Spatiotemporal Pattern of Acid Phosphatase Activity in Soils Cultivated With Maize Sensing to Phosphorus-Rich Patches

AimsAcid phosphatase (APase) secretion by roots allows plants to mobilize organic phosphorus (P) in low P soils. However, the spatiotemporal dynamics of soil APase activity in response to P-rich patches remain unclear.MethodsHere, we grew maize in rhizoboxes with two contrasting soil types and different localized P supplies. In situ soil zymography was applied to examine the spatial-temporal variation of APase activity.ResultsWe found P-rich patches can induce the secretion of APase from roots, indicating that even mineral P fertilizers were localized apply, mobilization of soil organic P by roots can also be enhanced; APase hotspot areas and APase activities in the rhizosphere and bulk soil of the same rhizobox showed opposite diurnal rhythms across the whole soil profile. The APase hotspot area was 10–140% larger at noon than at midnight in the rhizosphere, which is consistent with the diurnal rhythm of photosynthesis. In contrast, in bulk soil, the area was 18–200% larger at midnight than at noon, which led to spatiotemporal niche differentiation with regard to the utilization of soil organic P; this alleviated competition between plants and soil microorganisms.ConclusionOur findings showed that APase secretion of roots was plastic in P-rich patches and showed an opposite diurnal rhythm with soil microorganisms in bulk soil.

Phosphorus Release and Regeneration Following Laboratory Lysis of Bacterial Cells

The availability of phosphorus limits primary production in large regions of the oceans, and marine microbes use a variety of strategies to overcome this limitation. One strategy is the production of alkaline phosphatase (APase), which allows hydrolysis of larger dissolved organic phosphorus (DOP) compounds in the periplasm or at the cell surface for transport of orthophosphate into the cell. Cell lysis, driven by grazing and viral infection, releases phosphorus-containing cell components, along with active enzymes that could persist after lysis. The importance of this continued enzymatic activity for orthophosphate regeneration is unknown. We used three model bacteria – Escherichia coli K-12 MG1655, Synechococcus sp. WH7803, and Prochlorococcus sp. MED4 – to assess the impact of continued APase activity after cell lysis, via lysozyme treatment, on orthophosphate regeneration. Direct release of orthophosphate scaled with cell size and was reduced under phosphate-starved conditions where APase activity continued for days after lysis. All lysate incubations showed post-lysis orthophosphate generation suggesting phosphatases other than APase maintain activity. Rates of DOP hydrolysis and orthophosphate remineralization varied post-lysis among strains. Escherichia coli K-12 MG1655 rates of remineralization were 0.6 and 1.2 amol cell–1hr–1 under deplete and replete conditions; Synechococcus WH7803 lysates ranged from 0.04 up to 0.3 amol cell–1hr–1 during phosphorus deplete and replete conditions, respectively, while in Prochlorococcus MED4 lysates, rates were stable at 0.001 amol cell–1hr–1 in both conditions. The range of rates of hydrolysis and regeneration underscores the taxonomic and biochemical variability in the process of nutrient regeneration and further highlights the complexity of quantitatively resolving the major fluxes within the microbial loop.

Identification and Functional Analysis of Two Purple Acid Phosphatases AtPAP17 and AtPAP26 Involved in Salt Tolerance in Arabidopsis thaliana Plant

Tolerance to salinity is a complex genetic trait including numerous physiological processes, such as metabolic pathways and gene networks; thereby, identification of genes indirectly affecting, as well as those directly influencing, is of utmost importance. In this study, we identified and elucidated the functional characterization of AtPAP17 and AtPAP26 genes, as two novel purple acid phosphatases associated with high-salt tolerance in NaCl-stressed conditions. Here, the overexpression of both genes enhanced the expression level of AtSOS1, AtSOS2, AtSOS3, AtHKT1, AtVPV1, and AtNHX1 genes, involving in the K+/Na+ homeostasis pathway. The improved expression of the genes led to facilitating intracellular Na+ homeostasis and decreasing the ion-specific damages occurred in overexpressed genotypes (OEs). An increase in potassium content and K+/Na+ ratio was observed in OE17 and OE26 genotypes as well; however, lower content of sodium accumulated in these plants at 150 mM NaCl. The overexpression of these two genes resulted in the upregulation of the activity of the catalase, guaiacol peroxidase, and ascorbate peroxidase. Consequently, the overexpressed plants showed the lower levels of hydrogen peroxide where the lowest amount of lipid peroxidation occurred in these lines. Besides the oxidation resistance, the boost of the osmotic regulation through the increased proline and glycine-betaine coupled with a higher content of pigments and carbohydrates resulted in significantly enhancing biomass production and yield in the OEs under 150 mM NaCl. High-salt stress was also responsible for a sharp induction on the expression of both PAP17 and PAP26 genes. Our results support the hypothesis that these two phosphatases are involved in plant responses to salt stress by APase activity and/or non-APase activity thereof. The overexpression of PAP17 and PAP26 could result in increasing the intracellular APase activity in both OEs, which exhibited significant increases in the total phosphate and free Pi content compared to the wild-type plants. Opposite results witnessed in mutant genotypes (Mu17, Mu26, and DM), associating with the loss of AtPAP17 and AtPAP26 functions, clearly confirmed the role of these two genes in salt tolerance. Hence, these genes can be used as candidate genes in molecular breeding approaches to improve the salinity tolerance of crop plants.

The potential of corn-soybean intercropping to improve the soil health status and biomass production in cool climate boreal ecosystems

Intercropping (IC) is a promising approach used to improve soil health and sustainable crop production. However, it is unknown whether IC improve the soil health status and biomass productivity of crops cultivated in podzols under cool climate in boreal ecosystems. Two silage corn and three forage soybean genotypes were cultivated either as inter or monocrop (MC) treatments in a randomized complete block design. IC resulted in 28% increase in total forage production (FP). A reduction in rhizosphere soil pH (RS-pH) was observed in the IC treatments. Conversely, the rhizosphere soil acid phosphatase (RS-APase) activity was significantly higher (26–46%) in the IC treatments and occurred concomitant with a significant increase in available phosphorus (RS-Pavailable) (26–74%) in the rhizosphere. Furthermore, IC enhanced the active microbial composition and strong positive correlations were observed between RS-Pavailable, RS-APase, microbial biomass and FP; while RS-pH was negatively correlated with FP, RS-APase and RS-Pavailable. These findings suggested silage corn intercropped with forage soybean could be a viable approach to enhance FP through improved active microbial community structure, RS-APase activity and RS-Pavailable when cultivated on podzols in cool climate boreal ecosystem.

The effects of diversified phosphorus nutrition on the growth of oat (Avena sativa L.) and acid phosphatase activity

We studied the effect of differential phosphorus (P) supply on the development of oat seedlings (<em>Avena sativa</em> L. ‘Arab’) as well as localization and activity of acid phosphatases in tissues and root exudates. Plants were grown for 1–3 weeks on nutrient media with inorganic phosphate (+P, control), reduced Pi (0.1 P), phytic acid (PA) as organic P source, and without P addition (−P), in standard conditions or in a split-root culture system. Phosphate starvation reduced shoot growth but increased root elongation and root/shoot ratio, whereas 0.1 P and PA oat plants had similar growth parameters to +P plants. The growth on −P medium significantly decreased Pi content in all tissues, but only a slight Pi decrease was observed in plants grown on 0.1 P and PA media or various split-root system conditions. Pi starvation led to an increase in acid phosphatase (APase) activity in root exudates when compared to +P, 0.1 P, and PA plant samples. APase activity was especially intensive in root cross sections in rhizodermis and around/in vascular tissues of −P plants. For plants grown on 0.1 P medium and on phytic acid, APase activity did not change when compared to the control. Three major isoforms of APases were detected in plant tissues (similar in all studied conditions, with a higher activity of one isoform under Pi deficit). Generally, lowered Pi content (0.1 P) was not stressful to oat plants for up to 3 weeks of culture. Oat plants grew equally well on nutrient media with Pi and on media with phytate, although phytate was considered not available for other plants. The oat plants activated mainly extracellular APases, but also intracellular enzymes, rather via nonlocal signals, to acquire Pi from external/internal sources under Pi deficiency.

Response of arbuscular mycorrhizal fungi to soil phosphorus patches depends on context

Foraging strategies in arbuscular mycorrhizal fungi (AMF) for heterogeneously distributed resources in the soil remain to be explored. We used nylon-mesh bags of 30 μm to simulate patches of different phosphorus (P) supply levels (Expt 1) and P forms (organic v. inorganic, Expts 1 and 2). In Expt 1, host maize (Zea mays) was unfertilised; in each pot, five P-enriched bags were supplied with either Na-phytate or KH2PO4 at P rates of 0 (P0), 50 (P50), 100 (P100), 150 (P150) and 200 (P200) mg P kg–1. In Expt 2, maize plants were supplied with 20 (P20) or 50 (P50) mg P kg–1, and five P-enriched bags were supplied with different P forms (Na-phytate, lecithin, RNA, KH2PO4) and a nil-P control. Three fungal species (Funneliformis mosseae, Rhizophagus irregularis, and Glomus etunicatum) were compared in Expt 1, and the first two species in Expt 2. In Expt 1, the hyphal-length density (HLD) of G. etunicatum was not significantly different among different P levels when supplied with KH2PO4, whereas the HLD of R. irregularis tended to increase at higher P supply (above P50) in the Na-phytate treatment. The HLD of F. mosseae increased at P150 when supplied with KH2PO4, and increased at P100 and P150 in the Na-phytate treatment relative to P0. APase activity levels were more related to P supply level, in particular with F. mosseae inoculation and uninoculated control, showing that P200 significantly reduced APase relative to P0. In Expt 2, greater hyphal growth of both fungal species tended to occur with KH2PO4. At P20, the HLD of R. irregularis in treatments with KH2PO4 and lecithin, and of F. mosseae with KH2PO4, were higher than in P0. At P50, the HLD of F. mosseae was higher than of R. irregularis; but P form had no significant influence on HLD of F. mosseae, whereas the HLD of R. irregularis in the P-amended treatment (except with Na-phytate) was higher than in P0. APase activity did not differ significantly between the two fungal species. Highest APase activity generally occurred with lecithin, with no significant difference among the other P forms. Our results indicate that the response of AMF to P-enriched patches is complex, and both the form and amount of P supplied should be considered. Variations between AMF in the proliferation of hyphae to heterogeneous nutrient patches might be a mechanism by which these species can maintain diversity in intensive agricultural ecosystems.

Significance of intracellular and secreted acid phosphatase enzyme activities, and zinc and calcium interactions, on phosphorus efficiency in wheat, sunflower, chickpea, and lentil cultivars

Phosphorus efficiency (PE), and its relationship with intracellular (APase) and secreted (S-APase) acid phosphatases, anthocyanin accumulation, and calcium and zinc nutrition, were compared among 8 cultivars of each of wheat, sunflower, chickpea, and lentil grown under greenhouse conditions with low and high rates of P supply in a P-deficient calcareous soil. Except for the chickpea cultivars, deficiency of P resulted in significant decreases in shoot dry weight of all the crop cultivars and led to significant decreases in root dry weight in wheat and sunflower, significant increases in root dry weight in chickpea, and no significant difference in root dry weight in lentil. PE differed greatly among species and their cultivars. On average, shoot P concentration in cultivars of wheat, sunflower, chickpea, and lentil increased by 44%, 54%, 47%, and 8%, respectively, with P supply, and the increases in P concentration differed greatly among cultivars of all species. Intracellular leaf APase activity of wheat and lentil cultivars was slightly decreased by P supply, while it was unchanged in sunflower and chickpea cultivars. However, root-secreted acid phosphatase (S-APase) activity was significantly reduced by P supply in wheat, sunflower, and chickpea cultivars. Under low-P conditions, S-APase activities of all species except sunflower were negatively correlated with PE. Phosphorus deficiency increased the anthocyanin concentration of the cultivars of wheat and sunflower, whereas it was usually decreased in cultivars of the P-efficient species chickpea and lentil. In general, concentration of Ca was found to be lower, but Zn concentration was higher, in P-efficient cultivars than in P-inefficient cultivars. The results demonstrated that PE of the cultivars clearly depends on their ability to take up P and Zn, and on secretion of acid phosphatases from their roots under P deficiency. The results also suggest that characteristics of Zn and Ca nutrition should be taken into consideration when screening cultivars of crop species for their P efficiency.

Effect of Copper on Acid Phosphatase Activity in Yeast Yarrowia lipolytica

Acid phosphatase (APase) activity of the yeast Yarrowia lipolytica increased with increasing Cu2+ concentrations in the medium. Furthermore, the enzyme in soluble form was stimulated in vitro by Cu2+, Co2+, Ni2+, Mn2+ and Mg2+ and inhibited by Ag+ and Cd2+. The most effective ion was Cu2+, especially for the enzyme from cultures in medium containing Cu2+, whereas APase activity in wall-bound fragments was only slightly activated by Cu2+. The content of cellular phosphate involving polyphosphate was decreased by adding Cu2+, regardless of whether or not the medium was rich in inorganic phosphate. Overproduction of the enzyme stimulated by Cu2+ might depend on derepression of the gene encoding the APase isozyme.

UV irradiation of a B-cell hybridoma increases expression of alkaline phosphatase: involvement in apoptosis

Expression of alkaline phosphatase (APase) by 7TD1 B-cell hybridoma was amplified by ultraviolet irradiation; cell growth was inhibited and cell death by apoptosis was increased. Irradiation induced high levels of APase activity in cycling as well as in apoptotic cells. In contrast, APase activity faded with time in nonirradiated cells and was no longer expressed in spontaneous apoptotic cells appearing after several days in culture. This was demonstrated by cell morphology, DNA fragmentation, and flow cytometry after simultaneous staining of DNA with Hoechst 33342 and APase with naphthol AS-TR phosphate - fast red RC fluorescent reagent. Levamisole, a specific inhibitor of APase activity, almost totally abrogated apoptosis induced by ultraviolet irradiation at doses that failed to affect 7TD1 cell survival. These data suggest that APase could play a role in the signalling cascade that mediates apoptosis in irradiated cells. Key words: alkaline phosphatase, apoptosis, flow cytometry, levamisole, UV irradiation.