Mitsuru Osaki

Secretion of acid phosphatase by the roots of crop plants under phosphorus-deficient conditions and some properties of the enzyme secreted by lupin roots

Nine crop species were grown in P-sufficient and P-deficient nutrient solutions. The activity of acid phosphatase secreted by the roots increased under P-deficient conditions in all the species examined. That of lupin increased most remarkably. The properties of the enzyme secreted by the roots of lupin was investigated. Many isozymes existed in the roots and the leaves, but only one of them was secreted into the rhizosphere in a large amount. The molecular weight of the purified enzyme secreted was estimated to be 72 KD by SDS-PAGE and 140 KD by gel filtration; it was assumed to be a homo-dimer. The iso-electric point of the enzyme was 4.7 and the pH for optimum activity 4.3. When the enzyme was mixed with aqueous solution extracted from a P-deficient soil, its activity declined to 55% of its original activity after 14 days and to 9% after 21 days.

Structural principles of leucine-rich repeat (LRR) proteins

LRR‐containing proteins are present in over 2000 proteins from viruses to eukaryotes. Most LRRs are 20–30 amino acids long, and the repeat number ranges from 2 to 42. The known structures of 14 LRR proteins, each containing 4–17 repeats, have revealed that the LRR domains fold into a horseshoe (or arc) shape with a parallel β‐sheet on the concave face and with various secondary structures, including α‐helix, 310‐helix, and pII helix on the convex face. We developed simple methods to charactere quantitatively the arc shape of LRR and then applied them to all known LRR proteins. A quantity of 2Rsin(φ/2), in which R and φ are the radii of the LRR arc and the rotation angle about the central axis per repeating unit, respectively, is highly conserved in all the LRR proteins regardless of a large variety of repeat number and the radius of the LRR arc. The radii of the LRR arc with β‐α structural units are smaller than those with β‐310 or β‐pII units. The concave face of the LRR β‐sheet forms a surface analogous to a part of a Möbius strip. Proteins 2004;54:000–000.

Secretion of phytase from the roots of several plant species under phosphorus-deficient conditions

Phosphorus (P) deficiency increased the secretion of phytases from roots of various plant species. The secretory phytases were collected with a dialysis membrane tube for 24 hours from roots of sixteen plant species grown with low or adequate supply of P in nutrient solutions. The activity of not only secretory phytase, but also acid phosphatase, increased with the low P treatment in all of the plant species examined. Secretion of phytase by the roots under P-deficient conditions was highest in Brachiaria decumbens CIAT 606, Stylosanthes guianensis CIAT 184 and tomato, moderate in Brachiaria brizantha CIAT6780, Stylosanthes guianensis CIAT 2950, alfalfa, white clover and orchard grass, and lowest in Andropgon gayanus CIAT 621, Stylosanthes capitata CIAT 10280, upland rice, timothy, redtop, alsike clover, red clover and white lupin plants. An immunoreactive protein band that reacted with a polyclonal antibody raised against wheat bran phytase, corresponding to molecular weight 35–40 kD, could be detected in seven of the species tested. These results indicate that the secretory phytase may provide an efficient mechanism for certain plants to utilize inositol hexaphosphate in soil.

Beneficial effect of aluminum on growth of plants adapted to low pH soils

Abstract Plants in which growth was reduced by low and high Al applications were designated as Al-sensitive plant (Hordeum vulgare) and Al-medium tolerant plants (Leucaena leucocephala, Ischaemum barbatum, Stylosanthes guianensis, and Fagopyrum esculentum), respectively, while plants in which growth was not affected or was stimulated by Al application were designated as Al-tolerant plant (Brachiaria ruziziensis) and Al-stimulated plants (Melastoma malabathricum, Melaleuca cajuputi, Acacia mangium, Hydrangea macrophyila, Vaccinium macrocarpon, Polygonum sachalinense, and Oryza sativa), respectively. Plants tolerant to or stimulated by Al were further classified based on the criteria of Al accumulation: 1) Al-excluders such as M. cajuputi, A. mangium, L. leucocephala, I. barbatum, S. guianensis, and O. sativa, 2) Al root-accumulators such as V. macrocarpon, B. ruziziensis, and P. sachalinense, and 3) Al-accumulators such as M. malabathricum, H. macrophylla, and F. esculentum. The growth and N, P, and K upta...

Distribution and chemical speciation of aluminum in the Al accumulator plant, Melastoma malabathricum L.

The Al accumulation mechanisms in an Al accumulator plant, Melastoma malabathricum L. (Melastoma), was investigated. Al was located in the upper epidermal cells and also distributed in mesophyll cells in leaf sections. In root sections, Al was found in all the root tissues, particularly in the epidermis and endodermis. Al concentrations in young leaves, mature leaves, old leaves, and roots were 8.0, 9.2, 14.4, and 10.1 mg g1, respectively. Approximately 45% of total Al in oldest leaves, and approximately 60% of total Al in leaves of other positions and roots were extracted in Tris-HCl buffer (pH 7.0). Since Al in the residual parts was mostly dissolved in hot 0.5 M H2SO4 containing 2% cetyl trimethylammonium bromide, residual Al seemed to consist mainly of monomeric Al and Al bound to pectic substances and hemicellulose. Al in the Tris-HCl extract consisted of non-monomeric Al (complexed form). Oxalate concentration in the Tris-HCl extract in leaves was significantly higher in the +Al treatment than in the –Al treatment and there was a positive correlation between the Al concentration and oxalate concentration. 27Al NMR spectrum of fresh leaves indicated the presence in the order of monomeric Al, Al-oxalate, Al-(oxalate)2, and Al-(oxalate)3 in intact leaves.

Secreted acid phosphatase is expressed in cluster roots of lupin in response to phosphorus deficiency

The roots of white lupin (Lupinus albus L. cv. Kievskij mutant) secrete acid phosphatase, S-APase, when they grow under conditions of low available phosphorus (P). S-APases hydrolyze organic phosphate compounds in the rhizosphere and supply inorganic phosphate to the plants. Low phosphorus availability also induces vigorous growth of cluster roots. In this study, the function of cluster roots was investigated with reference to S-APase secretion. White lupins were grown in hydroponic culture in a greenhouse under P-deficient and P-sufficient conditions. S-APase in the excised roots after treatment was detected by staining with 4-methylumbelliferone phosphate (MUP). Gene expression of S-APase in cluster and normal roots was also investigated. Activity was greatest in the roots of plants grown under conditions of P -deficiency, particularly in cluster roots. S-APase gene expression was induced by a decrease in internal P concentrations, and was especially high in cluster roots formed under conditions of P -deficiency. It was suggested that decrease of internal P concentration stimulated both of the S-APase expression and cluster root formation.

Redistribution of carbon and nitrogen compounds from the shoot to the harvesting organs during maturation in field crops

Abstract The present paper alms at determining how carbon (C) and nitrogen (N) compounds are redistributed from leaves to harvesting organs during maturation in several major field crops. In order to illustrate these processes in the case of C and N compounds, 6 major crops in Hokkaido were grown and compared during maturation. The results obtained were as follows. 1) The N-redistribution rate during maturation was in the order of wheat, soybean and potato > maize > rice. The percentage of distributed nitrogen among the different nitrogenous fractions in leaves and stems was remained constant during maturation, suggesting that each nitrogen compound was equally decomposed. 2) The composition of protein amino acids was similar regardless of organs or crops. The composition of free amino acids differed widely among organs and crops, and differed from that of protein amino acids. Therefore during the translocation of amino acids from leaves to harvesting organs through stems, the composition of free amino ac...

Mechanisms of adaptation to high aluminum condition in native plant species growing in acid soils: a review

Various studies of the mechanisms of aluminum (Al) tolerance in crop plants have been conducted to improve crop productivity in acid soils. However, many native plant species vigorously grow in acid soils. These species have adapted well to high levels of Al in the medium. The mechanisms of this adaptation can be separated into Al exclusion and internal Al inactivation. In general, plant species that have developed mechanisms of the former type are called “Al excluders,” and those that have developed mechanisms of the latter type are called “Al accumulators.” Mechanisms of Al exclusion in excluder species have not been completely elucidated, and may involve some strong mechanisms in addition to the exudation of organic acid from roots. Al accumulator species are supposed to create an Al-ligand (mainly organic acids) complex for translocation from roots to shoots, and for accumulation in the leaves. Interestingly, the ligand of Al for translocation often differs from that for storage in the leaves. In addition, Al accumulator species seem to prevent the toxic effect of Al on the metabolism in leaves by means of isolating the Al in sites that are insensitive to Al (e.g., epidermal cells or vacuoles). Growth in some native plant species that have adapted to acid soils is enhanced by Al application. This Al-induced growth enhancement is considered to be caused by Al-induced stimulation of nutrient uptake in Al excluders (non-accumulators), and by physiological effects of Al itself in addition to the stimulation of nutrient uptake in accumulators. Elucidation of these mechanisms of adaptation to high levels of Al in medium will be useful for improved production of Al-tolerant crop cultivars and rehabilitation of forest areas.

Sequencing and analysis of approximately 40,000 soybean cDNA clones from a full-length-enriched cDNA library.

A large collection of full-length cDNAs is essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. We obtained a total of 39 936 soybean cDNA clones (GMFL01 and GMFL02 clone sets) in a full-length-enriched cDNA library which was constructed from soybean plants that were grown under various developmental and environmental conditions. Sequencing from 5′ and 3′ ends of the clones generated 68 661 expressed sequence tags (ESTs). The EST sequences were clustered into 22 674 scaffolds involving 2580 full-length sequences. In addition, we sequenced 4712 full-length cDNAs. After removing overlaps, we obtained 6570 new full-length sequences of soybean cDNAs so far. Our data indicated that 87.7% of the soybean cDNA clones contain complete coding sequences in addition to 5′- and 3′-untranslated regions. All of the obtained data confirmed that our collection of soybean full-length cDNAs covers a wide variety of genes. Comparative analysis between the derived sequences from soybean and Arabidopsis, rice or other legumes data revealed that some specific genes were involved in our collection and a large part of them could be annotated to unknown functions. A large set of soybean full-length cDNA clones reported in this study will serve as a useful resource for gene discovery from soybean and will also aid a precise annotation of the soybean genome.

Structural analysis of leucine-rich-repeat variants in proteins associated with human diseases

Abstract.A number of human diseases have been shown to be associated with mutation in the genes encoding leucine-rich-repeat (LRR)-containing proteins. They include 16 different LRR proteins. Mutations of these proteins are associated with 19 human diseases. The mutations occur frequently within the LRR domains as well as their neighboring domains, including cysteine clusters. Here, based on the sequence analysis of the LRR domains and the known structure of LRR proteins, we describe some features of different sequence variants and discuss their adverse effects. The mutations in the cysteine clusters, which preclude the formation of sulfide bridges or lead to a wrong paring of cysteines in extracellular proteins or extracellular domains, occur with high frequency. In contrast, missense mutations at some specific positions in LRRs are very rare or are not observed at all.