Ultrastructural study on scab resistance expressed in epidermal pectin layers of pear leaves

Proliferation and collapse of subcuticular hyphae of Venturia nashicola race 1 were studied ultrastructurally, after inoculation of susceptible Japanese pear cv. Kousui, resistant Japanese pear cv. Kinchaku, resistant Asian pear strain Mamenashi 12 and nonhost European pear cv. Flemish Beauty leaves, to understand the nature of the resistance mechanism. After cuticle penetration by the pathogen, the hyphae were observed at lower frequency in epidermal pectin layers and middle lamellae of leaves of the three resistant plants than in those of susceptible ones. This result suggested that fungal growth was suppressed in the incompatible interaction between pear and V. nashicola race 1. In the pectin layers of all inoculated plants, some hyphae had modifications such as breaks in the plasmalemma with plasmolysis, necrotic cytoplasm and degraded cell walls. More hyphae had collapsed in the leaves of the three resistant plants than in those of the susceptible cv. Kousui. In collapsed hyphae, the polymerized cell walls broke into numerous fibrous and amorphous pieces, showing that the scab resistance might be associated with cell wall-degrading enzymes from pear plants.     

Neutrophilic nodules in the intestinal walls of Japanese monkeys associated with the neutrophil chemotactic activity of larval extracts and secretions of Oesophagostomum aculeatum

High neutrophil chemotactic activity was detected in the culture medium from Oesophagostomum aculeatum larvae in vitro using blind-well chambers with Millipore filters, and guinea pig leucocytes as indicator cells. Neutrophil chemotactic activity was also detected in the extract from larval worms in a dose dependent fashion. This activity was detected in the low molecular weight fractions adjacent to a sodium chloride marker by gel filtration on Sephadex G200. These results were further confirmed with monkey neutrophils. The possible role of this activity in the formation of granulomatous lesions rich in neutrophils found in O aculeatum infections in the Japanese monkey is discussed.     

Cultivation of shiitake in sugi wood meal II: effects of seasoning treatment for wood meal on mycelial growth

The effects of seasoning treatment for fresh sugi wood meal on the mycelial growth of shiitake was investi-gated. The mycelial growth of shiitake in sugi wood meal increased to the same level as that in extract-free sugi wood meal on the 12th day and to approximately 70% of that in konara wood meal on the 28th day of the seasoning treatment. A drastic decrease in extracts of fresh sugi wood meal occurred during the early stage of treatment, and the neutral fraction of methanol extracts decreased to less than 2%. The drainage of sugi wood meal media significantly increased during the later stage of treatment. No changes in the chemical components of sugi wood meal were detected during the seasoning treatment. The mycelial growth of shiitake in the fully seasoned sugi wood meal, which was prepared for commercial mushroom cultivation, was at almost the same level as that in the extract-free sugi wood meal. All of the seasoned sugi wood meals contained a neutral fraction of less than 1%. These results suggested that seasoning treatment can promote mycelial growth of shiitake in sugi wood meal by eliminating inhibitors and improving the physical properties of sugi wood.Part of this paper was presented at the 49th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999     

Somatic embryo culture for propagation,artificial seed production,and conservation of sawara cypress (<Emphasis Type="Italic">Chamaecyparis pisifera</Emphasis> Sieb. et Zucc.)

 Somatic embryogenesis in Chamaecyparis pisifera Sieb. et Zucc. was initiated from immature seeds collected from the end of June to early July. Mass propagation through adventitious shoot bud production from somatic embryo culture on Woody Plant (WP) medium and artificial seed production using sodium alginate was achieved. A high bud forming index value (25.8) was obtained on medium supplemented with 1 μM 6-benzylaminopurine. The conversion rates from artificial seeds under aseptic and nonaseptic conditions were 60%–100% and 10%–12%, respectively. For germplasm conservation, somatic embryos and embryogenic cells were successfully stored at 4°C (medium-term storage) and in liquid nitrogen for long-term storage. Received: December 21, 2001 / Accepted: August 1, 2002 Acknowledgments This work was supported in part by the Japan Science and Technology Corporation and in part by a Grant for Research for the Future Program from the Japan Society for the Promotion of Science. Correspondence to:E. Maruyama     

Liquid culture and transformation of hinoki cypress (Chamaecyparis obtusa Sieb. et Zucc.)

Hinoki cypress (Chamaecyparis obtusa) is one of the most important timber resource forest trees in Japan. Because seed production from a seed orchard of hinoki cypress is not constant every year, micropropagation from a limited amount of material is useful. Up to now, the conventional tissue culture method using solid medium has been used. Here a new method using liquid culture in tubes rotated vertically is described. Shoot primordium of hinoki cypress was inoculated in Campbell and Durzan’s (CD) liquid medium containing different cytokinins (6-benzylaminopurine (BAP), Zeatin, thidiazurone (TDZ)), and the container tubes were rotated vertically around the axis at 2 times / min. Culture room temperature was 25°C and light condition was 16 h photoperiod per day of fluorescent lamps. Zeatin at 1μM concentration was the best for maintaining the shoot primordium production and TDZ induced callus on the surface of the shoot primordia. After shoot primordium multiplication in the liquid culture, they were transplanted to agar medium for shoot elongation. A high concentration of agar (up to 16 g/L) or AVF (anti vitrification factor from Dr. Nairn, 1995) was effective to prevent vitrification of the shoots. Transformation of shoot primordium was done using particle bombardment with vectors containingβ-glucuronidase (GUS) gene or herbicide resistance gene (bar). Positive result for transient transformation was observed with the histo-chemical study for transformation with GUS. Integration of a useful herbicidebar gene into the shoot primordium culture system was also tried and stably transformed plants were obtained. This is the first report of stable transformation of Japanese conifer using practically useful gene. The generous supply of AVF-B from Dr. B.J. Nairn, Tasman Forestry, NZ is also appreciated.     

Assignment of the 1H and 13C NMR spectra of 2-aminobenzamide-labeled galacto- and arabinooligosaccharides

1,4-Linked β-d-galactooligosaccharides with a degree of polymerization (DP) between 1 and 7 and 1,5-linked α-l-arabinooligosaccharides with a DP between 1 and 8 were labeled at their reducing ends with 2-aminobenzamide (2AB) in the presence of sodium cyanoborohydride. The 2AB-labeled oligosaccharides were shown to be homogeneous using high-performance anion-exchange chromatography (HPAEC) and by electrospray ionization mass spectrometry (ESI-MS). The signals in the ¹H and ¹³C nuclear magnetic resonance (NMR) spectra of the 2AB-labeled oligosaccharides were then assigned using one- and two-dimensional NMR spectroscopy. These NMR data will be useful for the structural analysis of enzymatically synthesized galactan and arabinan side chains derived from rhamnogalacturonan I.     

Safety of the tram car for slopes examined by stress analysis of the ground structure

For monorail systems, it is very important to ensure the safety of the ground structure. We developed a tram car for slopes that utilizes the mechanism of the monorail and analyzed the stress of the ground structure that had double rails. First, we divided the ground structure into seven components, and formulated a procedure for calculating the stress of the ground structure. As a result, the stress that acts upon the main rail and the main rail connecting part was near the allowable stress, and each calculated value was within the allowable stress. We measured the stress by running the vehicle, and compared the measured values with the calculated values. As a result, some measured values were larger than the calculated values. However, the ratio of measured value to allowable stress was 90% or less. Finally, we calculated the static loading coefficient for each part, results of which were as follows: main and subrail: 1.1; rail connection part and prop foundation part: 1.0; rail installation part: 1.3. In the recalculations, it became clear that the recalculated values exceeded the allowable stresses. These results suggest that we need to review the ground structure materials.     

Productivity of kayu putih (Melaleuca leucadendron LINN) tree plantation managed in non-timber forest production systems in Java,Indonesia

Kayu putih oil, extracted from the kayu putih tree (Melaleuca leucadendron), is an important non-timber forest product (NTFP) in Indonesia. We compared productivity of kayu putih tree plantations among three different sites in East, Central, and West Java, Indonesia. Leaf-twig, and branch production per tree differed significantly among the three sites. Tree biomass production was highest at the site in West Java practicing tumpangsari (a form of shifting cultivation) of kayu putih with rice (Oryza sativa) on alluvial soils. Tree biomass production was lowest at the site in East Java, practicing tumpangsari of kayu putih with maize (Zea mays) and cassava (Mani- hot esculenta). At this site, tree biomass production increased with increasing stand age and low production was compensated by greater biomass yield of companion crops. Intermediate tree biomass production was observed at the site in Central Java where intercropping is only practiced for the first two years after plantation establishment. Production of tree biomass and kayu putih oil did not increase with increasing stand density, indicating that at a given site, high tree stocking did not lead to high production. Stand-level productivity of the plantations increased with increasing stand age and maximum productivity was attained at 15 to 35 years, after which productivity declined. This suggested unsustainable stand productivity as plantations increase in age.     

Lignan production inDaphne odora cell cultures

Lignan production in callus and cell suspension cultures ofDaphne odora is reported for the first time. The cell suspension culture produced pinoresinol, lariciresinol, secoisolariciresinol, matairesinol, and wikstromol. The production of matairesinol in the cell suspension culture was much higher than that inDaphne odora stem tissues.Part of this report was presented at the 51th annual meeting of the Japan Wood Research Society, Tokyo, April 2001     

New lignan, isoactifolin, fromChamaecyparis obtusa cv. Breviramea

A new lignan isoactifolin was isolated from young shoots (with leaves) ofChamaecyparis obtusa cv. Breviramea. The structure of the compound was determined based on spectroscopic evidence.Parts of this report were presented at the 44th lignin symposium, Gifu, October 1999