Functional traits shape plant-plant interactions and recruitment in a hotspot of woody plant diversity.

Understanding and predicting recruitment in species-rich plant communities requires identifying functional determinants of both density-independent performance and interactions. In a common-garden field experiment with 25 species of the woody plant genus Protea, we varied the initial spatial and taxonomic arrangement of seedlings and followed their survival and growth during recruitment. Neighbourhood models quantified how six key functional traits affect density-independent performance, interaction effects and responses. Trait-based neighbourhood models accurately predicted individual survival and growth from the initial spatial and functional composition of species-rich experimental communities. Functional variation among species caused substantial variation in density-independent survival and growth that was not correlated with interaction effects and responses. Interactions were spatially restricted but had important, predominantly competitive, effects on recruitment. Traits increasing the acquisition of limiting resources (water for survival and soil P for growth) mediated trade-offs between interaction effects and responses. Moreover, resprouting species had higher survival but reduced growth, likely reinforcing the survival-growth trade-off in adult plants. Resource acquisition of juvenile plants shapes Protea community dynamics with acquisitive species with strong competitive effects suffering more from competition. Together with functional determinants of density-independent performance, this makes recruitment remarkably predictable, which is critical for efficient restoration and near-term ecological forecasts of species-rich communities.

Regulation and physiological function of proteins for heat tolerance in cowpea (Vigna unguiculata) genotypes under controlled and field conditions.

The expression of heat shock proteins is considered a central adaptive mechanism to heat stress. This study investigated the expression of heat shock proteins (HSPs) and other stress-protective proteins against heat stress in cowpea genotypes under field (IT-96D-610 and IT-16) and controlled (IT-96D-610) conditions. Heat stress response analysis of proteins at 72 h in the controlled environment showed 270 differentially regulated proteins identified using label-free quantitative proteomics in IT-96D-610 plants. These plants expressed HSPs and chaperones [BAG family molecular chaperone 6 (BAG6), Multiprotein bridging factor1c (MBF1C) and cold shock domain protein 1 (CSDP1) in the controlled environment]. However, IT-96D-610 plants expressed a wider variety of small HSPs and more HSPs in the field. IT-96D-610 plants also responded to heat stress by exclusively expressing chaperones [DnaJ chaperones, universal stress protein and heat shock binding protein (HSBP)] and non-HSP proteins (Deg1, EGY3, ROS protective proteins, temperature-induced lipocalin and succinic dehydrogenase). Photosynthesis recovery and induction of proteins related to photosynthesis were better in IT-96D-610 because of the concurrent induction of heat stress response proteins for chaperone functions, protein degradation for repair and ROS scavenging proteins and PSII operating efficiency (Fq'/Fm') than IT-16. This study contributes to identification of thermotolerance mechanisms in cowpea that can be useful in knowledge-based crop improvement.

Proteome changes and associated physiological roles in chickpea (Cicer arietinum) tolerance to heat stress under field conditions.

Interrogative proteome analyses are used to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in heat-stressed plants. The objectives of the study were to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in chickpea (Cicer arietinum L.) heat-tolerant (Acc#7) and sensitive genotype (Acc#8) from a field study. Proteomic and gene ontological analyses showed an upregulation in proteins related to protein synthesis, intracellular traffic, defence and transport in the heat-tolerant genotype compared to the susceptible one at the warmer site. Results from KEGG analyses indicate the involvement of probable sucrose-phosphate synthase (EC 2.4.1.14) and sucrose-phosphate phosphatase (EC 3.1.3.24) proteins, that were upregulated in the heat-tolerant genotype at the warmer site, in the starch and sucrose pathway. The presence of these differentially regulated proteins including HSP70, ribulose bisphosphate carboxylase/oxygenase activase, plastocyanin and protoporphyrinogen oxidase suggests their potential role in heat tolerance, at flowering growth stage, in field-grown chickpea. This observation supports unaltered physiological and biochemical performance of the heat-tolerant genotypes (Acc#7) relative to the susceptible genotype (Acc#8) in related studies (Makonya et al. 2019). Characterisation of the candidate proteins identified in the current study as well as their specific roles in the tolerance to heat stress in chickpea are integral to further crop improvement initiatives.

Clinical scribes and their association with patient experience in the otolaryngology clinic.

OBJECTIVES/HYPOTHESIS: Electronic health records have brought many advantages but also placed a documentation burden on the provider during and after the clinic visit. Some otolaryngologists have countered this challenge by employing clinical scribes. This project aimed to better understand the influence of scribes on patient experience in the otolaryngology clinic. STUDY DESIGN: Retrospective cohort survey study. METHODS: Patients presenting to the otolaryngology clinic for new and follow-up appointments were recruited to complete surveys about their experience. RESULTS: A total of 153 patients completed the survey, and 96 of those patients (62.7%) interacted with a scribe. Patient satisfaction was not significantly associated with participation of the scribe (P = .668). Similarly, patient rating of their physician on a scale of 1 to 10 was not associated with scribe involvement (P = .851). The patients who did interact with a scribe responded that the scribe positively impacted the visit 77.1% of the time. Participation of a resident, primary language other than English, and use of interpreter were associated with lower satisfaction (P = .004, P < .001, and P < .001, respectively). CONCLUSIONS: There are no published data on the effect of scribes on patient experience in the otolaryngology clinic. In other specialties, scribes have been demonstrated as having a positive effect on provider satisfaction, clinical productivity, and patient perception. These data demonstrate that patient satisfaction was neither impaired nor improved by the presence of the scribe in this clinic. In light of benefits demonstrated by prior studies, these findings support the conclusion that scribes are a useful adjunct in providing high-level otolaryngology care. LEVEL OF EVIDENCE: 4 Laryngoscope, 130:E134-E139, 2020.

Showing their mettle: extraradical mycelia of arbuscular mycorrhizae form a metal filter to improve host Al tolerance and P nutrition.

BACKGROUND: New evidence has shown that arbuscular mycorrhizal (AM) fungi can contribute to the aluminum (Al3+ ) tolerance of host plants growing in acidic soils with phytotoxic levels of Al3+ . The aim of this study was to investigate the role of AM fungi isolated from naturally occurring Al3+ acidic soils in conferring host tolerance to Al3+ toxicity in three wheat cultivars differing in Al3+ sensitivity. The experiment was conducted in a soilless substrate (vermiculite/perlite, 2:1 v/v) using two Al3+ -tolerant wheat genotypes and one Al3+ -sensitive wheat genotype. The wheat was colonized with a consortium of AM fungi isolated from an Andisol, with or without Al3+ at a concentration of 200 µmol L-1 . RESULTS: The response of wheat to Al3+ in the medium was dependent on both the plant genotype and AM colonization. The benefits of the AM fungi to the wheat cultivars included an increased P concentration and relatively low Al3+ accumulation in the plants. This was achieved through two mechanisms. First, the metal-chelating capacity of the AM fungi was clear in two of the cultivars ('Tukan' and 'Porfiado'), in which the enhanced extraradical mycelium development was able to retain Al3+ in the glomalin and hyphae. Second, the increased AM-induced acid phosphatase activity in the rhizosphere of the other cultivar ('Atlas 66') increased host nutrition possibly by hyphae-mediated nutrient uptake and glomalin-related soil protein. CONCLUSION: The results suggest that the role of AM fungi in cultivar-specific Al3+ detoxification can be achieved by increased extraradical mycelial filters and enhanced bioavailability of P in the host rhizosphere. © 2019 Society of Chemical Industry.

Glutamate dehydrogenase is essential in the acclimation of Virgilia divaricata, a legume indigenous to the nutrient-poor Mediterranean-type ecosystems of the Cape Fynbos.

Glutamate dehydrogenase (NAD(H)- GDH, EC 1.4.1.2) is an important enzyme in nitrogen (N) metabolism. It serves as a link between C and N metabolism, in its role of assimilating ammonia into glutamine or deaminating glutamate into 2-oxoglutarate and ammonia. GDH may also have a key in the N assimilation of legumes growing in P-poor soils. Virgilia divaricata is such a legume, growing in the nutrient limited soils of the mediterranean-type Cape fynbos ecosystem. In order to understand the role of GDH in the nitrogen nutrition of V. divaricata, the aim of this study was to identify the GDH gene transcripts, their relative expressions and enzyme activity in P-stressed roots and nodules during N metabolism. During P deficiency there was a reduction in total plant biomass as well as total plant P concentration. The analysis of the GDH cDNA sequences in V. divaricata revealed the presence of GHD1 and GHD2 subunits, these corresponding to the GDH1, GDH-B and GDH3 genes of legumes and non-legume plants. The relative expression of GDH1 and GDH2 genes in the roots and nodules, indicates that two the subunits were differently regulated depending on the organ type, rather than P supply. Although both transcripts appeared to be ubiquitously expressed in the roots and nodules, the GDH2 transcript evidently predominated over those of GDH1. Furthermore, the higher expression of both GDH transcripts in the roots than nodules, suggests that roots are more reliant on on GDH in P-poor soils, than nodules. With regards to GHD activity, both aminating and deaminating GDH activities were differently affected by P deficiency in roots and nodules. This may function to assimilate N and regulate internal C and N in the roots and nodules. The variation in GDH1 and GDH2 transcript expression and GDH enzyme activities, indicate that the enzyme may be regulated by post-translational modification, instead of by gene expression during P deficiency in V. divaricata.

Photosynthetic metabolism during phosphate limitation in a legume from the Mediterranean-type Fynbos ecosystem.

Phosphorus (P) is an essential mineral, required for crucial plant genetic, metabolic and signaling functions. Under P deficiency, normal physiological function can be disrupted, especially photosynthetic metabolism. The majority of photosynthetic studies of P stress has been on model organisms, and very little is known about plants that evolved on P deficient soils. Aspalathus linearis (Burm.f.) R.Dahlgren, a native to the Mediterranean ecosystem of South Africa was used to study the photosynthetic responses during short-term P limitation. A. linearis seedlings were cultured under glasshouse conditions and exposed to short-term P stress. Leaf photosynthetic gas exchange was coupled with metabolic analyses. In spite of the decline in leaf cellular Pi, the photosynthetic rates remained unchanged. These leaves also maintained their levels of light harvesting and reaction center pigments. The efficiency of the light reactions' utilization of ATP and NADPH increased during P-stress. Leaf glucose levels decreased during P-stress, while sucrose concentrations remained unaffected. These results show that during short-term P-stress, A. linearis can maintain its photosynthetic rates by altering the structural and functional components of the light reactions.

Differential patterns of nitrogen nutrition and growth cost of the indigenous Vachellia sieberiana and the introduced Chromolaena odorata in the savannah environment.

Vachellia sieberiana fixes atmospheric nitrogen (N) and distributes it back into ecosystems. We hypothesize that biological nitrogen fixation in this plant species is limited by competition from the invasive shrub, Chromolaena odorata. Competition would therefore result in the legume plant switching its limited nitrogen (N) sources in phosphorus-poor soils in savannah ecosystems when resources have to be shared. This study investigated the different patterns of N use and growth costs by a native and an introduced leguminous shrubby species. We propose that the two species sharing the same environment might result in competition. The competitive effect would induce in the indigenous legume to better utilize atmospheric-derived N modifying plant growth kinetics and plant mineral concentrations. Seedlings of V. sieberiana were cultivated in natural soil inoculum with low levels of phosphorus (mg L-1 ± SE) of 3.67 ± 0.88. The experiments were divided into two treatments where (i) seedlings of V. sieberiana were subjected to competition by cultivating them together with seedlings of C. odorata, and (ii) seedlings of V. sieberiana were cultivated independently. Although V. sieberiana was subjected to competition, the N2-fixing bacteria that occupied the nodules was Mesorhizobium species, similar to plants not subjected to competition. Total plant biomass was similar between treatments although V. sieberiana plants subjected to competition accumulated more below-ground biomass and showed higher carbon construction costs than plants growing individually. Total plant phosphorus and nitrogen decreased in seedlings of V. sieberiana under competition, whereas no differences were observed in percent N derived from the atmosphere (%NDFA) between treatments. The specific nitrogen utilization rate (SNUR) was higher in V. sieberiana plants subjected to competition while specific nitrogen absorption rate (SNAR) showed the opposite response. Vachellia sieberiana is highly adapted to nutrient-poor savannah ecosystems and can withstand competition from invasive shrubs by utilizing both atmospheric and soil nitrogen sources.

Chlorophyll fluorescence and carbohydrate concentration as field selection traits for heat tolerant chickpea genotypes.

Chickpea (Cicer arietinum L.), a cool season crop is severely affected by heat stress, predicted to increase due to warming climates. Research for identifying heat tolerance markers for potential chickpea genotype selection is imperative. The study assessed the response of four chickpea genotypes to a natural temperature gradient in the field using chlorophyll fluorescence, non-structural carbohydrate, chlorophyll concentrations, gas exchange and grain yield. Field experiments were carried out in two winter seasons at three locations with known differences in temperature in NE South Africa. Results showed two genotypes were tolerant to heat stress with an Fv/Fm of 0.83-0.85 at the warmer site, while the two sensitive genotypes showed lower Fv/Fm of 0.78-0.80. Both dark-adapted Fv/Fm and Fq'/Fm' (where Fq' = Fm' -F) measured at comparable high light levels correlated positively with grain yield. The two tolerant genotypes also showed higher photosynthetic rates, starch, sucrose and grain yield than the sensitive genotypes at the warmer site. However, these parameters were consistently higher at the cooler sites than at the warmer. These results were further validated by a climate chamber experiment, where higher Fv/Fm decline in the sensitive compared to tolerant genotypes was observed when they were exposed to short-term heat treatments of 30/25 °C and 35/30 °C. Tolerant genotypes had higher Fv/Fm (0.78-0.81) and grain yield plant-1(1.12-2.37g) compared to sensitive genotypes (0.74-0.75) and (0.32-0.89g plant-1) respectively in the 35/30 °C. It is concluded that chlorophyll fluorescence and leaf carbohydrates are suitable tools for selection of heat tolerant chickpea genotypes under field conditions, while the coolest site showed favourable conditions for chickpea production.

Roots and Nodules Response Differently to P Starvation in the Mediterranean-Type Legume Virgilia divaricata.

Virgilia divaricata is a tree legume that grows in the Cape Floristic Region (CFA) in poor nutrient soils. A comparison between high and low phosphate growth conditions between roots and nodules was conducted and evaluated for the plants ability to cope under low phosphate stress conditions in V. divaricata. We proved that the plant copes with low phosphate stress through an increased allocation of resources, reliance on BNF and enhanced enzyme activity, especially PEPC. Nodules had a lower percentage decline in P compared to roots to uphold its metabolic functions. These strategies partly explain how V. divaricata can sustain growth despite LP conditions. Although the number of nodules declined with LP, their biomass remained unchanged in spite of a plant decline in dry weight. This is achieved via the high efficiency of BNF under P stress. During LP, nodules had a lower % decline at 34% compared to the roots at 88%. We attribute this behavior to P conservation strategies in LP nodules that imply an increase in a metabolic bypass that operates at the PEP branch point in glycolysis. The enhanced activities of nodule PEPC, MDH, and ME, whilst PK declines, suggests that under LP conditions an adenylate bypass was in operation either to synthesize more organic acids or to mediate pyruvate via a non-adenylate requiring metabolic route. Both possibilities represent a P-stress adaptation route and this is the first report of its kind for legume trees that are indigenous to low P, acid soils. Although BNF declined by a small percentage during LP, this P conservation was evident in the unchanged BNF efficiency per weight, and the increase in BNF efficiency per mol of P. It appears that legumes that are indigenous to acid soils, may be able to continue their reliance on BNF via increased allocation to nodules and also due to increase their efficiency for BNF on a P basis, owing to P-saving mechanisms such as the organic acid routes.

Adaptive strategies for nitrogen metabolism in phosphate deficient legume nodules.

Legumes play a significant role in natural and agricultural ecosystems. They can fix atmospheric N2 and contribute the fixed N to soils and plant N budgets. In legumes, the availability of P does not only affect nodule development, but also N acquisition and metabolism. For legumes as an important source of plant proteins, their capacity to metabolise N during P deficiency is critical for their benefits to agriculture and the natural environment. In particular for farming, rock P is a non-renewable source of which the world has about 60-80 years of sustainable extraction of this P left. The global production of legume crops would be devastated during a scarcity of P fertiliser. Legume nodules have a high requirement for mineral P, which makes them vulnerable to soil P deficiencies. In order to maintain N metabolism, the nodules have evolved several strategies to resist the immediate effects of P limitation and to respond to prolonged P deficiency. In legumes nodules, N metabolism is determined by several processes involving the acquisition, assimilation, export, and recycling of N in various forms. Although these processes are integrated, the current literature lacks a clear synthesis of how legumes respond to P stress regarding its impact on N metabolism. In this review, we synthesise the current state of knowledge on how legumes maintain N metabolism during P deficiency. Moreover, we discuss the potential importance of two additional alterations to N metabolism during P deficiency. Our goals are to place these newly proposed mechanisms in perspective with other known adaptations of N metabolism to P deficiency and to discuss their practical benefits during P deficiency in legumes.

Extracellular matrix presentation modulates vascular smooth muscle cell mechanotransduction.

The development of atherosclerosis involves phenotypic changes among vascular smooth muscle cells (VSMCs) that correlate with stiffening and remodeling of the extracellular matrix (ECM). VSMCs are highly sensitive to the composition and mechanical state of the surrounding ECM, and ECM remodeling during atherosclerosis likely contributes to pathology. We hypothesized that ECM mechanics and biochemistry are interdependent in their regulation of VSMC behavior and investigated the effect of ligand presentation on certain stiffness-mediated processes. Our findings demonstrate that substrate stiffening is not a unidirectional stimulus-instead, the influence of mechanics on cell behavior is highly conditioned on ligand biochemistry. This "stiffness-by-ligand" effect was evident for VSMC adhesion, spreading, cytoskeletal polymerization, and focal adhesion assembly, where VSMCs cultured on fibronectin (Fn)-modified substrates showed an augmented response to increasing stiffness, whereas cells on laminin (Ln) substrates showed a dampened response. By contrast, cells on Fn substrates showed a decrease in myosin light chain (MLC) phosphorylation and elongation with increasing stiffness, whereas Ln supported an increase in MLC phosphorylation and no change in cell shape with increasing stiffness. Taken together, these findings show that identical cell populations exhibit opposing responses to substrate stiffening depending on ECM presentation. Our results also suggest that the shift in VSMC phenotype in a developing atherosclerotic lesion is jointly regulated by stromal mechanics and biochemistry. This study highlights the complex influence of the blood vessel wall microenvironment on VSMC phenotype and provides insight into how cells may integrate ECM biochemistry and mechanics during normal and pathological tissue function.

Balanced allocation of organic acids and biomass for phosphorus and nitrogen demand in the fynbos legume Podalyria calyptrata.

Podalyria calyptrata is from fynbos soils with low availability of phosphorus (P) and nitrogen (N). We investigated the physiological basis for tolerance of low P supply in nodulated P. calyptrata and examined responses to increased supply of combined-N as Ca(NO3)2 and P. It was hypothesized that increasing supply of combined-N would stimulate P-acquisition mechanisms and enhance plant growth with high P supply. Biomass, leaf [N] and [P], organic acid and phosphatase root exudates, and phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) activity in nodules and roots were examined in two N×P experiments. Low P supply decreased leaf [P] and limited growth, decreasing the nodule:root ratio but increasing nodular PEPC and MDH activity for enhanced P-acquisition or P-utilization. At low P supply, a N-induced demand for P increased root exudation of citrate and PEPC and MDH activity in roots. Greater combined-N supply inhibited nodulation more at low P supply than at high P supply. With a P-induced demand for N the plants nodulated prolifically and increased combined-N supply did not enhance plant growth. The physiological basis for N2-fixing P. calyptrata tolerating growth at low P supply and responding to greater P supply is through balanced acquisition of P and N for plant demand.

Nodules from Fynbos legume Virgilia divaricata have high functional plasticity under variable P supply levels.

Legumes have the unique ability to fix atmospheric nitrogen (N2) via symbiotic bacteria in their nodules but depend heavily on phosphorus (P), which affects nodulation, and the carbon costs and energy costs of N2 fixation. Consequently, legumes growing in nutrient-poor ecosystems (e.g., sandstone-derived soils) have to enhance P recycling and/or acquisition in order to maintain N2 fixation. In this study, we investigated the flexibility of P recycling and distribution within the nodules and their effect on N nutrition in Virgilia divaricata Adamson, Fabaceae, an indigenous legume in the Cape Floristic Region of South Africa. Specifically, we assessed tissue elemental localization using micro-particle-induced X-ray emission (PIXE), measured N fixation using nutrient concentrations derived from inductively coupled mass-spectrometry (ICP-MS), calculated nutrient costs, and determined P recycling from enzyme activity assays. Morphological and physiological features characteristic of adaptation to P deprivation were observed for V. divaricata. Decreased plant growth and nodule production with parallel increased root:shoot ratios are some of the plastic features exhibited in response to P deficiency. Plants resupplied with P resembled those supplied with optimal P levels in terms of growth and nutrient acquisition. Under low P conditions, plants maintained an increase in N2-fixing efficiency despite lower levels of orthophosphate (Pi) in the nodules. This can be attributed to two factors: (i) an increase in Fe concentration under low P, and (ii) greater APase activity in both the roots and nodules under low P. These findings suggest that V. divaricata is well adapted to acquire N under P deficiency, owing to the plasticity of its nodule physiology.

Expression of novel cytosolic malate dehydrogenases (cMDH) in Lupinus angustifolius nodules during phosphorus starvation.

During P deficiency, the increased activity of malate dehydrogenase (MDH, EC 1.1.1.37) can lead to malate accumulation. Cytosolic- and nodule-enhanced MDH (cMDH and neMDH, respectively) are known isoforms, which contribute to MDH activity in root nodules. The aim of this study was to investigate the role of the cMDH isoforms in nodule malate supply under P deficiency. Nodulated lupins (Lupinus angustifolius var. Tanjil) were hydroponically grown at adequate P (+P) or low P (-P). Total P concentration in nodules decreased under P deficiency, which coincided with an increase in total MDH activity. A consequence of higher MDH activity was the enhanced accumulation of malate derived from dark CO2 fixation via PEPC and not from pyruvate. Although no measurable neMDH presence could be detected via PCR, gene-specific primers detected two 1kb amplicons of cMDH, designated LangMDH1 (corresponding to +P, HQ690186) and LangMDH2 (corresponding to -P, HQ690187), respectively. Sequencing analyses of these cMDH amplicons showed them to be 96% identical on an amino acid level. There was a high degree of diversification between proteins detected in this study and other known MDH proteins, particularly those from other leguminous plants. Enhanced malate synthesis in P-deficient nodules was achieved via increased anaplerotic CO2 fixation and subsequent higher MDH activities. Novel isoforms of cytosolic MDH may be involved, as shown by gene expression of specific genes under P deficiency.

Short-term supply of elevated phosphate alters the belowground carbon allocation costs and functions of lupin cluster roots and nodules.

The legume Lupinus albus is able to survive under low nutrient conditions due to the presence of two specialized below ground organs for the acquisition of nitrogen and phosphate, respectively.In this regard, cluster roots increase phosphate uptake and root nodules acquire atmospheric N2via biological nitrogen fixation(BNF). Although these organs normally tolerate low phosphate conditions, very little is known about their physiological and metabolic flexibility during short-term changes in phosphate supply. The aim of this investigation was therefore to determine the physiological and metabolic flexibility of these organs during short-term supply of elevated phosphate nutrition. L. albus was cultivated in sand culture for 4 weeks at 0.1 mM phosphate supply, and then supplied with 2 mM phosphate for 2 weeks. Short-term elevated phosphate supply caused increased allocation of carbon and respiratory costs to nodules, at the expense of cluster root function. This alteration was also reflected in the increase in nodule enzyme activities related to organic acid synthesis, such as Phosphoenol-pyruvate Carboxylase (PEPC), Pyruvate Kinase (PK), Malate Dehydrogenase(NADH-MDH) and Malic Enzyme (ME). In cluster roots, elevated phosphate conditions caused a decline in these organic acid synthesizing enzymes. Phosphate recycling via Acid Phosphatase (APase),declined in nodules with elevated phosphate supply, but increased in cluster roots. Our findings suggest that during short-term elevated phosphate supply, there is a great degree of physiological and metabolic flexibility in lupin nutrient acquiring structures, and that these changes are related to the altered physiology of these organs [corrected].

Phosphorus deficiency affects the allocation of below-ground resources to combined cluster roots and nodules in Lupinus albus.

Lupins can rely on both cluster roots and nodules for P acquisition and biological nitrogen fixation (BNF), respectively. The resource allocation (C, N and P) between cluster roots and nodules has been largely understudied during P-deficient conditions. The aim of this investigation was therefore to determine the changes in resource allocation between these organs during fluctuations in P supply. Lupinus albus was cultivated in sand culture for 3 weeks, with either sufficient (2 mM high) or limiting (0.1 mM low) P supply. Although variation on P supply had no effect on the total biomass, there were significant differences in specialised below-ground organ allocation to cluster roots and nodule formation. Cluster root formation and the associated C-costs increased during low P supply, but at sufficient P-supply the construction and growth respiration costs of cluster roots declined along with their growth. In contrast to the cluster root decline at high P supply, there was an increase in nodule growth allocation and corresponding C-costs. However, this was not associated with an increase in BNF. Since cluster roots were able to increase P acquisition under low P conditions, this below-ground investment may also have benefited the P nutrition of nodules. These findings provide evidence that when lupins acquire N via BNF in their nodules, there may be a trade-off in resource allocation between cluster roots and nodules.

Increasing nitrogen supply stimulates phosphorus acquisition mechanisms in the fynbos species Aspalathus linearis.

We investigated the physiological basis for tolerance of limiting P supply and for enhanced growth with simultaneous addition of N and P in Aspalathus linearis (Burm. f.) R. Dahlgren. It was hypothesised that increasing N supply would stimulate P acquisition mechanisms and enhance plant growth with high P supply. In sand, plants received 100µM, 300µM, 500µM and 700µM N at a low P level of 10µM and a high P level of 100µM. In solution, plants received 200µM and 500µM N at a low P level of 5µM and a high P level of 15µM. Cluster roots formed only in plants with low P supply. Roots showed greater citrate and malate production and phosphatase activity at 5µM P than at 15µM P. At 10µM P, greater N supply enhanced cluster root formation to 60% of root biomass, and increased the phosphatase activity of noncluster roots and succinate release by both root types. At a high P supply of 15µM, greater N supply stimulated phosphatase activity of roots by 50%, increasing P uptake and plant growth. With increased resource partitioning towards P acquisition due to greater P demand, A. linearis is tolerant of low P supply and highly responsive to combined addition of N and P.

Infrared gas analysis technique for the study of the regulation of photosynthetic responses.

Homeostatic maintenance of physiological and biochemical processes is a key requirement for survival and adaptive responses of multicellular organisms such as plants. These important processes are in part mediated by various plant enzymes and hormones, many of which are in part, controlled by cyclic nucleotides and/or other signalling molecules. Infrared gas analysis (IRGA) technique is one of the modern methods which allows for rapid and accurate measurements of cyclic nucleotide mediated photosynthetic responses to plant hormones, and thus makes it a powerful and useful tool to study aspects of downstream cell signalling events in plants. In this chapter the basic protocols enabling the use of the IRGA technique to study signalling molecules, such as cyclic nucleotides on photosynthetic responses, are outlined.