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PubMed2005

Population genetic differentiation in taxa of Lotus (Fabaceae: Loteae) endemic to the Gran Canarian pine forest.

A survey of allozyme variation at 17 loci in 14 populations representing four taxonomically problematic Gran Canarian pine forest endemic taxa of Lotus (L. genistoides, L. holosericeus, L. spartioides and some taxonomically uncertain populations collected under the designation Lotus sp.) was conducted to examine their diversification and systematic relationships. All groups exhibited high values of genetic variation, although inbreeding was common within populations. Considerable among-population genetic homogeneity was detected, as inferred from low values of Gst within each of the groups. The high population sizes of these taxa and a lack of evidence for isolation by distance or genetic bottlenecks indicate that diversity has accumulated over a long period of environmental stability. The association of high genetic distances with low linear distances, and the substantial increase in the values of Gst when the taxa considered were merged in different combinations hint at an incipient (yet probably taxonomically insufficient) reproductive isolation. The genetic similarity between L. genistoides, L. holosericeus and L. spartioides, together with the different behaviour of the populations collected under the designation Lotus sp., may have important implications for the restructuring of the taxonomy of this group when the ongoing morphological studies are completed.

Atlantic Islands↗

PubMed2006

Genetics of symbiosis in Lotus japonicus: recombinant inbred lines, comparative genetic maps, and map position of 35 symbiotic loci.

Development of molecular tools for the analysis of the plant genetic contribution to rhizobial and mycorrhizal symbiosis has provided major advances in our understanding of plant-microbe interactions, and several key symbiotic genes have been identified and characterized. In order to increase the efficiency of genetic analysis in the model legume Lotus japonicus, we present here a selection of improved genetic tools. The two genetic linkage maps previously developed from an interspecific cross between L. japonicus Gifu and L. filicaulis, and an intraspecific cross between the two ecotypes L. japonicus Gifu and L. japonicus MG-20, were aligned through a set of anchor markers. Regions of linkage groups, where genetic resolution is obtained preferentially using one or the other parental combination, are highlighted. Additional genetic resolution and stabilized mapping populations were obtained in recombinant inbred lines derived by a single seed descent from the two populations. For faster mapping of new loci, a selection of reliable markers spread over the chromosome arms provides a common framework for more efficient identification of new alleles and new symbiotic loci among uncharacterized mutant lines. Combining resources from the Lotus community, map positions of a large collection of symbiotic loci are provided together with alleles and closely linked molecular markers. Altogether, this establishes a common genetic resource for Lotus spp. A web-based version will enable this resource to be curated and updated regularly.

Alleles↗

PubMed2007

Phytochelatin synthases of the model legume Lotus japonicus. A small multigene family with differential response to cadmium and alternatively spliced variants.

The biosynthesis of phytochelatins and homophytochelatins has been studied in nodulated plants of the model legume Lotus (Lotus japonicus). In the first 6 to 24 h of treatment with cadmium (Cd), roots started to synthesize elevated amounts of both polypeptides, with a concomitant increase of glutathione and a decrease of homoglutathione, indicating the presence of active phytochelatin synthase (PCS) genes. Screening of transformation-competent artificial chromosome libraries allowed identification of a cluster of three genes, LjPCS1, LjPCS2, and LjPCS3, which were mapped at 69.0 cM on chromosome 1. The genes differ in exon-intron composition and responsiveness to Cd. Gene structures and phylogenetic analysis of the three protein products, LjPCS1-8R, LjPCS2-7N, and LjPCS3-7N, are consistent with two sequential gene duplication events during evolution of vascular plants. Two sites for alternative splicing in the primary transcripts were identified. One of them, involving intron 2 of the LjPCS2 gene, was confirmed by the finding of the two predicted mRNAs, encoding LjPCS2-7R in roots and LjPCS2-7N in nodules. The amino acid sequences of LjPCS2-7R (or LjPCS2-7N) and LjPCS3-7N share 90% identity, but have only 43% to 59% identity with respect to the typical PCS1 enzymes of Lotus and other plants. The unusual LjPCS2-7N and LjPCS3-7N proteins conferred Cd tolerance when expressed in yeast (Saccharomyces cerevisiae) cells, whereas the alternatively spliced form, LjPCS2-7R, differing only in a five-amino acid motif (GRKWK) did not. These results unveil complex regulatory mechanisms of PCS expression in legume tissues in response to heavy metals and probably to other developmental and environmental factors.

Alternative Splicing↗

PubMed2004

[The construction and screening of genomic library based on transformable artificial chromosome (TAC) vector in Lotus japonicus].

Many vectors, especially artificial chromosome vectors, have been developed for genome-scale mapping and sequencing. As the first artificial chromosome, YAC has been extensively used in the genomic library construction and mapping. Shizuya et al. (1992) devised the bacterial artificial chromosome (BAC) originated from F factor of E.coli, which is much easy to handle and isolated with large harboring capacity. Liu et al. (1999) designed the transformation-competent artificial chromosome (TAC) vector, derived from PAC vector. TAC could shuttle a large-scale DNA fragment between bacteria and Agrobaceria. Further, it could integrate a large targetted DNA fragment into plant genome, as being documented in Arabidopsis and rice genome research (Liu et al. 2000, 2002). The time-saving virtue of TAC should be significant in genomics research in Lotus japonicus, a model plant of legume. Using a nuclei-based method of Liu and Whittier, high molecular weight DNA was isolated from Lotus japonicus (Gifu ecotype). The DNA was digested partially with Hind III and size-fractionated in the 10- to 20-kb size range as described (Liu and Whittier 1994). The partially digested and size selected DNA fragments were ligated with Hind III-digested pYLTAC7 and then used for transformation of E. coli DH10B by electroporation. Transformants carrying inserts were selected on LB agar plates containing 25 mg/L kanamycin and 5% sucrose. The library, 6 haploid genome equivalents, was pooled in 12 96-well microtiter plates at about 150 transformants per well. The TAC library was then arrayed in nylon membranes and subjected to screening. The probe, a homolog fragment of CEN gene controlling the structure of inflorescence Antirrhinum, was used for screening. 0.5 muL solution from the positive pool was titered and the secondary screening was conducted in the same way. Finally, these positive clonies were confirmed by Southern blot. These data showed that this genomic library was reliable for further molecular research in Lotus japonicus.

Chromosomes, Artificial, Bacterial↗

PubMed1990

Use of a Lotus 1-2-3 spreadsheet to plot the confidence and the prediction intervals in linear regression analysis.

Computer-derived spreadsheet programs such as Lotus 1-2-3 are being used by many investigators for individualized statistical calculations. Spreadsheet templates have been described for the performance of simple statistical tests by non-statisticians with relatively limited computer experience. This paper describes a template for use with Lotus 1-2-3 for automatic graphic display of confidence and prediction intervals of simple linear regression analysis. The graph can be printed using the Lotus printgraph program. The program also allows comparison of the slope and the intercept of two regression lines.

Computer Graphics↗

PubMed1987

Indocyanine green pharmacokinetics calculations using Lotus 123 spreadsheet.

Indocyanine green (ICG; Cardio Green, Hynson, Westcott & Dunning, Baltimore, MD) is a cholephilic dye that is commonly used as a liver function test. In order to obtain the pharmacokinetic values of ICG needed for liver function determinations, a linear and a log-linear regression must be done in addition to other calculations. Results can be obtained graphically, but are not as accurate as mathematical calculations. To obtain accurate results and expedite calculation time, a program was developed using the Lotus 123 spreadsheet program (Lotus 123, Lotus Development Corporation, Cambridge, MA) and an IBM compatible computer to do these calculations.

Animals↗

PubMed1982

Distribution of fucosyl-conjugates in rat cerebellar cells in vitro: binding patterns of Lotus tetragonolobus and Ulex europeus lectins.

Light microscope radioautographic procedures were employed to investigate the binding patterns of lectins derived from Lotus tetragonolobus (Lotus A) and Ulex europeus (UEA I) to the surfaces of rat cerebellar cells maintained in dispersed cell culture. Lotus A bound extensively to the surfaces of neuronal cell somata and neuronal processes at all time points investigated; the lectin also bound to nonneuronal cell surfaces, but to an extent much less than that for neurons. Lectin binding was completely inhibited by the presence of 0.1 M alpha-L-fucose in the reaction medium. Conversely, UEA I did not appear to bind to any significant extent to the surfaces of cerebellar cells in vitro at any stage of maintenance. The incorporation of [3H]fucose into fucosyl-conjugates by cerebellar neurons in culture was investigated using light microscope radioautography and polyacrylamide gel electrophoretic methods. Radioautographs indicated that the fucose came to be distributed throughout neuronal cell somata and processes and that, with increasing time, the grain density appeared to be highest over areas of neuropil and fascicles of cellular processes. The electrophoresis data demonstrated that, among others, the label was incorporated into prominent classes of polypeptides having nominal molecular weights of 400,000, 125,000, 91,000 and 46,000 daltons.

Animals↗

PubMed1977

A simple procedure for the isolation of L-fucose-binding lectins from Ulex europaeus and Lotus tetragonolobus.

L-Fucose-binding lectins from Ulex europeaus and Lotus tetragonolobus were isolated by affinity chromatography on columns of L-fucose-Sepharose 6B. L-Fucose was coupled to Sepharose 6B after divinyl sulfone-activation of the gel to give an affinity adsorbent capable of binding more than 1.2 mg of Ulex lextin/ml of gel, which could then be eluted with 0.1M or 0.05M L-fucose. Analysis of the isolated lectins by hemagglutination assay, by gel filtration, and polyacrylamide disc-electrophoresis revealed the presence of isolectins, or aggregated species, or both. The apparent mol. wt. of the major lectin fraction from Lotus was 35000 when determined on Sephadex G-200 or Ultrogel AcA 34. In contrast, the apparent mol. wt. of the major lectin fraction from Ulex was 68 000 when chromatographed on Sephadex G-200 and 45 000 when chromatographed on Ultrogel AcA 34. The yields of lectins were 4.5 mg/100 g of Ulex seeds and 394 mg/100 g of Lotus seeds.

ABO Blood-Group System↗

PubMed2004

The Lotus effect: superhydrophobicity and metastability.

To learn how to mimic the Lotus effect, superhydrophobicity of a model system that resembles the Lotus leaf is theoretically discussed. Superhydrophobicity is defined by two criteria: a very high water contact angle and a very low roll-off angle. Since it is very difficult to calculate the latter for rough surfaces, it is proposed here to use the criterion of a very low wet (solid-liquid) contact area as a simple, approximate substitute for the roll-off angle criterion. It is concluded that nature employs metastable states in the heterogeneous wetting regime as the key to superhydrophobicity on Lotus leaves. This strategy results in two advantages: (a) it avoids the need for high steepness protrusions that may be sensitive to breakage and (b) it lowers the sensitivity of the superhydrophobic states to the protrusion distance.

Letter↗

PubMed1986

The role of condensed tannins in the nutritional value of Lotus pedunculatus for sheep. 5. Effects on the endocrine system and on adipose tissue metabolism.

1. Three experiments were conducted using Lotus pedunculatus containing high concentrations of condensed tannins (CT), and utilizing the principle that polyethylene glycol (PEG) application (molecular weight 3350) will irreversibly bind a portion of the CT and thus reduce the dietary reactive (i.e. non-PEG bound) CT concentration. Lotus diets containing 95, 45 and 14 g total reactive CT/kg dry matter (DM), induced by spraying with three PEG rates, were given to sheep at hourly intervals (600 g DM/d) for 21 d (Expt 1). In Expts 2 and 3, lambs grazed areas oversown with either lotus (89 g CT/kg DM) or clovers (Trifolium repens and Trifolium pratense; less than 1 g CT/kg DM) for 42 and 92 d respectively. In Expt 2 half the animals grazing each forage received oral PEG (75 g/d), whilst in Expt 3 half the lambs were sired by rams selected respectively for low or high levels of subcutaneous fat deposition. 2. Hormone concentrations in plasma (Expt 1 only) were determined by radioimmunoassay. Rates of [U-14C]-acetate and D-[U-14C]glucose incorporation and oxidation by subcutaneous and abdominal adipose tissue removed at slaughter, together with rate of glycerol release, were determined during in vitro incubation in all three experiments. 3. Plasma concentration of growth hormone was positively and linearly related to dietary reactive CT concentration, whilst 3,5,3'-triiodothyronine (T3) concentration tended to be negatively and linearly related to dietary reactive CT concentration. Diet CT concentration had no effect on plasma concentrations of the other hormones measured.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

PubMed1987

The effect of condensed tannins on the site of digestion of amino acids and other nutrients in sheep fed on Lotus corniculatus L.

Sheep were used to evaluate the nutritional consequences of a low condensed-tannin concentration (22 g/kg dry matter (DM)) in lotus (Lotus corniculatus L.) (control group) compared with lotus given to sheep receiving intraruminal polyethylene glycol (PEG) infusion (PEG group). PEG selectively binds to tannins and prevents tannins from binding proteins. DM intakes (1430 (SE 28) g/d) and digestibility of energy (663 (SE 4.5) kJ/MJ intake) were similar for both groups but the apparent digestion of nitrogen was lower in the control sheep (0.70) than in the PEG sheep (0.78; P less than 0.001). The proportion of N apparently digested before the abomasum (i.e. in the rumen) was lower (P less than 0.05) in control sheep (0.12) than in PEG sheep (0.21; P less than 0.05). Rumen ammonia concentrations were lower (P less than 0.001) in control sheep than in PEG sheep. The proportion of neutral-detergent fibre (NDF) digested in the rumen was similar for both groups (0.48 (SE 0.012)) but less energy was digested in the rumen of the control (0.42) than of the PEG sheep (0.47; P less than 0.05). The flux of essential amino acids (EAA) through the abomasum of control sheep was 50% greater than that in PEG sheep; flux of non-essential amino acids (NEAA) was 14% higher in control than in PEG sheep. Apparent digestibility of EAA in the small intestine was similar for both treatments (0.67), but NEAA were less well digested in the control (0.55) than in the PEG sheep (0.69). The presence of tannins in the control group increased net apparent absorption of threonine (57%), valine (89%), isoleucine (94%), leucine (30%), tyrosine (41%), phenylalanine (93%), histidine (90%) and lysine (59%), and reduced NEAA absorption by 10%, compared with PEG sheep.

Abomasum↗

PubMed1994

The effect of condensed tannins in Lotus corniculatus on plasma metabolism of methionine, cystine and inorganic sulphate by sheep.

Fresh Lotus corniculatus containing 27 g extractable condensed tannin (CT)/kg dry matter (DM) and 8 g bound CT/kg DM was fed at hourly intervals to sheep held in metabolism cages to study the effects of CT on nutrient digestion and on metabolism of methionine, cystine and inorganic sulphate in plasma. Polyethylene glycol (PEG) was continuously infused into the rumen of half the sheep to remove the effects of CT. Principal measurements in the two groups were plasma irreversible loss (IRL) rate and interconversions of methionine, cystine and inorganic sulphate using 35S labelling. CT in Lotus corniculatus had no effects on the apparent digestion of cellulose and minerals, slightly depressed DM, organic matter and hemicellulose digestion and markedly reduced the apparent digestion of N (P < 0.01). The concentration of NH3 and molar proportions of iso-butyric acid, iso-valeric acid and n-valeric acid in rumen fluid were markedly increased by the PEG infusion (P < 0.01), whereas total volatile fatty acid concentration and molar proportions of acetic acid, propionic acid and n-butyric acid were not affected. PEG infusion temporarily increased rumen protozoa numbers. CT greatly increased the IRL of plasma cystine (13.1 v. 7.0 mumol/min; P < 0.05) and reduced IRL of plasma inorganic sulphate (36.8 v. 48.1 mumol/min; P < 0.01) but had no effect on methionine IRL. CT increased transulphuration of methionine to cystine (4.37 v. 1.24 mumol/min; P < 0.05), increased cystine entering the plasma from whole-body protein turnover plus absorption from the small intestine (9.34 v. 5.75 mumol/min; P < 0.05) and increased cystine flux to body synthetic reactions (11.89 v. 5.41 mumol/min; P < 0.05). CT had no effect on the proportion of methionine total flux transferred to sulphate (0.05 v. 0.06; P < 0.05), reduced the proportion of methionine flux transferred to body synthetic reactions (0.68 v. 0.86) and markedly reduced the proportion of cystine flux transferred to sulphate (0.09 v. 0.27; P < 0.01). It was concluded that CT in Lotus corniculatus reduced rumen protein degradation and markedly increased utilization of plasma cystine for body synthetic reactions.

Animal Feed↗

PubMed1997

Cell-specific expression of the promoters of two nonlegume hemoglobin genes in a transgenic legume, Lotus corniculatus.

The promoters of the hemoglobin genes from the nitrogen-fixing tree Parasponia andersonii and the related nonnitrogen-fixing Trema tomentosa both confer beta-glucuronidase reporter gene expression to the central zone of the nodules of a transgenic legume, Lotus corniculatus. beta-Glucuronidase expression was high in the uninfected interstitial cells and parenchyma of the surrounding boundary layer and was low in the Rhizobium-infected cells. This contrasts with the expression of both the P. andersonii hemoglobin protein in P. andersonii nodules and the endogenous Lotus leghemoglobins that are expressed in the infected cells at very high levels. The expression pattern of the P. andersonii and T. tomentosa hemoglobin promoters in L. corniculatus resembles that of a nonsymbiotic hemoglobin gene from Casuarina glauca, which was introduced into this legume, and suggests that only the nonsymbiotic functions of the P. andersonii promoter are being recognized. Deletion of the distal segments of both the P. andersonii and T. tomentosa promoters identified regions important for the control of their tissue-specific and temporal activity in Lotus. Potential regulatory elements, which enhance nodule expression and suppress nonnodule expression, were also identified and localized to a distal promoter segment. A proximal AAGAG motif is present in the P. andersonii, T. tomentosa, and nonsymbiotic Casuarina hemoglobin genes. Mutation of this motif in the P. andersonii promoter resulted in a significant reduction in both the nodule and root expression levels in L. corniculatus. Some of the regulatory motifs characterized are similar to, but different from, the nodulin motifs of the leghemoglobins.

Base Sequence↗

PubMed2004

Variation in nitrate uptake and assimilation between two ecotypes of Lotus japonicus and their recombinant inbred lines.

A large genetic variability was observed in the shoot NO(3) (-) content of recombinant inbred lines (RILs) of Lotus japonicus. To determine the cause of this variability, we have studied some aspects of nitrate uptake and assimilation in the two parental ecotypes (Gifu and Funakura) and four representatives of the RILs population differing both in their shoot biomass and shoot NO(3) (-) content. Higher shoot NO(3) (-)content was mainly due to an increase in the uptake of the ion regardless of the plant biomass production. The positive correlation observed between the shoot NO(3) (-) content and the steady state level of mRNA encoding high affinity NO(3) (-) transporters suggests that the higher NO(3) (-) influx is due to enhanced expression of the transporters. In contrast, neither the level of nitrate reductase mRNA, nor the potential enzyme activity in vivo in the different lines was correlated with the shoot NO(3) (-) content. This indicates that NO(3) (-) transport in Lotus is one of the main checkpoints controlling shoot NO(3) (-) accumulation. In addition, this study shows that at least in Lotus, it is possible, through breeding strategies, to lower the NO(3) (-) content without affecting biomass production.

Journal Article↗

PubMed1991

EELS data acquisition, processing and display for the Zeiss CEM 902 based on LOTUS 1-2-3: application examples from a biological system and inorganic transition metal compounds.

A personal computer combined with LOTUS 1-2-3 software, including the RS232 module of LOTUS MEASURE and a 12-bit ADC, has been used for data acquisition of electron energy-loss spectroscopy (EELS) spectra with the Zeiss CEM 902. The internal macro language of LOTUS 1-2-3 allows a menu-driven procedure. Macro-programs partly combined with external FORTRAN programs can be chosen from the menu for background subtraction, removal of multiple scattering effects by deconvolution, elemental quantification and several utilities. For special applications or conditions the macro programs can easily be modified. Spectra from crystals of two inorganic transition metal compounds, ruthenium trichloride and vanadium disulphide, and from a biological sample are presented as examples of the application of this software.

Animals↗

PubMed2026

Unraveling epigenetic and genetic variations in response to cold stress in two lotus ecotypes.

Genetic variations accumulate over long evolutionary timescales, whereas epigenetic modifications can arise rapidly and be inherited across generations. However, the interplay between genetic and epigenetic variations in shaping ecotype-specific phenotypic plasticity remains elusive. Focusing on two lotus ecotypes that evolved under distinct winter temperature regions and display divergent annual growth cycles under cold stress, we generated DNA methylation landscapes across three sequence contexts (CG, CHG, and CHH, where H = A, T, or C) and identified single-cytosine methylation polymorphisms (SMPs) and single-nucleotide polymorphisms (SNPs). Interestingly, only CG methylation patterns mirror population-level genetic variations. Using epigenetic genome-wide association analysis, we identified differentially methylated CG sites that are either cis- or trans-regulated by SNP. Notably, we constructed a multifactorial regulatory network centered on the NnMKK4-NnCYCD5 module, linking cold response with cell cycle regulation. Temperature stress experiments conducted on lotus ecotypes and transgenic Arabidopsis (OE-NnMKK4 and OE-NnCYCD5) confirmed that NnMKK4 acts as a cold receptor and that NnCYCD5 promotes cell cycle progression and growth under cold conditions. Collectively, our findings provide novel insights into the co-evolutionary dynamics of epigenetic and genetic variations that are associated with different growth cycles of lotus ecotypes in response to cold stress.

DNA methylation↗

PubMed2025

Cooperative contribution of multiple energy substrate pathways to floral thermogenesis in sacred lotus.

Floral thermogenesis in lotus (Nelumbo nucifera) is a highly energy-intensive process, requiring substantial metabolic reconfiguration and substrate input. However, the mechanisms coordinating energy substrate supply during this process remain unclear. Here, we integrated microscale proteomics, time-series transcriptomics, and mitochondrial feeding assays to elucidate the substrate provisioning strategies supporting thermogenesis in lotus receptacles. Proteomic analysis revealed a concerted upregulation of major energy metabolism pathways at the thermogenic initiation stage, accompanied by enhanced expression of energy dissipation-related proteins (alternative oxidase and uncoupling proteins), indicative of a metabolic shift favoring heat production over ATP synthesis. Our results highlight the cooperative contribution of multiple pyruvate sources to mitochondrial respiration. Both the mitochondrial pyruvate carrier (MPC)-mediated cytosolic pyruvate import and the NAD-dependent malic enzyme (NAD-ME)-derived intramitochondrial pyruvate flux were significantly elevated at the thermogenic stage. Notably, isotopic feeding experiments revealed that NAD-ME-derived pyruvate may contribute more substantially than MPC-derived pyruvate under thermogenic conditions, reflecting a highly flexible substrate utilization strategy. In addition, increased expression of alanine aminotransferase (AlaAT) and β-oxidation-related genes suggested that alanine transamination and fatty acid degradation may further expand the respiratory substrate pool. Collectively, this study uncovers a diverse and dynamic landscape of energy substrate supply that underpins heat production in thermogenic lotus tissues. These findings offer insights into how plants coordinate metabolic flexibility to meet the high energetic demands of floral thermogenesis.

Flowers↗

PubMed2006

Denitrification ability of rhizobial strains isolated from Lotus sp.

Ten rhizobial strains isolated from Lotus sp. have been characterized by their ability to denitrify. Out of the 10 strains, the five slow-growing isolates grew well under oxygen-limiting conditions with nitrate as a sole nitrogen source, and accumulated nitrous oxide in the growth medium when acetylene was used to inhibit nitrous oxide reductase activity. All five strains contained DNA homologous to the Bradyrhizobium japonicum nirK, norBDQ and nosZ genes. In contrast, fast-growing lotus rhizobia were incapable of growing under nitrate-respiring conditions, and did not accumulate nitrous oxide in the growth medium. DNA from each of the five fast-growing strains showed a hybridization band with the B. japonicum nirK gene but not with norBDQ and nosZ genes. Partial 16S rDNA gene sequencing revealed that fast-growing strains could be identified as Mesorhizobium loti species and the slow-growers as Bradyrhizobium sp.

Alphaproteobacteria↗