Comprehensive Analysis of Plant Gene Expression in Soybean Root Nodules at Different Growth Stages

Gene expression profiles in early (12 d after sowing, DAS), mature (35 DAS) and senescent (77 DAS) soybean ( Glycine max (L.) Merr. cv. Enrei) root nodules were analyzed using Lotus japonicus cDNA macroarray. The cDNA macroarray membranes contained 18,144 independent cDNAs of L. japonicus isolated from various organs. The membranes were hybridized to ³³P-labeled single-strand cDNAs transcribed from mRNA extracted from the soybean root nodules at three different growth stages. The results showed that over 75% of the cDNA clones gave a relative signal intensity (RSI) lower than 300, irrespective of the soybean growth stage. cDNA with an RSI value over 5,000 accounted for 2.1, 1.2 and 0.5% of the total cDNA clones at 12, 35 and 77 DAS, respectively. The highest RSI value was detected in a clone hybridized to putative pectinesterase (MWM097c10_r) at 12 DAS, while the RSI value of lycopene β-cyclase (MWL025d06_r) was the highest at 35 and 77 DAS. At 77 DAS, the percentage of transferase gene cDNA clones accounted for 20.5% of the total cDNA clones showing an RSI value over 500, a value clearly lower than that at 12 and 35 DAS. On the other hand, the percentage of hydrolase gene cDNA clones accounted for 45.2% at 77 DAS, a value around 1.5 times higher than that at 12 and 35 DAS. The chronological RSI profiles of individual cDNA clones were classified into nine patterns, and the profiles of the cDNA clones belonging to 15 homologous gene groups encoding enzymes of nitrogen and carbon metabolism, nodule specific proteins, enzymes of RNA and protein hydrolysis, and synthesis of jasmonic acid precursor were compared within and between groups. As a result, significant variations both within- and between-groups were revealed.     

Structural unit of xylans from sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) and hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>)

Arabinoglucuronoxylans (AGXs) isolated from the holocellulose of sugi (Cryptomeria japonica) and hinoki (Chamaecyparis obtusa) contained one 4-O-methyl-d-glucopyranosyluronic acid (4-O-Me-d-GlcAp) residue per 6.2 d-xylopyranose (d-Xylp) residues and one 4-O-Me-d-GlcAp residue per 3.8 d-Xylp residues. These AGXs were subjected to partial acid hydrolysis. Analyses by size exclusion chromatography and electrospray-ionization mass spectroscopy of the neutral sugar fractions in the hydrolysates showed the presence of xylooligosaccharides having a degree of polymerization of 2-8 in addition to d-Xyl, suggesting that the AGXs from sugi and hinoki contained unsubstituted chains consisting of at least eight d-Xyl residues. The acidic sugars in the hydrolysates were separated into two series of aldouronic acids composed of 4-O-Me-d-GlcAp and d-Xylp by ion-exchange chromatography. The first series included aldouronic acids from aldobiouronic acid (4-O-Me-d-GlcAp-Xyl) to aldopentaouronic acids (4-O-Me-d-GlcAp-Xyl₄). The second series were aldouronic acids composed of two 4-O-Me-d-GlcAp residues and 2-4 d-Xyl residues. In these acidic sugars, the uronic acid side chains were located on two contiguous d-Xyl residues. These facts indicated that AGXs from sugi and hinoki had a structural unit containing two 4-O-Me-d-GlcAp residues on two contiguous d-Xyl residues as well as AGXs from spruce and larch.     

Isolation and characterization of a novel immunomodulatory alpha-glucan-protein complex from the mycelium of Tricholoma matsutake in basidiomycetes

Tricholoma matsutake, a high-class edible mushroom in Japan, has been reported to have excellent biological activities, but difficulty in cultivating the fruit bodies and limited bulk availability have restricted detailed studies. We have developed a method of culturing in tanks, enabling the bulk supply of the mycelia. The preparation (CM6271) exerts modulative effects on the immune competence of mice and rats. In this study, a sodium hydroxide extract of CM6271 was defatted followed by fractionation with a combination of ion exchange chromatography and gel filtration in order to identify the components involved in the expression of the activity, and a single peak fraction (MPG-1) was obtained with reversed phase chromatography. MPG-1 was a glycoprotein (sugar:protein ratio, 94.3:5.7) with a relative molecular mass of 360 kDa, and the sugar moiety contained about 90% glucose. NMR spectra and methylation analysis revealed that the alpha-1,4-linkage was the predominant glucan linkage with alpha-1,6- and alpha-1,2-linkages in the minority. The amino acid composition in the protein moiety was rich in glutamine, alanine, asparagine, leucine, glycine, valine, serine, threonine, isoleucine, and proline. MPG-1 was resistant to degradation with amylase or protease. The oral administration of MPG-1 promoted, in a dose-dependent manner, the recovery of the mouse natural killer cell activity and serum IL-12 level that had been reduced by the loading of restraint stress. The dose of MPG-1 (25 mg/kg) required for the expression of the effect decreases to 1/12 of that of CM6271 (300 mg/kg). Furthermore, MPG-1 formed a complex with TGF-beta1 in vitro, modulating the biological activity of TGF-beta1 by binding to its active form. These results indicate that the mycelium of T. matsutake contains a novel alpha-glucan-protein complex with immunomodulatory activities.     

Displacement above the hypocenter of the 2011 Tohoku-Oki earthquake

The moment magnitude (M(w)) = 9.0 2011 Tohoku-Oki mega-thrust earthquake occurred off the coast of northeastern Japan. Combining Global Positioning System (GPS) and acoustic data, we detected very large sea-floor movements associated with this event directly above the focal region. An area with more than 20 meters of horizontal displacement, that is, four times larger than those detected on land, stretches several tens of kilometers long along the trench; the largest amount reaches about 24 meters toward east-southeast just above the hypocenter. Furthermore, nearly 3 meters of vertical uplift occurred, contrary to observed terrestrial subsidence.     

A network of superconducting gravimeters detects submicrogal coseismic gravity changes

With high-resolution continuous gravity recordings from a regional network of superconducting gravimeters, we have detected permanent changes in gravity acceleration associated with a recent large earthquake. Detected changes in gravity acceleration are smaller than 10(-8) meters seconds(-2) (1 micro-Galileo, about 10(-9) times the surface gravity acceleration) and agree with theoretical values calculated from a dislocation model. Superconducting gravimetry can contribute to the studies of secular gravity changes associated with tectonic processes.     

A closer look at water-related geologic activity on Mars

Water has supposedly marked the surface of Mars and produced characteristic landforms. To understand the history of water on Mars, we take a close look at key locations with the High-Resolution Imaging Science Experiment on board the Mars Reconnaissance Orbiter, reaching fine spatial scales of 25 to 32 centimeters per pixel. Boulders ranging up to approximately 2 meters in diameter are ubiquitous in the middle to high latitudes, which include deposits previously interpreted as finegrained ocean sediments or dusty snow. Bright gully deposits identify six locations with very recent activity, but these lie on steep (20 degrees to 35 degrees) slopes where dry mass wasting could occur. Thus, we cannot confirm the reality of ancient oceans or water in active gullies but do see evidence of fluvial modification of geologically recent mid-latitude gullies and equatorial impact craters.     

Antioxidant constituents of radish sprout (Kaiware-daikon), Raphanus sativus L

Methanol extracts of 11 kinds of commonly available vegetables were examined for hydroxyl radical scavenging potency using the bleomycin-Fe method. In this method, the iron ion and bleomycin in water form hydroxyl radicals, and the scavenging activity is monitored by the modified thiobarbituric acid method. All extracts showed scavenging capacity, even though the activity of some of them was lower than that of l-ascorbic acid. Those vegetables were classified into three groups according to their activity, groups showing strong activity, moderate activity, and weak activity, as compared to the activity of l-ascorbic acid at the same concentration. Among them, the methanol extract of radish sprout (Japanese name "kaiware-daikon") exhibited the highest potency (1.8 times as l-ascorbic acid). Then, we investigated the constituents of the methanol extract of radish sprout and the contribution to the overall activity of each compound by examining their activity. As the result, several kinds of sinapinic acid esters and flavonoids were isolated with high radical scavenging potency, which must contribute substantially to the activity.     

Precipitation chemistry in Japan 1989–1993

Precipitation chemistry in Japan was discussed on a wet-only sample database obtained in a nationwide survey from April 1989 to March 1993. Wet-only samples were collected at 29 stations over Japan on a biweekly basis. Commonly determined chemical parameters were measured in laboratories. The volume-weighted annual mean pH at each site ranged from 4.50 to 5.83 with a mean of 4.76. Concentration ranges and means (parenthesized) on an equivalent basis for major ions were as follows: nss-SO₄ ^2–; 5.2–58.9 (38.6), NO₃ ^–; 1.8–25.0 (14.1), NH₄ ⁺; 0.55–29.8 (18.3), nss-Ca²⁺; 2.0–34.5(14.2), Na⁺; 6.4–275.3 (49.1), Cl^–; 13.7–322.4 (63.5) eq L^–1. Acid-base relationships for Phase-II records were quantitatively discussed in terms of three measures: pH, fractional acidity, and our proposed pA_i.     

N-acetylglucosaminyltransferase I activity of bovine oviduct epithelial cells: stimulation by luteinizing hormone, vascular endothelial growth factor and tumor necrosis factor alpha

N-acetylglucosaminyltransferase I (GnT I; EC 2.4.1.101), which catalyzes the first step in the conversion of oligomannose to complex or hybrid N-glycans of glycoproteins, was found in media cultured with bovine oviduct epithelial cells (BOEC) obtained from non-pregnant cows during the follicular phase. Combined treatment with specific hormones increased GnT I release from BOEC. Luteinizing hormone (LH; 10 ng/ml) alone slightly, but together with 17beta-estradiol (E2; 1 ng/ml), synergistically increased GnT I activity. Vascular endothelial growth factor (VEGF) and tumor necrosis factor (TNF) alpha, which have been shown to have their highest activities in the bovine oviduct during the periovulatory period, also increased in GnT I activity. This study provides the first evidence of an increase of GnT I release from BOEC in vitro, and shows that endocrine as well as local factors such as LH, VEGF and TNFalpha increase this activity. The results suggest that GnT I activity in the bovine oviduct may contribute to the induction of glycosylation and thereby contributing to the provision of the optimal microenvironment for fertilization and early development of the embryos.