Low Pi Research Articles

Comprehensive Evolution and Expression anaLysis of PHOSPHATE 1 Gene Family in Allotetraploid Brassica napus and Its Diploid Ancestors.

The members of PHOSPHATE 1 (PHO1) family play important roles in plant phosphate (Pi) transport and adaptation to Pi deficiency. The functions of PHO1 family proteins have been reported in several plant species, with the exception of Brassica species. Here, we identified 23, 23, and 44 putative PHO1 family genes in Brassica rapa, Brassica oleracea, and Brassica napus by whole genome analysis, respectively. The phylogenetic analysis divided PHO1 family proteins into eight groups, which represented the orthologous relationships among PHO1 members. The gene structure and the conserved motif analysis indicated that the most PHO1 family genes had similar gene structures and the PHO1 proteins shared mutual conserved motifs. The chromosome distribution analysis showed that the majority of BnPHO1 family genes distributed analogously at chromosomes with BrPHO1 and BoPHO1 family genes. The data showed that PHO1 family genes were highly conserved during evolution from diploid to tetraploid. Furthermore, the expression analysis showed that PHO1 family genes had different expression patterns in plant tissues, suggesting the diversity of gene functions in Brassica species. Meanwhile, the expression analysis also revealed that some PHO1 family genes were significantly responsive to Pi deficiency, suggesting that PHO1 family genes play critical roles in Pi uptake and homeostasis under low Pi stress. Altogether, the characteristics of PHO1 family genes provide a reliable groundwork for further dissecting their functions in Brassica species.

Chloroplastic Sec14-like proteins modulate growth and phosphate deficiency responses in Arabidopsis and rice.

Phosphorus is an essential nutrient acquired from soil as phosphate (Pi), and its deficiency severely reduces plant growth and crop yield. Here, we show that single nucleotide polymorphisms (SNPs) at the PHOSPHATIDYLINOSITOL TRANSFER PROTEIN7 (AtPITP7) locus, which encodes a chloroplastic Sec14-like protein, are associated with genetic diversity regarding Pi uptake activity in Arabidopsis (Arabidopsis thaliana). Inactivation of AtPITP7 and its rice (Oryza sativa) homolog (OsPITP6) through T-DNA insertion and CRISPR/Cas9-mediated gene editing, respectively, decreased Pi uptake and plant growth, regardless of Pi availability. By contrast, overexpression of AtPITP7 and OsPITP6 enhanced Pi uptake and plant growth, especially under limited Pi supply. Importantly, overexpression of OsPITP6 increased tiller number and grain yield in rice. Targeted metabolome analysis of glycerolipids in leaves and chloroplasts revealed that inactivation of OsPITP6 alters phospholipid contents, independent of Pi availability, diminishing the reduction in phospholipid content and increase in glycolipid content induced by Pi deficiency; meanwhile, overexpression of OsPITP6 enhanced Pi deficiency induced metabolic alterations. Together with transcriptome analysis of ospitp6 rice plants and phenotypic analysis of grafted Arabidopsis chimeras, these results suggest that chloroplastic Sec14-like proteins play an essential role in growth modulations in response to changes in Pi availability, although their function is critical for plant growth under any Pi condition. The superior traits of OsPITP6-overexpressing rice plants also highlight the potential of OsPITP6 and its homologs in other crops as additional tools for improving Pi uptake and plant growth in low Pi environments.

Systemic adaptation of rice plants under low phosphate conditions and interaction with endophytic bacteria

Phosphate (Pi) is essential for plants. Plants have adapted mechanisms to overcome Pi deficiencies. This study examined the interaction of two contrasting rice varieties (G22 and G299) and two endophytic bacterial strains. Four different culture media were established: full Pi (P0), Pi starvation (P*), insoluble Pi with Pi-solubilizing Burkholderia sp. strain 205 (P+205), or Pi-insolubilizing strain 113 (P+113). We investigated the responses of rice to these media. Root length and weight and the number of crown roots were higher in the P* and P+113 medium than the two other media. However, shoot length, and weight were lower. Most amino acid families were higher in the P+113 medium than in the other media. The roots of G299 plants in the P+113 medium showed the highest relative expression of all phosphate-analyzed genes; however, these genes were expressed at low levels in the leaves of both rice varieties. Notably, the jasmonic acid gene OsJAZ5 showed the highest expression in the roots of G299 plants in the P+113 medium. Our results demonstrate the strong effects of the different genetic backgrounds of bacteria and rice plants on the response to low Pi. We also demonstrate the involvement of jasmonic acid in low Pi and soluble-phosphate-bacteria interaction in G299 plants. A positive interaction between Burkholderia sp. strain 205 and rice plants has been noticed in the promotion of plant growth. Further studies under field conditions should be undertaken to develop this potential strain as a biofertilizer.

Frequency and Risk Factors of Hypophosphatemia in Patients Admitted to Emergency Intensive Care Unit in Zagazig University Hospitals.

Inorganic phosphate is a major electrolyte that participates in many functional and integral processes in the human body. Low Pi levels may lead to multiple organ dysfunction. It is estimated to occur in 40-80% of intensive care unit (ICU) patients. However, it may be ignored during the initial evaluation in ICU. This prospective cross-sectional study included 500 adult ICU cases in two groups; a group with normal Pi levels and a group with hypophosphatemia. All admitted patients were subjected to full history taking, and clinical, laboratory, and radiological evaluation. Collected data were coded, processed, and analyzed using statistical package for social sciences (SPSS) software. Among 500 adult ICU patients; 56.8% had normal phosphate levels while the remaining 43.2% had low phosphate levels. Patients in the hypophosphatemia group were associated with a significantly higher Acute Physiological and Chronic Health Evaluation (APACHE II) score, a longer hospital and ICU stay, a higher incidence of mechanical ventilation use with a longer duration on it, and a significantly higher mortality rate. Risk factors for hypophosphatemia include a higher APACHE II score, longer stay in the hospital and ICU, a higher ratio of mechanical ventilation, and a higher mortality rate. El-Sayed Bsar AEM, El-Wakiel SAR, El-Harrisi MAH, Elshafei ASH. Frequency and Risk Factors of Hypophosphatemia in Patients Admitted to Emergency Intensive Care Unit in Zagazig University Hospitals. Indian J Crit Care Med 2023;27(4):277-282.

Clinical and immunological evaluation of peri-implant tissues around ultra-polished and conventional smoothness zirconia abutments. A 1-year follow-up randomized clinical trial.

This is a randomized clinical trial to compare the clinical and immunological performance of ultrasmooth versus regular-smooth zirconia abutments placed subgingivally after a period of 1 year. Sixty-two bone level platform-switched implants (NobelParallel CC) were placed epicrestally in the mandibular molar or premolar region in 62 patients. After osseointegration, implants were restored with auto polymerizing acrylic resin crowns and subsequently randomly allocated to 2 groups according to the type of screw-retained zirconia crown prescribed. The control group received custom zirconia restoration with the subgingival zirconia part conventionally polished, while the test group implants were restored with ultra-polished zirconia abutments. Periodontal parameters (PD, PI, BOP) and marginal bone level changes (MBLC) were recorded for each implant 2 months after insertion (T0), 1 month after final delivery of the crown (T2) and at the 1-year follow-up (T3). Immunological mediators from GCF (IL-1α, IL-1ra and TNF-α) were inspected at 1 month after provisional (T1) and accordingly at T2 and T3. Data was analyzed statistically, and significance level was set to α = 0.05. After 1-year, there were no significant changes in PD control - 2.18 ±0.89 mm and test - 2.5 ±0.72 mm (p = 0.073). PD between T2 and T3 dropped significantly in the test group (p = 0.037), and remained stable in the control group. PI was not different in both groups at T0 (p = 0.518) and T2 (p = 0.817). At T3, the test group (0.9 ±1.01) had a significantly lower PI than the control group (1.55 ±1.23) (p = 0.035). There was no difference in BOP positive cases between groups after 1 year (control - 61.3%, test - 51.7%, p = 0.455). The amount of IL-1ra decreased significantly in the test group (41.75 ±57.58) (p = 0.001) but not in the control group (59.59 ± 70.43) (p = 0.177). MBLC for the control and test groups after 1 year were 0.68 ±0.7 mm and 0.94 ±0.65 mm (p = 0.061). PD dynamics, PI, BOP and IL-1ra revealed better outcomes around ultra-polished zirconia abutments than around conventionally polished zirconia abutments. This article is protected by copyright. All rights reserved.

Comparative venom toxin analyses of Nigerian viperidae and elapidae snakes

Envenoming by snakebite is a serious health problem that maims and kills a large number of people, primarily in rural areas of developing African countries. The first comparative venom proteomic analyses of four snakes from the viperidae (E. ocellatus and B. arietans) and elapidae (N. haje and N. katiensis) families are presented in this study. Two-dimensional electrophoresis was combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry to analyze the venoms. Proteins were identified by comparing mass spectrometry spectra to those in the reviewed Uniprot-Serpentes database. A protein spot was considered differentially present between samples at a p-value of < 0.05 and a fold change of >2. Viper venoms contained cytotoxic-inducing proteins such as SVMPs, SVSPs, and cytotoxins, whereas elapid snake venoms contained neurotoxic proteins such as PLA2, 3-FTx, and neurotoxins. The PDQuest annotated protein spots on the 2-DE gels showed that the proteins in these snakes' venoms were differentially expressed between snake families and species. The elapid venoms were predominantly acidic (low pI) with low molecular masses, whereas the viperid venoms had high molecular masses and a pI in the region of 7. Venom phosphodiesterase, L-amino acid oxidase and cysteine-rich venom protein were common in the venoms of these snakes, while an uncommon protein actiflagelin was detected in the Naja venoms. Our findings show that there is significant variation in the toxin profiles of these snakes, both at the species and family levels. This has an impact on the clinical manifestations of envenomation. A thorough understanding of the various toxins found in venomous snakes may aid in the development of new and improved therapeutic strategies.

Heat-killed Malassezia pachydermatis suspension modulates the activity of macrophages challenged with Encephalitozoon cuniculi.

Phagocytic responses are critical for effective host defense against opportunistic fungal pathogens, such as Encephalitozoon cuniculi, an obligate intracellular fungus that causes emerging encephalitozoonosis in humans and other animals. Malassezia has immunomodulatory effects and can modulate the production of pro- and anti-inflammatory cytokines via keratinocytes and human monocytes. In this study, we evaluated the modulatory effects of heat-killed Malassezia pachydermatis suspension on macrophages challenged with Encephalitozoon cuniculi. Macrophages were treated with heat-killed M. pachydermatis suspension before being infected with spores of E. cuniculi. The cultures were stained with calcofluor, and the spores, internalized or not, were counted to determine their phagocytic capacity and index (PC and PI, respectively). Microbicidal and phagocytic activities were evaluated by transmission electron microscopy (TEM). The untreated macrophages had higher PC and PI and number of phagocytosed spores than treated macrophages. However, TEM revealed that treated macrophages had higher microbicidal activity because there were few spores in different degrees of degeneration and amorphous materials in the phagocytic vacuoles. Macrophages treated with heat-killed M. pachydermatis suspension had lower PC and PI and incipient presence of E. cuniculi in phagosomes. Treated macrophages had a mixed pattern of cytokine release with Th1, Th2, and Th17 profiles, with emphasis on interleukin (IL)-10, IL-4, IL-17, IL-6, and interferon (IFN)-γ secretion, and particularly high production of anti-inflammatory cytokines. Our results suggest that treatment with heat-killed M. pachydermatis suspension increases the release of cytokines and decreases the phagocytic activity of macrophages challenged with E. cuniculi.

PHOSPHATE STARVATION RESPONSE1 (PHR1) interacts with JASMONATE ZIM-DOMAIN (JAZ) and MYC2 to modulate phosphate deficiency-induced jasmonate signaling in Arabidopsis.

Phosphorus (P) is a macronutrient necessary for plant growth and development. Inorganic phosphate (Pi) deficiency modulates the signaling pathway of the phytohormone jasmonate in Arabidopsis thaliana, but the underlying molecular mechanism currently remains elusive. Here, we confirmed that jasmonate signaling was enhanced under low Pi conditions, and the CORONATINE INSENSITIVE1 (COI1)-mediated pathway is critical for this process. A mechanistic investigation revealed that several JASMONATE ZIM-DOMAIN (JAZ) repressors physically interacted with the Pi signaling-related core transcription factors PHOSPHATE STARVATION RESPONSE1 (PHR1), PHR1-LIKE2 (PHL2), and PHL3. Phenotypic analyses showed that PHR1 and its homologs positively regulated jasmonate-induced anthocyanin accumulation and root growth inhibition. PHR1 stimulated the expression of several jasmonate-responsive genes, whereas JAZ proteins interfered with its transcriptional function. Furthermore, PHR1 physically associated with the basic helix-loop-helix (bHLH) transcription factors MYC2, MYC3, and MYC4. Genetic analyses and biochemical assays indicated that PHR1 and MYC2 synergistically increased the transcription of downstream jasmonate-responsive genes and enhanced the responses to jasmonate. Collectively, our study reveals the crucial regulatory roles of PHR1 in modulating jasmonate responses and provides a mechanistic understanding of how PHR1 functions together with JAZ and MYC2 to maintain the appropriate level of jasmonate signaling under conditions of Pi deficiency.

Root-Expressed Rice PAP3b Enhances Secreted APase Activity and Helps Utilize Organic Phosphate.

Phosphate (Pi) deficiency leads to the induction of purple acid phosphatases (PAPs) in plants, which dephosphorylate organic phosphorus (P) complexes in the rhizosphere and intracellular compartments to release Pi. In this study, we demonstrate that OsPAP3b belongs to group III low-molecular weight PAP and is low Pi-responsive, preferentially in roots. The expression of OsPAP3b is negatively regulated with Pi resupply. Interestingly, OsPAP3b was found to be dual localized to the nucleus and secretome. Furthermore, OsPAP3b is transcriptionally regulated by OsPHR2 as substantiated by DNA-protein binding assay. Through in vitro biochemical assays, we further demonstrate that OsPAP3b is a functional acid phosphatase (APase) with broad substrate specificity. The overexpression (OE) of OsPAP3b in rice led to increased secreted APase activity and improved mineralization of organic P sources, which resulted in better growth of transgenics compared to the wild type when grown on organic P as an exogenous P substrate. Under Pi deprivation, OsPAP3b knock-down and knock-out lines showed no significant changes in total P content and dry biomass. However, the expression of other phosphate starvation-induced genes and the levels of metabolites were found to be altered in the OE and knock-down lines. In addition, in vitro pull-down assay revealed multiple putative interacting proteins of OsPAP3b. Our data collectively suggest that OsPAP3b can aid in organic P utilization in rice. The APase isoform behavior and nuclear localization indicate its additional role, possibly in stress signaling. Considering its important roles, OsPAP3b could be a potential target for improving low Pi adaptation in rice.

NtPIN3 positively regulates low phosphorus tolerance by changing root elongation, Pi concentration and antioxidant capacity in tobacco

Phosphorus (P) is a critical nutrient for plants, and inorganic orthophosphate (Pi) deficiency results in declining crop quality and yields. Auxin plays a key regulatory effect on plant growth in response to Pi deficiency, yet the regulatory mechanisms remain poorly understood. In this study, we cloned and identified an auxin efflux transporter, NtPIN3, involved in the low phosphate response of tobacco (Nicotiana tabacum). NtPIN3 is located on the cytoplasmic membrane, and high expression levels of NtPIN3 were observed under phosphorus deficiency conditions. Moreover, NtPIN3 overexpression in tobacco could dramatically enhance low Pi tolerance by increasing the root length, which was consistent with the GUS activity and the auxin concentration in the root. Notably, Pi concentration and antioxidant capacity in overexpression transgenic tobacco were significantly increased compared with the wild type (WT) under low Pi stress conditions due to Pi uptake (NtPT1 and NtPT2) and antioxidant capacity (superoxide dismutase, peroxidase, and catalase). In contrast, compared with WT, the ntpin3 mutant significantly reduced root length, auxin concentration, Pi concentration, and antioxidant capacity under low Pi stress conditions, suggesting that NtPIN3 is a positive regulator of low phosphate in tobacco. In addition, the yeast-two-hybrid and luciferase complementation imaging assay confirmed that NtPIN3 interacted with NtPIN2. Altogether, the NtPIN3 involved in low Pi response and overexpression of NtPIN3 contributed to enhanced tolerance to low Pi stress, which proved an underlying gene for breeding low Pi-stress tolerant plants.

Transcription factor OsNAC016 negatively regulates phosphate-starvation response in rice

Phosphate (Pi), the main form of inorganic phosphorus that can be absorbed by plants, is one of the most limiting macro-nutrients in plants. However, the underlying molecular mechanism determining how plants sense external Pi levels and reprogram transcriptional and adaptive responses is incompletely understood. At present, few rice NAC members have been reported to be involved in the signaling pathways of Pi homeostasis in plants. Here, our research demonstrated that OsNAC016, a Pi-starvation responsive gene in rice, was regulated by PHOSPHATE STARVATION RESPONSE protein 1 (OsPHR1) and OsPHR4. Under Pi-starvation stress, the root growth of OsNAC016-overexpression lines was inhibited more severely, and overexpression plants had lower Pi content than wild type, while osnac016 mutant was hyposensitive to Pi starvation, indicating that OsNAC016 negatively modulates rice Pi-starvation response. Chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) analysis and transient transactivation assays indicated that OsNAC016 could activate the SPX-domain-containing protein 2 (OsSPX2) gene through binding to its promoter. Further, we found that Pi starvation enhanced OsNAC016 binding to the OsSPX2 promoter, thus strongly promoting OsSPX2 expression. At the same time, Pi starvation induced OsNAC016 protein accumulation in plants. Moreover, similar to OsSPX2, OsNAC016 negatively regulates leaf inclination by repressing the cell elongation in lamina joint in rice under Pi-starvation stress. Together, our findings demonstrate that OsNAC016 negatively regulates rice phosphate-starvation response and leaf inclination by activating OsSPX2 expression under Pi-starvation conditions. These data provide a strategy to create smart crops with ideal shoot architecture and high phosphorus utilization efficiency.

Mutation of OsLPR3 Enhances Tolerance to Phosphate Starvation in Rice.

Low Phosphate Root (LPR) encodes a protein localized to the endoplasmic reticulum (ER) and cell wall. This gene plays a key role in responding to phosphate (Pi) deprivation, especially in remodeling the root system architecture (RSA). An identification and expression analysis of the OsLPR family in rice (Oryza sativa) has been previously reported, and OsLPR5, functioning in Pi uptake and translocation, is required for the normal growth and development of rice. However, the role of OsLPR3, one of the five members of this family in rice, in response to Pi deficiency and/or in the regulation of plant growth and development is unknown. Therefore, in this study, the roles of OsLPR3 in these processes were investigated, and some functions were found to differ between OsLPR3 and OsLPR5. OsLPR3 was found to be induced in the leaf blades, leaf sheaths, and roots under Pi deprivation. OsLPR3 overexpression strongly inhibited the growth and development of the rice but did not affect the Pi homeostasis of the plant. However, oslpr3 mutants improved RSA and Pi utilization, and they exhibited a higher tolerance to low Pi stress in rice. The agronomic traits of the oslpr3 mutants, such as 1000-grain weight and seed length, were stimulated under Pi-sufficient conditions, indicating that OsLPR3 plays roles different from those of OsLPR5 during plant growth and development, as well as in the maintenance of the Pi status of rice.

Functional Analysis of Bna-miR399c-PHO2 Regulatory Module Involved in Phosphorus Stress in Brassica napus

Phosphorus stress is one of the important factors restricting plant growth and development, and the microRNA (miRNA) family is involved in the regulation of the response to plant nutrient stress by repressing the expression of target genes at the post-transcriptional or translational level. miR399 is involved in the transportation of phosphate in multiple plants by improving tolerance to low Pi conditions. However, the effect of miR399 on the response of low Pi stress in rapeseed (Brassica napus L.) is unclear. The present study showed a significant increase in taproot length and lateral root number of plants overexpressing Bna-miR399c, while the biomass and Pi accumulation in shoots and roots increased, and the anthocyanin content decreased and chlorophyll content improved under low Pi stress. The results illustrate that Bna-miR399c could enhance the uptake and transportation of Pi in soil, thus making B. napus more tolerant to low Pi stress. Furthermore, we confirmed that BnPHO2 is one of the targets of Bna-miR399c, and the rejection of Pi in rapeseed seedlings increased due to the overexpression of BnPHO2. Hence, we suggest that miR399c-PHO2 module can effectively regulate the homeostasis of Pi in B. napus. Our study can also provide the theoretical basis for germplasm innovation and the design of intelligent crops with low nutrient input and high yield to achieve the dual objectives of income and yield increase and environmental protection in B. napus.

A transcriptional activator from Rhizophagus irregularis regulates phosphate uptake and homeostasis in AM symbiosis during phosphorous starvation.

Phosphorus (P) is one of the most important nutrient elements for plant growth and development. Under P starvation, arbuscular mycorrhizal (AM) fungi can promote phosphate (Pi) uptake and homeostasis within host plants. However, the underlying mechanisms by which AM fungal symbiont regulates the AM symbiotic Pi acquisition from soil under P starvation are largely unknown. Here, we identify a HLH domain containing transcription factor RiPho4 from Rhizophagus irregularis. To investigate the biological functions of the RiPho4, we combined the subcellular localization and Yeast One-Hybrid (Y1H) experiments in yeasts with gene expression and virus-induced gene silencing approach during AM symbiosis. The approach during AM symbiosis. The results indicated that RiPho4 encodes a conserved transcription factor among different fungi and is induced during the in planta phase. The transcription of RiPho4 is significantly up-regulated by P starvation. The subcellular localization analysis revealed that RiPho4 is located in the nuclei of yeast cells during P starvation. Moreover, knock-down of RiPho4 inhibits the arbuscule development and mycorrhizal Pi uptake under low Pi conditions. Importantly, RiPho4 can positively regulate the downstream components of the phosphate (PHO) pathway in R. irregularis. In summary, these new findings reveal that RiPho4 acts as a transcriptional activator in AM fungus to maintain arbuscule development and regulate Pi uptake and homeostasis in the AM symbiosis during Pi starvation.

Effect of argon plasma pre-treatment of healing abutments on peri-implant microbiome and soft tissue integration: a proof-of-concept randomized study

PurposeBiofilm-free implant surface is ultimate prerequisite for successful soft and bone tissue integration. Objective of the study was to estimate the effects of argon plasma healing abutment pre-treatment (PT) on peri-implant soft-tissue phenotype (PiSP), inflammation, plaque accumulation and the microbiome (PiM) between non-treated (NPT) and treated (PT) abutments following 3-months healing period. The hypothesis was that cell-conductive and antimicrobial properties of PT would yield optimal conditions for soft tissue integration.Material and MethodsTwo months following second-phase surgery, microbiological and clinical parameters were assessed around thirty-six healing abutments with two types of microtopography, smooth surface (MACHINED) and ultrathin threaded microsurface (ROUGH). A two level randomization schema was used to achieve equal distribution and abutments were randomly divided into rough and machined groups, and then divided into PT and NPT groups. PiM was assessed using next-generation DNA sequencing.ResultsPiM bacterial composition was highly diverse already two months post-implantation, consisting of key-stone pathogens, early and late colonizers, while the mycobiome was less diverse. PT was associated with lower plaque accumulation and inflammation without significant impact on PiSP, while in NPT clinical parameters were increased and associated with periopathogens. NPT mostly harbored late colonizers, while PT exerted higher abundance of early colonizers suggesting less advanced plaque formation. Interaction analysis in PT demonstrated S. mitis co-occurrence with pro-healthy Rothia dentocariosa and co-exclusion with Parvimonas micra, Porphyromonas endodontalis and Prevotella oris. PiSP parameters were generally similar between the groups, but significant association between PiM and keratinized mucosa width was observed in both groups, with remarkably more expressed diversity in NPT compared to PT. PT resulted in significantly lower BOP and PI around rough and machined abutments, respectively, without specific effect on PiM and PiSP.ConclusionsPT contributed to significantly the less advanced biofilm accumulation and inflammation without specific effects on PiSP.

The revealing of a novel lipid transfer protein lineage in green algae

BackgroundNon-specific lipid transfer proteins (nsLTPs) are a group of small and basic proteins that can bind and transfer various lipid molecules to the apoplastic space. A typical nsLTP carries a conserved architecture termed eight-cysteine motif (8CM), a scaffold of loop-linked helices folding into a hydrophobic cavity for lipids binding. Encoded by a multigene family, nsLTPs are widely distributed in terrestrial plants from bryophytes to angiosperms with dozens of gene members in a single species. Although the nsLTPs in the most primitive plants such as Marchantia already reach 14 members and are divergent enough to form separate groups, so far none have been identified in any species of green algae.ResultsBy using a refined searching strategy, we identified putative nsLTP genes in more than ten species of green algae as one or two genes per haploid genome but not in red and brown algae. The analyses show that the algal nsLTPs carry unique characteristics, including the extended 8CM spacing, larger molecular mass, lower pI value and multiple introns in a gene, which suggests that they could be a novel nsLTP lineage. Moreover, the results of further investigation on the two Chlamydomonas nsLTPs using transcript and protein assays demonstrated their late zygotic stage expression patterns and the canonical nsLTP properties were also verified, such as the fatty acids binding and proteinase resistance activities.ConclusionsIn conclusion, a novel nsLTP lineage is identified in green algae, which carries some unique sequences and molecular features that are distinguishable from those in land plants. Combined with the results of further examinations of the Chlamydomonas nsLTPs in vitro, possible roles of the algal nsLTPs are also suggested. This study not only reveals the existence of the nsLTPs in green algae but also contributes to facilitating future studies on this enigmatic protein family.

The utilization and molecular mechanism of arbuscular mycorrhizal symbiosis in vegetables

Vegetables are an important food and many of them can form AM (arbuscular mycorrhiza) symbiosis with AMF (AM fungi) that belongs to the sub-phylum Glomeromycota. The symbiosis with AM in vegetables enhances their tolerance to various stresses, such as low Pi (phosphate), salinity and soil-borne diseases. In the past decades, the molecular mechanism of AM symbiosis in vegetables has begun to emerge. Here, we review the key studies characterizing the molecular mechanism of AM symbiosis and highlights the huge potential of AMF in vegetable cultivation.

Endoplasmic reticulum calnexins participate in the primary root growth response to phosphate deficiency.

Accumulation of incompletely folded proteins in the endoplasmic reticulum (ER) leads to ER stress, activates ER protein degradation pathways, and upregulates genes involved in protein folding. This process is known as the unfolded protein response (UPR). The role of ER protein folding in plant responses to nutrient deficiencies is unclear. We analyzed Arabidopsis (Arabidopsis thaliana) mutants affected in ER protein quality control and established that both CALNEXIN (CNX) genes function in the primary root response to phosphate (Pi) deficiency. CNX1 and CNX2 are homologous ER lectins promoting protein folding of N-glycosylated proteins via the recognition of the GlcMan9GlcNAc2 glycan. Growth of cnx1-1 and cnx2-2 single mutants was similar to that of the wild type under high and low Pi conditions, but the cnx1-1 cnx2-2 double mutant showed decreased primary root growth under low Pi conditions due to reduced meristematic cell division. This phenotype was specific to Pi deficiency; the double mutant responded normally to osmotic and salt stress. Expression of CNX2 mutated in amino acids involved in binding the GlcMan9GlcNAc2 glycan failed to complement the cnx1-1 cnx2-2 mutant. The root growth phenotype was Fe-dependent and was associated with root apoplastic Fe accumulation. Two genes involved in Fe-dependent inhibition of primary root growth under Pi deficiency, the ferroxidase LOW PHOSPHATE 1 (LPR1) and P5-type ATPase PLEIOTROPIC DRUG RESISTANCE 2 (PDR2) were epistatic to CNX1/CNX2. Overexpressing PDR2 failed to complement the cnx1-1 cnx2-2 root phenotype. The cnx1-1 cnx2-2 mutant showed no evidence of UPR activation, indicating a limited effect on ER protein folding. CNX might process a set of N-glycosylated proteins specifically involved in the response to Pi deficiency.

Genome wide association studies for acid phosphatase activity at varying phosphorous levels in Brassica juncea L.

Acid phosphatases (Apases) are an important group of enzymes that hydrolyze soil and plant phosphoesters and anhydrides to release Pi (inorganic phosphate) for plant acquisition. Their activity is strongly correlated to the phosphorus use efficiency (PUE) of plants. Indian mustard (Brassica juncea L. Czern & Coss) is a major oilseed crop that also provides protein for the animal feed industry. It exhibits low PUE. Understanding the genetics of PUE and its component traits, especially Apase activity, will help to reduce Pi fertilizer application in the crop. In the present study, we evaluated 280 genotypes of the diversity fixed foundation set of Indian mustard for Apase activity in the root (RApase) and leaf (LApase) tissues at three- low (5µM), normal (250µM) and high (1mM) Pi levels in a hydroponic system. Substantial effects of genotype and Pi level were observed for Apase activity in both tissues of the evaluated lines. Low Pi stress induced higher mean RApase and LApase activities. However, mean LApase activity was relatively more than mean RApase at all three Pi levels. JM06016, IM70 and Kranti were identified as promising genotypes with higher LApase activity and increased R/S at low Pi. Genome-wide association study revealed 10 and 4 genomic regions associated with RApase and LApase, respectively. Annotation of genomic regions in the vicinity of peak associated SNPs allowed prediction of 15 candidates, including genes encoding different family members of the acid phosphatase such as PAP10 (purple acid phosphatase 10), PAP16, PNP (polynucleotide phosphorylase) and AT5G51260 (HAD superfamily gene, subfamily IIIB acid phosphatase) genes. Our studies provide an understanding of molecular mechanism of the Apase response of B. juncea at varying Pi levels. The identified SNPs and candidate genes will support marker-assisted breeding program for improving PUE in Indian mustard. This will redeem the crop with enhanced productivity under restricted Pi reserves and degrading agro-environments.

Establishing the Prevalence of Osteomalacia in Arab Adolescents Using Biochemical Markers of Bone Health.

Nutrition-acquired osteomalacia is a bone mineralization disorder associated with dietary calcium and/or solar vitamin D deficiency, risk factors considered common in the Middle Eastern region. Establishing less invasive, cheap, and widely available diagnostic markers for this underdiagnosed entity is essential, in particular for screening in high-risk groups. This study assessed the prevalence of biochemical osteomalacia in Arab adolescents. In this cross-sectional study performed between September 2019 and March 2021, adolescents aged 12-17 years from 60 different secondary and preparatory year schools in Riyadh, Saudi Arabia were included. Anthropometrics and fasting blood samples were collected. Biochemical osteomalacia was defined as any two of the following four serum markers of hypomineralization, namely low 25 hydroxyvitamin D (25OHD &lt; 30 nmol/L), high alkaline phosphatase (ALP), low calcium (Ca), and/or inorganic phosphorous (Pi). A total of 2938 Arab adolescents [1697 girls; mean age (years) 14.8 ± 1.8; 1241 boys; mean age 15.1 ± 1.6] were recruited. Vitamin D deficiency was noted in 56.2% (n = 953) of girls and 27.1% (n = 336) of boys (p &lt; 0.001). The overall prevalence of biochemical osteomalacia was 10.0% (n = 295/2938) and was higher in girls than boys (14.7% vs. 3.6%, p &lt; 0.001). The prevalence of low serum Ca and/or Pi was also higher in girls than in boys (24.2% vs. 12.5%, respectively, p &lt; 0.001), as well as elevated ALP (5.1% vs. 1.5%, p &lt; 0.001). Overall, girls were 4.6 times (95% CI 3.3-6.4) more likely to have biochemical osteomalacia than boys. Screening of apparently healthy Arab adolescents revealed a high prevalence of deranged mineralization markers suggestive of biochemical osteomalacia, which was significantly more common in girls than boys and was likely associated with Arab traditional clothing and diet. The proposed combination of typically altered mineralization markers for the diagnosis of osteomalacia is, at best, suggestive until further comparisons with established diagnostic tools (histological analysis of bone biopsies) are conducted.