Interspecific facilitation of micronutrient uptake between cluster-root-bearing trees and non-cluster rooted-shrubs in a Banksia woodland.

Background and aims: Belowground interspecific plant facilitation is supposed to play a key role in enabling species co-existence in hyperdiverse ecosystems in extremely nutrient-poor, semi-arid habitats, such as Banksia woodlands in southwestern-Australia. Manganese (Mn) is readily mobilised by Banksia cluster root activity in most soils and accumulates in mature leaves of native Australian plant species without significant remobilisation during leaf senescence. We hypothesised that neighbouring shrubs are facilitated in terms of Mn uptake depending on distance to surrounding cluster root-forming Banksia trees. Methods: We mapped all Banksia trees and selected neighbouring shrubs within a study site in Western Australia. Soil samples were collected and analysed for physical properties and nutrient concentrations. To assesses the effect of Banksia tree proximity on leaf Mn concentrations [Mn] of non-cluster-rooted woody shrubs, samples of similarly aged leaves were taken. We used multiple linear models to test for factors affecting shrub leaf [Mn]. Results: None of the assessed soil parameters showed a significant correlation with shrub leaf Mn concentrations. However, we observed a significant positive effect of very close Banksia trees (2 m) on leaf [Mn] in one of the understorey shrubs. We found additional effects of elevation and shrub size. Conclusions: Leaf micronutrient concentrations of understorey shrubs were enhanced when growing within 2 m of tall Banksia trees. Our model predictions also indicate that belowground facilitation of Mn uptake was shrub size-dependent. We discuss this result in the light of plant water relations and shrub root system architecture. Supplementary Information: The online version contains supplementary material available at 10.1007/s11104-023-06092-6.

Phosphorus absorption kinetics and exudation strategies of roots developed by three lupin species to tackle P deficiency.

MAIN CONCLUSION: Different lupin species exhibited varied biomass, P allocation, and physiological responses to P-deprivation. White and yellow lupins had higher carboxylate exudation rates, while blue lupin showed the highest phosphatase activity. White lupin (Lupinus albus) can produce specialized root structures, called cluster roots, which are adapted to low-phosphorus (P) soil. Blue lupin (L. angustifolius) and yellow lupin (L. luteus), which are two close relatives of white lupin, do not produce cluster roots. This study characterized plant responses to nutrient limitation by analyzing biomass accumulation and P distribution, absorption kinetics and root exudation in white, blue, and yellow lupins. Plants were grown in hydroponic culture with (64 µM NaH2PO4) or without P for 31 days. Under P limitation, more biomass was allocated to roots to improve P absorption. Furthermore, the relative growth rate of blue lupin showed the strongest inhibition. Under + P conditions, the plant total-P contents of blue lupin and yellow lupin were higher than that of white lupin. To elucidate the responses of lupins via the perspective of absorption kinetics and secretion analysis, blue and yellow lupins were confirmed to have stronger affinity and absorption capacity for orthophosphate after P-deprivation cultivation, whereas white lupin and yellow lupin had greater ability to secrete organic acids. The exudation of blue lupin had higher acid phosphatase activity. This study elucidated that blue lupin was more sensitive to P-scarcity stress and yellow had the greater tolerance of P-deficient condition than either of the other two lupin species. The three lupin species have evolved different adaptation strategies to cope with P deficiency.

Effects of Phosphate-solubilizing Bacteria on Soil Phosphorus Fractions and Supply to Maize Seedlings Grown in Lateritic Red Earths and Cinnamon Soils.

Phosphorus (P) is often the limiting factor for plant growth because of its low mobility and availability in soils. Phosphate-solubilizing bacteria (PSB) have been shown to increase the availability of soil P fractions, thereby promoting plant growth. We herein investigated the effects of PSB on P availability in two important Chinese soil types: Lateritic red earths (La) and Cinnamon soils (Ci). We initially isolated 5 PSB strains and assessed their effects on soil P fractions. PSB mainly increased moderately labile P in La and labile P in Ci. We then selected the most promising PSB isolate (99% similarity with Enterobacter chuandaensis) and examined its effects on P accumulation in maize seedlings. The results obtained showed that plant P accumulation increased in response to a PSB inoculation in both soil types and the combination of the PSB inoculation and tricalcium phosphate fertilization in La significantly enhanced P accumulation in plant shoots. The present study demonstrated that the PSB isolates tested differed in their ability to mobilize P from distinct P fertilizers and that PSB isolates have potential as a valuable means of sustainably enhancing seedling growth in Chinese agricultural soils.

Lipidome Profiling of Phosphorus Deficiency-Tolerant Rice Cultivars Reveals Remodeling of Membrane Lipids as a Mechanism of Low P Tolerance.

Plants have evolved various mechanisms for low P tolerance, one of which is changing their membrane lipid composition by remodeling phospholipids with non-phospholipids. The objective of this study was to investigate the remodeling of membrane lipids among rice cultivars under P deficiency. Rice (Oryza sativa L.) cultivars (Akamai, Kiyonishiki, Akitakomachi, Norin No. 1, Hiyadateine, Koshihikari, and Netaro) were grown in 0 (-P) and 8 (+P) mg P L-1 solution cultures. Shoots and roots were collected 5 and 10 days after transplanting (DAT) in solution culture and subjected to lipidome profiling using liquid chromatography-mass spectrometry. Phosphatidylcholine (PC)34, PC36, phosphatidylethanolamine (PE)34, PE36, phosphatidylglycerol (PG)34, phosphatidylinositol (PI)34 were the major phospholipids and digalactosyldiacylglycerol (DGDG)34, DGDG36, 1,2-diacyl-3-O-alpha-glucuronosylglycerol (GlcADG)34, GlcADG36, monogalactosyldiacylglycerol (MGDG)34, MGDG36, sulfoquinovosyldiacylglycerol (SQDG)34 and SQDG36 were the major non-phospholipids. Phospholipids were lower in the plants that were grown under -P conditions than that in the plants that were grown under +P for all cultivars at 5 and 10 DAT. The levels of non-phospholipids were higher in -P plants than that in +P plants of all cultivars at 5 and 10 DAT. Decomposition of phospholipids in roots at 5 DAT correlated with low P tolerance. These results suggest that rice cultivars remodel membrane lipids under P deficiency, and the ability of remodeling partly contributes to low P tolerance.

Multiple analysis of root exudates and microbiome in rice (Oryza sativa) under low P conditions.

Plants release various metabolites from roots and root exudates contribute to differences in stress tolerance among plant species. Plant and soil microbes have complex interactions that are affected by biotic and abiotic factors. The purpose of this study was to examine the differences in metabolites in root exudates of rice (Oryza sativa) cultivars and their correlation with bacterial populations in the rhizosphere. Two rice cultivars (O. sativa cv. Akamai and O. sativa cv. Koshihikari) were grown in soils fertilized with 0 g P kg-1 (- P) or 4.8 g P kg-1 (+ P). Root exudates and root-attached soil were collected at 13 and 20 days after transplanting (DAT) and their metabolites and bacterial community structure were determined. The exudation of proline, serine, threonine, valine and 4-coumarate were increased under low P conditions in both cultivars. There was a positive correlation between the concentration of pantothenate in root exudates and the representation of members of the genera Clostridium and Sporosarcina, which were negatively correlated with root dry weight. Gracilibacter, Opitutus, Pelotomaculum, Phenylobacterium and Oxobacter were positively correlated with root dry weight and presence of allantoin, 2-aminobtyrate and GlcNac. This study provides new information about the response of plants and rhizosphere soil bacteria to low P conditions.

Recent insights into the metabolic adaptations of phosphorus-deprived plants.

Inorganic phosphate (Pi) is an essential macronutrient required for many fundamental processes in plants, including photosynthesis and respiration, as well as nucleic acid, protein, and membrane phospholipid synthesis. The huge use of Pi-containing fertilizers in agriculture demonstrates that the soluble Pi levels of most soils are suboptimal for crop growth. This review explores recent advances concerning the understanding of adaptive metabolic processes that plants have evolved to alleviate the negative impact of nutritional Pi deficiency. Plant Pi starvation responses arise from complex signaling pathways that integrate altered gene expression with post-transcriptional and post-translational mechanisms. The resultant remodeling of the transcriptome, proteome, and metabolome enhances the efficiency of root Pi acquisition from the soil, as well as the use of assimilated Pi throughout the plant. We emphasize how the up-regulation of high-affinity Pi transporters and intra- and extracellular Pi scavenging and recycling enzymes, organic acid anion efflux, membrane remodeling, and the remarkable flexibility of plant metabolism and bioenergetics contribute to the survival of Pi-deficient plants. This research field is enabling the development of a broad range of innovative and promising strategies for engineering phosphorus-efficient crops. Such cultivars are urgently needed to reduce inputs of unsustainable and non-renewable Pi fertilizers for maximum agronomic benefit and long-term global food security and ecosystem preservation.

AtALMT3 is Involved in Malate Efflux Induced by Phosphorus Deficiency in Arabidopsis thaliana Root Hairs.

Under phosphorus (P)-deficient conditions, organic acid secretion from roots plays an important role in P mobilization from insoluble P in the soil. In this study, we characterized AtALMT3, a homolog of the Arabidopsis thaliana aluminum-activated malate transporter family gene. Among the 14 AtALMT family genes, only AtALMT3 was significantly up-regulated in P-deficient roots. AtALMT3 promoter::ß-glucuronidase is expressed in the epidermis in roots, especially in root hair cells. AtALMT3 protein was localized in the plasma membrane and in small vesicles. Fluorescence of AtALMT3::GFP was not observed on the vacuole membrane of protoplast after lysis, indicating that AtALMT3 localizes mainly in the plasma membrane. Compared with the wild-type (WT) line, malate exudation in the AtALMT3-knockdown line (atalmt3-1) and overexpression line (atalmt3-2) under P deficiency were, respectively, 37% and 126%. In contrast, no significant difference was found in citrate exudation among these lines. The complementation of the atalmt3-1 line with AtALMT3 recovered the malate exudation to the level of the WT. Taken together, these results suggest that AtALMT3 localized in root hair membranes is involved in malate efflux in response to P deficiency.

Molecular mechanisms underpinning phosphorus-use efficiency in rice.

Orthophosphate (H2 PO4- , Pi) is an essential macronutrient integral to energy metabolism as well as a component of membrane lipids, nucleic acids, including ribosomal RNA, and therefore essential for protein synthesis. The Pi concentration in the solution of most soils worldwide is usually far too low for maximum growth of crops, including rice. This has prompted the massive use of inefficient, polluting, and nonrenewable phosphorus (P) fertilizers in agriculture. We urgently need alternative and more sustainable approaches to decrease agriculture's dependence on Pi fertilizers. These include manipulating crops by (a) enhancing the ability of their roots to acquire limiting Pi from the soil (i.e. increased P-acquisition efficiency) and/or (b) increasing the total biomass/yield produced per molecule of Pi acquired from the soil (i.e. increased P-use efficiency). Improved P-use efficiency may be achieved by producing high-yielding plants with lower P concentrations or by improving the remobilization of acquired P within the plant so as to maximize growth and biomass allocation to developing organs. Membrane lipid remodelling coupled with hydrolysis of RNA and smaller P-esters in senescing organs fuels P remobilization in rice, the world's most important cereal crop.

Ancient rice cultivar extensively replaces phospholipids with non-phosphorus glycolipid under phosphorus deficiency.

Recycling of phosphorus (P) from P-containing metabolites is an adaptive strategy of plants to overcome soil P deficiency. This study was aimed at demonstrating differences in lipid remodelling between low-P-tolerant and -sensitive rice cultivars using lipidome profiling. The rice cultivars Akamai (low-P-tolerant) and Koshihikari (low-P-sensitive) were grown in a culture solution with [2 mg l-1 (+P)] or without (-P) phosphate for 21 and 28 days after transplantation. Upper and lower leaves were collected. Lipids were extracted from the leaves and their composition was analysed by liquid chromatography/mass spectrometry (LC-MS). Phospholipids, namely phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and phosphatidylinositol (PI), lysophosphatidylcholine (lysoPC), diacylglycerol (DAG), triacylglycerol (TAG) and glycolipids, namely sulfoquinovosyl diacylglycerol (SQDG), digalactosyldiacylglycerol (DGDG), monogalactosyldiacylglycerol (MGDG) and 1,2-diacyl-3-O-alpha-glucuronosyl glycerol (GlcADG), were detected. GlcADG level was higher in both cultivars grown in -P than in +P and the increase was larger in Akamai than in Koshihikari. DGDG, MGDG and SQDG levels were higher in Akamai grown in -P than in +P and the increase was larger in the upper leaves than in the lower leaves. PC, PE, PG and PI levels were lower in both cultivars grown in -P than in +P and the decrease was larger in the lower leaves than in the upper leaves and in Akamai than in Koshihikari. Akamai catabolised more phospholipids in older leaves and synthesised glycolipids in younger leaves. These results suggested that extensive phospholipid replacement with non-phosphorus glycolipids is a mechanism underlying low-P-tolerance in rice cultivars.

Identification and distribution of cellobiose 2-epimerase genes by a PCR-based metagenomic approach.

Cellobiose 2-epimerase (CE) catalyzes the reversible epimerization of cellobiose to 4-O-ß-D-glucopyranosyl-D-mannose. By using a PCR-based metagenomic approach, 71 ce-like gene fragments were obtained from wide-ranging environmental samples such as sheep rumen, soils, sugar beet extracts, and anaerobic sewage sludge. The frequency of isolation of the fragments similar to known sequences varied depending on the nature of the samples used. The ce-like genes appeared to be widely distributed in environmental bacteria belonging to the phyla Bacteroidetes, Chloroflexi, Dictyoglomi, Firmicutes, Proteobacteria, Spirochaetes, and Verrucomicrobia. The phylogenetic analysis suggested that the cluster of CE and CE-like proteins was functionally and evolutionarily separated from that of N-acetyl-D-glucosamine 2-epimerase (AGE) and AGE-like proteins. Two ce-like genes containing full-length ORFs, designated md1 and md2, were obtained by PCR and expressed in Escherichia coli. The recombinant mD1 and mD2 exhibited low K m values and high catalytic efficiencies (k cat/K m) for mannobiose compared with cellobiose, suggesting that they should be named mannobiose 2-epimerase, which is involved in a new mannan catabolic pathway we proposed.

The mannobiose-forming exo-mannanase involved in a new mannan catabolic pathway in Bacteroides fragilis.

We have proposed a new mannan catabolic pathway in Bacteroides fragilis NCTC 9343 that involves a putative mannanase ManA in glycoside hydrolase family 26 (BF0771), a mannobiose and/or sugar transporter (BF0773), mannobiose 2-epimerase (BF0774), and mannosylglucose phosphorylase (BF0772). If this hypothesis is correct, ManA has to generate mannobiose from mannans as the major end product. In this study, the BF0771 gene from the B. fragilis genome was cloned and expressed in Escherichia coli cells. The expressed protein was found to produce mannobiose exclusively from mannans and initially from manno-oligosaccharides. Production of 4-O-ß-D-glucopyranosyl-D-mannose or 4-O-ß-D-mannopyranosyl-D-glucose from mannans was not detectable. The results indicate that this enzyme is a novel mannobiose-forming exo-mannanase, consistent with the new microbial mannan catabolic pathway we proposed.

Structure of novel enzyme in mannan biodegradation process 4-O-ß-D-mannosyl-D-glucose phosphorylase MGP.

The crystal structure of a novel component of the mannan biodegradation system, 4-O-ß-D-mannosyl-D-glucose phosphorylase (MGP), was determined to a 1.68-Å resolution. The structure of the enzyme revealed a unique homohexameric structure, which was formed by using two helices attached to the N-terminus and C-terminus as a tab for sticking between subunits. The structures of MGP complexes with genuine substrates, 4-O-ß-D-mannosyl-D-glucose and phosphate, and the product D-mannose-1-phosphate were also determined. The complex structures revealed that the invariant residue Asp131, which is supposed to be the general acid/base, did not exist close to the glycosidic Glc-O4 atom, which should be protonated in the catalytic reaction. Also, no solvent molecule that might mediate a proton transfer from Asp131 was observed in the substrate complex structure, suggesting that the catalytic mechanism of MGP is different from those of known disaccharide phosphorylases.

Phytate degradation by fungi and bacteria that inhabit sawdust and coffee residue composts.

Phytate is the primary source of organic phosphorus, but it cannot be directly utilized by plants and is strongly adsorbed by the soil, reducing bioavailability. Composting is a process used to improve the bioavailability of phytate in organic wastes through degradation by microorganisms. In this study, we aimed to investigate the phytate-degrading ability of fungi and bacteria that inhabit sawdust compost and coffee residue compost, and their contribution to the composting process. In the plate assay, the fungi that formed clear zones around their colonies belonged to the genera Mucor, Penicillium, Galactomyces, Coniochaeta, Aspergillus, and Fusarium, while the bacteria belonged to the genera Pseudomonas, Enterobacter, Chitinophaga, and Rahnella. Eight fungal isolates (genera Mucor, Penicillium, Galactomyces, and Coniochaeta) and four bacterial isolates (genera Pseudomonas, Enterobacter, and Rahnella) were selected to evaluate phytase activity in their liquid culture and their ability to degrade phytate in organic materials composed of mushroom media residue and rice bran. The selected fungi degraded phytate in organic materials to varying degrees. Penicillium isolates showed the highest degradation ability and Coniochaeta isolate exhibited relatively high degradation ability. The clear zone diameters of these fungal isolates displayed significantly positive and negative correlations with inorganic and phytate phosphorus contents in the organic materials after incubation, respectively; however, none of the selected bacteria reduced phytate phosphorus in organic materials. It is therefore possible that fungi are major contributors to phytate degradation during composting.

Isolation and characterization of cellulose-decomposing bacteria inhabiting sawdust and coffee residue composts.

Clarifying the identity and enzymatic activities of microorganisms associated with the decomposition of organic materials is expected to contribute to the evaluation and improvement of composting processes. In this study, we examined the cellulolytic and hemicellulolytic abilities of bacteria isolated from sawdust compost (SDC) and coffee residue compost (CRC). Cellulolytic bacteria were isolated using Dubos mineral salt agar containing azurine cross-linked (AZCL) HE-cellulose. Bacterial identification was performed based on the sequence analysis of 16S rRNA genes, and cellulase, xylanase, ß-glucanase, mannanase, and protease activities were characterized using insoluble AZCL-linked substrates. Eleven isolates were obtained from SDC and 10 isolates from CRC. DNA analysis indicated that the isolates from SDC and CRC belonged to the genera Streptomyces, Microbispora, and Paenibacillus, and the genera Streptomyces, Microbispora, and Cohnella, respectively. Microbispora was the most dominant genus in both compost types. All isolates, with the exception of two isolates lacking mannanase activity, showed cellulase, xylanase, ß-glucanase, and mannanase activities. Based on enzyme activities expressed as the ratio of hydrolysis zone diameter to colony diameter, it was suggested that the species of Microbispora (SDCB8, SDCB9) and Paenibacillus (SDCB10, SDCB11) in SDC and Microbispora (CRCB2, CRCB6) and Cohnella (CRCB9, CRCB10) in CRC contribute to efficient cellulolytic and hemicellulolytic processes during composting.

New microbial mannan catabolic pathway that involves a novel mannosylglucose phosphorylase.

The consecutive genes BF0771-BF0774 in the genome of Bacteroides fragilis NCTC 9343 were found to constitute an operon. The functional analysis of BF0772 showed that the gene encoded a novel enzyme, mannosylglucose phosphorylase that catalyzes the reaction, 4-O-ß-d-mannopyranosyl-d-glucose+Pi→mannose-1-phosphate+glucose. Here we propose a new mannan catabolic pathway in the anaerobe, which involves 1,4-ß-mannanase (BF0771), a mannobiose and/or sugar transporter (BF0773), mannobiose 2-epimerase (BF0774), and mannosylglucose phosphorylase (BF0772), finally progressing to glycolysis. This pathway is distributed in microbes such as Bacteroides, Parabacteroides, Flavobacterium, and Cellvibrio.

Evaluation of cellulolytic and hemicellulolytic abilities of fungi isolated from coffee residue and sawdust composts.

This study focused on the evaluation of cellulolytic and hemicellulolytic fungi isolated from sawdust compost (SDC) and coffee residue compost (CRC). To identify fungal isolates, the ITS region of fungal rRNA was amplified and sequenced. To evaluate enzyme production, isolates were inoculated onto wheat bran agar plates, and enzymes were extracted and tested for cellulase, xylanase, ß-glucanase, mannanase, and protease activities using different azurine cross-linked (AZCL) substrates. In total, 18 isolates from SDC and 29 isolates from CRC were identified and evaluated. Four genera (Aspergillus, Galactomyces, Mucor, and Penicillium) and five genera (Aspergillus, Coniochaeta, Fusarium, Penicillium, and Trichoderma/Hypocrea) were dominant in SDC and CRC, respectively. Penicillium sp., Trichoderma sp., and Aspergillus sp. displayed high cellulolytic and hemicellulolytic activities, while Mucor isolates exhibited the highest ß-glucanase and mannanase activities. The enzyme analyses revealed that Penicillium, Aspergillus, and Mucor isolates significantly contributed to the degradation of SDC, whereas Penicillium, Aspergillus, and Trichoderma isolates had a dominant role in the degradation of CRC. Notably, isolates SDCF5 (P. crustosum), CRCF6 (P. verruculosum), and CRCF2 and CRCF16 (T. harzianum/H. lixii) displayed high activity regarding cellulose and hemicellulose degradation, which indicates that these species could be beneficial for the improvement of biodegradation processes involving lignocellulosic materials.

Influence of arsenic stress on synthesis and localization of low-molecular-weight thiols in Pteris vittata.

The roles of low-molecular-weight thiols (LMWTs), such as glutathione and phytochelatins, in arsenic (As) tolerance and hyperaccumulation in Pteris vittata an As-hyperaccumulator fern remain to be better understood. This study aimed to thoroughly characterize LMWT synthesis in P. vittata to understand the roles played by LMWTs in As tolerance and hyperaccumulation. LMWT synthesis in P. vittata was induced directly by As, and not by As-mediated oxidative stress. Expression of PvECS2, one of the putative genes of γ-glutamylcysteine synthetase (γECS), increases in P. vittata shoots at 48h after the onset of As exposure, almost corresponding to the increase in the concentrations of γ-glutamylcysteine and glutathione. Furthermore, localization of As showed similar trends to those of LMWTs in fronds at both whole-frond and cellular levels. This study thus indicates the specific contribution of LMWTs to As tolerance in P. vittata. γECS may be responsible for the As-induced enhancement of LMWT synthesis.

Recent progress in plant nutrition research: cross-talk between nutrients, plant physiology and soil microorganisms.

Mineral nutrients taken up from the soil become incorporated into a variety of important compounds with structural and physiological roles in plants. We summarize how plant nutrients are linked to many metabolic pathways, plant hormones and other biological processes. We also focus on nutrient uptake, describing plant-microbe interactions, plant exudates, root architecture, transporters and their applications. Plants need to survive in soils with mineral concentrations that vary widely. Describing the relationships between nutrients and biological processes will enable us to understand the molecular basis for signaling, physiological damage and responses to mineral stresses.

Identification of the cellobiose 2-epimerase gene in the genome of Bacteroides fragilis NCTC 9343.

Cellobiose 2-epimerase (CE, EC 5.1.3.11) catalyzes the reversible epimerization of cellobiose to 4-O-beta-D-glucopyranosyl-D-mannose. In this study, we found a CE gene in the genome sequence of non-cellulolytic Bacteroides fragilis NCTC 9343. The recombinant enzyme, expressed in Escherichia coli cells, catalyzed a hydroxyl stereoisomerism at the C-2 positions of the reducing terminal glucose and at the mannose moiety of cello-oligosaccharides, lactose, beta-mannobiose (4-O-beta-D-mannopyranosyl-D-mannose), and globotriose [O-alpha-D-galactopyranosyl-(1-->4)-O-beta-D-galactopyranosyl-(1-->4)-D-glucose]. The CE from B. fragilis showed less than 40% identity to reported functional CEs. It exhibited 44-63% identities to N-acyl-D-glucosamine 2-epimerase-like hypothetical proteins of unknown function in bacterial genome sequences of the phyla Firmicutes, Bacteroidetes, Proteobacteria, Chloroflexi, and Verrucomicrobia. On the other hand, it showed less than 26% identity to functional N-acyl-D-glucosamine 2-epimerases. Based on the amino acid homology and phylogenetic positions of the functional epimerases, we emphasize that many genes for putative N-acyl-D-glucosamine 2-epimerases and related hypothetical proteins of unknown function reported to date in the bacterial genomes should be annotated as CE-like proteins or putative CEs.

Effects of epilactose on calcium absorption and serum lipid metabolism in rats.

Epilactose (4-O-beta-galactopyranosyl-D-mannnose) is a rare disaccharide in cow milk that can be synthesized from lactose by the cellobiose 2-epimerase of Ruminococcus albus. In this study, we examined the biological activities of epilactose using male Wistar-ST rats. The apparent rates of calcium and magnesium absorption of rats fed epilactose and fructooligosaccharide diets were greater than those fed control and lactose diets, accompanied by greater weight gain of the cecal wall and higher levels of short-chain fatty acids and other organic acids. Epilactose also increased the calcium absorption in everted small intestinal sacs. In addition, the levels of plasma total cholesterol and nonhigh-density lipoprotein cholesterol were lower in epilactose-fed rats. These results indicate that epilactose promotes calcium absorption in the small intestine and possibly lowers the risk of arteriosclerosis. Cecal microbes may efficiently utilize epilactose and contribute to these biological activities.