Comparative studies of the changes in enzymatic activities in hardy and less hardy cultivars of winter wheat in late fall and in winter under snow

Abstract

The synthetic activity of protein and changes in the activity profiles of enzymes during cold acclimation were studied in leaves, stems, and crowns of a hardy winter wheat cultivar ( Triticum aestivum L. cv. Horoshirikomugi) and a less hardy winter wheat cultivar (Triticum aestivum L. cv. Norin 61). The results showed that one of the genetic differences between the two cuitivars is found in the activities of protein synthesis under low temperatures. The profiles of the changes in enzymatic activities showed that the adjustment of enzymatic activities was a tissue-specific event to accommodate environmental changes. The synthesis of the enzymes involved in a peroxide scavenging system such as hexokinase, glucose-6-phosphate dehydrogenase, dehydroascorbate reductase, ascorbate peroxidase, and so on started at the acclimation stage, while the phosphoglucose isomerase, which appears to be a maintenance-type enzyme in the wintering wheat, showed a small increase in the activity at later stages of acclimation period in their stems of both cuitivars. When the field was covered with snow, the catalase activity in the leaves started to decrease abruptly, while catalase activities in the stems and crowns did not exhibit appreciable changes during the life under snow. The life-cycle dependent changes in enzymatic activities appear to occur in late March or early April under snow.     

Isolation of endophytic streptomycetes from above- and belowground organs of Quercus serrata

The occurrence and localization of endophytic actinomycetes within diverse organs of host plants provide ecological information that can be used to evaluate the significance of their spatial habitats. The aim of this study was to isolate and characterize endophytic actinomycetes in different organs of Quercus serrata. For this purpose, actinomycete isolates were obtained from surface-sterilized tissues of both symptomless seedlings and aged trees of Q. serrata and rhizosphere soil of the sampled seedlings. Thirty-five isolates with the ability to sporulate, including 4 from leaves of the aged trees, 10, 6, and 15 from leaves, stems, and roots of the seedlings, respectively, and 8 soil-derived isolates, were selected and characterized. The 16S rDNA nucleotide sequence analyses revealed that all of them belonged to the genus Streptomyces. According to a neighbor-joining phylogenetic tree constructed based on the results, the isolates of plant origin were divided into three major clades with high bootstrap values of 98 or 99 %, whereas eight soil-derived isolates were located at different positions from those of the endophyte isolates. Moreover, two larger clades were formed, one of which contained isolates derived only from aboveground parts, while the other contained isolates from all of the organs. These results suggest that the endophytic streptomycetes in Q. serrata may differ in their habitat positions (i.e., either above- or belowground parts).     

Development of CAPS markers and their application in breeding for mango, Mangifera indica L.

To develop cleaved amplified polymorphic sequence (CAPS) markers for the identification of true hybrids in F1 progeny, we cloned sucrose synthase (SS) and sucrose phosphate synthase (SPS) genes from 19 mango cultivars. Through comparison of amino acid sequences in a BLASTX search, cloned SS and SPS genes were found to be homologs of Citrus unshiu CitSUS1 (AB022092) and CitSPS1 (O22060), respectively. Therefore, we designated the SS and SPS genes as MiSUS1 and MiSPS1, respectively. There were 11 genotypes in MiSUS1 and 10 in MiSPS1 sequences. In the MiSUS1 nucleotide sequences, 46 SNPs and 5 in/dels were detected. Twenty-six out of the 46 SNPs mapped on the restriction enzyme recognition sites were successfully converted to 25 CAPS markers. In the MiSPS1 nucleotide sequences, 99 SNPs and 6 in/dels were detected. Thirty out of 99 SNPs and 2 out of 6 in/dels mapped on the recognition sites were also converted to 32 CAPS markers. By using two CAPS markers, SS-2798 and SPS-5745, we identified 37 true hybrids that had different genotypes to ‘Irwin’ from 82 F1 progeny obtained through a cross between ‘Irwin’, the maternal parent, and paternal parents by using flies as pollinators. The method developed here to identify true hybrids is a simple, rapid and highly reliable tool for efficient mango breeding.     

Responses of Riverine Macroinvertebrates to Zinc in Natural Streams: Implications for the Japanese Water Quality Standard

We conducted a field survey in Japanese streams and rivers to generate useful information about whether the Japanese water quality standard for zinc (Zn) in the freshwater environment (30 μg/L) is over- or underprotective of the populations and communities of lotic benthic macroinvertebrates. Many macroinvertebrate populations were reduced and community characteristics were altered at two polluted sites with Zn concentrations above 1,000 μg/L as compared with control sites with Zn concentrations below 30 μg/L. In contrast, at least one site among three polluted sites with Zn concentrations of 80–126 μg/L experienced noncritical effects (i.e., no significant difference was observed between the sample mean at a polluted site and that at the control sites, and the sample mean was within the minimum–maximum range of those at the control sites) for most of the ten community metrics for diversity and abundance. In addition, the abundances of 95% of the dominant taxa were not always significantly lower at more than one polluted site (Zn concentrations, ≥80 μg/L). Our results suggest that Zn concentrations below approximately 100 μg/L are likely to protect most aspects of richness and abundance of the riverine macroinvertebrate communities in the study areas. This study shows a case that the Japanese Zn standard is likely overprotective of the macroinvertebrate populations and communities.     

An econometric estimation of Japanese shrimp supply and demand during the 1990s

Abstract

A simultaneous‐equation model of the supply and demand of shrimp in Japan is constructed, and sensitivity analyses are conducted to measure the effects of changing exchange rates and the stock holdings of shrimp. Using monthly statistics from January 1990 to December 1997, the Japanese own‐price elasticity of supply is found to be inelastic and has a less important effect than the beginning stocks factor. The Japanese demand for shrimp is also found to be price‐and income‐inelastic. The beginning of stock holdings is more sensitive to Japanese shrimp demand than the import price and income. The result of seasonality analyses shows that Japanese shrimp demand also depends on cultural factors.     

A genetically engineered derivative of Salmonella Enteritidis as a novel live vaccine candidate for salmonellosis in chickens

To construct a novel live Salmonella Enteritidis (SE) vaccine candidate, SE was genetically engineered using the allelic exchange method to delete two virulence genes, lon and cpxR. The lon gene deletion is essential to impair Salmonella replication and avoid overwhelming systemic disease in the host. The cpxR gene deletion is needed to enhance the ability of bacteria to adhere and invade the host cell. Scanning electron microscopy revealed that the derivatives JOL917 (Δlon), JOL918 (ΔcpxR), and JOL919 (Δlon/ΔcpxR) had increased surface fimbrial filamentous structures. Significant elevations of extracellular polysaccharide and FimA expression were observed for the derivatives compared to the parental wild type JOL860, while biochemical properties of the derivatives were not altered. In the safety examination by inoculation of the derivatives in chickens, gross lesion scores of the liver, spleen, kidney, small intestine and caecal tonsils were moderate in the JOL917 and JOL918 groups, and significantly lower in the JOL919 group than those of the JOL860. Bacterial counts from the spleen and caeca of the JOL917 and JOL918 groups were moderate, and significantly reduced in the JOL919 group compared to the JOL860 group. In addition, only the JOL919 group showed significantly lower bacterial counts in the faecal samples than those of the JOL860 group. Significant elevations of IgG and secretory IgA levels observed in the derivative groups, while the JOL919 and JOL860 groups showed a potent lymphocyte proliferation response as compared to those of the control group. In the protection efficacy examination, JOL919 immunized group showed significantly lower depression, lower gross lesion in the liver and spleen, and lower number of the SE positive internal organs than those of the control group against a virulent wild type SE challenge.     

Molecular identification and characterization of the intervening sequences (IVSs) within 23S ribosomal RNA (rRNA) genes of Taylorella asinigenitalis isolated in France

In the helix 25 region, 32 French Taylorella asinigenitalis isolates carried at least one 23S rRNA gene not containing intervening sequences (IVSs). No IVSs in the region were identified in three isolates and the other remaining 29 isolates carried one or more IVSs (UCD-1(T)IVS1A, UCD-1(T)IVS1B and UK-1IVS1B) described already and two new kinds of IVS (TaIVS1C and TaIVS1D). In the helix 45 region, no T. asinigenitalis isolates not carrying any IVSs were identified. UK-1IVS2B was identified in the region from 26 isolates. Five new kinds of IVSs (TaIVS2D, E, F, G and H) occurred in the region in the 13 isolates. Distinctly different tandem repeat units (RS48 and RS32 and RS-A, -B and -C) were evident in both regions, respectively, from the French (n=32) and American (n=3) T. asinigenitalis isolates. Thus, several different kinds of tandem repeat units and their combinations in IVSs in both regions within the gene were shown in 32 French isolates.     

Differential spread of discoloured and non‐conductive sapwood among four Fagaceae species inoculated with Raffaelea quercivora

To reveal the relationship between the susceptibility of Fagaceae species to Raffaelea quercivora Kubono et Shin‐Ito and the tangential expansion of regions of discoloured and non‐conductive sapwood among the species, we inoculated branches of three Quercus species and one Castanopsis species with the fungus. The sapwood around the inoculation hole in all four species became non‐conductive in response to the infection before the discolouration. The expansion of the region of non‐conductive sapwood ceased within 2 weeks after the inoculation. The region of non‐conductive sapwood in Q. crispula and Q. serrata was larger than that in Q. glauca and C. cuspidata var. sieboldii. These results suggest that the region of non‐conductive sapwood expanded soon after the infection by R. quercivora. It was also clear that the transverse non‐conductive sapwood has close relationship with the susceptibility among Fagaceae species to R. quercivora.