Microbiological Studies on the Humification Process (Part 2) on the Aerobic Decomposition of the Fresh Plant Residue in the Case of Single Inoculation

The humification of several constituents of plant was studied by Trussow¹⁾ and Waksmans²⁾ in the Past, or by Kononova,³⁾ Tepper⁴⁾ and Laatsch⁵⁾ in recent years. In the previous paper, we experimented the microbial action on humus separated from an uncultivated soil, and showed that the inoculation of Trichoderma koningi and soil extract prqmoted the humification of the sample, but such tendency was not revealed in the case of single inocolation of two strains of Actinomyces and four strains of aerobic bacteria, and that when the humification of the soil humus was promoted it occurred, in the first place, as the transformation of the fulvic acid fraction, not as that of the precipitation part (the humic acid fraction). Following the previous report, we studied the humification process of fresh plant residue (upland rice straw) by a similar procedure.     

The process of manganese depositton in paddy soils

Abstract

The changes in quality and quantity of phenolic substances in the decaying process of rice straw in a soil were compared under moist and flooded conditions for 200 days. The amounts of phenolic substances divided into fractions of humic acid and fulvic acid, ether- and butanol-extractable and organic solvent-unextractable fractions, then the amounts of individual phenolic acids were determined. The following results were obtained.

1) Alkali-extractable total phenolics as well as individual phenolic acids decreased more rapidly under moist, than under flooded, conditions as rice straw decayed in the soil. The phenolics present were mainly attributable to the straw, not to the soil.

2) The decrease in the level of total phenolics in the early stage of the decaying process was mainly due to the decrease in ether-extractable phenolic compounds in the fulvic acid fraction, and in the later stage, was mainly due to the decrease in butanol-extractable phenolics in the humic acid fraction.

3) The amounts of butanol-extractable phenolics and organic solvent-unextractable phenolics were larger in humic acid than in fulvic acid. On the other hand, a larger amount of organic solvent-extractable phenolics, especially ether-extractable phenolics, was present in fulvic acid.

4) The degradation patterns and pathways of individual phenolic acids in the decaying process of rice straw in soil were found to be the lame as those of decaying straw without soil which were reported previously.

5) The level of phenolic substances in the humic acid was not greatly changed during the decaying process, but the phenolic substances in fulvic acid rapidly increased for 30 days and then rapidly decreased to a constant level.     

Cloning of nicotianamine synthase genes from Arabidopsis thaliana

Nicotianamine synthase (NAS) catalyzes the trimerization of S-adenosylmethionine to form one molecule of nicotianamine (NA). NA is present in all the plants; it chelates metal cations, and is considered to play a role in metal homeostasis in plants. Moreover, in graminaceous monocotyledonous plants, NA is an essential intermediate in the biosynthesis of mugineic acid family phytosiderophores (MAs). In order to identify the gene encoding NAS in dicotyledonous plants, Arabidopsis thaliana databases were searched using the nucleotide sequence of the NAS gene from barley (HvNAS), which we have recently isolated. We found several ESTs and three genomic sequences highly homologous to HvNAS in the databases. Based on these nucleotide sequences and that of HvNAS, we designed 2 sets of primers to isolate the NAS orthologues in Arabidopsis and succeeded in obtaining three DNA clones encoding AtNAS (AtNAS1, 2, and 3). These clones were expressed in Escherichia coli and their protein products displayed the NAS activity. The expression of AtNASl was detected in both shoots and roots of A. thaliana by RT-PCR; AtNAS3 expression was only detected in the shoots. In contrast, AtNAS2 expression was not detected in any organs.     

Influences of organic debris and rice root on the nitrogen fixation in the submerged soil

Abstract

A series of experiments has been conducted on the N2 fixation in the paddy soils by the authors (1–4). The amount of organic substrates for microorganisms and the degree of reduction of the soil are found to be two major factors affecting the N₂-fixing activity of the heterotrophic microorganisms in the submerged soil. Organic debris, rice root and their neighboring soils are identified to be the important micro-sites for the heterotrophic N₂ fixers. The organic debris and the rice root are considered to play dual roles by supplying the organic substances; (1) increase of the population of the heterotrophic N₂ fixers—the amount of nitrogenase, (2) preparation of the reduced conditions favorable for the nitrogenase activity.

However, it is not yet clearly known which of these two roles of the organic substrates is more essential to the N₂-fixing activity in the paddy soil. In addition, it is expected that there must be some differences between the organic debris and rice root in their contribution to the N₂ fixation in the paddy soil.

An experiment was carried out to clarify these problems. Moist soil sample was collected from the plough layer of the paddy field at Central Agric. Exp. Sta. in Konosu City, Saitama Pref., passed through a 5 mm sieve and placed in pots (3 kg moist soil/pot). Ammonium sulfate, calcium superphosphate, and potassium chloride at the rate of 0.4-0.4-0.4 (N-P₂O₅-K₂0) g/pot were incorporated into the soil 7 days before transplanting. Split application of ammonium sulfate at the rates of 0.2 and 0.4 g N/pot were also incorporated at 30 and 41 days after transplanting respectively. These pots were divided into three series; planted (P-series), non-planted (N-series), and non-planted and applied with organic manure (O-series). In case of O-series, 60 g of fairly rotted organic manure was applied to each pot. Each pot of P-series was planted with two 4O-day-old seedlings of rice plant at 7 days after submergence. The Nseries was regarded as a control. Each series was not replicated in this preliminary experiment.     

Stress-wave velocity of trees and dynamic Young’s modulus of logs of 4-year-old Eucalyptus camaldulensis trees selected for pulpwood production in Thailand

Eucalyptus camaldulensis Dehnh. is extensively planted in Thailand to produce wood chips used as raw material for pulp and paper. To promote the utilization of the wood from plantation-grown E. camaldulensis for solid lumber, stress-wave velocity of trees and dynamic Young’s modulus of logs were investigated for 4-year-old trees of eight half-sib families selected for pulpwood production on the basis of the growth characteristics in the previous tree breeding program. For the eight families, the mean stem diameter at 1.3 m above ground level and mean tree height were 7.6 cm and 11.9 m, respectively. The mean stress-wave velocity of eight families was 3.45 km/s. Dynamic Young’s modulus of logs ranged from 7.88 to 17.64 GPa, and the mean value for the eight families was 11.72 GPa. Stress-wave velocity of trees was significantly correlated with dynamic Young’s modulus of logs, suggesting that dynamic Young’s modulus of wood can be evaluated nondestructively by stress-wave velocity of trees. Significant differences in stress-wave velocity and dynamic Young’s modulus of logs were obtained among families. Thus, to promote the utilization of E. camaldulensis wood for solid lumber production, selection of trees with high Young’s modulus should be applied to trees already selected for the growth characteristics in the previous tree breeding program.     

Variation in tree growth characteristics, stress-wave velocity, and Pilodyn penetration of 24-year-old teak (Tectona grandis) trees originating in 21 seed provenances planted in Indonesia

Growth characteristics [stem diameter (D), tree height (H), and bole volume (V)], stress-wave velocity (SWV), and Pilodyn penetration (P) were measured for 21 seed provenances of 24-year-old teak trees planted in Indonesia to characterize variation in tree growth characteristics, SWV, and P among seed provenances. Broad-sense heritability and correlations between the measured characteristics were also determined. Significant differences for all measured characteristics were observed among provenances, indicating that these characteristics are genetically controlled. Broad-sense heritabilities of growth characteristics, SWV, and P are moderate values. These results indicate potential for improving growth characteristics and wood properties of teak trees with the help of breeding programs. Highly significant positive correlations were observed among the growth characteristics, suggesting that they are closely related. In contrast, no significant correlations were observed between the growth characteristics and SWV, indicating that they are independent. We conclude that mechanical properties are also important criteria for selecting plus trees in tree breeding programs. Principal component analysis revealed that seed provenances from Indonesia (Bangilan, Deling, and Randublatung) and India (Malabar and Central Province) have high scores of growth characteristics and SWV.     

Virulence of Raffaelea quercivora isolates inoculated into Quercus serrata logs and Q. crispula saplings

This study was conducted to compare the virulences of various isolates of Raffaelea quercivora—a fungus that causes Japanese oak wilt disease—towards Japanese oak trees. Five isolates were collected from a wide range of Japan and inoculated into Quercus serrata logs and Q. crispula saplings. The tangential length of the discolored sapwood in the Q. serrata logs differed significantly among the isolates. The trend in isolate virulence was similar for the Q. serrata logs and the Q. crispula saplings. This is the first report suggesting that there is variability in virulence among isolates of R. quercivora.     

No evidence of horizontal infection in horses kept in close contact with dogs experimentally infected with canine influenza A virus (H3N8)

Background

Since equine influenza A virus (H3N8) was transmitted to dogs in the United States in 2004, the causative virus, which is called canine influenza A virus (CIV), has become widespread in dogs. To date, it has remained unclear whether or not CIV-infected dogs could transmit CIV to horses. To address this, we tested whether or not close contact between horses and dogs experimentally infected with CIV would result in its interspecies transmission.

Methods

Three pairs of animals consisting of a dog inoculated with CIV (10^8.3 egg infectious dose₅₀/dog) and a healthy horse were kept together in individual stalls for 15 consecutive days. During the study, all the dogs and horses were clinically observed. Virus titres in nasal swab extracts and serological responses were also evaluated. In addition, all the animals were subjected to a gross pathological examination after euthanasia.

Results

All three dogs inoculated with CIV exhibited clinical signs including, pyrexia, cough, nasal discharge, virus shedding and seroconversion. Gross pathology revealed lung consolidations in all the dogs, and Streptococcus equi subsp. zooepidemicus was isolated from the lesions. Meanwhile, none of the paired horses showed any clinical signs, virus shedding or seroconversion. Moreover, gross pathology revealed no lesions in the respiratory tracts including the lungs of the horses.

Conclusions

These findings may indicate that a single dog infected with CIV is not sufficient to constitute a source of CIV infection in horses.     

Molecular typing of VapA-positive Rhodococcus equi isolates from Jeju native horses, Korea

We recently demonstrated the presence of virulence-associated protein antigen (VapA)-positive Rhodococcus equi in Jeju Island, Korea. These bacteria contained one of two distinct plasmid types, a 90-kb type II plasmid, which has been found in isolates from the native Kiso horses of Japan, and a new variant, a 90-kb type V plasmid. However, the genotypic characters of the VapA-positive R. equi from Jeju native horses and their environments are poorly understood. Ninety-eight isolates from soil samples and 89 isolates from fecal samples were collected from five farms that breed or have bred Jeju native horses, and were tested for the presence of VapA by immunoblotting and PCR. Of the 98 soil isolates and 89 fecal isolates, seven and 13 were VapA-positive R. equi, respectively. In 2003, two Jeju foals died suddenly and were brought to the Faculty of Veterinary Medicine, Cheju National University, for postmortem examination. Pure cultures of R. equi were isolated from the lung lesions of both foals. Of the 16 clinical isolates, 14 were VapA-positive R. equi. Of the 34 VapA-positive clinical and environmental isolates, 16 contained the 90-kb type II plasmid and 18 contained a 90-kb type V plasmid. Pulsed-field gel electrophoresis (PFGE) patterns of the VapA-positive isolates from Jeju horses and Kiso horses, containing the 90-kb type II plasmid, were compared and formed two distinct groups. Furthermore, 18 virulent isolates containing the 90-kb type V plasmid formed two distinct PFGE groups (of 16 and two isolates). These results demonstrate that two virulence plasmid types are widespread in R. equi in Jeju native horses. However, there is little diversity in the PFGE patterns of virulent isolates, suggesting the clonal spread of virulent R. equi. The PFGE pattern of the virulent R. equi isolates from Jeju native horses in Korea is not identical to those of Kiso native horses in Japan.