Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

Electroencephalogram of Japanese Black calves affected with cerebrocortical necrosis

Electroencephalogram (EEG) examinations were carried out on three Japanese Black calves (1, 2 and 14 week old) affected with cerebrocortical necrosis (CCN) in order to investigate the relationship between EEG abnormalities and encephalic lesions. Some neurologic signs such as circling, astasia, convulsion and opisthotonus appeared in two of the calves (case 1 and 3). The other calf (case 2), however, showed no neurologic signs. The EEGs obtained from the two calves (case 1 and 2), in the awake state, showed constant high amplitude (50-160 microV) and slow (1-4 Hz) activity (HASA). One of these (case 1) had necrosis mainly in the right hemisphere and showed a markedly asymmetric sharp wave. On the other hand, in case 3, the EEG showed diffuse lowered activity, and almost flat in some leads. A decrease in fast activity observed in common with all three animals. These findings suggest that one of the characteristic EEG patterns of CCN patient may be HASA and decreased fast activity. Histopathologic diagnoses of the calves were CCN in various degrees. The degrees of abnormality of the EEGs seemed to accord with the seriousness of the lesions. Because the EEG obtained from the calf having no obvious neurologic signs showed an abnormal pattern, it was considered that EEG examinations in the early stages of disorders are valuable to diagnose CCN.     

Biosynthesis of a syringyl 8-O-4′ neolignan in Eucommia ulmoides: formation of syringylglycerol-8-O-4′-(sinapyl alcohol) ether from sinapyl alcohol

To investigate the biosynthesis and stereochemistry of syringylglycerol-8-O-4′-(sinapyl alcohol) ether (SGSE), a syringyl 8-O-4′ neolignan, feeding experiments and enzyme assays using Eucommia ulmoides were carried out. Diastereoselective formation of erythro-SGSE was found. When [8-¹⁴C]sinapyl alcohol was administered to excised shoots of E. ulmoides, ¹⁴C was incorporated into free SGSE and SGSE glucosides. In stems, incorporation into (+)-erythro-[¹⁴C]SGSE (0.037%) with 9.1% enantiomeric excess (% e.e.) was found; incorporation into the threo isomer was not detectable. Erythro-[¹⁴C]SGSE glucosides (0.047%) dominated over threo forms (0.007%) with 74.0% diastereomeric excess (% d.e.); both diastereomers were levorotatory with 32.0% e.e. and 18.3% e.e., respectively. In leaves, higher incorporation into (−)-erythro-[¹⁴C]SGSE (0.500%, 15.9% e.e.) than into the threo isomer (0.206%, 7.4% e.e.) was observed (41.6% d.e.). (−)-Erythro-[¹⁴C]SGSE glucosides (1.692%, 25.0% e.e.) were produced at higher rates than threo isomers (0.177%, 16.4% e.e.) with 81.0% d.e. In incubations of a mixture of [8-¹⁴C]sinapyl and [8-¹⁴C]coniferyl alcohols with an insoluble enzyme preparation from stems of E. ulmoides, erythro-SGSE was preferentially produced. The highest % d.e. (82.8) was observed at 60 min with the (+)-erythro isomer (21.4% e.e.) and the (−)-threo form (4.3% e.e.).Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002, and the 47th Lignin Symposium, Fukuoka, October 2002     

Assessing raindrop impact energy at the forest floor in a mature Japanese cypress plantation using continuous raindrop-sizing instruments

The raindrop size distribution of throughfall and open rainfall was monitored continuously during a rainfall event using laser raindrop-sizing instruments (LD gauges), in order to calculate the raindrop impact energy in a plantation of mature Japanese cypress (Chamaecyparis obtusa), where surface erosion at the forest floor had been a problem. Data from two rainfall events were analyzed. The LD gauges recorded qualitative raindrop size distribution, and the capture rate during each rainfall event was used to manipulate raindrop data quantitatively. Throughfall and open rainfall comparisons revealed several important differences. First, throughfall raindrops were fewer in number and larger in size than open rainfall drops. In one rainfall event, for example, throughfall raindrops were less than one-fifth as frequent as open rainfall raindrops; in addition, the maximum throughfall raindrop diameter was 6.35mm compared to 3.31mm for open rainfall raindrops. Second, throughfall raindrops that were larger than the largest open rainfall raindrops comprised 63.8% of the throughfall precipitation by volume. Third, total raindrop impact energy from throughfall was over twice that of open rainfall. Moreover, comparison of throughfall events implied that throughfall raindrops did not always have a uniform distribution between different events or among different periods of time in one rainfall event, in contrast to findings in previous studies which showed that throughfall raindrops had a uniform size distribution independent of rainfall intensity. It is possible that an abrupt transition of throughfall intensity from low to high changes the distribution of throughfall raindrops.     

Relationship between fracture toughness and acoustic emission during cleavage failure in adhesive joints

Summary Fracture toughness in mode I was determined for wood bonded with various adhesive systems. Tapered double-cantilever beam (TDCB) specimens were used, with attached transducer connected to instrumentation for monitoring acoustic emission. Adhesives used fro bonding the TDCB specimens included urea-formaldehyde resin, phenol-resorcinol resin, and emulsion polymer isocyanate, among others. Test parameters measured included critical load for cleavage fracture and number of events (stress waves) emitted, from which fracture toughness and cumulative count of acoustic emission were computed. It was found that there was a well defined, linear relation between fracture toughness and cumulative counts of acoustic emission per fracture area. The fracture toughness values of systems containing polyvinyl acetate and of epoxy resin were larger than those of thermosetting resins such as urea-or phenol formaldehyde. The high fracture toughness imparted by polyvinyl acetate was attributed to viscous movement of linear molecular chain segments, as compared with the more brittle behavior of the crosslinked molecular structure of such resins as ureaformaldehyde.     

Vertical profiles of environmental factors within tropical rainforest, Lambir Hills National Park, Sarawak, Malaysia

Environmental factors, such as global solar radiation, wind speed, air temperature, humidity, and CO₂ concentration, were measured above and within the canopy of a tropical rainforest in Lambir Hills National Park, Sarawak, Malaysia. Few data concerning the environment of this forest have been reported. Intensive observations were carried out in 1998, 1999, and 2000 with the following results: (1) The fraction of global solar radiation reaching the upper layer of the canopy varied with global solar radiation above the canopy. Even though the global solar radiation above the canopy fluctuated, the fraction of that reaching the lower canopy and the ground was constantly approximately 5%. (2) The fraction of wind speed reaching each layer of the canopy increased with wind speed above the canopy. Little wind was usually present at the lower canopy. (3) The daytime air temperature at the canopy top was higher than that near the ground. The maximum difference between the air temperature at the canopy top and that at the ground was about 5°C, and the diurnal temperature ranges at the canopy top and those at the ground were about 8°C and about 5°C, respectively. The highest daytime water vapor pressure occurred within the canopy and particularly near the ground. Vertical gradients of water vapor pressure during the day were steep, probably because of high transpiration. (4) In the 1998 observation the minimum and the maximum CO₂ concentrations were 360 ppm in the day and 450 ppm at night, while in the 2000 observation the minimum and the maximum CO₂ concentrations were 350 ppm in the day and 540 ppm at night. The higher CO₂ concentration in the daytime and the lower concentration at night observed during the 1998 observation period were probably due to reduced photosynthesis and soil respiration caused by exceptional dry conditions during the observation period.     

Stereochemistry and biosynthesis of 8-O-4′ neolignans in Eucommia ulmoides: diastereoselective formation of guaiacylglycerol-8-O-4′-(sinapyl alcohol) ether

Stereochemistry and biosynthesis of guaiacylglycerol-8-O-4′-(sinapyl alcohol) ether (GGSE), an 8-O-4′ neolignan, which consists of coniferyl and sinapyl alcohol moieties, in Eucommia ulmoides were investigated. Four 8-O-4′ neolignans, GGSE, syringylglycerol-8-O-4′-(coniferyl alcohol) ether (SGCE), guaiacylglycerol-8-O-4′-(coniferyl alcohol) ether (GGCE), and syringylglycerol-8-O-4′-(sinapyl alcohol) ether (SGSE), were synthesized. Their erythro and threo diastereomers were separated through acetonide derivatives, intermediates of the synthesis, and identified by means of nuclear magnetic resonance (NMR) spectroscopy. All of the erythro-acetonide derivatives have larger coupling constants (ca 9 Hz) for the C₇-H resonances than those of the threo ones (1.5–2 Hz). In the case of the four 8-O-4′ neolignans, the C₇-H coupling constants of the threo-isomers are not smaller than those of the erythro ones. GGSE isolated previously from this plant was identified as the erythro isomer by comparison of the ¹³C-NMR data with synthetic erythro-GGSE and threo-GGSE and the other 8-O-4′ neolignans mentioned as above. Administration of a mixture of [8-¹⁴C]coniferyl alcohol and [8-¹⁴C]sinapyl alcohol to excised shoots of E. ulmoides was carried out and the incorporation of ¹⁴C into erythro-[¹⁴C]GGSE was found to be higher than that in threo-[¹⁴C]GGSE. The occurrence of diastereoselective formation of erythro-GGSE by cross coupling of coniferyl and sinapyl alcohols is suggested.Part of this paper was presented at the 47th Lignin Symposium, Fukuoka, October 2002 and the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, April 2003     

Effect of particle shape on linear expansion of particleboard

Hinoki (Japanese cypress) strand with 0.6mm thickness was hammer-milled to prepare three types of particles that had the same thickness but different lengths and widths. Screen analysis and image analysis were conducted to evaluate the shape and distribution of the particle dimensions. Laboratory-scale particleboards were fabricated at three density levels using these particles. To determine the true effect of particle shape on the linear expansion (LE) of the board, these boards exhibited almost the same temperature behavior during pressing, the same density profile, and the same bending properties at each board density level. LE at 40°C and 90% relative humidity of the board was found to be affected by the particle shape. The board composed of small particles showed a larger LE at the same density level. It was considered that the out-of-plane orientation angle of the particles affected the LE of the boards. Thickness swelling and internal bond strength were also affected by the particle shape.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000     

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