<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.     

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.     

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.     

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.     

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     

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.     

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.     

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.     

Effect of seed coverings and seed pretreatments on the germination response of <Emphasis Type="Italic">Alnus glutinosa</Emphasis> and <Emphasis Type="Italic">Betula pubescens</Emphasis> seeds

The germination of common alder (Alnus glutinosa (L.) Gaertn.) and downey birch (Betula pubescens Ehrh.) seeds is often poor in bare-root nurseries. The effect of a variety of seed coverings and a few seed pretreatments on seedling emergence was examined in this study in an attempt to address this problem. Seeds of each species were sown in trays containing nursery soil, covered with grit, gravel, sand, combinations of these coverings, a hydromulch or a sealed plastic cloche and then incubated for 6 weeks at 17–20°C. The grit combined with sand or gravel, the hydromulch and the cloche increased seedling emergence when compared with the standard grit. In another experiment, seeds of each species were fully imbibed (FI) (>50% moisture content, MC), as per standard practice, or adjusted to target MC (TMC) (30–35% MC) levels, and then chilled to release dormancy. Some seeds of each MC treatment were primed at 20°C for 2 days following chilling, after which all seeds were evaluated in laboratory tests and a nursery trial. Germination potential of the FI seeds declined in the lab tests by the second test date, which was reflected in low seedling emergence in the nursery in birch. The primed FI seeds of alder germinated most rapidly in the nursery, but other effects were not significant. Seedling emergence was better in the nursery in response to the TMC than the FI pretreatment in birch.

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.     

Toxic effects of annonaceous acetogenins on <Emphasis Type="Italic">Oncopeltus fasciatus</Emphasis>

From an Argentine collection of Annona cherimolia and a Bolivian collection of Annona montana (Annonaceae), the acetogenins, squamocin, molvizarin, itrabin, almuñequin, cherimolin-1, annonacin, annonacin-A, densicomacin-1, cis-annonacin-10-one and murihexocin-A, were obtained to study their toxicity against the cotton pest Oncopeltus fasciatus (Hemiptera: Lygaeidae). Topical ventral application of O. fasciatus nymphs with solutions of the mentioned acetogenins produced acute and delayed mortality.     

Antifeedant activities of secondary metabolites from <Emphasis Type="Italic">Ajuga nipponensis</Emphasis> against adult of striped flea beetles, <Emphasis Type="Italic">Phyllotreta striolata</Emphasis>

Antifeedant activities of the isolated chemical compounds from Ajuga nipponensis, were studied against adult of striped leaf beetles. The methanol, petroleum ether, ethyl acetate and n-butanol extracts at 1.0 mg/ml, were used in this study. All four extracts exhibited more than 65 antifeedant index at 24 h and ethyl acetate extract showed significant activity against striped leaf beetles with 83.12 antifeedant index. Six compounds and one fraction were isolated by chromatography and their structures were identified by NMR, MS and FTIR spectra. At 2.0 mg/ml for 24 h the three compounds 20-hydroxyecdysone, acacetin and apigenin showed considerable activities with antifeedant indexes 59.29, 51.22 and 61.55, respectively. In contrast to this the antifeedant indexes of acacetin and apigenin, were sharply reduced as the time extended and that of 20-hydroxyecdysone remained unchanged. In addition, the synergistic effects of two mixtures of secondary metabolites, were studied and no sharp difference was observed.     

Biological performance of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> leaf oils from Thailand against the subterranean termite <Emphasis Type="Italic">Coptotermes formosanus</Emphasis> Shiraki

The antitermitic activities of leaf oils and their constituents, taken from three clones of Eucalyptus camaldulensis Dehnh. in Thailand, against Coptotermes formosanus Shiraki were investigated in contact and noncontact tests. The termiticidal mechanism was also examined. Antitermitic tests demonstrated that E. camaldulensis leaf oils were both contact toxicants and fumigants to C. formosanus with LC₅₀ values ranging between 12.68 and 17.50 mg/g by the contact method, and between 12.65 and 17.50 mg/petri dish (100 cm³) by the noncontact method. p-Cymene and γ-terpinene were primarily responsible for the contact toxicity and 1,8-cineole was responsible for fumigation. From the investigation of termiticidal mechanism, E. camaldulensis leaf oils exhibited the inhibition of acetylcholinesterase activity and showed the common symptoms of a neurotoxic mode of action against C. formosanus. Part of this report was presented at the 58th Annual Meeting of the Japan Wood Research Society, Tsukuba, March 2008     

Distinction of resin compounds between the healthy bark and the resinous stem canker of <Emphasis Type="Italic">Thujopsis dolabrata</Emphasis> var. <Emphasis Type="Italic">hondae</Emphasis>

Methyl esters of higher fatty acids were detected from the healthy bark of Thujopsis dolabrata var. hondae but not from the bark of the resinous stem canker of T. dolabrata var. hondae. This difference enabled us to distinguish healthy trees from diseased ones. Fourteen diterpenes were also isolated from the n-hexane extracts of the bark-glued resin taken from the resinous stem canker of T. dolabrata var. hondae. Of these diterpenes, abietane diterpenes [abieta-7,13-diene (1), abietinol (2), dehydroabietinol (4)], pimarane diterpene [sandaracopimaric acid (8)], and labdane diterpenes [manool (10), torulosol (11), torulosal (12), cupressic acid (13)] were first isolated from T. dolabrata var. hondae.Part of this paper was presented at the 47th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999