Decoupling of P from C, N, and K Elements in Cucumber Leaves Caused by Nutrient Imbalance under a Greenhouse Continuous Cropping System

There is insufficient information regarding the stoichiometric variation and coupling status of carbon (C), nitrogen (N), phosphorus (P), and potassium (K) in the leaves of nutrient-enriched greenhouse agroecosystems with increasing planting time. Therefore, we assessed the variation in elemental stoichiometry ratios in soil and cucumber (Cucumis sativus L.) leaves, and the coupling status of elemental utilization in the leaves under continuous cropping systems using natural (only soil; i.e., control soil, CO) and artificial (soil + straw + chicken + urea; i.e., straw mixture soil, ST) soil via monitoring studies for 11 years in a solar greenhouse. Soil organic C, total N, and total P concentrations increased by 63.4%, 72.7%, and 144.3% in the CO, respectively, after 11 years of cultivation (compared to the first year), and by 18.1%, 24.3%, and 117.7% in the ST under continuous cropping conditions, respectively. Total K concentrations remained unchanged in both soils. Moreover, the availability of these soil elements increased to different degrees in both soils after 11 years of planting. Additionally, the leaf P concentration increased by 9.8% in the CO, while leaf N and K concentrations did not change, suggesting decoupling of P utilization from that of N and K in leaves under a continuous cropping system. These findings suggest that imbalanced soil nutrients under continuous cropping conditions results in decoupling of P from N and K in the utilization of leaf nutrients.

A novel dominant selection system for plant transgenics based on phosphite metabolism catalyzed by bacterial alkaline phosphatase

Selective markers are generally indispensable in plant genetic transformation, of which the frequently used are of antibiotic or herbicide resistance. However, the increasing concerns on transgenic biosafety have encouraged many new and safe selective markers emerging, with an eminent representative as phosphite (Phi) in combination to its dehydrogenase (PTDH, e.g. PtxD). As bacterial alkaline phosphatase (BAP) can resemble PtxD to oxidatively convert toxic Phi into metabolizable phosphate (Pi), herein we harnessed it as the substitute of PtxD to develop an alternative Phi-based selection system. We first validated the Escherichia coli BAP (EcBAP) did own an extra enzymatic activity of oxidizing Phi to Pi. We further revealed EcBAP could be used as a dominant selective marker for Agrobacterium-mediated tobacco transformation. Although the involved Phi selection for transformed tobacco cells surprisingly required the presence of Pi, it showed a considerable transformation efficiency and dramatically accelerated transformation procedure, as compared to the routine kanamycin selection and the well-known PtxD/Phi system. Moreover, the EcBAP transgenic tobaccos could metabolize toxic Phi as a phosphorus (P) fertilizer thus underlying Phi-resistance, and competitively possess a dominant growth over wild-type tobacco and weeds under Phi stress. Therefore, this novel BAP/Phi-coupled system, integrating multiple advantages covering biosafe dominant selective marker, plant P utilization and weed management, can provide a PTDH-bypass technological choice to engineer transgenic plant species, especially those of great importance for sustainable agriculture.

Transcriptome Analysis of Zygophyllum xanthoxylum Adaptation Strategies to Phosphate Stress

Soil phosphate (Pi) deficiency is a global issue and a major constraint on plant growth. Plants typically acclimatize to low Pi by enhancing their P utilization and/or P acquisition efficiencies; however, different species have variable preferred strategies. RNA sequencing analysis was performed on the shoots and roots of Zygophyllum xanthoxylum, under 1 day and 10 days of Pi stress, to investigate their adaptation strategies to P deprivation. A total of 364,614 unigenes and 9,270 differentially expressed genes (DEGs) were obtained via transcriptome sequencing. An analysis of the DEGs revealed that under the 10D treatment, anthocyanin synthesis genes were upregulated under Pi stress, whereas gibberellin, ethylene, and cytokinins synthesis genes were upregulated, and abscisic acid synthesis genes were downregulated. Genes related to organic acid synthesis, encoding for purple acid phosphatases (APase) and nucleases (RNase) were upregulated under the 1D and 10D treatments, respectively. Furthermore, genes associated with Pi transport were induced by Pi stress. Zygophyllum xanthoxylum has special P adaptation strategies, the variation trends of genes involved in external P mobilization and acquisition, which were different from that of most other species; however, the expression levels of organophosphorus mobilization related genes, such as APases and RNases, were significantly increased. Meanwhile, PHT2s and TPTs, which distributed Pi to effective sites (e.g., chloroplast), played critical roles in the maintenance of photosynthesis. We speculated that these were economic and energy saving strategies, and there are critical adaptive mechanisms that Z. xanthoxylum employs to cope with deficits in Pi.

Responses of Root Endophytes to Phosphorus Availability in Peach Rootstocks With Contrasting Phosphorus-Use Efficiencies

Phosphorus (P) is an important macronutrient for all lives, but it is also a finite resource. Therefore, it is important to understand how to increase the P availability and plant uptake. The endophytes can help host plants to improve P uptake and will be apparently affected by plant genotypes. To investigate the mechanism of root endophytes in promoting P uptake of peach rootstocks, we analyzed the variations of the root endophytic fungal and bacterial communities of peach rootstocks with different P efficiencies under high or low level of P addition. Results showed that Proteobacteria was the dominant bacterial phylum in the roots of all rootstocks under the two levels of P addition. At low P level, the abundance of Actinoplanes in phosphorus-inefficiency root system was apparently higher than that at high P level. Actinoplanes produced important secondary metabolites, improving the stress resistance of plants. Under high P condition, the abundance of Ferrovibrio was higher in Qing Zhou Mi Tao than in Du Shi. Fe oxides considerably reduced the availability of applied P, which partially explained why the P utilization in Qing Zhou Mi Tao is inefficient. Further, Ascomycota was the dominant fungal phylum in the roots of all rootstocks under different levels of P addition. The fungi community of roots varied in different rootstocks at each P level, but was similar for the same rootstock at different P levels, which indicated that genotype had a greater effect than P addition on the fungal community of peach rootstocks.

Periphytic microbial response to environmental phosphate bioavailability – relevance to P management in paddy fields

Periphyton occurs widely in shallow-water ecosystems such as paddy fields and plays critical parts in regulating local phosphorus cycling. As such, understanding the mechanisms of the biofilm’s response to environmental P variability may lead to better perceptions of P utilization and retention in rice farms. Present study aims at exploring the biological and biochemical processes underlying periphyton’s P buffering capability through examining changes in community structure, phosphorus uptake and storage, and molecular makeup of exometabolome at different levels of P availability. Under stressed (both excessive and scarce) phosphorus conditions, we found increased populations of the bacterial genus capable of transforming orthophosphate to polyphosphate, as well as mixotrophic algae who can survive through phagotrophy. These results were corroborated by observed polyphosphate buildup under low and high P treatment. Exometabolomic analyses further revealed that periphytic organisms may substitute S-containing lipids for phospholipids, use siderophores to dissolve iron (hydr)oxides to scavenge adsorbed P, and synthesize auxins to resist phosphorus starvation. These findings not only shed light on the mechanistic insights responsible for driving the periphytic P buffer but attest to the ecological roles of periphyton in aiding plants such as rice to overcome P limitations in natural environment. Importance The ability of periphyton to buffer environmental P in shallow aquatic ecosystems may be a natural lesson on P utilization and retention in paddy fields. This work revealed the routes and tools through which periphytic organisms adapt to and regulate ambient P fluctuation. The mechanistic understanding further implicates that the biofilm may serve rice plants to alleviate P stress. Additional results from extracellular metabolite analyses suggest the dissolved periphytic exometabolome can be a valuable nutrient source for soil microbes and plants to reduce biosynthetic costs. These discoveries have the potential to improve our understanding of biogeochemical cycling of phosphorus in general and to refine P management strategies for rice farm in particular.

Evaluation of Potato Varieties Grown in Hydroponics for Phosphorus Use Efficiency

Global phosphate mineral resources are nonrenewable and are inevitably depleting. Exploiting elite varieties has become imperative for the efficient use of phosphorus (P) for sustainable crop production. Three potato varieties were hydroponically evaluated for P mobilization, uptake, and utilization efficiencies at different P levels and sources during 28 d seedling growth. ‘Harley Blackwell’, ‘La Chipper’, and ‘Red LaSoda’ were selected from a previous study and grown in modified Hoagland solution, with different P concentrations of soluble high P as NaH2PO4 (10 mg L−1 P), soluble low P (1 mg L−1 P), and 286 mg L−1 sparingly soluble P as tri-calcium phosphate [TCP, Ca3(PO4)2] with 2286 mg L−1 CaSO4. ‘Harley Blackwell’ and ‘La Chipper’ had significantly greater biomass than ‘Red LaSoda’ in the low P or TCP treatments. In low-P stress, P utilization efficiency was significantly greater for ‘Harley Blackwell’ than that of the other two varieties. ‘Red LaSoda’ was more efficient in P mobilization from TCP as compared to the other two cultivars. The holistic score analysis indicated that ‘Harley Blackwell’ was the most P-efficient while ‘Red LaSoda’ was the least P-efficient. The results of this study show that the TCP solution was successful for screening P-efficient potato varieties.

Genetic Dissection of Phosphorous Uptake and Utilization Efficiency Traits Using GWAS in Mungbean

Mungbean (Vignaradiata L. Wilczek) is an early maturing legume grown predominantly in Asia for its protein-rich seeds. P deficiency can lead to several physiological disorders which ultimately result in a low grain yield in mungbean. The genetic dissection of PUpE (Puptake efficiency) and PUtE (P utilization efficiency) traits are essential for breeding mungbean varieties with a high P uptake and utilization efficiency. The study involves an association mapping panel consisting of 120 mungbean genotypes which were phenotyped for total dry weight, P concentration, total P uptake, and P utilization efficiency under low P (LP) and normal P (NP) conditions in a hydroponic system. A genotyping-by-sequencing (GBS) based genome-wide association study (GWAS) approach was employed to dissect the complexity of PUpE and PUtE traits at the genetic level in mungbean. This has identified 116 SNPs in 61 protein-coding genes and of these, 16 have been found to enhance phosphorous uptake and utilization efficiency in mungbeans. We identified six genes with a high expression (VRADI01G04370, VRADI05G20860, VRADI06G12490, VRADI08G20910, VRADI08G00070 and VRADI09G09030) in root, shoot apical meristem and leaf, indicating their role in the regulation of P uptake and utilization efficiency in mungbean. The SNPs present in three genes have also been validated using a Sanger sequencing approach.

Effects of Exogenous 6-Phytase (EC 3.1.3.26) Supplementation on Performance, Calcium and Phosphorous Digestibility, and Bone Mineralisation and Density in Weaned Piglets

Phosphorus (P) is an essential mineral for growing piglets, which is poorly accessible in vegetable feedstuffs as it is stored as phytates. Thus, phytase supplementation is essential to increase P availability. Two experiments were conducted to evaluate a novel 6-phytase (EC 3.1.3.26) in weaned pigs fed low-P diets. In each experiment, one hundred and twenty piglets were fed a positive control (PC; adequate in Ca and P), a negative control (NC; limiting in Ca and P), or NC supplemented with 125, 250, or 500 FTU/kg of phytase (NC125, NC250, and NC500, respectively). P content was lower in diets of Experiment 1 than diets of Experiment 2. In Experiment 1, piglets offered PC or phytase diets had higher growth and efficiency compared with NC diets. In Experiment 2, similar effects were obtained, but the effects were less significant. In both experiments, P and Ca ATTD and bone density were significantly increased with phytase supplementation. Moreover, PC and NC500 had higher P concentrations and lower alkaline phosphatase activity in plasma than NC. To conclude, supplementation with the new 6-phytase at doses up to 500 FTU/kg enhanced P utilization, growth performance, and bone density in piglets fed P-limiting diets.

Mineral Phosphorus Supply in Piglets Impacts the Microbial Composition and Phytate Utilization in the Large Intestine

A sufficient supply of phosphorus (P) to pigs in livestock farming is based on the optimal use of plant-based phytate and mineral P supplements to ensure proper growth processes and bone stability. However, a high P supplementation might bear the risk of higher environmental burden due to the occurrence of excess P and phytate degradation products in manure. In this context, the intestinal microbiota is of central importance to increase P solubility, to employ non-mineral P by the enzymatic degradation of phytate, and to metabolize residual P. A feeding experiment was conducted in which piglets were fed diets with different P levels, resulting in three groups with low, medium (covering requirements), and high concentrations of available P. Samples from caecum and colon digesta were analysed for microbial composition and phytate breakdown to estimate the microbial contribution to metabolize P sources. In terms of identified operational taxonomic units (OTU), caecum and colon digesta under the three feeding schemes mainly overlap in their core microbiome. Nevertheless, different microbial families correlate with increased dietary P supply. Specifically, microbes of Desulfovibrionaceae, Pasteurellaceae, Anaerovoracaceae, and Methanobacteriaceae were found significantly differentially abundant in the large intestine across the dietary treatments. Moreover, members of the families Veillonellaceae, Selenomonadaceae, and Succinivibrionaceae might contribute to the observed phytate degradation in animals fed a low P diet. In this sense, the targeted manipulation of the intestinal microbiota by feeding measures offers possibilities for the optimization of intestinal phytate and P utilization.

Formulation of diets for pigs based on a ratio between digestible calcium and digestible phosphorus results in reduced excretion of calcium in urine without affecting retention of calcium and phosphorus compared with formulation based on values for total calcium

An experiment was conducted to test the hypothesis that formulating diets for pigs based on a ratio between standardized total tract digestible (STTD) Ca and STTD P instead of total Ca and STTD P does not decrease Ca retention, but increases P utilization. Forty barrows (59.4 ± 3.8 kg) were individually housed in metabolism crates and allotted to 4 corn-soybean meal-based diets in a randomized complete block design with 2 blocks and 5 pigs per diet in each block. Diets were formulated using a 2 × 2 factorial design with 2 diet formulation principles (total Ca or STTD Ca) and 2 inclusion levels of microbial phytase (0 or 500 units per kg of feed). Phytase was assumed to release 0.11% STTD P and 0.16% total Ca. Diets were formulated based on requirements for total Ca and STTD P or a ratio between STTD Ca and STTD P of 1.25:1. Diets were fed for 11 d and fecal and urine samples were collected from feed provided on d 6 to 9. Interactions (P < 0.05) between diet formulation principle and phytase level were observed for Ca intake, Ca in feces, Ca absorbed, Ca retained, P digestibility, P absorbed, and P in urine. Phytase increased (P < 0.05) the digestibility of Ca in both total Ca and STTD Ca diets. Without phytase, Ca intake, Ca in feces, and Ca absorbed was greater (P < 0.05) from pigs fed total Ca diets than from pigs fed STTD Ca diets, but P absorbed, P digestibility, and P in urine was greater (P < 0.05) from pigs fed STTD Ca diets than from pigs fed total Ca diets. However, in the presence of phytase, no differences between diet formulation principles were observed in these variables. Regardless of phytase, Ca in urine was lower (P < 0.05) from pigs fed STTD Ca diets than from pigs fed total Ca diets. There were no differences in Ca retention between pigs fed STTD Ca diets and total Ca diets, but pigs fed total Ca diets retained less (P < 0.05) Ca if diets contained phytase. No differences in P retention were observed between diet formulation principles, but pigs fed non-phytase diets retained more (P < 0.05) P than pigs fed diets with phytase. In conclusion, because diets formulated based on STTD Ca contain less Ca than total Ca diets, pigs fed STTD Ca diets excreted less Ca in urine, but retention of Ca was not affected. Formulating non-phytase diets based on STTD Ca instead of total Ca increased P absorption, which confirms the detrimental effect of excess Ca on P digestibility. However, P retention was not improved if pigs were fed STTD Ca diets.