water (W), full strength Hoagland’s nutrient solution (H) or Hoagland without P (H-P). At the end of the experiment, height, total dry biomass, number of cluster roots (CR) per plant, CR /total root weight, were measured. Also acid exudation of CR was assayed using bromocresol purple on sterile agar plates. Treatments significantly affected growth and proportion of CR, the highest growth was observed with H. Under all treatments plants produced a similar number of CR. However, the proportion of CR biomass was higher with W and H-P than with H. Plants under W exhibited the lowest growth and low shoot/root ratio. Acid exudation of CR was not detectable in our experiment. These results are discussed comparing CR formation in low P conditions on Lupinus albus and other Proteaceae species, and the possible role of CR formation in E. coccineum considering its wide geographical distribution.

Cluster Roots of Leucadendron laureolum (Proteaceae) and Lupinus albus (Fabaceae) Take Up Glycine Intact: An Adaptive Strategy to Low Mineral Nitrogen in Soils?

South African soils are not only low in phosphorus (P) but most nitrogen (N) is in organic form, and soil amino acid concentrations can reach 2.6 g kg(-1) soil. The Proteaceae (a main component of the South African Fynbos vegetation) and some Fabaceae produce cluster roots in response to low soil phosphorus. The ability of these roots to acquire the amino acid glycine (Gly) was assessed. Uptake of organic N as 13C-15N-Gly was determined in cluster roots and non-cluster roots of Leucadendron laureolum (Proteaceae) and Lupinus albus (Fabaceae) in hydroponic culture, taking account of respiratory loss of 13CO2. Both plant species acquired doubly labelled (intact) Gly, and respiratory losses of 13CO2 were small. Lupin (but not leucadendron) acquired more intact Gly when cluster roots were supplied with 13C-15N-Gly than when non-cluster roots were supplied. After treatment with labelled Gly (13C : 15N ratio = 1), lupin cluster roots had a 13C : 15N ratio of about 0.85 compared with 0.59 in labelled non-cluster roots. Rates of uptake of label from Gly did not differ between cluster and non-cluster roots of either species. The ratio of C : N and 13C : 15N in the plant increased in the order: labelled roots < rest of the root < shoot in both species, owing to an increasing proportion of 13C translocation. Cluster roots of lupin specifically acquired more intact Gly than non-cluster roots, whereas Gly uptake by the cluster and non-cluster roots of leucadendron was comparable. The uptake capacities of cluster roots are discussed in relation to spatial and morphological characteristics in the natural environment.

Microbial community composition and functioning in the rhizosphere of three Banksia species in native woodland in Western Australia

Annual plant species differ in their rhizosphere microbial community composition. However, rhizosphere communities are often investigated under controlled conditions, and it is unclear if perennial plants growing in the field also have rhizosphere communities that are specific to a particular plant species. The aim of our study was to determine the bacterial community composition of three species of Banksia ( B. attenuata R. Brown, B. ilicifolia R. Brown and B. menziesii R. Brown) growing in close proximity in a native woodland in Western Australia and to relate community structure to function. All three species are small trees that produce cluster roots in the field following winter rains. Cluster roots and rhizosphere soil were sampled in early spring (August 2001) and again four weeks later (September 2001). Many new cluster roots were formed in the period between the August and the September sampling. Rhizosphere soil pH, percent soil moisture and C and N content did not differ significantly among species or sampling times. However, the bacterial community composition on the cluster roots and in the rhizosphere soil, studied by denaturing gradient gel electrophoresis (DGGE), differed among the three species, with cluster root age class (young or mature to senescing) and also between sampling times. These changes in community composition were accompanied by changes in the activity of some of the enzymes studied. The activities of β-glucosidase and protease increased over time. The three species differed in asparaginase activity, but not in the activity of acid and alkaline phosphatase in the rhizosphere. These results suggest a relationship between the changes in composition and function of bacterial communities.

Cluster-root production and organic anion exudation in a group of old-world lupins and a new-world lupin

We examined the capacity of several Old-World lupin species (Lupinus luteus L., L. hispanicus Boiss. et Reuter and L. angustifolius L.) and one species of a New-World lupin (L. mutabilis Sweet) to form cluster roots under a range of conditions in solution culture. The effect of the synthetic auxin, IBA (indole-3-butyric acid), on cluster-root development in L. luteus and L. albus L. provided with an adequate phosphorus (P) supply was also investigated. In addition, the effect of a high nitrate-N (NO3-N) supply on the efflux of citrate and malate from roots of L. angustifolius was examined to determine if specific regions of the root system exuded these organic anions. When P-deficient, L. hispanicus, L. luteus and L. mutabilis formed cluster roots that secreted organic anions. Citrate was generally the dominant organic anion exuded, although succinate was also exuded in large quantities from L. luteus. Citrate efflux by L. hispanicus and L. luteus was at least comparable to that reported for P-deficient L. albus[up to 1.092 nmol g−1 fresh weight (FW) s−1], but was over an order of magnitude lower in L. mutabilis (0.036 nmol g−1 FW s−1). Citrate and malate were not detected in significant amounts from either the lateral roots or the root tips of any species grown under P-sufficient or -deficient conditions. Citrate efflux from cluster roots of L. luteus showed a diurnal pattern, similar to that reported for L. albus, with maximum efflux during the day, and declining to a minimum before dawn. IBA added to the nutrient solution induced cluster-root formation on both L. albus and L. luteus at concentrations of P that would normally suppress the production of these roots. However, the IBA-induced cluster roots did not exude significant amounts of citrate. Although L. angustifolius did not produce cluster roots when P-deficient, it produced cluster-like root structures that exuded citrate (0.053 nmol g−1 FW s−1) when grown at a high nitrate-N (NO3-N) supply. L. angustifolius did not exude significant citrate or malate from lateral roots or root tips when grown at either high or low NO3-N supply. Our findings for L. hispanicus and L. luteus are the first reports of cluster-root formation in response to P deficiency for these Old-World species, and for L. mutabilis, it is the first report of cluster roots for a New-World lupin species. These reports indicate that evolutionary and biogeographical aspects of cluster-root formation in the genus Lupinus need to be revised. Furthermore, investigation is warranted to determine the capacity of species of the large group of New-World lupins to form cluster roots in soils of their native habitats.

A comparison of the effects of auxin on cluster root initiation and development in Grevillea robusta Cunn. ex R. Br. (Proteaceae) and in the genus Lupinus (Leguminosae).

The effect of NAA (naphthaleneacetic acid) on the development of cluster roots in members of the Proteaceae and Leguminosae was investigated. The exogenous addition of NAA led to initiation of cluster roots in phosphate conditions normally inhibitory for their development, but initiation took place within the limits of the cluster pattern under −P conditions. There was no change in spacing within the cluster root nor between cluster roots in Grevillea robusta Cunn. ex R. Br. or in rootlet length or cluster root length. In Lupinus albus L., change in rootlet length and cluster root length was noted at 10-10 and 1012 M NAA. In L. albus, the length of time that roots were exposed to NAA does not appear to be important, with similar levels of cluster root initiation after 48 h and 7 days. Cluster root production in G. robusta differed from that in L. albus in terms of the concentration of NAA needed to induce initiation, and in the effects of extremely low levels of NAA on rootlet numbers and lengths. L. arboreus L. does not produce cluster roots under −P conditions. Furthermore, neither L. arboreus L., L. angustifolius L., L. luteus L. nor L. mutabilis L. were induced to produce cluster roots under −P conditions, nor under +P conditions in the presence of exogenous NAA. Thus, exogenous NAA only leads to the induction of cluster roots, at levels of P normally inhibitive of their development, in species of Lupinus that produce them under −P conditions. Auxin-induced cluster roots develop within the same constraints as those developing under −P conditions. NAA does not induce cluster roots in species of Lupinus that do not produce them under −P conditions.