Mapping of EST-derived CAPS markers in Italian ryegrass (Lolium multiflorum Lam.)

New molecular markers derived from expressed sequence tag (EST) sequences were mapped on linkage maps of Italian ryegrass by a two-way pseudo-testcross strategy. cDNA sequences were obtained from various tissues of Italian ryegrass ( Lolium multiflorum ) and converted into cleaved amplified polymorphic sequence (CAPS) markers. Of 260 EST primer pairs that amplified a single band, 74 generated bands that showed clear polymorphisms among individuals of an F₁ mapping family. Of the 74 polymorphic marker loci, 69 were mapped on an Italian ryegrass linkage map previously constructed using amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), and simple sequence repeat (SSR) markers. The newly-developed EST-CAPS markers would be useful as an efficient tool to identify genetic markers and to identify candidate genes for quantitative trait loci (QTLs) associated with important traits in Italian ryegrass.     

Dwarfing effect associated with the threshability gene Q on wheat chromosome 5A

A dwarfing effect of the 44.1 cM chromosomal region between the threshability gene Q and Xfba068 on the long arm of hexaploid wheat chromosome 5A has been reported. To clarify whether Q or its adjacent region is responsible for regulating culm elongation, two precise genetic stocks of near‐isogenic lines (NIL), a single chromosome substitution line, ‘Chinese Spring’ (CS; ‘Cappelle‐Desprez’ 5A) (NIL‐Q) and a single chromosome recombinant substitution line (NIL‐q) were used. The target segment of NIL‐q included the q allele and QEet.ocs‐5A.1, an earlinessper se gene, from spelt wheat in the CS genetic background. They were grown under 16‐h day length with and/ or without vernalization treatment. Being independent of heading date, NIL‐Q showed shorter elongation in lower internodes and decreased internode differentiation in comparison with NIL‐q. The culm‐length reduction associated with Q was confirmed in the recombinant F5 population derived from the cross between NIL‐Q and NIL‐q. Vernalization promotion had a tendency to reduce this dwarfing effect.     

Genetic Control of Vernalization, Day-length Response, and Earliness per se by Homoeologous Group-3 Chromosomes in Wheat

To identify homoeologous group-3 chromosomes that carry genes for vernalization, day-length responses, and earliness per se, a series of aneuploid lines (mono-somics and tetrasomics) and chromosome-substitution lines in ‘Chinese Spring’ (CS) were surveyed under different vernalization and day-length regimes in controlled environments. The results indicated that genes on all three chromosomes of group 3 can have striking effects on ear-emergence time. The replacement of CS 3B by its homologues in ‘Lutescens 62’ and ‘Cheyenne’ produced an increased insensitivity to vernalization, while 3B homologues from ‘Ceska Presivka’ gave CS a remarkable sensitivity to vernalization. This provided evidence for multiple allelism at a new Vrn locus on chromosome 3B. A negative association between gene dosage and day-length response was found in CS 3D which was thought to carry a gene for promoting insensitivity to day-length. The behaviour of CS monosomic 3A and CS (Timstein 3A), in reducing numbers of days to heading independently of environmental stimuli, suggested the presence of earliness per se genes on this chromosome.     

Evaluation of maturity of poultry manure compost by phospholipid fatty acids analysis

To study the influence of the physical properties of compost feedstock on some characteristics associated with maturity, two types of compost were made from poultry manure, rice husk, and rice bran. The bulk density of one type (PMC) was always higher than that of another type (NMC) during composting. In the case of PMC, the change in temperature, decrease in NH₄⁺, appearance of NO₃^–, and increase in germination indices (GI) with Japanese Komatsuna (Brassica campestris cv. Osome) were all more delayed than in NMC. As the composting process progressed, the proportion of branched (iso-, anteiso-, 10Me-) and saturated phospholipid fatty acids (PLFA) [BRANCHED FAMES (fatty acid methyl esters), biomarkers for gram-positive bacteria] gradually increased, then reached plateau. The high proportion of BRANCHED FAMES was maintained over a long storage period. The straight hydroxyl and saturated PLFAs (SOH-FAMES) initially increased, then disappeared with the progress of composting. The increase in BRANCHED FAMES and the decrease in SOH-FAMES were more delayed in PMC than NMC. The day on which the proportion of BRANCHED FAMES reached plateau and the proportion of SOH-FAMES dipped below 2 mol% coincided with the maturity stage based on the changes of physicochemical characteristics and GI. The composition of BRANCHED FAMES showed highly positive and negative correlation with GI and NH₄⁺, respectively. In the case of SOH-FAMES, inverse correlations were observed. This indicates that the proportion of BRANCHED FAMES and/or SOH-FAMES can be used as a tool for evaluating the maturity of poultry manure compost.     

Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays)

Polymerase Chain Reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. In this paper, recombinant DNAs introduced into the seven lines of GM maize, such as Event 176, Bt11, T25, MON810, GA21, DLL25, and MON802, are sequenced. On the basis of the obtained sequence, 14 primer pairs for the detection of the segments, such as promoter, terminator regions, and construct genes, were designed. To confirm the specificities of the designed primer pairs, PCR was performed on genomic DNAs extracted from GM and non-GM maize, GM and non-GM soy, and other cereal crops. Because the presence of the corresponding DNA segments was specifically detected in GM crops by the designed primer pairs, it was concluded that this method is useful for fast and easy screening of GM crops including unauthorized ones.     

Elemental affinity for siderite found in a Japanese paddy subsoil

ABSTRACT

Siderite (FeCO₃), a type of carbonate mineral, is very occasionally recognized as a nodule in anoxic soils and sediments. During siderite formation, elements are expected to be accumulated or excluded between siderite and bulk soil. Therefore, we verified the affinity of 40 elements for siderite found in the gley horizon of a smectite-rich paddy field in northeastern Japan from the elemental concentration of the soil and the siderite dissolved in 2.88 mol L^?1 hydrochloric acid (10% HCl). The concentration ratio of the target element of the soil to the siderite was expressed by the product of the dimensionless R₀ value and ratio of titanium (Ti) concentration of the soil to the siderite (C_siderite /C_soil) = R₀ (Ti_siderite /Ti_soil). The affinity of each element for the siderite was analyzed by comparing the R₀ values. In comparison with Ti used as the reference element, P, Cr, Co and Mo were accumulated (R₀ > 1.0), and alkali metals, such as Li, Na and K, and chalcophile elements, such as Cu, Zn, Cd and Pb, tended to be equal or excluded from the siderite (R₀ ≦1.0). Of the group 2 elements, Ca, Sr and Ba were accumulated, while Mg was excluded. The affinity of Mn for siderite exceeded that of Fe as the main component of the siderite, which reflects the fact that siderite was formed in a fresh water condition. The siderite size did not influence the elemental affinity. Observation of micromorphology using a scanning electron microscope showed that the siderite nodules were aggregations of siderite particles and clays, suggesting that the siderite grew by taking in clay around it. The rare earth element (REE) distribution pattern showed a tendency not to increase linearly but to increase with forming a concave curve at the intervals of the four elements with the increase of atomic number (i.e. tetrad effects). Although these findings are from the lower horizons of anoxic soils, the elemental affinity for siderite of this study is useful in order to elucidate the complex elemental dynamics in the anoxic environment.     

Method for efficiently obtaining fertilized eggs from the black scraper Thamnaconus modestus by natural spawning in captivity

To develop an efficient and stable method for enhancing spawning of black scrapers in captivity, we examined the following three factors: (1) different kinds of spawning substrate, (2) the area of the spawning substrate, (3) the grain size of the spawning substrate. The results demonstrated that sand is the optimum spawning substrate when compared to a hard polyvinyl chloride sheet, a piece of shading net, or artificial spawning grass. No difference was observed in the spawning frequency between a small sand bed of area 400?cm² and sand spread over the bottom of the tank to an area of 7085?cm², since the egg masses were spawned on the sand in a small area approximately 15?cm in diameter. In addition to the spawned eggs, the small movable sand bed is useful for subsequent harvest. No differences were observed in either the spawning frequency or the hatching rate when using sand beds with different grain sizes. In large-scale egg collection trials, an average of 33 egg masses from which an average of 403,000 larval fish were obtained. We can therefore conclude that the procedure developed in the present study is applicable to the commercial-scale seed production of black scraper.     

Quantification of waste feed and fish faeces in sediments beneath yellowtail pens and possibility to reduce waste loading by co‐culturing with sea cucumbers: an isotopic study

To evaluate environmental impacts of yellowtail culture and to examine the efficiency of the sea cucumber Apostichopus japonicus in reduction of waste loading, surface sediments were sampled and A. japonicus were cultured at a yellowtail farm in Owase Bay, Japan. Waste feed‐ and faeces‐derived organic matter (WOM, FOM) in sedimentary organic matter (SOM) was estimated based on the δ¹³C and δ¹⁵N of the fish feed (?21.7‰, 9.2‰), yellowtail faeces (?20.6‰, 6.2‰) and SOM reference (?24.4‰, 4.4‰). Small WOM (0–22% in SOM) but substantial FOM (28–61%) loadings in the fish farm area and high sulphide concentrations in the sediments (1.0 mg g^?1) suggest that reduction in the fish stocks or mitigation of the faeces should be considered. A. japonicus juveniles were cultured in three cages deployed below a pen and growth was assessed after three different periods (62, 107, 181 days). A. japonicus grew well during the first 107 days (daily specific growth rate, 3.7%) and their survivorship was high (80–90%). Growth ceased after 107 days, probably due to fouling on the cages. The δ¹³C and δ¹⁵N of their hypothetical diet (–19.7‰, 5.5‰) were close to the FOM values, suggesting assimilation of FOM.