Two-year dynamics of foliage labelling in 8-year-old <Emphasis Type="Italic">Pinus pinaster</Emphasis> trees with <Superscript>15</Superscript>N, <Superscript>26</Superscript>Mg and <Superscript>42</Superscript>Ca—simulation of Ca transport in xylem using an upscaling approach

• Introduction   

Atmospheric deposition is an important input of major nutrients into forest ecosystems. The long-term goal of this work was to apply stable isotope methodology to assess atmospheric nutrient deposition in forest systems.     

Effect of major inorganic nutrients on <Emphasis Type="Italic">β</Emphasis>-thujaplicin production in a suspension culture of <Emphasis Type="Italic">Cupressus lusitanica</Emphasis> cells

 The optimum conditions for β-thujaplicin production in a Cupressus lusitanica cell suspension culture were investigated. The conditions required for β-thujaplicin production were clearly different from the conditions for cell growth. The initial phosphate concentration and pH did not affect β-thujaplicin production. A total nitrogen source concentration higher than 3.2 mM suppressed production due to the presence of the ammonium ion. β-Thujaplicin production was observed at 95 mg/l without adding the ammonium ion to the medium. Strict control of major inorganic nutrients was not necessary to produce β-thujaplicin. This finding seems to be favorable for future automated production of β-thujaplicin in commercial cell culture plants.     

AFLP fingerprinting of <Emphasis Type="Italic">Populus deltoides</Emphasis> and <Emphasis Type="Italic">Populus </Emphasis>×<Emphasis Type="Italic"> canadensis</Emphasis> elite accessions

Amplified fragment length polymorphism (AFLP) was used in genetic fingerprinting of 22 elite Chinese Populus deltoides and Populus × canadensis accessions. The results indicated that each of the nine AFLP primer pairs selected generated fingerprint profiles that were unique to each of the accessions. Therefore, each accession was definitively identified by any of the nine primer pairs. Furthermore, the inter-accession genetic relationships inferred based on 461 polymorphic fragments from the nine AFLP primer pairs were largely consistent with phylogenetic relationships based on morphologic traits. Bootstrap analysis showed that three AFLP primer pairs were required to obtain genetic similarity values with a maximum CV of 10% while 10 AFLP primer pairs could give a maximum CV of 5%. Thus, AFLP can readily be applied for a rapid and accurate evaluation of the degree of similarity between poplar cultivars. In this study, the number of AFLP fragments used was sufficient to establish a reliable estimate of genetic similarity among accessions, with a maximum CV of 5.12%. Therefore, the information on the genetic relationships among the poplar accessions generated in this study in connection with knowledge on agronomic traits may have an impact on poplar breeding and planting in China.     

Effect of botanical insecticide Nimbecidine<Superscript>®</Superscript> on food consumption and egg hatchability of the terrestrial snail <Emphasis Type="Italic">Monacha obstructa</Emphasis>

In the laboratory, a commercial neem-based insecticide—Nimbecidine^®—was evaluated as a potential pest management tool for the terrestrial snail, Monacha obstructa (Pfeiffer, 1842) (Hygromiidae). Effects of different concentrations of the botanical insecticide on food consumption and egg hatchability of the terrestrial snail were studied. Generally, food consumption of immature and adult snails decreased as the concentrations of Nimbecidine^® increased. At the highest concentration (10 ml/l), the snails avoided contacting with food completely. The food intake of immature individuals was significantly (p < 0.05) more affected by the Nimbecidine^® treatment (at 1.25 ml/l) than that of adults. LC₅₀ of Nimbecidine^® for the treated eggs was 2.18 ml/l, and eggs failed to hatch at concentration of 10 ml/l. Nimbecidine^® showed sufficient biological activity against the food consumption and eggs viability of M. obstructa, thus the preparation has a potential to protect field crops from this pest snails.     

Quantitative genetics of cold hardiness and growth in <Emphasis Type="Italic">Eucalyptus</Emphasis> as estimated from <Emphasis Type="Italic">E.</Emphasis><Emphasis Type="Italic">urophylla</Emphasis> × <Emphasis Type="Italic">E. tereticornis</Emphasis> hybrids

Interspecific hybrids of Eucalyptus urophylla S. T. Blake × E. tereticornis Smith were used for quantitative genetic analysis of cold hardiness (CH) and growth traits height (H), diameter at breast height (D) and volume (V) at 0.5, 1.5, 2.5 and 4 years of age. The effect of E. tereticornis male variance was significant (P < 0.05) for CH as well as 2.5- and 4-year-old growth except for H ₄, and E. urophylla female × E. tereticornis male interaction was significant for 0.5- and 1.5-year-old growth and CH except for CH _0.5, whereas E. urophylla female effect was not significant for the majority of traits analyzed. Both additive and dominance gene action were present in the genetics of all the traits measured at most ages, indicating that those traits may be improved by hybrid breeding. The σ _D ²/σ _A ² ratios tended to decline after 1.5 years of age in CH and growth traits, implying that breeding value predictions derived from older performance might be more reliable. Estimates of narrow-sense heritability (h ² ) for CH and growth ranged from 0.03 ± 0.05 to 0.28 ± 0.13, depending on trait and age. Additive genetic correlations between CH and V were 0.28 ± 0.48 and 0.69 ± 0.30 at ages 1.5 and 2.5, respectively, suggesting that selection for growth would cause a favorable increase in cold hardiness in E. urophylla × E. tereticornis hybrids.     

Conjugation of δ-endotoxin from <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> with abamectin of <Emphasis Type="Italic">Streptomyces avermitilis</Emphasis> as a new type of biocide,GCSC-BtA,for control of agricultural insect pests

Conjugation of δ-endotoxin from Bacillus thuringiensis with abamectin, a toxin of Streptomyces avermitilis, was carried out to form a new type of biocide, GCSC-BtA based on “Germany-China Scientific Cooperation” research, for the control of agricultural insect pests. The strategy for biochemical linkage was designed by conjugating an amino group in B.t. protoxin with a carboxyl group in carboxylated abamectin under the treatment of conjugator EDC [1-Ethyl-3-(3-dimethylaminopropyl carbodiimide Hydrochloride)]. The formation of B.t. protoxin was processed by solubilizing B.t. crystal in 25?mM dithiothreitol (DTT) at 37?°C for 2?h. The carboxylated abamectin was formed by carboxylating the NaH-activated abamectin with 10?mg/ml butyric anhydride at 111?°C in a water-circumfluent condensation device for 2?h. The conjugating reaction, consisting of 5?mg/ml B.t. protoxin, 10?mg/ml carboxylated abamectin and 19.17?mg/ml EDC, was successfully conducted at room temperature for 24?h. Significant differences were found between pure abamectin, carboxylated abamectin and the conjugated BtA by means of UV-photo absorptions recorded at wavelengths 354, 438, 518, 600?nm (P?<?0.01). LT₅₀ of the conjugated GCSC-BtA to the 3^rd instar larvae of Plutella xylostella (L.) (Lep., Plutellidae) was 35.27 μg a.i./ml, about 62?% and 76?% of that caused by the B.t. protoxin and the caxboxylated abamectin, respectively. The conjugated GCSC-BtA caused 87.14?% mortalities in larvae of P. xylostella, 93.75?% in adult Myzus persicae (Sulzer) (Hom., Aphididae) and 89.33?% in adult Phyllotreta vittata Fabricius (Col., Chrysomelidae) as compared to 48.33?% by the B.t. crystal only in P. xylostella. The symptoms caused by conjugated GCSC-BtA in the 3^rd instar of P. xylostella were black color in the head part and white-yellow in the abdomen of dead larvae, which differed from the black color or the white-yellow all along the body caused by either the B.t. crystal or the abamectin, respectively. It was concluded that the conjugated GCSC-BtA biocide had a broader host spectrum and a faster killing speed than either the B.t. crystal or abamectin alone for the control of agricultural pests.     

Effect of the addition of <Emphasis Type="Italic">Acacia mangium</Emphasis> bark on thermosetting of phenol–formaldehyde resin

The effect of addition of Acacia mangium bark powder on the thermosetting processes of two commercial phenol resins, PF-A and PF-B, was examined by bond strength test, torsional braid analysis, and differential scanning calorimetry. When the bark powder was added to PF-A, the bond strength of plywood pressed at 110°C increased and was comparable to that of plywood pressed at 120 and 130°C. However, when the bark powder was added to PF-B, the bond strength of plywood pressed at 110°C was still lower than that of plywood pressed at 120 and 130°C. The relative rigidity and loss tangent of PF-A cured with the bark powder obtained by heating at 100°C were comparable to those at 120 and 140°C, and the reaction enthalpy was increased by bark powder addition. In contrast, chemical reactions for cured PF-B were not enhanced by bark powder addition.     

Comparative analysis of diterpene composition in the bark of the hybrid larch F<Subscript>1</Subscript>, <Emphasis Type="Italic">Larix gmelinii</Emphasis> var. <Emphasis Type="Italic">japonica × L. kaempferi</Emphasis> and their parent trees

The diterpene compositions in the bark of branches were investigated for two families of the F₁ hybrid, Kurile larch (Larix gmelinii var. japonica Pilg.) × Japanese larch [Larix kaempferi (Lamb.) Carr.] (hereafter F₁) and their parents clones. 13-Epimanool, larixol, larixyl acetate, 13-epitorulosyl acetate (not detected in L. gmelinii var. japonica), isopimaric acid, abietic acid, dehydroabietic acid, and neoabietic acid were detected. Larixol and abietic acid represented more than 50% of the diterpene content in L. gmelinii var. japonica and L. kaempferi, respectively. Larixol and abietic acid were the predominant diterpene components in the F₁, and the proportions of these diterpenes were between those of the parental species. Therefore, the diterpene compositions in the F₁ were hereditarily infl uenced by their parents. The ratios of labdane, pimarane, and abietane diterpenes suggested that the main diterpene biosynthesis pathway in L. gmelinii var. japonica was from copalyl diphosphate (CDP) to labdane-type diterpenes, and that in L. kaempferi was from CDP to abietane-type diterpenes via pimarane type. Furthermore, linear discriminant analysis suggested that the diterpene contents are effective indices for the discrimination of the hybrid seedlings. Part of this article was presented at the Annual Meetings of the Japan Society for Bioscience, Biotechnology, and Agrochemistry in Kyoto, March 2006, and in Tokyo, March 2007     

Influence of stem rot pathogen <Emphasis Type="Italic">Fomitiporia</Emphasis> sp. on “Sanbu-sugi” cultivar of the Japanese cedar <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

An unidentified Fomitiporia sp. causes severe white-rot on stems of a cultivar “Sanbu-sugi” of the Japanese cedar, Cryptomeria japonica. The influence of the fungus on tree health and wood properties was determined. Bark from dead branches and xylem from living branches contained more glucose than bark from living branches and xylem from dead branches. Tree heights at which annual rings were disconnected were 2, 4 and 6 m at ages 37, 15 and 24 years old, respectively. The pH values of damaged parts were lower than those of non-damaged parts, and the damaged parts were clearly identified using bromocresol green solution. Weight loss of sapwood during 60 days of fungal degradation was 1.4 %, which was less than that by a saprophyte, Trametes versicolor. The amount of polyphenols in the heartwood from damaged tree stems was less than that from non-damaged stems. Degraded parts were less stiff than the non-degraded sapwood as measured with a wood-decay tester, Pilodyn. Our observations indicate that damaged stems are chemically and physically inferior to non-damaged stems.     

Soil microorganisms alleviate the allelopathic effect of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E</Emphasis>. <Emphasis Type="Italic">urophylla</Emphasis> leachates on <Emphasis Type="Italic">Brassica chinensis</Emphasis>

Soil microbes may be critical players in determining the allelopathic potential of some plants. Low levels of plant community biodiversity in Eucalyptus plantations have been attributed to the allelopathic potential of these tree species. The role of soil microbes in the allelopathic effect of leaf leachates of the hybrid tree Eucalyptus grandis × E. urophylla, was tested in Petri dish assays with Brassica chinensis as a receiver plant. Soils were collected from either a local garden (soil A) or a Eucalyptus plantation (soil B) and half of each soil was sterilized to remove microbes. These soils were then treated with E. grandis × E. urophylla leachates for 0–72 h. Seed germination of B. chinensis was significantly inhibited in soils treated with leaf leachates relative to untreated soils. The inhibitory effect of the leaf leachates was more pronounced in sterilized soils. Total phenolic content was obvious lower in nonsterile leachate-treated soils than in sterile soils. Biomass of B. chinensis was negatively correlated with the total phenolic content in soils. Our findings suggest that soil microbes can alleviate the allelopathic potential of Eucalyptus and thereby its negative impact on plant growth.     

Fitness of heat and desiccation tolerant hybrid strains of <Emphasis Type="Italic">Heterorhabditis</Emphasis> <Emphasis Type="Italic">bacteriophora</Emphasis> (Rhabditidomorpha: Heterorhabditidae)

Entomopathogenic nematodes of the genus Heterorhabditis are a suitable alternative to chemical insecticides because of their high control effect against insects in cryptic environments and their environmental safety. The availability of H. bacteriophora hybrid strains with increased tolerance to environmental extremes could be a driving force for a more widespread use of nematode-based plant protection products. However, increase in heat and desiccation tolerance of hybrid strains could compromise their fitness regarding virulence, host penetration and reproductive capacity. The fitness of heat and desiccation tolerant hybrid strains was compared to the commercial strain EN 01. Only the heat tolerant strains were superior or similar in fitness to strain EN 01. The strains with increased desiccation tolerance were usually less fit, a possible result of a trade-off effect of selection for desiccation tolerance. Hybrid strains selected for enhanced tolerance after an adaptation to stress were generally better in fitness ranking compared to those for which adaptation prior to stress exposure was excluded. This might be a result of pleitropy. Host penetration and virulence was not correlated. The commercial strain had the highest reproduction per mean number of nematodes penetrating the host insect, which is a result of automatic selection of inbred lines with high reproduction potential during the commercial production process in liquid culture.     

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from <Emphasis Type="Italic">in vitro</Emphasis> cultured stem of <Emphasis Type="Italic">Populus alba</Emphasis>×<Emphasis Type="Italic">P. berolinensis</Emphasis>

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba~P berolinensis. Stems ofPopulus alba~P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.     

Micropropagation of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E. urophylla</Emphasis> AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.     

Biological performance,water absorption,and swelling of wood treated with nano-particles combined with the application of Paraloid B72<Superscript>®</Superscript>

We evaluated fungal decay and mold resistance, leaching, and water absorption of nano-compounds and Paraloid B72^® (PB72) in treated wood specimens to develop new methods of consolidation by combining nano-particles and consolidants. Scots pine wood specimens were treated with dispersions of nano-CuO, nano-ZnO, nano-B₂O₃, nano-TiO₂, and nano-CeO₂. PB72 treatments of nano-particle-treated wood specimens were then carried out by either vacuum or immersion for 24 h. Previously, decayed wood specimens were also consolidated with the nano-compounds and PB72. PB72 treatments reduced element release from treated wood specimens. Nearly all nano-compounds + PB72 treatments increased the biological performance of treated wood specimens against decay fungi tested. PB72-only treated wood specimens had the highest weight losses in decay tests. No improvements were obtained in mold resistance tests when the nano-compounds and PB72 were combined. In nano-compound-only treatments, unleached specimens showed slightly lower water absorption values compared to untreated control specimens. Incorporation of PB72 into nano-compound-treated wood specimens resulted in considerably lower water absorption and volumetric swell. In previously decayed specimens treated with the nano-compounds and PB72 solution, water absorption after 2-h immersion declined compared to control specimens.     

Lethal and sub-lethal effects from short-term exposure of <Emphasis Type="Italic">Rhyzopertha dominica</Emphasis> on wheat treated with Storicide II<Superscript>®</Superscript>

Hard red winter wheat was treated at 0 (untreated control), 25, 50, 75, and 100% of the label rate of the insecticide Storicide II ^® , which is chlorpyrifos-methyl and deltamethrin applied at label rates of 3 and 0.5 ppm, respectively. Paired male and female Rhyzopertha dominica F., the lesser grain borer, were exposed at 27°C and 60% RH on wheat treated at each of the five rates above for 2, 4, 8, 16, or 32 h, and then transferred to untreated wheat and held for 1 week at the same environmental conditions. After this 1-week holding period, the parental adults were removed, mortality was assessed, and the wheat was then held for 7 weeks at the same environmental conditions to determine progeny production. As the concentration and exposure interval increased, mortality of both sexes approached 100%, but at the intermediate concentration–time combinations male mortality was greater than female mortality. Progeny production also decreased with increasing concentration of Storicide II^® as the exposure time increased, with non-linear patterns of decrease at the lower concentrations and time combinations and linear decline at the higher levels of concentration and time. Results seem to indicate greater susceptibility of males to Storicide II^®, and also show delayed parental mortality from the insecticide exposure and sub-lethal effects of reduced progeny production.     

5<Emphasis Type="Italic">Α</Emphasis>-Reductase inhibitory tannin-related compounds isolated from <Emphasis Type="Italic">Shorea laeviforia</Emphasis>

Five tannin-related compounds – gallic acid, flavogallonic acid dilactone, valoneic acid dilactone, gallagyldilactone, ellagic acid – were isolated from the heartwood of Shorea laeviforia, and the inhibitory activity of each against rat liver 5-reductase was evaluated. Valoneic acid dilactone and gallagyldilactone exhibited positive inhibitory activity, but gallic acid and ellagic acid did not. Flavogallonic acid dilactone stimulated 5-reductase activity, even though this compound is structurally similar to valoneic acid dilactone. The kinetic study of valoneic acid dilactone and gallagyldilactone indicated that the inhibitory behavior of 5-reductase were not competitive against the substrate (testosterone) and were partially competitive against the cofactor (NADPH). Additionally, double inhibition analysis of valoneic acid dilactone and NADP⁺ showed synergetic inhibition. These results suggested that neither valoneic acid dilactone nor gallagyldilactone can affect the binding of testosterone but that either compound could interact with an enzyme–NADP⁺ complex to inhibit 5-reductase.     

Seasonal variation of selected trace elements in rare endemic species <Emphasis Type="Italic">Thuriferous Juniper</Emphasis> growing in the <Emphasis Type="Italic">Aurès</Emphasis> Mountains of Algeria

We used energy dispersive X-ray fluorescence (EDXRF) to determine the seasonal variation of selected trace elements (Cr, Mn, Zn, Cu, Se and Fe) and some potential toxic elements (Cd, Pb and Br) in Juniperus thurifera subsp. africana Maire (Cupressaceae) a rare medicinal tree, growing indigenously in Aurès Mountains of Algeria. The precision of the results was assessed by analyzing the certified reference material IPE44 (WEPAL) grass leaves. Results showed J. thurifera was characterized by high Cr and Mn contents. During autumn and winter mineral concentrations were higher in general. The levels of Cr, Se and Mn were higher during autumn and winter than during spring and summer. Zn contents were higher during summer than in other seasons. Cu content did not vary by season. The potential toxic elements in J. thurifera (Pb > Cd > Br) were below the permissible limits recommended by the Joint WHO/FAO guidelines except for Pb in autumn and winter.     

Ethanol production with <Emphasis Type="Italic">β</Emphasis>-xylosidase,xylose isomerase,and <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> from the hydrolysate of Japanese beech after hot-compressed water treatment

Ethanol was produced from the hydrolysate collected as the water-soluble (WS) portion after hot-compressed water (HCW) treatment of Japanese beech. The process involved saccharification with β-xylosidase followed by isomerization with xylose isomerase and fermentation with Saccharomyces cerevisiae. Several process schemes were compared to investigate the effect of process integration of saccharification, isomerization, and fermentation. Higher ethanol yields were obtained for the processes that integrated isomerization and fermentation or saccharification and isomerization. Integration of isomerization and fermentation was effective in converting xylose into ethanol. Similarly, integration of saccharification and isomerization was effective in converting xylooligosaccharides into xylulose. It is presumed that the saccharification reaction toward xylose and the isomerization reaction toward xylulose were linked and therefore each reaction was enhanced.