<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

At5g54160 gene encodes <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> 5-hydroxyconiferaldehyde <Emphasis Type="Italic">O</Emphasis>-methyltransferase

The function of an Arabidopsis thaliana gene, At5g54160 annotated as a caffeic acid O-methyltransferase CAOMT gene was characterized. The recombinant enzyme of this gene (AtOMT1) catalyzed the O-methylation of phenylpropanoid and flavonoid substrates. The specificity constants (k _cat/K _m) for 5-hydroxyconiferaldehyde (5-HCAld) and quercetin were both 0.11 μM⁻¹·min⁻¹. On the other hand, lignins of At5g54160-knockout Arabidopsis mutants lacked syringyl units. In addition, we showed that the gene silencing also resulted in significant accumulation of caffeyl alcohol (CaAlc). These results strongly suggested that At5g54160 gene is involved in syringyl lignin synthesis for the methylation of both 5-hydroxyconiferaldehyde and 3,4-dihydroxyphenyl compound(s). Part of this work was presented at the Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, March 24–27, 2007     

<Emphasis Type="Italic">Bursaphelenchus mucronatus</Emphasis> (Nematoda: Aphelenchoididae) vectored by <Emphasis Type="Italic">Monochamus </Emphasis><Emphasis Type="Italic">urussovi</Emphasis> (Coleoptera: Cerambycidae) in Hokkaido,Japan

Bursaphelenchus mucronatus Mamiya et Enda has been recovered for the first time from adults of the cerambycid beetle, Monochamus urussovi (Fischer), in Hokkaido, Japan. The nematode was also recovered from the inner bark of Picea jezoensis (Siebold et Zuccarini) Carrière and Abies sachalinensis (Fr Schmidt) Masters infested with M. urussovi larvae. PCR–RFLP analysis indicated that B. mucronatus in Hokkaido is the European type.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Cultivation of <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> on umbrella plant (<Emphasis Type="Italic">Cyperus alternifolius</Emphasis>) substrate

Mycelial growth and mushroom yields of three strains of Pleurotus eryngii produced on wheat bran-supplemented umbrella plant (Cyperus alternifolius) substrate were assessed using surface brightness, bromophenol blue color reactions, ergosterol and glucosamine contents, and water potential as indicators of strain performance. Mycelial growth was 31%–46% greater, depending on strain, on the umbrella plant substrate compared with the mushroom industry standard sugi (Cryptomeria japonica) substrate. Mushroom yields on the first flush were 20%–23% higher, depending on strain, on the plastic bottle-contained umbrella plant substrate. However, yields on the second break were lower from the umbrella plant substrate. Because many growers in Japan only harvest one flush, production of P. eryngii on umbrella plant substrate may offer commercial producers an alternative basal ingredient to diminishing supplies of sugi sawdust.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002.     

Effects of side-chain hydroxyl groups on pyrolytic <Emphasis Type="Italic">β</Emphasis>-ether cleavage of phenolic lignin model dimer

Effects of side chain hydroxyl groups on pyrolytic β-ether cleavage of phenolic model dimers were studied with various deoxygenated dimers under pyrolysis conditions of N₂/400°C/1 min. Although phenolic dimer with hydroxyl groups at the C _α ⁻ and C _γ ⁻positions was much more reactive than the corresponding nonphenolic type, deoxygenation at the C _γ -position substantially reduced the reactivity up to the level of the nonphenolic type. These results are discussed with the cleavage mechanism via quinone methide intermediate formation, which is activated through intramolecular hydrogen bonds between C _α ⁻ and C _γ ⁻ hydroxyl groups.     

Recessive inheritance of morphological characteristics of utsukushimatsu,<Emphasis Type="Italic">Pinus densiflora</Emphasis> f.<Emphasis Type="Italic">umbraculifera</Emphasis>

Pinus densiflora f.umbraculifera, commonly known as utsukushimatsu, is a distinctively shaped form of Japanese red pine whose growth is restricted to a forest stand in Shiga Prefecture, Japan. The inheritance mode of morphological characteristics of utsukushimatsu was studied to preserve the genetic resource of this pine. As previously reported, F₁ trees grown from open-pollinated seeds harvested from trees inhabiting the native stand showed two phenotypes: one resembling utsukushimatsu, which produces multiple trunks, and the other resembling normalP. densiflora, which produces one or a few trunks. In the present study, controlled pollination was carried out using F₁ and normalP. densiflora trees. Segregation ratios of the two phenotypes observed in the F₂ population showed that the morphological characteristics of utsukushimatsu are inherited recessively. This suggests that the mutation of one gene or a few closely linked genes controls the morphological characteristics of utsukushimatsu. Since multiple trunk formation of utsukushimatsu might be related to a loss of lateral bud inhibition, it follows that a simple gene mutation breaks apical dominance inP. densiflora.     

Structural changes in lignin of tropical woods during digestion by termite, <Emphasis Type="Italic">Cryptotermes brevis</Emphasis>

Wood samples of apitong (Dipterocarpus grandiflorua) and ilang-ilang (Ilang-Ilang C. dadloyi) and feces of termites [Cryptotermes brevis (Walker)] fed on these woods were collected from University of the Philippines, Los Baňos. Lignin of each sample was isolated by Björkman’s procedure. There was no significant difference in ¹H nuclear magnetic resonance (NMR) spectra or in the methoxyl content between Björkman lignins from original woods and termite feces. Differences were detected in the contents of aliphatic and unconjugated phenolic hydroxyl groups, suggesting minor structural changes of lignin during digestion by termites. In addition, the ratio of syringyl to guaiacyl nuclei of Björkman lignin from termite feces determined by ¹H NMR spectra was higher than those from the original woods. The molar ratio of syringyl to guaiacyl nuclei of termite feces was higher than those from the original woods as determined by alkaline nitrobenzene oxidation. These results suggest that the structural changes of lignin in the termite gut are due to the insignificant formation of C-C linkages in guaiacyl nuclei. It was concluded that there were minor changes in the structural features of lignin under mostly anaerobic conditions, in contrast to the significant changes that occur through biological modification under aerobic conditions.     

Radial variations of wood properties of an endangered species, <Emphasis Type="Italic">Pinus armandii</Emphasis> var. <Emphasis Type="Italic">amamiana</Emphasis>

A dead tree of Pinus armandii Franch. var. amamiana (Koidz.) Hatusima (abbreviated to PAAm) was obtained from a natural habitat on Tanega-shima Island and various properties of its wood were investigated. Grain angle was measured and soft X-ray analysis was undertaken to obtain the density in each annual ring. Unit shrinkage and dynamic properties were measured by shrinkage, bending, and compression tests. Variations of wood properties in the radial direction, relationships of wood properties to density, and annual ring width were examined. Roughly speaking, variations in the radial direction of the grain angle, twist angle by drying, Young’s modulus and strength in static bending, absorbed energy in impact bending, compressive Young’s modulus, compressive strength, and compressive proportional limit corresponded to the variation of annual ring width. As a result, it was determined that if PAAm is afforested artificially for the purposes of lumber production and conservation, the annual rings of logs should not be too widely spaced. Wood properties of PAAm were similar to those of Japanese black pine (Pinus thunbergii Parl.), which is another representative pine on Tanegashima Island. This study was presented in part at the 56th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007     

The monomer composition controls the Σ<Emphasis Type="Italic">β-O</Emphasis>-4/Σ<Emphasis Type="Italic">O</Emphasis>-4 end monomer ratio of the linear lignin fraction

Lignins are cell wall phenolic heteropolymers that result from the oxidative coupling of three monolignols bearing p-coumaryl (H), coniferyl (G), and sinapyl (S) units, in a reaction mediated by peroxidases. Here, we report the existence of a relationship between the Σβ-O-4/ΣO-4 end monomer ratio of the linear lignin fraction, released through the specific cleavage of the alkyl ether linkages by thioacidolysis, and the G/S ratio of lignins, when this was estimated in differentially evolved vascular land plants. Most importantly, in the case of angiosperms, Gnetales, and lycopods, the Σβ-O-4/ΣO-4 end monomer ratio was apparently predictable from the proportions at which the G and S units were mixed. In the case of G lignins (present in basal gymnosperms and ferns), the Σβ-O-4/ΣO-4 end monomer ratio decayed exponentially to increase the O-4-linked dihydroconiferyl alcohol (DHCA) content. The results obtained suggest that the Σβ-O-4/ΣO-4 end monomer ratio of the linear lignin fraction depends intimately on the lignin monomer composition, and, therefore, on the chemical nature of the radicals derived from three monolignols (coniferyl, dihydroconiferyl, and sinapyl alcohols), whose gain have been finely tuned during land plant evolution.     

Aphidicidal activity of some indigenous plant extracts against bean aphid <Emphasis Type="Italic">Aphis </Emphasis><Emphasis Type="Italic">craccivora</Emphasis> Koch (Homoptera: Aphididae)

Aphidicidal activity of hot and cold water extracts of some indigenous plants, Azadirachta indica A. Juss (neem), Calotropis procera (Aiton) W.T. Aiton (akanda), Polygonum hydropiper L. (biskatali) and Ipomoea sepiaria J. Koenig ex Roxb. (bankalmi), were tested against the bean aphid, Aphis craccivora Koch. Hot water extract of P. hydropiper and A. indica was found to be the most effective (87.6–94.5 and 80.47–89.6% mortality respectively, P < 0.01) among all the extracts. Other hot and cold water extracts also appeared to be useful (59.5–87.5% mortality) as pesticides for A. craccivora. The highest yield (3.25 kg per plant) was obtained using hot water extract of P. hydropiper followed by hot water extract of A. indica (3.15 kg per plant). The lowest yield (0.32 kg per plant) was recorded from the control block. All the phytoproduct treatments had significantly (P < 0.01) better yield than the control block.     

Laboratory and field evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for controlling Mulberry whitefly <Emphasis Type="Italic">Pealius mori</Emphasis> Takahashi (Homoptera: Aleyrodidae) in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> Linn)

In a laboratory screening of 12 isolates of entomopathogenic fungi against nymphs of the mulberry whitefly (Pealius mori Takahashi), Beauveria bassiana (Balsamo-Crivelli) Vuillemin CKB-048 was the most virulent, causing 87 ± 3% mortality at 1 × 10⁶ conidia/ml. Infection was confirmed by growth of the fungus from cadavers and by scanning electron microscopy of treated nymphs. Beauveria bassiana CKB-048 was formulated as a wettable powder (1 × 10⁹ conidia/g) and tested in two mulberry (Morus alba Linn) plantations in central and northeastern Thailand. In both locations, two spray applications of B. bassiana CKB-048 at 3.75 × 10¹² to 6.25 × 10¹² conidia/ha and at 14 day intervals provided good control of whitefly nymphs; control with B. bassiana CKB-048 was comparable to that with the pesticide buprofezin at 250 g of active ingredient/ha. In addition, no mortality of silkworm larvae occurred when the larvae were fed with mulberry leaves sprayed with B. bassiana CKB-048 7, 14, or 21 days earlier.     

Effectiveness of Neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) insecticides against Brassica pod midge (<Emphasis Type="Italic">Dasineura brassicae</Emphasis> Winn<Emphasis Type="Italic">.</Emphasis>)

The efficiency of the botanical insecticide (BI) NeemAzal T/S (containing 1% Azadirachtin A), on the basis of azadirachtin applied in a dose of 20 g a.i. ha⁻¹ against Brassica pod midge (Dasineura brassicae), has been monitored for 4 years. The biological efficiency of BI was compared with the efficiency of some synthetic insecticides. It was ascertained that BI was very efficient in decreasing the number of damaged oilseed rape pods (ranging from 56.5 to 85.9% compared to untreated plants) and its efficiency was comparable with synthetic insecticides based on Chloronicotinyl (Thiacloprid) and Neonicotinoid (Acetamiprid). BI’s efficiency was, in some years, even significantly higher compared to pyrethroid (λ-cyhalothrin). The high biological efficiency of azadirachtin that we ascertained was significantly reflected in increasing potential crop yields. The yield increase of azadirachtin ranged between 9.3 and 19.4% compared to the control sample. Azadirachtin showed the highest yield for the whole time of experimentation, and in some years the yield increase was significantly higher compared to some synthetic agents.     

Structural characteristics and eco-adaptability of heteromorphic leaves of <Emphasis Type="Italic">Populus euphratica</Emphasis>

The microstructural and ultrastructural traits of three kinds of typical leaves ofPopulus euphratica Olive, including lanceolate, broad-ovate and dentate broad-ovate leaves, were studied by using electron microscope and optical microscope. The resuits showed that with the leaves changing from lanceolate shape to dentate broad-ovate shape, their structure obviously tended to be xeromorph: developed palisade tissue, undeveloped spongy tissue, thick cutin layer and sunken stomas. The amount of mitochondria tended to be increased, and the shape of chloroplasts varied from regular spindle to irregular rotundity or oval. The leaves were covered with wax without cilium, and the stomas on the upper and lower epidermis of the leaves opened unevenly. The stomas on the lower epidermis were deeper than those on the upper epidermis under the scanning electron microscope. The results implied that the structural characteristics of the diversiform-leaves ofP euphratica are related to its eco-adaptability.     

Spatial pattern of beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.) and oak (<Emphasis Type="Italic">Quercus pubescens</Emphasis> Mill.) seedlings in natural pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) woodlands

Spatial pattern of recruitment is an important factor influencing population dynamics of plant communities. The spatial pattern is determined by seed dispersal and by the spatial variability of germination and initial survival. In the process of forest expansion following farmland abandonment, mid- and late-successional species are often dispersed in pioneer forests by birds. This could result in an aggregated spatial pattern of seeds that could influence the dynamics of these species in mixed pioneer forests. In the sub-Mediterranean area, mid- and late-successional species such as Quercus pubescens (downy oak) and Fagus sylvatica (European beech) are expected to replace pioneer Pinus species. Using a point sampling method we demonstrated that beech and oak seedlings (height <2 m) have a clumped distribution in the understorey of pine. This could result from an aggregated dispersal by jays (dispersal effect) or from preferential recruitment in particular habitats (habitat effect). To test these hypotheses we proposed a statistical analysis of spatial patterns of regeneration of beech and oak. Ground cover variables (i.e. cover by rock outcrops, herbs, box shrubs, mosses or pine) did not differ significantly around seedlings as compared with random sample plots. Likewise, clumped seedlings had growth similar to isolated seedlings, thus refuting the hypothesis of preferential location in the most favourable microsites. Aggregated dispersal seems to be involved in the process of regeneration. Since beech and oak seedlings have contrasting ecological demands, we discuss the implication of this pattern for the replacement dynamics of pine by these species.     

Production of <Emphasis Type="Italic">Pinus halepensis</Emphasis> seedlings inoculated with the edible fungus <Emphasis Type="Italic">Lactarius deliciosus</Emphasis> under nursery conditions

Inoculation with edible fungi bestows an added value on mycorrhized plants since production of mushrooms as a secondary crop can be an economically-valuable resource in forestry management. In order to establish a protocol for controlled mycorrhization of Pinus halepensis with the edible fungus Lactarius deliciosus, several factors such as fertilisation (35, 60 or 120 mg total N and 13.5, 27 or 54 mg total P), potting substrate (sphagnum peat or sphagnum peat/vermiculite) and fungal inoculum (mycelial slurry produced in a bioreactor, alginate beads or peat/vermiculite) were assessed. The most effective inoculum was mycelial slurry at a dose of 10 ml/plant. The two potting substrates assessed were compatible with mycorrhiza formation. The greatest number of mycorrhized seedlings was obtained with a moderate level of N (35 mg/plant) or P fertilisation (27 mg/plant). Inoculation did not produce a consistent growth effect on P. halepensis, but seedlings met the quality requirements, making them suitable for planting out.     

Estimation of stand attributes in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> and <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> stands using QuickBird panchromatic data

This study aims to estimate stand density and stand volume in Cryptomeria japonica and Chamaecyparis obtusa stands from high-resolution satellite data and verify the reliability and uncertainty of the data. Sixty circular sample plots of 0.04 ha each were established. Their stand densities were estimated from the number of tree crowns derived from high-resolution satellite data using the watershed method. Stand densities derived from field surveys in the sample plots were compared with those obtained from high-resolution satellite data by stand age class. As a result, there was a positive correlation between them for sample plots of 41 years of age and over (R = 0.82); however, there was no correlation between them for sample plots of 40 years and under. Individual diameters at breast height (DBH) were estimated from crown areas obtained from high-resolution satellite data for the two species. Using the estimated DBH, individual tree heights were predicted from the height–diameter curves. Stand volumes were estimated from the sum of individual volumes, which were derived from volume formulas having two variables, i.e., DBH and height. Stand volumes derived from the field survey were compared with those obtained from high-resolution satellite data. The correlation coefficient between them for stands of 41 years of age and over was 0.78.     

Effect of rhizobial inoculation methods and host plant provenances on nodulation and growth of <Emphasis Type="Italic">Acacia senegal</Emphasis> and <Emphasis Type="Italic">Acacia nilotica</Emphasis>

The purpose of this work was to determine the most efficient methods of inoculation to significantly improve nodulation and growth of Acacia senegal and Acacia nilotica, grown under greenhouse conditions. Our results showed that inoculation using dissolved alginate beads containing rhizobia significantly improved the growth of both acacia species better than the growth of plants in other treatments. The experiment with A. nilotica was conducted in two unsterilised soils from different areas. Plants grown in soil from Bel Air were well-nodulated and showed better growth than plants grown in soil from Sangalkam. However, no difference between these soils was shown between the several methods of inoculation and their effect on the nodulation and growth of plants. An interaction between A. senegal and A. nilotica provenances and the effect of inoculation with rhizobia was also demonstrated. Bel Air provenance of A. senegal, Dahra and RIM provenances of A. nilotica grew best of several provenances tested. These results suggest that (1) it may be possible to improve growth and yield of A. senegal and A. nilotica by careful selection of each symbiotic partner; and (2) nursery-grown seedlings of A. senegal and A. nilotica should be inoculated, just after sowing, with dissolved alginate beads containing a mixture of selected rhizobia. Résumé. Lobject de notre travail était de déterminer la méthodologie dinoculation la plus efficace pour améliorer significativement la nodulation et la croissance dAcacia senegal et dAcacia nilotica poussant dans des conditions de serre. Nos résultats ont montré que linoculation utilisant des billes dalginate dissoutes améliorait significativement la croissance des deux espèces dAcacia par rapport à celle des plantes des autres traitements. Lexpérience avec A. nilotica a été faite dans deux types de sol non stériles. Nous avons observé que les plants poussant dans le sol de Bel Air étaient plus nodulés et mieux développés que ceux poussant dans le sol de Sangalkam. Cependant, aucune différence significative entre les deux sols na été mise en évidence en termes deffet des différentes méthodologies dinoculation sur la nodulation et la croissance des plantes. Il a aussi été démontré lexistence dune interaction entre les provenances dA. senegal et dA. nilotica et leffet de linoculation avec rhizobium. Cest ainsi que les provenances Bel Air dA. senegal, Dahra et RIM dA. nilotica ont le mieux poussé parmi les différentes provenances testées. Lensemble de ces résultats suggèrent: ⁽¹⁾ il devrait être possible daméliorer la croissance et la production dA. senegal et dA. nilotica par une sélection des deux partenaires symbiotiques; ⁽²⁾ Les jeunes plantules dA. senegal et dA. nilotica produites en pépinière devraient être inoculées juste après le semis avec des billes dalginate dissoutes contenant un cocktail de souches sélectionnées.