Effects of environmentally relevant concentrations of oxytetracycline and sulfadiazine on the bacterial communities, antibiotic resistance genes, and functional genes are different between maize rhizosphere and bulk soil.

Antibiotic contamination in soil has become a major concern worldwide. At present, it is not clear how two co-existed antibiotics with environmentally relevant concentrations would affect soil bacterial community structure, the abundances of antibiotic resistance genes (ARGs) and functional genes, and whether the effects of antibiotics would differ between rhizosphere and bulk soil. We conducted a greenhouse pot experiment to grow maize in a loess soil treated with oxytetracycline (OTC) or sulfadiazine (SDZ) or both at an environmentally relevant concentration (1 mg kg-1) to investigate the effects of OTC and SDZ on the rhizosphere and bulk soil bacterial communities, abundances of ARGs and carbon (C)-, nitrogen (N)-, and phosphorus (P)-cycling functional genes, and on plant growth and plant N and P nutrition. The results show that the effects of environmentally relevant concentrations of OTC and SDZ on bacterial communities and abundances of ARGs and functional genes differ between maize rhizosphere and bulk soil. The effects of two antibiotics resulted in a higher absolute abundances of accA, tet(34), tnpA-04, and sul2 in the rhizosphere soil than in the bulk soil and different bacterial community compositions and biomarkers in the rhizosphere soil and the bulk soil. However, OTC had a stronger inhibitory effect on the abundances of a few functional genes in the bulk soil than SDZ did, and their combination had no synergistic effect on plant growth, ARGs, and functional genes. The role of co-existed OTC and SDZ decreased shoot height and increased root N concentration. The results demonstrate that environmentally relevant concentrations of antibiotics shift soil microbial community structure, increase the abundances of ARGs, and reduce the abundances of functional genes. Furthermore, soil contamination with antibiotics can diminish agricultural production via phytotoxic effects on crops, and combined effects of antibiotics on plant growth and nutrient uptake should be considered.

Mini-drone assisted tree canopy sampling: a low-cost and high-precision solution.

The collection of tree canopy samples in forest ecosystems has been challenging for researchers and managers during the past decades. Various methods, including pole pruner, tree climber, shooter, throw-line launcher, hydraulic lift (e.g., tower crane) and UAV (unmanned aerial vehicle)-based devices, have been used, however, they are limited by sampling height restrictions, safety hazards to a climber, low retrieving accuracy, high equipment costs, and transportation inconvenience. This study proposed a novel method for collecting tree canopy samples using a portable mini-drone. The mini-drone is operated to pull a traction line across the target branch, drag the retrieving rope to the selected cutting point of the branch, and carry the equipped wire saw or chain saw to cut the canopy sample off. Through on-site testing and field trials, this method was feasible for lower- and middle-canopy sampling (up to 30 meters tall) across most temperate broad-leaved and coniferous tree species. This technique would have great potential in plantation and old-growth forests. Adopting this low-cost mini-drone technique, researchers can collect tree canopy samples safely and efficiently, leading to improvements in relevant physiological and ecological studies focusing on functional traits of branches, leaves, and seeds.

Precipitation pattern alters the effects of nitrogen deposition on the growth of alien species Robinia pseudoacacia.

Aims: Nitrogen (N) supply and precipitation pattern (amount and frequency) both affect plant growth. However, N deposition is increasing and precipitation regimes are changing in the context of global change. An experiment was conducted to access how the growth of Robinia pseudoacacia, a widely distributed and cultivated N2-fixing alien species, is affected by both the pattern of precipitation and N supplies. Methods: Seedlings were grown in a glasshouse at four different N levels combined with different precipitation regimes, including three precipitation amounts, and two precipitation frequencies. After treatment for 75 days, plant height, biomass allocation, leaf and soil nutrient concentrations were measured. Results: Plants under high precipitation frequency had greater biomass compared with plants lower precipitation frequency, despite receiving the same amount of precipitation. Higher N supply reduced biomass allocation to nodules. Under low precipitation level, nodule growth and N2 fixation of R. pseudoacacia was more inhibited by high N deposition compared with plants under higher precipitation level. Even slightly N deposition under higher precipitation inhibited N2 fixation but it was insufficient to meet the N needs of the plants. Conclusions: Even at low levels, N deposition might inhibit N2 fixation of plants but low N in soil cannot meet the N requirements of plants, and caused N2 fixation limitation in plants during seedling stage. There was likely a transition from N2 fixation to acquisition of N from soil directly with root when N supply was increased.

New methylene blue-mediated photodynamic inactivation of multidrug-resistant Fonsecaea nubica infected chromoblastomycosis in vitro.

Chromoblastomycosis is a fungal disease presented with local warty papule, plaque, and verrucous nodules. In addition, the incidence and drug resistance of chromoblastomycosis are increasing each year worldwide. Photodynamic therapy is a promising method to treat mycoses. The purpose of this study was to evaluate the effect of new methylene blue (NMB)-induced PDT on multidrug-resistant chromoblastomycosis in vitro. We isolated one wild-type strain pathogen from one clinical patient diagnosed with chromoblastomycosis for over 27 years. The pathogen was identified by histopathology, the morphology of fungal culture, and genetic testing. Drug susceptibility testing was performed on the isolate. It was cultured with logarithmic growth phase spore in vitro and incubated with different concentrations of NMB for 30 min, and received illumination by red light-emitted diode with different light doses. After photodynamic treatment, the scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were conducted. The pathogen was Fonsecaea nubica, and it was resistant to itraconazole, terbinafine, amphotericin B, voriconazole andcaspofungin. At the same NMB concentration, the sterilization efficiency of NMB-photodynamic therapy (PDT) on F. nubica increased with increasing light intensity; F. nubica was completely killed at 25 µmol/L NMB with a light dose of 40 J/cm2 or 50 µmol/L NMB and light doses of ≥ 30 J/cm2. SEM and TEM observed ultrastructural changes after PDT. NMB-PDT inactivates the survival of multidrug-resistant F. nubica in vitro; it therefore has the potential to become an alternative or adjuvant treatment for refractory chromoblastomycosis.

Enhanced nodulation and phosphorus acquisition from sparingly-soluble iron phosphate upon treatment with arbuscular mycorrhizal fungi in chickpea.

The coordination/trade-off among below-ground strategies for phosphorus (P) acquisition, including root morphology, carboxylate exudation and colonisation by arbuscular mycorrhizal fungi (AMF), is not well understood. This is the first study investigating the relationships between root nodulation, morphology, carboxylates and colonisation by an indigenous community of AMF under varying P levels and source. Two chickpea genotypes with contrasting amounts of rhizosheath carboxylates were grown in pots at six P levels (from 0 to 160 µg g-1 ) as KH2 PO4 (KP, highly soluble) or FePO4 (FeP, sparingly soluble), with or without AMF (±AMF) treatment. Under both FeP and KP, the presence of AMF inhibited shoot growth and shoot branching, decreased total root length and specific root length, increased mean root diameter and root tissue density and reduced carboxylates. However, the role of AMF in acquiring P differed between the two P sources, with the enhanced P acquisition under FeP while not under KP. Co-inoculation of AMF and rhizobia enhanced nodulation under FeP, but not under KP. Our results suggest that the effects of AMF on shoot branching were mediated by cytokinins as the reduced shoot branching in FeP40 and KP40 under +AMF relative to -AMF coincided with a decreased concentration of cytokinins in xylem sap for both genotypes.

Genome-wide association studies and transcriptome sequencing analysis reveal novel genes associated with Al tolerance in wheat.

Aluminum (Al) toxicity is a major threat to the productivity and quality of wheat on acid soil. Identifying novel Al tolerance genes is crucial for breeders to pyramid different tolerance mechanisms thus leading to greater Al tolerance. We aim to identify novel quantitative trait loci (QTL) and key candidate genes associated with Al tolerance in wheat. Herein, we investigated the genotypic variation in Al tolerance among 334 wheat varieties using an acid soil assay. Genome-wide association study (GWAS) and transcriptome were carried out to identify key genes for Al tolerance. GWAS identified several QTL associated with acid soil tolerance including one major QTL on chromosome 1A, in addition to the QTL on 4D where TaALMT1 is located. The four significant markers around the newly identified QTL explained 27.2% of the phenotypic variation. With the existence of reported markers for TaALMT1, more than 97% of the genotypes showed tolerance to Al. For those genotypes with the existence of the novel QTL on 1A but without TaALMT1, more than 90% of genotypes showed medium or high tolerance to Al, confirming the existence of the Al tolerance gene(s) on chromosome 1A. By combining GWAS and RNA-seq analysis, we identified 11 candidate genes associated with Al tolerance. The results provide new insights into the genetic basis of Al tolerance in wheat. The identified genes can be used for the breeding of Al tolerant accessions.

Differences in foliar phosphorus fractions, rather than in cell-specific phosphorus allocation, underlie contrasting photosynthetic phosphorus use efficiency among chickpea genotypes.

Although significant intraspecific variation in photosynthetic phosphorus (P) use efficiency (PPUE) has been shown in numerous species, we still know little about the biochemical basis for differences in PPUE among genotypes within a species. Here, we grew two high PPUE and two low PPUE chickpea (Cicer arietinum) genotypes with low P supply in a glasshouse to compare their photosynthesis-related traits, total foliar P concentration ([P]) and chemical P fractions (i.e. inorganic P (Pi), metabolite P, lipid P, nucleic acid P, and residual P). Foliar cell-specific nutrient concentrations including P were characterized using elemental X-ray microanalysis. Genotypes with high PPUE showed lower total foliar [P] without slower photosynthetic rates. No consistent differences in cellular [P] between the epidermis and mesophyll cells occurred across the four genotypes. In contrast, high PPUE was associated with lower allocation to Pi and metabolite P, with PPUE being negatively correlated with the percentage of these two fractions. Furthermore, a lower allocation to Pi and metabolite P was correlated with a greater allocation to nucleic acid P, but not to lipid P. Collectively, our results suggest that a different allocation to foliar P fractions, rather than preferential P allocation to specific leaf tissues, underlies the contrasting PPUE among chickpea genotypes.

Genome-wide association study reveals quantitative trait loci for waterlogging-triggered adventitious roots and aerenchyma formation in common wheat.

Waterlogging severely affects wheat growth and development. Limited availability of oxygen in the root zone negatively affects the metabolism of plants. The formation of adventitious roots (ARs) and root cortical aerenchyma (RCA) are the most important adaptive trait contributing to plants' ability to survive in waterlogged soil conditions. This study used a genome-wide association study (GWAS) approach with 90K single nucleotide polymorphisms (SNPs) in a panel of 329 wheat genotypes, to reveal quantitative trait loci (QTL) conferring ARs and RCA. The wheat genotypes exposed to waterlogging were evaluated for ARs and RCA in both field and glasshouse over two consecutive years. Six and five significant marker-trait associations (MTAs) were identified for ARs and RCA formation under waterlogging, respectively. The most significant MTA for AR and RCA was found on chromosome 4B. Two wheat cultivars with contrasting waterlogging tolerance (tolerant: H-242, sensitive: H-195) were chosen to compare the development and regulation of aerenchyma in waterlogged conditions using staining methods. Results showed that under waterlogging conditions, H2O2 signal generated before aerenchyma formation in both sensitive and tolerant varieties with the tolerant variety accumulating more H2O2 and in a quicker manner compared to the sensitive one. Several genotypes which performed consistently well under different conditions can be used in breeding programs to develop waterlogging-tolerant wheat varieties.

Root acid phosphatases and rhizobacteria synergistically enhance white lupin and rice phosphorus acquisition.

The rhizosheath is a belowground area that acts as a communication hub at the root-soil interface to promote water and nutrient acquisition. Certain crops, such as white lupin (Lupinus albus), acquire large amounts of phosphorus (P), owing partially to exudation of acid phosphatases (APases). Plant growth-promoting rhizobacteria also increase soil P availability. However, potential synergistic effects of root APases and rhizosheath-associated microbiota on P acquisition require further research. In this study, we investigated the roles of root purple APases (PAPs) and plant growth-promoting rhizobacteria in rhizosheath formation and P acquisition under conditions of soil drying (SD) and P treatment (+P: soil with P fertilizer; -P: soil without fertilizer). We expressed purple acid phosphatase12 (LaPAP12) in white lupin and rice (Oryza sativa) plants and analyzed the rhizosheath-associated microbiome. Increased or heterologous LaPAP12 expression promoted APase activity and rhizosheath formation, resulting in increased P acquisition mainly under SD-P conditions. It also increased the abundance of members of the genus Bacillus in the rhizosheath-associated microbial communities of white lupin and rice. We isolated a phosphate-solubilizing, auxin-producing Bacillus megaterium strain from the rhizosheath of white lupin and used this to inoculate white lupin and rice plants. Inoculation promoted rhizosheath formation and P acquisition, especially in plants with increased LaPAP12 expression and under SD-P conditions, suggesting a functional role of the bacteria in alleviating P deficit stress via rhizosheath formation. Together, our results suggest a synergistic enhancing effect of LaPAP12 and plant growth-promoting rhizobacteria on rhizosheath formation and P acquisition under SD-P conditions.

Inhibition of efflux pump encoding genes and biofilm formation by sub-lethal photodynamic therapy in methicillin susceptible and resistant Staphylococcus aureus.

BACKGROUND: Photodynamic therapy (PDT) is an effective method to inactivate microorganisms which is based on reactive oxygen species (ROS) generated by photosensitizer and light at certain wavelength. Exposure to sub-lethal dose of PDT (sPDT) could activate the regulatory systems in the surviving bacteria in response to oxidative stress. This study aimed to evaluate the effect of sPDT on efflux pump and biofilm formation in Staphylococcus aureus (S. aureus), which are two important virulence related factors. METHODS: Different light irradiation time and toluidine blue O (TBO) concentrations were tested to select a sPDT in methicillin-susceptible and methicillin-resistant S. aureus (MSSA and MRSA). Efflux function was evaluated with EtBr efflux experiment. Biofilm formation was evaluated by crystal violet staining. Gene expressions of norA, norB, sepA, mepA and mdeA following sPDT were analyzed with real-time PCR. RESULTS: Sub-lethal PDT was set at 40 J/cm2 associated with 0.5 µM TBO. Efflux function was significantly inhibited in both strains. The average expression levels of mdeA and mepA in MSSA and MRSA were increased by (3.09, 1.77, 1.57) and (3,44, 1.59, 6.29) fold change respectively, norB and sepA were decreased by (3.77, 6.14) and (3.02, 3.47) fold change respectively. Expression level of norA was decreased by 5.44-fold change in MSSA but increased by 2.80-fold change in MRSA. Biofilm formation in both strains was impeded. CONCLUSIONS: TBO-mediated sPDT could inhibit efflux pump function, alter efflux pump encoding gene expression levels and retard biofilm formation in MSSA and MRSA. Therefore, sPDT is proposed as a potential adjuvant therapy for infections.

Salt­responsive transcriptome analysis of canola roots reveals candidate genes involved in the key metabolic pathway in response to salt stress.

Salinity is a major constraint on crop growth and productivity, limiting sustainable agriculture in arid regions. Understanding the molecular mechanisms of salt-stress adaptation in canola is important to improve salt tolerance and promote its cultivation in saline lands. In this study, roots of control (no salt) and 200 mM NaCl-stressed canola seedlings were collected for RNA-Seq analysis and qRT-PCR validation. A total of 5385, 4268, and 7105 DEGs at the three time points of salt treatment compared to the control were identified, respectively. Several DEGs enriched in plant signal transduction pathways were highly expressed under salt stress, and these genes play an important role in signaling and scavenging of ROS in response to salt stress. Transcript expression in canola roots differed at different stages of salt stress, with the early-stages (2 h) of salt stress mainly related to oxidative stress response and sugar metabolism, while the late-stages (72 h) of salt stress mainly related to transmembrane movement, amino acid metabolism, glycerol metabolism and structural components of the cell wall. Several families of TFs that may be associated with salt tolerance were identified, including ERF, MYB, NAC, WRKY, and bHLH. These results provide a basis for further studies on the regulatory mechanisms of salt stress adaptation in canola.

Alkaline Salt Inhibits Seed Germination and Seedling Growth of Canola More Than Neutral Salt.

Salinity is a major constraint to crop growth and productivity, limiting sustainable agriculture production. Planting canola (Brassica napus L.) variety with salinity-alkalinity tolerance as a green manure on the large area of salinity-affected land in Xinjiang could alleviate feed shortage. To investigate the differential effects of neutral and alkaline salt stress on seed germination and seedling growth of canola, we used two salts at varying concentrations, i.e., NaCl (neutral salt at 100, 150, and 200 mM) and Na2CO3 (alkaline salt at 20, 30, and 40 mM). To further explore the effects of Na+ and pH on seed germination, we included combined of NaCl (0, 100, 150, and 200 mM) and pH (7.1, 8.0, 9.0, 10.0, and 11.0). Shoot growth was promoted by low concentrations of NaCl and Na2CO3 but inhibited at high salt concentrations. Given the same Na+ concentration, Na2CO3 inhibited seed germination and seedling growth more than NaCl. The results showed that the main factor affecting seed germination and seedling growth is not pH alone, but the interaction between pH and salt ions. Under NaCl stress, canola increased the absorption of K+, Ca2+, and Mg2+ in roots and K+ in leaves. However, under Na2CO3 stress, canola maintained a high K+ concentration and K+/Na+ ratio in leaves and increased Ca2+ and Mg2+ in roots. Our study showed that alkaline salts inhibit canola seed germination and seedling growth more significantly than neutral salts and salt species, salt concentration, and pH significantly affected on seed germination and seedling growth. However, pH affected seed germination and seedling growth mainly through an interaction with salt ions.

Interactive effects of phosphorus fertilization and salinity on plant growth, phosphorus and sodium status, and tartrate exudation by roots of two alfalfa cultivars.

BACKGROUND AND AIMS: Soil phosphorus (P) deficiency and salinity are constraints to crop productivity in arid and semiarid regions. Salinity may weaken the effect of P fertilization on plant growth. We investigated the interactive effects of soil P availability and salinity on plant growth, P nutrition and salt tolerance of two alfalfa (Medicago sativa) cultivars. METHODS: A pot experiment was carried out to grow two cultivars of alfalfa in a loess soil under a combination of different rates of added P (0, 40, 80 and 160 mg P kg-1 soil as monopotassium phosphate) and sodium chloride (0, 0.4, 0.8 and 1.6 g NaCl kg-1 soil). Plant biomass, concentrations of P ([P]), sodium ([Na]) and potassium ([K]) were determined, and rhizosheath carboxylates were analysed. KEY RESULTS: There were significant interactions between soil P availability and salinity on some, but not all, of the parameters investigated, and interactions depended on cultivar. Plant growth and P uptake were enhanced by P fertilization, but inhibited by increased levels of salinity. Increasing the salinity resulted in decreased plant P-uptake efficiency and [K]/[Na]. Only soil P availability had a significant effect on the amount of tartrate in the rhizosheath of both cultivars. CONCLUSIONS: Increased salinity aggravated P deficiency. Appropriate application of P fertilizers improved the salt tolerance of alfalfa and increased its productivity in saline soils.

Co-infection by Soil-Borne Fungal Pathogens Alters Disease Responses Among Diverse Alfalfa Varieties.

Fusarium oxysporum f. sp. medicaginis (Fom) and Rhizoctonia solani (Rs) are the major soil-borne fungal pathogens that pose severe threats to commercial alfalfa production in China. However, the effects of Fom and Rs co-infection on alfalfa and whether co-infection alters disease resistance responses among diverse varieties remain unknown. A collection of 80 alfalfa varieties (Medicago sativa) originated from seven countries were used to study the effects of Fom and Rs co-infection on alfalfa and host resistance responses. The co-infection resulted in more severe disease and reductions in growth and biomass allocation across varieties in comparison with either single infection by Fom or Rs; in addition, root morphology was much more strongly altered by the co-infection. Principal component analysis based on all plant traits showed that varieties under the co-infection were related to the single infection by Rs, being separated from Fom, and hierarchical clustering found differential response patterns among varieties upon co-infection compared with either single infection, with most varieties being highly susceptible to the co-infection. Furthermore, varieties that were most resistant to either single infection were not effective to co-infection, and there was no individual variety with resistance to both pathogens singly and co-infected. This study reveals for the first time that the co-infection by Fom and Rs alters disease resistance responses among diverse alfalfa varieties and provides useful information for developing alfalfa varieties with resistance to the co-occurrence of different soil-borne pathogens.

Integrated transcriptomics and metabolomics analysis to characterize alkali stress responses in canola (Brassica napus L.).

BACKGROUND: Soil salinization is a major constraint limiting agricultural development and affecting crop growth and productivity, especially in arid and semi-arid regions. Understanding the molecular mechanism of the adaptability of canola to salt stress is very important to improve the salt tolerance of canola and promote its cultivation in saline alkali soil. RESULTS: To identify the metabolomic and transcriptomic mechanisms of canola under alkaline salt stress, we collected roots of control (no salt treatment) and 72 h Na2CO3-stressed canola seedlings (hydroponics) for metabolic profiling of metabolites, supplemented with RNA-Seq analysis and real-time quantitative PCR validation. Metabolomic analysis showed that the metabolites of amino acids and fatty acids were higher accumulated under alkaline salt stress, including L-proline, L-glutamate, L-histidine, L-phenylalanine, L-citrulline, L-tyrosine, L-saccharopine, L-tryptophan, linoleic acid, dihomo gamma linolenic acid, alpha linolenic acid, Eric acid, oleic acid and neuronic acid, while the metabolism of carbohydrate (sucrase, alpha, alpha trehalose), polyol (ribitol), UDP-D-galactose, D-mannose, D-fructose and D-glucose 6-phosphate decreased. Transcriptomic and metabolomic pathway analysis indicated that carbohydrate metabolism may not play an important role in the resistance of canola to alkaline salt stress. Organic acid metabolism (fatty acid accumulation) and amino acid metabolism are important metabolic pathways in the root of canola under alkaline salt stress. CONCLUSIONS: These results suggest that the genes and metabolites involved in fatty acid metabolism and amino acids metabolism in roots of canola may regulate salt tolerance of canola seedlings under alkaline salt stress, which improves our understanding of the molecular mechanisms of salt tolerance in canola.

Novel Genes and Genetic Loci Associated With Root Morphological Traits, Phosphorus-Acquisition Efficiency and Phosphorus-Use Efficiency in Chickpea.

Chickpea-the second most important grain legume worldwide-is cultivated mainly on marginal soils. Phosphorus (P) deficiency often restricts chickpea yields. Understanding the genetics of traits encoding P-acquisition efficiency and P-use efficiency will help develop strategies to reduce P-fertilizer application. A genome-wide association mapping approach was used to determine loci and genes associated with root architecture, root traits associated with P-acquisition efficiency and P-use efficiency, and any associated proxy traits. Using three statistical models-a generalized linear model (GLM), a mixed linear model (MLM), and a fixed and random model circulating probability unification (FarmCPU) -10, 51, and 40 marker-trait associations (MTAs), respectively were identified. A single nucleotide polymorphism (SNP) locus (Ca1_12310101) on Ca1 associated with three traits, i.e., physiological P-use efficiency, shoot dry weight, and shoot P content was identified. Genes related to shoot P concentration (NAD kinase 2, dynamin-related protein 1C), physiological P-use efficiency (fasciclin-like arabinogalactan protein), specific root length (4-coumarate-CoA ligase 1) and manganese concentration in mature leaves (ABC1 family protein) were identified. The MTAs and novel genes identified in this study can be used to improve P-use efficiency in chickpea.

Foliar nutrient allocation patterns in Banksia attenuata and Banksia sessilis differing in growth rate and adaptation to low-phosphorus habitats.

BACKGROUND AND AIMS: Phosphorus (P) and nitrogen (N) are essential nutrients that frequently limit primary productivity in terrestrial ecosystems. Efficient use of these nutrients is important for plants growing in nutrient-poor environments. Plants generally reduce foliar P concentration in response to low soil P availability. We aimed to assess ecophysiological mechanisms and adaptive strategies for efficient use of P in Banksia attenuata (Proteaceae), naturally occurring on deep sand, and B. sessilis, occurring on shallow sand over laterite or limestone, by comparing the allocation of P among foliar P fractions. METHODS: We carried out pot experiments with slow-growing B. attenuata, which resprouts after fire, and faster growing opportunistic B. sessilis, which is killed by fire, on substrates with different P availability using a randomized complete block design. We measured leaf P and N concentrations, photosynthesis, leaf mass per area, relative growth rate and P allocated to major biochemical fractions in B. attenuata and B. sessilis. KEY RESULTS: The two species had similarly low foliar total P concentrations, but distinct patterns of P allocation to P-containing fractions. The foliar total N concentration of B. sessilis was greater than that of B. attenuata on all substrates. The foliar total P and N concentrations in both species decreased with decreasing P availability. The relative growth rate of both species was positively correlated with concentrations of both foliar nucleic acid P and total N, but there was no correlation with other P fractions. Faster growing B. sessilis allocated more P to nucleic acids than B. attenuata did, but other fractions were similar. CONCLUSIONS: The nutrient allocation patterns in faster growing opportunistic B. sessilis and slower growing B. attenuata revealed different strategies in response to soil P availability which matched their contrasting growth strategy.

Rapid nitrogen fixation contributes to a similar growth and photosynthetic rate of Robinia pseudoacacia supplied with different levels of nitrogen.

Robinia pseudoacacia L. is a legume species that is widely used in afforestation, which has high N2 fixation capacity and rapid growth rate. Both nitrogen (N) supply and phenology affect plant growth, photosynthesis and leaf senescence. The aim of this study was to determine how N supply affects N2 fixation, leaf photosynthesis and senescence of R. pseudoacacia at different phenological stages. Seedlings of R. pseudoacacia were supplied with different levels of 15N-labelled NH4NO3 solution, with seedlings of Sophora japonica Linn. as reference plants to calculate the percentage of N derived from the atmospheric N2 (%Ndfa). Compared with plants supplied with a high N level, those with a low N supply had a higher %Ndfa at an early developmental stage. Nitrogen fixation compensated the effect of a low N supply on plant growth in R. pseudoacacia. A high N supply decreased biomass allocation to lateral roots and nodules, and increased the relative growth rate of plant height as well as specific leaf area. The eighth mature compound leaf of R. pseudoacacia tended to have a higher net photosynthetic rate than the fourth leaf, and the leaves still maintained a moderate photosynthetic rate in early autumn. Plants tended to allocate more biomass to leaves at an early developmental stage and to stems and roots at a later developmental stage (3 months old). The N level did not affect leaf photosynthesis at different phenological stages, primarily due to (i) a high %Ndfa under low N supply at early growing stage, and a similar high %Ndfa under all N supplies at a late growing stage, and (ii) the delayed greening phenotype of expanding leaves to save nutrients for mature leaves.

Accumulation of phosphorus and calcium in different cells protects the phosphorus-hyperaccumulator Ptilotus exaltatus from phosphorus toxicity in high-phosphorus soils.

Ptilotus exaltatus accumulates phosphorus (P) to > 40 mg g-1 without toxicity symptoms, while Kennedia prostrata is intolerant of increased P supply. What physiological mechanisms underlie this difference and protect P. exaltatus from P toxicity? Ptilotus exaltatus and K. prostrata were grown in a sandy soil with low-P, high-P and P-pulse treatments. Both species hyperaccumulated P (>20 mg g-1) under high-P and P-pulse treatments; shoot dry weight was unchanged for P. exaltatus, but decreased by >50% for K. prostrata. Under high-P, in young fully-expanded leaves, both species accumulated P predominantly as inorganic P. However, P. exaltatus preferentially allocated P to mesophyll cells and stored calcium (Ca) as occasional crystals in specific lower mesophyll cells, separate from P, while K. prostrata preferentially allocated P to epidermal and spongy mesophyll cells, but co-located P and Ca in palisade mesophyll cells where granules with high [P] and [Ca] were evident. Mesophyll cellular [P] correlated positively with [potassium] for both species, and negatively with [sulfur] for P. exaltatus. Thus, P. exaltatus tolerated a very high leaf [inorganic P] (17 mg g-1), associated with P and Ca allocation to different cell types and formation of Ca crystals, thereby avoiding deleterious precipitation of Ca3(PO4)2. It also showed enhanced [potassium] and decreased [sulfur] to balance high cellular [P]. Phosphorus toxicity in K. prostrata arose from co-location of Ca and P in palisade mesophyll cells. This study advances understanding of leaf physiological mechanisms for high P tolerance in a P-hyperaccumulator and indicates P. exaltatus as a promising candidate for P-phytoextraction.