IL‐12p40 gene expression in lung and hilar lymph nodes of MPS‐resistant pigs

Mycoplasma pneumonia of swine (MPS) is caused by Mycoplasma hyopneumoniae (M.hp) and is a common chronic respiratory disease of pigs. Recently, a genetically selected variant of the Landrace pig (Miyagino L2) has a lower incidence of pulmonary MPS lesions. We investigated the pathological and immunological characteristics of MPS resistance in these pigs (n = 24) by comparing with the normal landrace pig (control: n = 24). The pathological MPS lung lesion score in MPS‐selected landrace pigs was significantly lower than in the control. The gene expression of interleukin (IL)‐12p40, which acts as a chemoattractant and a component of the bioactive cytokines IL‐12 and IL‐23, was significantly higher at the hilar lymph nodes, lung, and spleen in MPS‐selected landrace pigs than in control landrace pigs, and these were negatively correlated with the macroscopic MPS lung lesion score. In summary, we demonstrate that resistance against MPS in Miyagino L2 pigs is associated with IL‐12p40 up‐regulation, in comparison with normal landrace pigs without the MPS vaccine. In addition, a comparative study of macroscopic MPS lung lesions and IL‐12p40 gene expression in lung and hilar lymph nodes may lead to beneficial selection traits for the genetic selection for MPS resistance in pigs.     

Application of NaLA,a fishing net configuration and loading analysis system,to bottom gill nets

ABSTRACT:   The net-shape and loading analysis system (NaLA) was developed to determine fishing net configuration and load in a previous study. The system has since been applied to general gill nets and aquaculture nets, and its validity has been proven through model experiments in tanks. In this study, the system was applied to estimate the dynamic behavior of a bottom gill net for walleye pollock, to test the system's applicability of the system to gear operations in the field. To obtain in situ data, four bottom gill net operations were performed in February 2004 off the coast of Sawara, Hokkaido, Japan. During operations, vertical displacements of the bottom gill net's float and sinker lines were measured as representative values of gear behavior, and ocean current direction, and speed at the gear position were observed simultaneously. Then, bottom gill net behavior was simulated using NaLA, incorporating observed environmental conditions and gear specifications. The resulting calculated behavior was compared to measured behavior in terms of the relationship between net height and environmental or setting conditions. Agreement between the calculated and measured net behavior was found. Thus, it is believe that our NaLA calculation model has the potential to simulate the dynamic behavior of bottom gill nets in situ .     

Effect of burning on soil organic matter content and N mineralization under shifting cultivation system of Karen people in Northern Thailand

In the traditional shifting cultivation system practiced by the Karen people in northern Thailand, the effects of burning on the content of extractable organic matter, microbial biomass, and N mineralization process of the soils were studied. Five plots (5×5 m² quadrat) with 0, 10, 20, 50, and 100 Mg ha^-1 of slashed materials were arranged and burned. Ten to 20 Mg ha^-1 of slashed biomass corresponded to the amount commonly burned by the Karen people. During the burning process, the soil temperature at the depth of 2.5 cm in the 100 Mg ha^-1 plot almost evenly increased to 300°C while the temperature in the 10 to 50 Mg ha^-1 plots increased with large variations from 50 to 300°C. Burning caused a conspicuous increase in the contents of organic C and (organic + mineral)-N extracted at room temperature and a simultaneous decrease in the contents of microbial biomass C and N, especially in the soil of the 100 Mg ha^-1 plot. In the rainy season, the values of the changes induced by burning reverted to the values recorded before burning, except for the microbial biomass in the 100 Mg ha^-1 plot, which still remained lower. Based on an incubation experiment, N mineralization rate was higher in the soils taken just after burning, especially in the 100 Mg ha^-1 plot, than in the soils taken during the rainy season. However, the soil in the 100 Mg ha^-1 plot was considered to have the lowest ability to supply mineral N among the soils in the rainy season. Burning of 10 to 20 Mg ha^-1 biomass corresponding to the values recorded in Karen peoples' shifting cultivation system was more compatible with soil ecology in terms of N supply at the initial stage of crop growth and of microbial biomass recovery during the rainy season, compared to the burning of 100 Mg ha^-1 biomass corresponding to the value recorded in a natural forest. Thus, the shifting cultivation system implemented by the Karen people can be considered to be a well-balanced agricultural system.     

A binding protein for fungal signal molecules in the cell wall of Pisum sativum

In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules.     

Identification of genes expressed during spore germination of Mycosphaerella pinodes

Mycosphaerella blight, caused by Mycosphaerella pinodes, is one of the major diseases of cultivated pea (Pisum sativum L.). To isolate the genes that are up- and down-regulated during spore germination, suppression subtraction hybridization (SSH) was performed between ungerminated and germinated spores. The 232 and 128 clones from forward and reverse libraries, respectively, were collected, sequenced, and analyzed with a BLASTX homology search. About 95% of the 32 selected clones were expressed during spore germination on a paper sheet and during infection of pea leaves. We discuss the applicability of the SSH libraries for analyzing M. pinodes genes involved in the early stage of infection.     

Harpinpsta from Pseudomonas syringae pv. tabaci Is Defective and Deficient in Its Expression and HR-inducing Activity

To elucidate the role of harpins produced by Pseudomonas syringae, the corresponding hrpZ gene was isolated from P. s. pv. tabaci. The sequence information revealed that this gene carries a serious mutation with 326 bp lacking in the central region and potentially encodes only 140 N-terminal amino acids because of a frame shift. The investigation of biological properties using recombinant harpin indicated harpin_psta was incapable of inducing HR in both host and nonhost plants. Based on an immunoblot analysis to detect harpin from P. s. pathovars in hrp-inducing medium, the truncated harpin_psta was neither expressed nor secreted into the culture medium. These results suggest that harpin is not the sole determinant of the host-parasite specificity in P. s. pv. tabaci. Received 10 August 2000/ Accepted in revised form 21 December 2000     

Chemical forms and stability of selenium in soil

The chemical forms of selenium (Se) in soils derived from different parent rocks, and the stability of Se species in soils were investigated. Around 40% of the total Se occurred as organic Se on the average. Total selenite content was higher than that of total selenate. The content of organic Se decreased with heating of the soils at high temperatures, and most of it disappeared at heating temperatures above 500°C. However, the results of incubation experiments indicated that Se species were stable in soil, and that their chemical forms did not change appreciably with the variation of soil conditions. In soluble Se, organic Se accounted for about 50%, and in contrast to the total Se, the selenate content was higher than that of selenite.     

Expression of Medicago truncatula ecto-apyrase MtAPY1;1 in leaves of Nicotiana benthamiana restricts necrotic lesions induced by a virulent fungus

Ecto-apyrase(s) participates in cell-wall-associated defense through ATP hydrolysis. Here we analyzed Medicago truncatula genes through cDNA screening and in silico analyses against known databases. This study revealed seven genes, five of which (MtAPY1;1 to MtAPY1;5) are members of a legume-specific family, whereas two genes (MtAPY2;1 and MtAPY2;2) are close to those in other plants. Agrobacterium-based transient expression in Nicotiana benthamiana, combined with a c-myc epitope tag technology, confirmed that the MtAPY1;1 is a secreted protein. Transient expression of MtAPY1;1 in leaves of N. benthamiana restricted disease development by a virulent fungus, suggesting a role in disease resistance.