Effect of salinity on growth and accumulation of organic and inorganic solutes in the leguminous plants Alhagi pseudoalhagi and Vigna radiata

The response of two leguminous plants Alhagi pseudoalhagi and Vigna radiata to seawater salinity was studied over a period of 30 d. The growth of Vigna radiata was markedly and gradually reduced by increasing salinity levels, whereas that of Alhagi pseudoalhagi was promoted at 9.1 and 16.2 dS m^-1 salinity but then was slightly reduced at 28.2 dS m^-1 salinity. These results indicate that Alhagi pseudoalhagi belongs to the group of halophytic plants. Seawater salinity caused changes in the membrane permeability measured as electrolyte leakage in both plants. Alhagi pseudoalhagi maintained a lower membrane permeability than Vigna radiata. With increasing salinity levels, the membrane permeability decreased in Alhagi pseudoalhagi, whereas, in Vigna radiata it slightly increased at 9.1 dS m^-1. The leaf water potential and the osmotic potential decreased in both plants along with the seawater salinity levels. However, the turgor potential and osmotic adjustment in Alhagi pseudoalhagi were maintained at a higher level than in Vigna radiata. The contributions of organic and inorganic solutes to the osmotic adjustment differed: Alhagi pseudoalhagi achieved osmotic adjustment through Cl^- and Na⁺ uptake from the substrate, while the contribution of K⁺, Ca²⁺, and organic solutes to the osmotic adjustment was limited. These results suggest that the differences in salt tolerance between Alhagi pseudoalhagi and Vigna radiata can not be due to differences in specific-ion effects, but may be related to some factors involved in membrane permeability and osmotic adjustment.     

Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms

Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.     

Simultaneous detection of benzimidazole-resistant strains of Fusarium head blight using the loop-mediated isothermal amplification-fluorescent loop primer method

The loop-mediated isothermal amplification (LAMP)-fluorescent loop primer (FLP) method detects genetic polymorphisms by using a LAMP amplicon and measuring the peak temperatures of fluorescence resonance energy transfer between an FLP and a quencher probe, which is specifically hybridized to a sequence including a single nucleotide polymorphism (SNP). In the present study, the LAMP-FLP method was used to detect mutant genotypes F167Y, E198Q, and F200Y in the β2-tubulin gene region of causal pathogens of Fusarium head blight of wheat that result in methyl benzimidazole carbamate (MBC) resistance, proving its usefulness for monitoring strains with SNPs in target regions of MBC resistance.     

Iron-binding compounds produced by white-rot fungusPhanerochaete sordida YK-624

Iron-binding compounds were isolated from a culture ofPhanerochaete sordida YK-624 and were found to bind to Fe(III) preferentially compared with Fe(II). Two iron-binding compounds were purified to near-homogeneity with gel permeation chromatography. Hydrolysis of the iron-binding compounds with 6N hydrochloric acid gave ninhydrin-negative products. The molecular weight of these compounds was 3–5 kDa. These compounds may play an important role in the reduction of extracellular manganese dioxide to Mn(II) by intracellular ferrireductases for lignin degradation by manganese peroxidase.     

Evaluation of the extent of the oxidation reaction during chlorine bleaching of pulp

A modified method was developed to evaluate how much chlorine is consumed by the oxidation reaction during the chlorine bleaching process. This evaluation is, in principle, based on the sum of chloride produced during the chlorination stage (C-stage) and produced during alkali treatment of both the C-stage effluent and the chlorinated pulp. Results obtained by this method proved that about 50%–75% of chlorine was consumed by the oxidation reaction during chlorine bleaching, depending on the reaction condition of chlorination. Even under a reaction condition that is not favorable to an oxidation reaction (low pH), approximately three electrons were abstracted from one lignin structural unit by chlorine bleaching. This result provides additional evidence for our recent observation that lignin is extensively oxidized during chlorine bleaching even when pure chlorine without any chlorine dioxide substitution was used.Part of this paper was presented at the 40th Lignin Symposium. Tsukuba, Japan, October 12, 1995     

Studies on microörganisms and their activities in soil Part. 2. Virgin and cultivated soil of the same origin

The nature of soil is modified differently depending upon the artificial condition such as its utilization or management. It is therefore expected that the microbiological characteristic of soil is changed also. Greaves¹⁾ and Williams²⁾ reported that the reclamation of virgin soil brought about a change of bacterial count. Suzuki et al³⁾ observed that the kind of fungi differed between a virgin and a cultivated soil and that the vegetative mycelium was numerous in the former than in the latter. On the other hand, according to W aksman and Starkey⁴⁾, the bacterial count differed depending upon the fertility of soil. Singh⁵⁾ reported also that the number of fungi and actinomycetes was higher in a fertile than in an infertile soil. Lochhead⁶⁾, and Lochhead and Chasen studied the bacterial flora of a fertile (long-continued application of manure) and an infertile (no fertilizer for many years) soil and found that a certain difference could be observed when morphological, physiological and nutritional classification are tried.     

Comparative proteomic analysis of liver mitochondrial proteins derived from cloned adult pigs reconstructed with Meishan pig fibroblast cells and European pig enucleated oocytes

Somatic cell nuclear transfer (SCNT) has been exploited in efforts to clone and propagate valuable animal lineages. However, in many instances, recipient oocytes are obtained from sources independent of donor cell populations. As such, influences of potential nuclear-cytoplasmic incompatibility, post SCNT, are largely unknown. In the present study, alterations in mitochondrial protein levels were investigated in adult SCNT pigs produced by microinjection of Meishan pig fetus fibroblast cells into enucleated matured oocytes (maternal Landrace genetic background). Mitochondrial fractions were prepared from liver samples by mechanical homogenization and differential centrifugation. Liver mitochondria were then subjected to two-dimensional difference gel electrophoresis (2-D DIGE). Protein expression changes were confirmed with a volume ratio greater than 2 fold (P<0.05). 2-D DIGE analysis further revealed differential expression of three proteins between the Meishan (n=3) and Landrace (n=3) breeds. Differential expression patterns of 16 proteins were detected in SCNT pig liver tissue (n=3) when compared with Meishan control samples. However, none of the 16 proteins correlated with the three differentially expressed Meishan and Landrace liver mitochondrial proteins. In summary, alteration of mitochondrial protein expression levels was observed in adult SCNT pigs that did not reflect the breed difference of the recipient oocytes. Comparative proteomic analysis represents an important tool for further studies on SCNT animals.