Acid Phosphatase Gene Research Articles

Analysis and profiling of the purple acid phosphatase gene family in wheat (Triticum aestivum L.).

Purple acid phosphatases (PAPs) play a vital role in plant phosphorus nutrition, serving as a crucial family of metallo-phosphoesterase enzymes. This research aimed to identify the PAP genes from the A/B/D genomes of Triticum aestivum to elucidate evolutionary mechanisms of the gene family in plants and provide genomic information for subsequent research on phosphorous-use efficiency in wheat crops. In total, 105 PAP genes (TaPAPs) were identified from the A/B/D genomes by using the Arabidopsis thaliana and Oryza sativa PAP protein sequences as queries for BLASTP against the wheat protein database. The TaPAPs were grouped into six subfamilies, Ia (17), Ib (26), IIa (11), IIb (30), IIIa (12), and IIIb (9), based on their similarities in the structure of genes and the presence of conserved protein motifs. A majority of TaPAPs were derived from tandemly (20) or segmentally (87) duplicated, with the homoeologous chromosomes 5A/B/D harboring the most duplicated PAP genes. Further analysis indicated that TaPAPs were responsible for the modulation of seed, root, and leaf development and hormone synthesis and signaling, as well as plant responses to abiotic stresses, including low temperatures, drought, and anaerobic conditions. Nine TaPAPs (TaPAP9-4A/4B/4D, TaPAP24-6A/6B/6D, and TaPAP28-7A/7B/7D) were constitutively expressed in diverse tissues such as root, shoot, leaf, spike, and seed, while the remaining genes exhibited tissue-specific expression patterns. Concerning the response to phosphate (Pi) deprivation, 57 TaPAPs were highly expressed in roots under Pi stress, including TaPAP31-4A, 4B, and 4D homeologs from the subfamily IIIb. A TaPAP31-4A transgene in A. thaliana promoted plant growth and development while increasing plant resistance to Pi-deficiency stress by enhancing the secretion of phosphatase. These discoveries provide a scientific foundation for comprehending the role of TaPAPs, offering valuable insights for identifying additional candidate genes and fostering the development of new wheat varieties with enhanced tolerance to low phosphorus conditions.

Metabolism of isodecyl diphenyl phosphate in rice and microbiome system: Differential metabolic pathways and underlying mechanisms

Isodecyl diphenyl phosphate (IDDP) is among the emerging aromatic organophosphate esters (aryl-OPEs) that pose risks to both human beings and other organisms. This study aims to investigate the translocation and biotransformation behavior of IDDP in rice and the rhizosphere microbiome through hydroponic exposure (the duration of hydroponic exposure was 10 days). The rhizosphere microbiome 9-FY was found to efficiently eliminate IDDP, thereby reducing its uptake in rice tissues and mitigating the negative impact of IDDP on rice growth. Furthermore, this study proposed the first-ever transformation pathways of IDDP, identifying hydrolysis, hydroxylation, methylation, methoxylation, carboxylation, and glucuronidation products. Notably, the methylation and glycosylation pathways were exclusively observed in rice, indicating that the transformation of IDDP in rice may be more complex than in microbiome 9-FY. Additionally, the presence of the product COOH-IDDP in rice suggested that there might be an exchange of degradation products between rice and rhizobacteria, implying their potential interaction. This finding highlights the significance of rhizobacteria's role which cannot be overlooked in the accumulation and transformation of organic pollutants in grain crops. The study revealed active members in 9-FY during IDDP degradation, and metagenomic analysis indicated that most of the active populations contained IDDP-degrading genes. Moreover, transcriptome sequencing showed that cytochrome P450, acid phosphatase, glucosyltransferase, and methyltransferases genes in rice were up-regulated, which was further confirmed by RT-qPCR. This provides insight into the intermediate products identified in rice, such as hydrolysis, hydroxylated, glycosylated, and methylated products. These results significantly contribute to our understanding of the translocation and transformation of organophosphate esters (OPEs) in plants and the rhizosphere microbiome, and reveal the fate of OPEs in rice and microbiome system to ensure the paddy yield and rice safety.

N6-methyladenosine RNA methyltransferase CpMTA1 mediates CpAphA mRNA stability through a YTHDF1-dependent m6A modification in the chestnut blight fungus.

In eukaryotic cells, N6-methyladenosine (m6A) is the most prevalent RNA epigenetic modification that plays crucial roles in multiple biological processes. Nevertheless, the functions and regulatory mechanisms of m6A in phytopathogenic fungi are poorly understood. Here, we showed that CpMTA1, an m6A methyltransferase in Cryphonectria parasitica, plays a crucial role in fungal phenotypic traits, virulence, and stress tolerance. Furthermore, the acid phosphatase gene CpAphA was implicated to be a target of CpMTA1 by integrated analysis of m6A-seq and RNA-seq, as in vivo RIP assay data confirmed that CpMTA1 directly interacts with CpAphA mRNA. Deletion of CpMTA1 drastically lowered the m6A level of CpAphA and reduced its mRNA expression. Moreover, we found that an m6A reader protein CpYTHDF1 recognizes CpAphA mRNA and increases its stability. Typically, the levels of CpAphA mRNA and protein exhibited a positive correlation with CpMTA1 and CpYTHDF1. Importantly, site-specific mutagenesis demonstrated that the m6A sites, A1306 and A1341, of CpAphA mRNA are important for fungal phenotypic traits and virulence in C. parasitica. Together, our findings demonstrate the essential role of the m6A methyltransferase CpMTA1 in C. parasitica, thereby advancing our understanding of fungal gene regulation through m6A modification.

GWAS unravels acid phosphatase ACP2 as a photosynthesis regulator under phosphate starvation conditions through modulating serine metabolism in rice

Inorganic phosphorus (Pi) deficiency significantly impacts plant growth, development, and photosynthetic efficiency. This study evaluated 206 rice accessions from a MiniCore population under both Pi-sufficient (Pi+) and Pi-starvation (Pi−) conditions in the field to assess photosynthetic phosphorus use efficiency (PPUE), defined as the ratio of AsatPi− to AsatPi+. A genome-wide association study and differential gene expression analyses identified an acid phosphatase gene (ACP2) that responds strongly to phosphate availability. Overexpression and knockout of ACP2 led to a 67% increase and 32% decrease in PPUE, respectively, compared with wild type. Introduction of an elite allele A, by substituting the v5 SNP G with A, resulted in an 18% increase in PPUE in gene-edited ACP2 rice lines. The phosphate-responsive gene PHR2 was found to transcriptionally activate ACP2 in parallel with PHR2 overexpression, resulting in an 11% increase in PPUE. Biochemical assays indicated that ACP2 primarily catalyzes the hydrolysis of phosphoethanolamine and phospho-L-serine. In addition, serine levels increased significantly in the ACP2v8G-overexpression line, along with a concomitant decrease in the expression of all nine genes involved in the photorespiratory pathway. Application of serine enhanced PPUE and reduced photorespiration rates in ACP2 mutants under Pi-starvation conditions. We deduce that ACP2 plays a crucial role in promoting photosynthesis adaptation to Pi starvation by regulating serine metabolism in rice.

Interactions between halotolerant nitrogen-fixing bacteria and arbuscular mycorrhizal fungi under saline stress.

Soil salinity negatively affects crop development. Halotolerant nitrogen-fixing bacteria (HNFB) and arbuscular mycorrhizal fungi (AMF) are essential microorganisms that enhance crop nutrient availability and salt tolerance in saline soils. Studying the impact of HNFB on AMF communities and using HNFB in biofertilizers can help in selecting the optimal HNFB-AMF combinations to improve crop productivity in saline soils. We established three experimental groups comprising apple plants treated with low-nitrogen (0 mg N/kg, N0), normal-nitrogen (200 mg N/kg, N1), and high-nitrogen (300 mg N/kg, N2) fertilizer under salt stress without bacteria (CK, with the addition of 1,500 mL sterile water +2 g sterile diatomite), or with bacteria [BIO, with the addition of 1,500 mL sterile water +2 g mixed bacterial preparation (including Bacillus subtilis HG-15 and Bacillus velezensis JC-K3)]. HNFB inoculation significantly increased microbial biomass and the relative abundance of beta-glucosidase-related genes in the rhizosphere soil under identical nitrogen application levels (p < 0.05). High-nitrogen treatment significantly reduced AMF diversity and the relative abundance of beta-glucosidase, acid phosphatase, and urea-related genes. A two-way analysis of variance showed that combined nitrogen application and HNFB treatment could significantly affect soil physicochemical properties and rhizosphere AMF abundance (p < 0.05). Specifically, HNFB application resulted in a significantly higher relative abundance of Glomus-MO-G17-VTX00114 compared to that in the CK group at equal nitrogen levels. The impact of HNFB on the AMF community in apple rhizospheres is influenced by soil nitrogen levels. The study reveals how varying nitrogen levels mediate the relationship between exogenous HNFB, soil properties, and rhizosphere microbes.

Comparative Response of Arbuscular Mycorrhizal Fungi versus Endophytic Fungi in Tangor Citrus: Photosynthetic Efficiency and P-Acquisition Traits

Citrus plants are prone to phosphorus (P) deficiency, especially in acidic soil, making them more dependent on root-associated endophytic fungi for growth and development. Beni-Madonna, a hybrid of Citrus nanko × C. amakusa, is a citrus known as “tangor” that is highly popular in China and other parts of the world due to its deep red color and jelly-like flesh. In this study, the inoculation response to two arbuscular mycorrhizal fungi (Diversispora versiformis, Dv; Funneliformis mosseae, Fm) and an endophytic fungus (Serendipita indica, Si) with regard to the growth, leaf gas exchange, light energy efficiency, P levels, acid phosphatase activity, and expression of the purple acid phosphatase (PAP) and phosphate transporter (PT) genes through a potted experiment using Beni-Madonna (tangor) citrus plants grafted on trifoliate orange (Poncirus trifoliata) was studied. Two years following inoculation, the root fungal colonization rates of inoculated plants were significantly increased, accompanied by an increase in plant height and stem diameter, with Si presenting the best effect. Fungal inoculations also significantly increased the leaf chlorophyll index, nitrogen-balance index, photosynthesis rate, stomatal conductance, transpiration rate, photosynthetic efficiency, and quantum yield of PSII, while it reduced photoinhibition and heat dissipation, with Si having the best effect on light energy efficiency. Si significantly increased leaf and root P levels, as well as root CsPAP1‒3, CsPT2, CsPT3, and CsPT7 expression and soil acid phosphatase activity; Fm in mycorrhizal fungi significantly increased leaf and root P levels, as well as root CsPT1 and CsPT7 expression and root/soil acid phosphatase activity. These observations suggested an advantage of Si over the other two AMF in terms of improved plant growth and P acquisition, while Fm displayed prominent effects on increased photosynthetic efficiency.

SlPHL1 positively modulates acid phosphatase in response to phosphate starvation by directly activating the genes SlPAP10b and SlPAP15 in tomato.

Increased acid phosphatase (APase) activity is a prominent feature of tomato (Solanum lycopersicum) responses to inorganic phosphate (Pi) restriction. SlPHL1, a phosphate starvation response (PHR) transcription factor, has been identified as a positive regulator of low Pi (LP)-induced APase activity in tomato. However, the molecular mechanism underlying this regulation remains to be elucidated. Here, SlPHL1 was found to positively regulate the LP-induced expression of five potential purple acid phosphatase (PAP) genes, namely SlPAP7, SlPAP10b, SlPAP12, SlPAP15, and SlPAP17b. Furthermore, we provide evidence that SlPHL1 can stimulate transcription of these five genes by binding directly to the PHR1 binding sequence (P1BS) located on their promoters. The P1BS mutation notably weakened SlPHL1 binding to the promoters of SlPAP7, SlPAP12, and SlPAP17b but almost completely abolished SlPHL1 binding to the promoters of SlPAP10b and SlPAP15. As a result, the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 was substantially diminished. In addition, not only did transient overexpression of either SlPAP10b or SlPAP15 in tobacco leaves increase APase activity, but overexpression of SlPAP15 in Arabidopsis and tomato also increased APase activity and promoted plant growth. Subsequently, two SPX proteins, SlSPX1 and SlSPX4, were shown to physically interact with SlPHL1. Moreover, SlSPX1 inhibited the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 and negatively regulated the activity of APase. Taken together, these results demonstrate that SlPHL1-mediated LP signaling promotes APase activity by activating the transcription of SlPAP10b and SlPAP15, which may provide valuable insights into the mechanisms of tomato response to Pi-limited stress.

Engineering Yarrowia lipolytica for sustainable ricinoleic acid production: A pathway to free fatty acid synthesis

Ricinoleic acid (C18:1-OH, RA) is a valuable hydroxy fatty acid with versatile applications. The current industrial source of RA relies on the hydrolysis of castor bean oil. However, the coexistence of the toxic compound ricin and the unstable supply of this plant have led to an exploration of promising alternatives: generating RA in heterologous plants or microorganisms. In this study, we engineered the oleaginous yeast Yarrowia lipolytica to produce RA in the form of free fatty acids (FFA). First, we overexpressed fungal Δ12 oleate hydroxylase gene (CpFAH12) from Claviceps purpurea while deleting genes related to fatty acid degradation (MEF1 and PEX10) and oleic acid desaturation (FAD2). Since Δ12 oleate hydroxylase converts oleic acid (C18:1) located at the sn-2 position of phosphatidylcholine (PC), we next focused on increasing the PC pool containing oleic acid. This objective was achieved thorough implementing metabolic engineering strategies designed to enhance the biosynthesis of PC and C18 fatty acids. To increase the PC pool, we redirected the flux towards phospholipid biosynthesis by deleting phosphatidic acid phosphatase genes (PAH1 and APP1) and diacylglycerol acyltransferase gene (DGA1), involved in the production of diacylglycerol and triacylglycerol, respectively. Furthermore, the PC biosynthesis via the CDP-DAG pathway was enhanced through the overexpression of CDS1, PSD1, CHO2, and OPI3 genes. Subsequently, to increase the oleic acid content within PC, we overexpressed the heterologous fatty acid elongase gene (MaC16E) involved in the conversion of C16 to C18 fatty acids. As RA production titer escalated, the produced RA was mainly found in the FFA form, leading to cell growth inhibition. The growth inhibition was mitigated by inducing RA secretion via Triton X-100 treatment, a process that simultaneously amplified RA production by redirecting flux towards RA synthesis. The final engineered strain JHYL-R146 produced 2.061 g/L of free RA in a medium treated with 5% Triton X-100, constituting 74% of the total FFAs produced. Generating free RA offers the added benefit of bypassing the hydrolysis stage required when employing castor bean oil as an RA source. This achievement represents the highest level of RA synthesis from glucose reported thus far, underscoring the potential of Y. lipolytica as a host for sustainable RA production.

Effective gene silencing using type I-E CRISPR system in the multiploid, radiation-resistant bacterium Deinococcus radiodurans.

The extremely radiation-resistant bacterium, Deinococcus radiodurans, is a microbe of importance, both, for studying stress tolerance mechanisms and as a chassis for industrial biotechnology. However, the molecular tools available for use in this organism continue to be limiting, with its multiploid genome presenting an additional challenge. In view of this, the clustered regularly interspaced short palindromic repeat (CRISPR)-Cas tools provide a large repertoire of applications for gene manipulation. We show the utility of the type I-E Cascade system for knocking down gene expression in this organism. A single-vector system was designed for the expression of the Cascade components as well as the crRNA. The type I-E Cascade system was better tolerated than the type II-A dCas9 system in D. radiodurans. An assayable acid phosphatase gene, phoN integrated into the genome of this organism could be knocked down to 10% of its activity using the Cascade system. Cascade-based knockdown of ssb, a gene important for radiation resistance resulted in poor recovery post-irradiation. Targeting the Radiation and Desiccation Response Motif (RDRM), upstream of the ssb, prevented de-repression of its expression upon radiation exposure. In addition to this, multi-locus targeting was demonstrated on the deinococcal genome, by knocking down both phoN and ssb expression simultaneously. The programmable CRISPR interference tool developed in this study will facilitate the study of essential genes, hypothetical genes, and cis-elements involved in radiation response as well as enable metabolic engineering in this organism. Further, the tool can be extended for implementing high-throughput approaches in such studies. IMPORTANCE Deinococcus radiodurans is a microbe that exhibits a very high degree of radiation resistance. In addition, it is also identified as an organism of industrial importance. We report the development of a gene-knockdown system in this organism by engineering a type I-E clustered regularly interspaced short palindromic repeat (CRISPR)-Cascade system. We used this system to silence an assayable acid phosphatase gene, phoN to 10% of its activity. The study further shows the application of the Cascade system to target an essential gene ssb, that caused poor recovery from radiation. We demonstrate the utility of CRISPR-Cascade to study the role of a regulatory cis-element in radiation response as well as for multi-gene silencing. This easy-to-implement CRISPR interference system would provide an effective tool for better understanding of complex phenomena such as radiation response in D. radiodurans and may also enhance the potential of this microbe for industrial application.

Fertilization and cultivation management promotes soil phosphorus availability by enhancing soil P-cycling enzymes and the phosphatase encoding genes in bulk and rhizosphere soil of a maize crop in sloping cropland

Fertilization and cultivation managements exert significant effects on crop growth and soil-associated nutrients in croplands. However, there is a lack of knowledge regarding how these practices affect soil phosphorus-cycling enzymes and functional genes involved in regulating global P-cycling, especially under intense agricultural management practices in sloping croplands. A long-term field (15-year) trial was conducted in a 15° sloping field based on five treatments: no fertilizer amendments + downslope cultivation (CK); mixed treatment of mineral fertilizer and organic manure + downslope cultivation (T1); mineral fertilizer alone + downslope cultivation (T2); 1.5-fold mineral fertilizer + downslope cultivation (T3); and mineral fertilizer + contour cultivation (T4). Bulk and rhizosphere soil samples were collected after the maize crop was harvested to determine the P fraction, P-cycling enzymes, and phosphatase-encoding genes. Results indicated that fertilization management significantly increased the inorganic (Pi) and organic soil (Po) P fractions compared to CK, except for NaOH-extractable Po in T1 and T3 in bulk and rhizosphere soils, respectively. For the cultivation treatments, the content of Pi pools in the downslope cultivation of T1 and T3 was significantly larger than that in the contour cultivation of T4 in bulk and rhizosphere soils. However, the content of NaOH-extractable Po in T1 and T3 was lower compared to T4 in bulk soil and vice versa for the NaHCO3-P and HCl-Po fractions in the rhizosphere. We also found that fertilization and cultivation managements significantly increased the activity of acid phosphatase (ACP), alkaline phosphatase (ALP), phytase, phosphodiesterases (PDE), and phoC and phoD gene abundance in bulk and rhizosphere soils, with a larger effect on the activity of ALP and the phosphatase encoding phoD gene, especially in T1 and T3 in the rhizosphere. Soil organic carbon (SOC) and microbial biomass C and P (MBC and MBP) were the main predictors for regulating P-cycling enzymes and phoC- and phoD gene abundance. A strong association of P-cycling enzymes, especially ALP and phytase, and the abundance of phoD genes with the P fraction indicated that the soil P cycle was mainly mediated by microbial-related processes. Together, our results demonstrated that an adequate amount of mineral fertilizer alone or combined with organic fertilizer plus downslope cultivation is more effective in promoting soil P availability by enhancing the activity of ALP, phytase, and phoD genes. This provides valuable information for sustaining soil microbial-regulated P management practices in similar agricultural lands worldwide.

Molecular Mechanism Underlying the Action of a Celastrol-Loaded Layered Double Hydroxide-Coated Magnesium Alloy in Osteosarcoma Inhibition and Bone Regeneration.

Osteosarcoma (OS) is a malignant bone tumor that threatens human health. Surgical removal of the tumor and followed by implantation with a graft is the golden standard for its clinical treatment. However, avoiding recurrence by enhancing the antitumor properties of the implants and improving osteogenesis around the implants remain a challenge. Here, we developed a layered double hydroxide (LDH)-coated magnesium (Mg) alloy and loaded it with celastrol. The celastrol-loaded Mg alloy exhibited enhanced corrosion resistance and sustained release of celastrol. In vitro cell culture suggested that the modified Mg alloy loaded with an appropriate amount of celastrol significantly inhibited the proliferation and migration of bone tumor cells while having little influence on normal cells. A mechanistic study revealed that the celastrol-loaded Mg alloy upregulated reactive oxygen species (ROS) generation in bone tumor cells, resulting in mitochondrial dysfunction due to reduced membrane potential, thereby inducing bone tumor cell apoptosis. Furthermore, it was found that celastrol-induced autophagy in tumor cells inhibited cell apoptosis in the initial 6 h. After ≥12 h of culture, inhibition of the PI3K-Akt-mTOR signaling pathway was noted, resulting in excessive autophagy in tumor cells, finally causing cell apoptosis. The celatsrol-loaded Mg alloy also exhibited effective antitumor properties in a subcutaneous tumor model. In vitro tartrate-resistant acid phosphatase (TRAP) staining and gene expression results revealed that the modified Mg alloy reduced the viability of osteoclasts, inducing a potential pathway for the increased bone regeneration around the modified Mg alloy seen in vivo. Together, the results of our study show that the celatsrol-loaded Mg alloy might be a promising implant for treating OS.

Nontarget Site-Based Resistance to Fenoxaprop-P-ethyl and Candidate Genes Involved in Alopecurus japonicus

Nontarget-site resistance (NTSR) is a complex multigenic trait that is associated with the potential mechanisms of herbicide resistance which pose a serious threat to global crop protection. However, the NTSR mechanisms of Alopecurus japonicus, a malignant weed infesting wheat fields, are less characterized. In this study, we used RNA-sequencing transcriptome and enzyme activity detection to investigate the NTSR mechanisms and candidate genes involved in fenoxaprop-P-ethyl (FE) in a previously identified resistant population compared to the sensitive population of A. japonicus. Transcriptome analysis identified nine upregulated genes, which were constitutively overexpressed and upregulated by FE application in the resistant population, and the results were validated using quantitative real-time PCR. These genes including one cytochrome P450 monooxygenase (P450) gene (CYP75B4), one ATP-binding cassette (ABC) transporter gene (ABCG36), one laccase (LAC) gene (LAC15), one 9-cis-epoxycarotenoid dioxygenase (NCED) gene (NCED5), two purple acid phosphatase (PAP) genes (PAP4, PAP15), one sucrose phosphate synthase (SPS) gene (SPS3), one protein related to disease resistance gene (RGA3) and one immune protein gene (R1B-17). The activity assay of LAC, NCED, PAP and SPS revealed that the activities of these enzymes in the resistant population were significantly higher than those in the sensitive population at 0 h and after FE application at 12 h, 24 h and 72 h. Nevertheless, whether LAC, NCED, PAP and SPS genes were involved in herbicide metabolism needs to be further validated. Our results revealed that CYP, ABC transporter and LAC genes may participate in A. japonicus resistance. These genes identified in the present study provide new insights into the resistance mechanism of weeds in response to herbicide. Our study also implies the complexity of the NTSR mechanisms of weeds.

Genome-wide and candidate gene association studies identify BnPAP17 as conferring the utilization of organic phosphorus in oilseed rape

Phosphorus (P) is essential for living plants, and P deficiency is one of the key factors limiting the yield in rapeseed production worldwide. As the most important organ for plants, root morphology traits (RMTs) play a key role in P absorption. To investigate the genetic variability of RMT under low P availability, we dissected the genetic structure of RMTs by genome-wide association studies (GWAS), linkage mapping and candidate gene association studies (CGAS). A total of 52 suggestive loci were associated with RMTs under P stress conditions in 405 oilseed rape accessions. The purple acid phosphatase gene BnPAP17 was found to control the lateral root number (LRN) and root dry weight (RDW) under low P stress. The expression of BnPAP17 was increased in shoot tissue in P-efficient cultivars compared to root tissue and P-inefficient cultivars in response to low P stress. Moreover, the haplotype of BnPAP17Hap3 was detected for the selective breeding of P efficiency in oilseed rape. Over-expression of the BnPAP17Hap3 could promote the shoot and root growth with enhanced tolerance to low P stress and organic phosphorus (Po) utilization in oilseed rape. Collectively, these findings increase our understanding of the mechanisms underlying BnPAP17-mediated low P stress tolerance in oilseed rape.

Two different strategies of Diversispora spurca-inoculated walnut seedlings to improve leaf P acquisition at low and moderate P levels.

Walnut (Juglans regia) is an important nut tree species in the world, whereas walnut trees often face inadequate phosphorus (P) levels of soil, negatively limiting its growth and yield. Arbuscular mycorrhizal fungi (AMF) can colonize walnut roots, but whether and how AMF promotes walnut growth, physiological activities, and P acquisition is unclear. The present study aimed to evaluate the effects of Diversispora spurca on plant growth, chlorophyll component concentrations, leaf gas exchange, sugar and P concentrations, and expression of purple acid phosphatase (PAP) and phosphate transporter (PT) genes in leaves of J. regia var. Liaohe 1 seedling under moderate (100 μmol/L P) and low P (1 μmol/L P) levels conditions. Three months after inoculation, the root mycorrhizal colonization rate and soil hyphal length were 45.6-53.2% and 18.7-39.9 cm/g soil, respectively, and low P treatment significantly increased both root mycorrhizal colonization rate and soil hyphal length. Low P levels inhibited plant growth (height, stem diameter, and total biomass) and leaf gas exchange (photosynthetic rate, transpiration rate and stomatal conductance), while AMF colonization significantly increased these variables at moderate and low P levels. Low P treatment limited the level of chlorophyll a, but AMF colonization did not significantly affect the level of chlorophyll components, independent on soil P levels. AMF colonization also increased leaf glucose at appropriate P levels and leaf fructose at low P levels than non-AMF treatment. AMF colonization significantly increased leaf P concentration by 21.0-26.2% than non-AMF colonization at low and moderate P levels. Low P treatment reduced the expression of leaf JrPAP10, JrPAP12, and JrPT3;2 in the inoculated plants, whereas AMF colonization up-regulated the expression of leaf JrPAP10, JrPAP12, and JrPT3;2 at moderate P levels, although AMF did not significantly alter the expression of JrPAPs and JrPTs at low P levels. It is concluded that AMF improved plant growth, leaf gas exchange, and P acquisition of walnut seedlings at different P levels, where mycorrhizal promotion of P acquisition was dominated by direct mycorrhizal involvement in P uptake at low P levels, while up-regulation of host PAPs and PTs expressions at moderate P levels.

Molecular characterization and functional analysis of tartrate-resistant acid phosphatase (ACP5) gene in red drum (Sciaenops ocellatus)

Tartrate-resistant acid phosphatase (ACP5) plays an important biological function in immune defense and is highly expressed in activated macrophages, osteoclasts and dendritic cells. In teleost, the functionality of ACP5 remains to be revealed. In this study, we cloned and identified SoACP5 from red drum (Sciaenops ocellatus) and analyzed its function in vivo and in vitro. The open reading frame of SoACP5 is 1002 bp in length, encoding 333 amino acids. SoACP5 shares high sequence identities (96.70%–49.25%) with ACP5 of other species. The SoACP5 mRNA was widely distributed in collected tissues of healthy red drum, and with the maximum in gills. The expression of SoACP5 increased significantly in vivo following challenge with Edwardsiella tarda. Moreover, the recombinant SoACP5 protein (rSoACP5) was purified with his-tag band resin columns, and confirmed to have phosphatase activity which was optimal at pH 5 and 55 °C. Various metal ions (K+, Zn2+, Mn2+, Mg2+, Ca2+, Cu2+, Fe2+ and Fe3+) have different effects on phosphatase activity. rSoACP5 induced the cellular proliferation of peripheral blood leukocytes. The over-expression and knockdown of SoACP5 in vivo had a significant effect on bacterial proliferation. Furthermore, both of the antibacterial activity and phosphatase activity were decreased when the reducedSoACP5 was oxidized by H2O2. In summary, the present study indicated that SoACP5 is likely involved in host defense against bacterial infection in S. ocellatus.

Metabolism balance regulation for squalene production by disturbing triglyceride (TAG) synthesis in Schizochytrium sp.

Schizochytrium sp., a unicellular, eukaryotic heterotrophic marine microorganism, is rich in lipids that can serve as a potential cell factory for producing high-value compounds through the mevalonate (MVA) pathway, which competes with triglyceride (TAG) synthesis for precursors. In this study, TAG synthesis was disturbed to shift the precursors for squalene production by the MVA pathway, and the changes in cell metabolism of Schizochytrium limacinum SR21 were explored. Compared with the wild strain, the Δelo1 (elongase gene for fatty acid synthesis) and Δpap (phosphatidic acid phosphatase gene for triglyceride synthesis) strains provided more precursors for squalene synthesis, showing an increase of 35.6 % and 37.1 %, respectively. Considering the adverse effect of Δpap on TAG synthesis, the elo1 knockout and pap interference were regulated combinedly in the mutant strain (named the MIX strain) to improve squalene synthesis. The biomass and lipid contents of the MIX strain decreased by 13.6 % and 18.5 %, while the squalene titer increased by 41.4 %, reaching a value of 10.1 mg/L. The pap interference induced the flow of more glyceraldehyde 3-phosphate during pyruvate synthesis to produce more acetyl-coenzyme A (CoA). In contrast, the combined Δelo1 redistributed acetyl-CoA, promoting the production of adequate adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide phosphate (NADPH) by the tricarboxylic acid (TCA) cycle flux, and increased the flow of the precursors into the MVA pathway. This study provides a novel strategy for exploring lipid metabolism and lays a foundation for establishing cell factories for terpenoid synthesis by Schizochytrium sp.

Bovine lactoferricin on non-specific immunity of giant freshwater prawns, Macrobrachium rosenbergii

This study aimed to investigate the effects of dietary Bovine lactoferricin (LFcinB) on the growth performance and non-specific immunity in Macrobrachium rosenbergii. Five experimental diets were 1.0‰ Bovine lactoferricin (LCB1); 1.5‰ Bovine lactoferricin (LCB1.5); 2.0‰ Bovine lactoferricin (LCB2); 2.5‰ Bovine lactoferricin (LCB2.5); the control group, basal diet without Bovine lactoferricin. A total of 600 prawns were randomly assigned to 5 groups in triplicate in 15 tanks for an 8-week feeding trial. The results showed the final weight, weight gain rate, specific growth rate and survival rate of prawns in the treatment groups were significantly improved versus the control (P < 0.05). The feed conversion ratio was reduced significantly in treatment groups compared to the control (P < 0.05). Compared with the control, alkaline phosphatase (AKP), acid phosphatase (ACP), lysozyme (LZM), catalase (CAT), superoxide dismutase (SOD) activities in the hepatopancreas of the treatment groups were significantly enhanced, and malondialdehyde (MDA) content was reduced significantly (P < 0.05). Compared with the control, the relative expression levels of AKP, ACP, LZM, CAT, SOD, Hsp70, peroxiredoxin-5, Toll, dorsal and relish genes were significantly higher among treatment groups, except for the AKP gene in the LCB1 group and the Hsp70 gene in the LCB1.5 group (P < 0.05). Compared with the control, the relative expression levels of TOR, 4E-BP, eIF4E1α and eIF4E2 genes were significantly enhanced in the LCB1.5 group (P < 0.05). When resistance against Vibrio parahaemolyticus in prawn is considered, higher doses of Bovine lactoferricin show better antibacterial ability. The present study indicated that dietary Bovine lactoferricin could significantly improve the growth performance and improve the antioxidative status of M. rosenbergii. The suitable addition level is 1.5 g/kg. LFcinB has great potential as a new feed additive without the threat of drug resistance.

Watermelon responds to organic fertilizer by enhancing root-associated acid phosphatase activity to improve organic phosphorus utilization

Organic fertilizer is commonly used to increase crop yields and improve soil quality. However, it is unclear whether crops adapt to organic fertilizer by regulating metabolic pathways that are involved in nutrient utilization. In this study, we focused on the organic phosphorus (Po) in organic fertilizer and, using watermelon, investigated changes in gene expression and metabolic pathways in response to organic fertilizer and the combination of chemical fertilizer and organic fertilizer (chemical fertilizer 70% and organic fertilizer 30%, based on phosphorus supply). Purple acid phosphatase (PAP) gene expression was upregulated in leaves and roots of watermelon grown in organic fertilizer alone and in the combination of chemical/organic fertilizer, resulting in enhanced phosphatase activity in roots. When the ratio of chemical to organic fertilizer was 85/15, root-associated acid phosphatase (APase) activity increased over chemical fertilizer alone. This formulation also resulted in increased inorganic phosphate (Pi) concentration in roots and leaves, and the upregulation of the secretory APase genes ClaPAP10/12/15/26, and ClaPAP18 in roots. In conclusion, watermelon responds to organic fertilizer by upregulating expression of secretory ClaPAP genes, subsequently enhancing root-associated APase activity further improving the hydrolysis of phosphomonoesters, and ultimately facilitating Po utilization by roots. The mechanisms of P utilization by roots comprise the enhancement of APase and phytase activity, absorption of small Po molecules, uptake of Pi, and the increase of lateral root number when organic fertilizer is applied to the plants. These findings help to establish the mechanisms by which plants respond to organic fertilizer by regulating metabolic pathways at the transcriptional level.

Changes in rhizosphere phosphorus fractions and phosphate-mineralizing microbial populations in acid soil as influenced by organic acid exudation

Interactions were explored between organic acids, phosphate-mineralizing microorganisms, and phosphorus fractions in rhizosphere soil. Bulk soil, rhizosphere soil, and root samples were randomly collected from a mature lemon tree plantation. Composition of acid (phoC) and alkaline (phoD) phosphatase gene harboring bacterial populations in rhizosphere soil were different. Whereas only soil properties affected the phoC-harboring bacterial population, soil properties and organic acids shaped the phoD-harboring bacterial population. Organic acids, especially oxalic acid, rather than phosphatase significantly directly and indirectly affected phosphorus fractions. When oxalic acid concentration increased, abundances of phoD-harboring Pseudomonas and Bacillus also increased. Those genera then released alkaline phosphatase, which reduced the contents of calcium-bound organophosphorus, insoluble organophosphorus, and residual phosphorus. By contrast, when oxalic acid concentration decreased, abundance of phoC-harboring Xanthomonas increased. The genus released acid phosphatase, which reduced calcium-bound organophosphorus content. In addition, alkaline phosphatase released by phoD-harboring bacteria was more important than acid phosphatase in acid soil. In conclusion, oxalic acids have essential roles in directly and indirectly changing phosphorus fractions in lemon tree rhizosphere soil.

PtoWRKY40 interacts with PtoPHR1-LIKE3 while regulating the phosphate starvation response in poplar.

Plants usually suffer from phosphorus starvation because of the low inorganic phosphate (Pi) status of most soils. To cope with this, plants have evolved an adaptive phosphate starvation response (PSR) which involves both developmental and metabolic changes regulated mainly by PHOSPHATE STARVATION RESPONSE1 (PHR1) and its homologs. Here, we elucidated how perennial woody plants, such as poplars (Populus spp.), respond to low-Pi stress. We first performed RNA-seq analysis of low-Pi-treated poplars and identified PtoWRKY40 is rapidly downregulated and protein degraded after stress. Overexpressing and knocking-down PtoWRKY40 downregulated and upregulated the expression of Pi starvation signaling genes, respectively, such as PHOSPHATE TRANSPORTER1 (PHT1)-type genes and PURPLE ACID PHOSPHATASE genes. PtoWRKY40 bound to the W box in the promoter of several PtoPHT1s and repressed their expression. Moreover, PtoWRKY40 interacted with PtoPHR1-LIKE3 (PtoPHL3), a PHR1 homolog in poplar, to inhibit the latter binding to the P1BS element and thus reduced PtoPHT1s' transcription under Pi-sufficient conditions. However, Pi deficiency decreased PtoWRKY40 abundance and therefore released its inhibition on PHT1s. In conclusion, we have uncovered a PSR mechanism mediated by PtoWRKY40 and PtoPHL3 which regulates Pi content in poplars, deepening our understanding of how poplars adapt to diverse Pi conditions and regulate appropriate responses to maintain Pi homeostasis.