Comparison of terpenes in extracts from the resin and the bark of the resinous stem canker of Chamaecyparis obtusa and Thujopsis dolabrata var. hondae

A monoterpene and 15 diterpenes were isolated from the ethyl acetate extracts of the bark-glued resin from the resinous stem canker ofThujopsis dolabrata var.hondae Makino. A monoterpene (nezukone20) and 4 diterpenes (acetyl torulosol5, acetyl isocupressic acid8, acetyl abietinol11, and 7-methoxytotarol18) were characteristic constituents of the ethyl acetate extracts but were absent in then-hexane extracts from the resinous stem canker ofT. dolabrata var.hondae. These terpenes were first isolated fromT. dolabrata var.hondae andT. dolabrata. The available literature suggests that diterpene18 is a new compound. The resinous stem canker ofChamaecyparis obtusa Endlicher contained larger amounts of manool1,trans-communic acid6, and ferruginol12 and smaller amounts of isocupressic acid7 and abietinol10 than the resinous stem canker ofT. dolabrata var.hondae. The concentration of18 was less than 2% in the extracts, and the resinous stem canker ofC. obtusa lacked this compound. The resinous stem cankers ofC. obtusa andT. dolabrata var.hondae provided extracts 15.6 and 4.96 times, respectively, heavier than the healthy ones. Large differences in the ratios and compositions of terpenes were also observed between the resinous stem canker and the healthy trees. Terpenes isolated from the extracts contained many kinds of diterpene, especially the labdane-type diterpenes, in these diseased trees. These results suggest that the presence of labdane-type diterpenes is closely associated with the resinous stem canker or the causal fungi of this disease.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000     

Constituents from the roots of <Emphasis Type="Italic">Taxus cuspidata</Emphasis>

The known propelargonidin, afzelechin-(48)-afzelechin (1), the known lignans 7-hydroxynortrachelogenin (2), epinortrachelogenin (3), nortrachelogenin (4), hydroxymatairesinol (5), allohydroxymatairesinol (6), matairesinol (7), oxomatairesinol (8), and isotaxiresinol (9), and the known taxoids taxinine M (10), taxayuntin (11), and 10-deacetyltaxol (12), and 10-deacetylbaccatin III (13) were isolated from the roots of Taxus cuspidata (Japanese yew, Taxaceae). The propelargonidin was isolated from Taxus spp. for the first time, and was detected in the roots, bark, and twigs.     

<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.     

<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.     

Phoretic mites of three bark beetles (<Emphasis Type="Italic">Pityokteines</Emphasis> spp.) on Silver fir

The species composition and abundance of phoretic mites of the bark beetles Pityokteines curvidens, P. spinidens, and P. vorontzowi on Silver fir (Abies alba) were investigated in 2003 at two locations (Trakoscan and Litoric) in Croatia. Stem sections and branches from A. alba trees infested by Pityokteines ssp. were collected and incubated in rearing cages. Bark beetles emerging from the stem sections and branches were examined for photetic mites. A total of ten mite species were documented for the first time as associates of Pityokteines spp. on A. alba. These included Dendrolaelaps quadrisetus, Ereynetes scutulis, Histiostoma piceae, Paraleius leontonychus, Pleuronectocelaeno japonica, Proctolaelaps hystricoides, Schizostethus simulatrix, Tarsonemus minimax, Trichouropoda lamellose, and Uroobovella ipidis. T. minimax was the most frequent phoretic mite of all the three scolytines and U. ipidis was also common, whereas, the other mite species occurred less frequently. The species spectrum and relative abundance of mite associates were similar for all three Pityokteines species. Another species, Pleuronectocelaeno barbara was commonly found phoretic on P. curvidens, captured in pheromone traps in 2005 at the location Litoric. Furthermore, two previously collected mite specimens from Switzerland, phoretic on P. curvidens, were identified as Nanacarus sp. and Bonomia sp. The records from Croatia and Switzerland in the present study increase the number of known mite associates of Pityokteines spp. from one previously documented species to 14 species. None of the phoretic mites found in the survey in Croatia appear to have the potential to be used for biological control of Pityokteines spp., although the feeding habits are unknown for many species recorded.     

Extractives of <Emphasis Type="Italic">Quercus crispula</Emphasis> sapwood infected by the pathogenic fungi <Emphasis Type="Italic">Raffaelea quercivora</Emphasis> I: comparison of sapwood extractives from noninfected and infected samples

The extracts of Quercus crispula infected by the ambrosia fungus, Raffaelea quercivora, were investigated. Phenol and tannin analyses indicated that normal sapwood (NS) contained a considerable amount of hydrolysable tannins, while infected colored sapwood (IS) contained less hydrolysable tannins and more phenols than NS. In treating pentagalloyl glucose (PGG), which is a model compound of hydrolysable tannins, with a culture medium of R. quercivora, PGG was rapidly hydrolyzed to produce gallic acid. The resulting gallic acid decreased in concentration over the subsequent cultivation period eventually disappeared. Measuring tannase and laccase activities of the culture medium of R. quercivora, tannase activity increased gradually from the beginning, while laccase activity increased rapidly at 5 days of incubation and disappeared at 8 days. An oxidative product from gallic acid treated with laccase was isolated by preparative high performance liquid chromatography, and was identified as purprogallincarboxylic acid (PGCA) by nuclear magnetic resonance spectroscopy and electron-impact mass spectrometry. PGCA was present in a 70% aqueous acetone extract of IS, and showed slight growth inhibition against R. quercivora. Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, 2007     

Heartwood extractives from the Amazonian trees <Emphasis Type="Italic">Dipteryx odorata,Hymenaea courbaril</Emphasis>, and <Emphasis Type="Italic">Astronium lecointei</Emphasis> and their antioxidant activities

Heartwood extracts from Amazonian trees cumaru-ferro (Dipteryx odorata), jatoba (Hymenaea courbaril), and guarita (Astronium lecointei) exhibit antioxidant activities comparable with that of α-tocopherol, a well-known antioxidant. This article reports the characterization of the antioxidant compounds in the extracts of the three heartwoods. Silica gel column chromatography of the cumaru-ferro EtOAc extract yielded (−)-(3R)-7,2′,3′-trihydroxy-4′-methoxyisoflavan and (+)-(3R)-8,2′,3′-trihydroxy-7,4′-dimethoxyisoflavan. Silica gel column chromatography followed by preparative high-performance liquid chromatography of the jatoba EtOAc extract yielded (−)-fisetinidol and (+)-trans-taxifolin. Chemical structures were assigned using electron-ionization mass spectrometry, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy including nuclear Overhauser effect spectroscopy (NOESY), as well as optical rotation and circular dichroism. Gas chromatography-mass spectrometry demonstrated that the isolated compounds were predominant in the EtOAc extracts. In the guarita EtOAc extract, catechin and gallic acid were identified by comparing their retention times and mass fragmentation patterns with those of authentic samples. Antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl assay demonstrated that all these compounds had activities comparable with that of α-tocopherol. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007     

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     

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.     

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.     

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.     

Performance of two <Emphasis Type="Italic">Trichogramma</Emphasis><Emphasis Type="Italic">brassicae</Emphasis> strains under greenhouse and field conditions for biocontrol of the silver Y moth in spinach cultures

The suitability of Trichogramma brassicae Bezd. (Hymenoptera: Trichogrammatidae) to control the silver Y moth, Autographa gamma (L.) (Lepidoptera: Noctuidae) in spinach was investigated under greenhouse and field conditions. Two strains of T. brassicae were selected to study host searching efficiency and dispersal ability of the wasps on spinach. The experiments were conducted with defined release densities. The results show that T. brassicae strain I failed to locate host eggs in all experiments. In contrast, T. brassicae strain II females were able to locate 37% of the exposed egg clusters in the greenhouse within 48 h. Individuals colonised rapidly at least an area of 0.25 m². At distances up to 26 cm to the release point of the T. brassicae females, parasitization rate was 100%. Moreover, under field conditions silver Y moth eggs were exposed two times for 48 h on 12 experimental plots of 100 m² in two spinach fields. During the first exposure interval, i.e. 4 days after mass release of T. brassicae strain II, overall parasitization rates reached 20% on both fields. Furthermore parasitization rates still reached 16 and 19% during the second exposure interval, i.e. 9 days after natural enemy release, indicating a constant Trichogramma activity over time. Parasitization rates were highest in the close vicinity of the release point, i.e. 86%, whereas 6.5 m away the mean parasitization rate decreased to 3%. In conclusion results indicate a high potential of T. brassicae strain II as antagonist of A. gamma. Nevertheless the dispersal ability of T. brassicae strain II was limited, but adjustment of release densities and techniques might compensate this disadvantage.     

Structure identification for compound I separated and purified fromtaxoids-produced endophytic fungi （Alternaria. alternata var. taxi 1011）

Endophytic fungi are widely found in almost all kinds of plants. Many endophytic fungi can produce some physio-logical active compounds, which are same to or analog to those isolated from their hosts. Producing physiological active compounds through microbial fermentation can give a new way to resolve resource limitation and to find out alternative source.Through the methods of organic solvent extraction, thin layer chromatography (TLC) and column chromatography, compound I was isolated, purified from the liquid fermentation metabolites of the taxoids-produced endophytic fungi (Altemaria. altemata var.taxi 1011 Y. Xiang et LU An-guo) that was screened from the bark of Taxus. cuspidata Sieb.et Zucc.. Compound I was identified as one kind of taxoids type Ⅲ, based on the analyzing results by using the methods of ultraviolet spectroscopy (UV), infrared spectroscopy (IR), mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR). This study provides a completed method for separation and purification of the endophytic fungi as well as structure identification of its fermentation metabolite     

A new diterpene dimer from the bark ofChamaecyparis obtusa

This paper describes characterization of diterpenes of the bark of Japanese cypress,Chamaecyparis obtusa (S. and Z.) Endl, without the resinous stem canker to learn the difference between the cypress bark affected with the canker. A diterpene dimer and two diterpenes, 6,12-dihydroxyabieta-8,11,13-trien-7-one and 6,12-dihydroxyabieta-5,8,11, 13-tetraen-7-one, were firstly isolated from Japanese cypress. The dimer, 6-(abieta-6,8,11, 13-tetraenyl-12-oxy)-7-methoxyabieta-8,11,13-trien-12-ol, was a new compound. It is a terpene indicative ofC. obtusa not infected with the resinous stem canker. Five known diterpenes were also isolated.     

Isolation of phenylisoserine methyl ester from the leaves of Taxus cuspidata var. nana

From methanolic extracts of leaves of kyaraboku, Taxus cuspidata var. nana, phenylisoserine methyl ester (3) was isolated along with taxinine (1), taxol (2), sciadopitysin (4), ginkgetin (5), isorhamnetin (6), and quercetin (7). This is the first time that phenylisoserine methyl ester has been isolated from T. cuspidata var. nana. Compound 3 was also isolated from the ethanolic extracts of leaves of T. cuspidata var. nana. Furthermore, compound 3 was identified in methanolic extracts from the bark of this tree.     

Antioxidant compounds from <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> biomass by subcritical liquid water extraction

The effectiveness of subcritical liquid water processing for extraction of antioxidants from the leaves of Eucalyptus grandis biomass was evaluated by determining the capability of the processed samples to scavenge peroxynitrite radicals in vitro, as compared with the extracts obtained by conventional extraction. Pyrogallol (1), 5-hydroxymethyl-2-furaldehyde (2), and 4,4,6,6-tetramethyl-3,5-dioxo-cyclohex-1-enecarboxylic acid (3) were identified as the major products obtained from the subcritical liquid water extracts. Among these, pyrogallol (1) exhibited stronger antioxidant activity than butylated hydroxytoluene, which was used as a standard for the antioxidant activity measurement. This research was presented at the conference of the Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA) in Tokyo, March 2007.     

Interactions between<Emphasis Type="Italic"> Orthotomicus erosus</Emphasis> (Woll.) (Col., Scolytidae) and the Argentine ant<Emphasis Type="Italic"> Linepithema humile</Emphasis> (Mayr) (Hym., Formicidae)

Interactions occurring between Orthotomicus erosus, an economically important pine bark beetle in Portugal, and the aggressive Argentine ant Linepithema humile were investigated. Tri-trophic experimental enclosures consisting of Pinus pinaster logs, O. erosus adults and L. humile workers were set up in the field and in the laboratory. Interactions between ants and scolytids were observed and quantified in terms of the bark beetles success in colonizing the logs. The presence of L. humile did not influence the ability of O. erosus to colonize the logs, apparently due to the difficulties met by the ants in handling the bark beetles and to their inability to injure them. Thus, although abundant in many pine stands in Portugal and in spite of its aggressiveness, L. humile does not seem to affect populations of O. erosus.     

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     

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     

Inhibitory effects of <Emphasis Type="SmallCaps">l</Emphasis>-pipecolic acid from the edible mushroom, <Emphasis Type="Italic">Sarcodon aspratus</Emphasis>, on angiotensin I-converting enzyme

The aqueous extract of the edible mushroom Sarcodon aspratus showed inhibitory effects against angiotensin I-converting enzyme (ACE). l-Pipecolic acid (l-piperidine-2-carboxylic acid) was isolated from a hot-water extract in a 0.02% yield as an active principle. The mode of inhibition of l-pipecolic acid was found to be competitive, whereas its d-isomer showed no significant inhibitory effects against ACE, suggesting that the configuration of the carboxyl group in the molecule plays an important role in the enzyme inhibition.