Introduction of mitochondrial DNA from <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> into <Emphasis Type="Italic">Pleurotus pulmonarius</Emphasis> by interspecific protoplast fusion

Interspecific protoplast fusion between two monokaryotic strains (a methionine auxotrophic and chloramphenicol-resistant Pleurotus ostreatus strain and a wild-type strain of Pleurotus pulmonarius) was carried out to introduce mitochondrial DNA (mtDNA) from P. ostreatus into P. pulmonarius. Because mycelial colonies regenerated on minimum medium containing chloramphenicol only after the treatment of protoplast electrofusion, the regenerants were regarded as protoplast fusants. The fusants isolated from regenerated colonies were uninucleate, and their resistance of chloramphenicol seemed to be a stable trait. The fusants mated with P. pulmonarius but not with P. ostreatus, and showed almost identical random amplified polymorphic DNA (RAPD) patterns to that of the parental P. pulmonarius monokaryon. The sizes of mitochondrial genomes were estimated to be 81.5 kb for P. ostreatus monokaryon, 54.9 kb for P. pulmonarius monokaryon, and 84.5 or 86.0 kb for the fusants. BamHI and EcoRI digest patterns of the fusant mtDNAs were almost the same as the parental P. ostreatus monokaryon. From the above results, the fusants seemed to carry nuclei derived from the monokaryon of P. pulmonarius and mtDNA including the chloramphenicol-resistance gene from the P. ostreatus monokaryon, suggesting that the P. ostreatus mtDNA had been introduced into P. pulmonarius cells by interspecific protoplast fusion. Contribution No. 383 of the Tottori Mycological Institute     

Effects of deer browsing on the survival and growth of <Emphasis Type="Italic">Abies homolepis</Emphasis> and <Emphasis Type="Italic">Picea jezoensis</Emphasis> var. <Emphasis Type="Italic">hondoensis</Emphasis> seedlings after fencing on Mt. Ohdaigahara

We evaluated the survival and growth of Abies homolepis and Picea jezoensis var. hondoensis seedlings on Mt. Ohdaigahara, where the population of sika deer (Cervus nippon) is high and an experimental fence has been in place for 13 years. No significant differences were detected in the survival of small seedlings between fenced and unfenced plots. The growth of A. homolepis was significantly higher in the fenced plot, but growth of P. jezoensis var. hondoensis did not show significant differences between fenced and unfenced plots. Seedlings of height ≤5 cm in the forest floor vegetation of the unfenced plot were probably too small for deer to find and browse, so they survived.     

Evaluation of <Emphasis Type="Italic">Serangium parcesetosum</Emphasis> (Col.: Coccinellidae) for biological control of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> under greenhouse conditions

The cotton whitefly, Bemisia tabaci (Genn.) (Hom., Aleyrodidae) is increasingly a very important pest on many vegetables, field crops and ornamental plants. Therefore, controlling of this pest is still needed especially under glasshouse conditions. The specialist whiteflies’ predator, Serangium parcesetosum Sicard (Col., Coccinellidae) appears to have a great potential for the cotton whitefly control. In this study, the dynamic changes in B. tabaci populations in glasshouse cabins in response to S. parcesetosum were monitored. B. tabaci were introduced to cotton plants in three cabins in average of 50 adults per plant. One and two weeks later, adult females and males of S. parcesetosum were introduced at a rate of one female and one male per plant in the first and second cabins, respectively. The third cabin was considered as a control. The results showed that the mean number of whiteflies in the control cabin was found significantly higher than that of either when S. parcesetosum was introduced 1 or 2 weeks after the infestation with the whitefly. Also, the mean number of B. tabaci was significantly higher when the predator was introduced 2 weeks rather than 1 week after B. tabaci infestation. The maximum mean weekly number of whiteflies/plant was 192.3 in the second week, whereas it was 294.6 in the third week and 1136.4 in the fifth week, in first, second and control cabins, respectively. In the last experimental week, the mean weekly numbers were 74.7, 122.9 and 684.7 whiteflies/plant in the three cabins, respectively. S. parcesetosum has been successfully fed, reproduced and established its population on B. tabaci on cotton plants. The mean weekly number of the predatory individuals increased gradually with the progress of the experimental time. The results demonstrated that the maximum reduction percentage in B. tabaci population was 90.7 and 86.5% in the fifth week after B. tabaci infestation, when the predator was introduced 1 and 2 weeks after the infestation with the whiteflies, respectively. Nevertheless, it is speculated that an earlier release of S. parcesetosum would be more effective in the biological control of B. tabaci.     

Intraspecific variation in <Emphasis Type="Italic">Trifolium pratense</Emphasis>: impact on feeding and host location by <Emphasis Type="Italic">Sitona lepidus</Emphasis> (Coleoptera,Curculionidae)

Red clover (Trifolium pratense L.) is becoming increasingly important in grassland systems because of its high productivity, protein content and nitrogen-fixing ability, but its use has been constrained by losses due to pests and diseases, and this contributed to the decline of red clover usage allowing white clover (T. repens L.) to become the dominant legume of UK grasslands. One of the major pests of red clover is the clover root weevil (Sitona lepidus Gyllenhal, Coleoptera, Curculionidae) which attacks both the shoots and roots, particularly the N-fixing root nodules, of clover plants. This current work investigates the feeding preferences of S. lepidus with respect to 11 varieties (Pawera, Kuhn, Astra, Norseman, Norseman low, Norseman high, Marcom, Merviot, Milvus, Britta, Sabtoron) and 5 lines (AA30, AA31, AA4493, AA4494 and AA4495) of red clover in order to identify relationships between the variation in attractiveness of different red clover varieties for both adult and larval stages of the weevil. Of those tested Norseman high showed potential resistance, being less favoured by both adults and larvae of the weevil. This may be attributed to the potentially high phyto-oestrogen levels in this variety.     

Introduced <Emphasis Type="Italic">Eucalyptus</Emphasis> psyllids in Brazil

In the last 10 years, four species of psyllid were detected in Brazil on eucalypts: Ctenarytaina spatulata in 1994, Blastopsylla occidentalis in 1997, Ctenarytaina eucalypti in 1998 and Glycaspis brimblecombei in 2003. The latter two are serious pests in several countries. In Brazil, G. brimblecombei caused significant damage to the eucalypt plantations in the first years of its introduction. Now this pest is under control due to the programmes of integrated pest management, where the parasitoid is the principal control agent. The four eucalypt psyllid species introduced into Brazil are presented with information on distribution, hosts, biology and control.     

Resistance of gamma-irradiated sapwood of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> to biological attacks

Any means helpful for the promotion of termite feeding activity has potential for use in a matrix in termite bait application. Therefore, energy transfer by gamma irradiation is worthy of consideration for converting wood into termite-accessible material. Wood specimens gamma-irradiated at 100 kGy and at lower levels were tested for their degrees of polymerization (DP) of cellulose and biological resistance. The DP of cellulose adversely decreased with increased doses of gamma irradiation. Termite wood consumption rates, which were determined by laboratory tests using undifferentiated larvae (workers) of Coptotermes formosanus Shiraki, were significantly higher at 100 kGy than at other doses. On the other hand, the decay resistance of gamma-irradiated wood against the fungi Fomitopsis palustris (Berkeley et Curtis) Murrill and Trametes versicolor (L. ex Fr.) Quel did not vary by irradiation dose.     

The role of competitors for <Emphasis Type="Italic">Chrysomela lapponica</Emphasis>, a north Eurasian willow pest,in pioneering a new host plant

The Palaearctic leaf beetle Chrysomela lapponica usually feeds upon willows in the northern region of its occurrence. However, in Central Europe, some populations are known that have specialised on birch. In this study, we investigated the significance of other herbivores occurring together on the same host plants as possible exploitative competitors of C. lapponica. Two populations were studied: a population from Finland specialised on the willow Salix borealis, and a population from the Czech Republic, specialised on the birch Betula pubescens. Abundances of folivorous and suctivorous insects on both host plants were recorded at both population sites. The willow leaf beetle Phratora vitellinae was the most abundant herbivorous insect at both study sites on willow. A field study was conducted to examine the effects of P. vitellinae on the performance of C. lapponica. The presence of P. vitellinae larvae on the same twig upon which C. lapponica larvae were feeding did not affect increase of body weight in C. lapponica larvae. Thus, the high resource availability of both willows and birches suggest that interspecific competition is unlikely to be a selection factor driving the evolution of host shift in C. lapponica.     

Toxic effects of annonaceous acetogenins from <Emphasis Type="Italic">Annona cherimolia</Emphasis> (Magnoliales: Annonaceae) on <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis> (Lepidoptera: Noctuidae)

Plants belonging to the family Annonaceae have been commonly described in traditional medicine as remedies against head lice, and for their insecticidal properties. Characteristic constituents from a few genera of these plants are the annonaceous acetogenins. Fourteen annonaceous acetogenins have been isolated from our Argentine collection of the seeds of A. cherimolia. We report herein the antifeedant and insecticidal effects of nine of those acetogenins on Spodoptera frugiperda (J. E. Smith). The acetogenin squamocin, one of the major constituents of the extract, displayed toxic effects on early larval instars when incorporated to the larval diet at a dose of 50 μg per g of diet. The remaining annonaceous acetogenins tested, itrabin, asimicin, neoanonin, cherimolin-1, cherimolin-2, almuñequin, motrilin, and tucumanin produced pupal mortality and adult malformations leading to death, when incorporated to the larval diet at the same dose. The evaluation of indices of food consumption, growth, and food utilization indicated that squamocin was the only tested acetogenin to produce significant decrease in the growth rate and to reduce the efficiency with which larvae converted ingested food into biomass. All the acetogenins produced more than 80% pupal mortality with no dependence on the position of the THF rings or the number and location of the OH groups.     

Pathogenicity of entomopathogenic fungi to <Emphasis Type="Italic">Thaumetopoea pityocampa</Emphasis> (Schiff.) (Lepidoptera: Thaumatopoeidae) larvae in laboratory conditions

Pathogenicity of 13 strains of entomopathogenic fungi belonging to genera Paecilomyces, Tolypocladium, Beauveria, Metarhizium, and Lecanicillium (=Verticillium) was evaluated on fourth instar larvae of Thaumetopoea pityocampa (Schiff.) (Lep.: Thaumatopoeidae). Larvae were treated individually by dipping into conidial suspension with the concentration of 1.0 × 10⁵ conidia ml⁻¹. All the tested isolates resulted in various levels of mortality (16–100%), and eleven of them were able to grow and sporulate on dead larvae. Mortality caused by three isolates of P. fumosoroseus, one isolate of Beauveria bassiana and one isolate of Metarhizium anisopliae were significantly higher than that of the other treatments. Three isolates of P. fumosoroseus, one isolate of P. farinosus, and one isolate of M. anisopliae showed mycelial growth and sporulation on significantly more cadavers compared to other treatments. Dose–mortality test was conducted with P. fumosoroseus (isolate no: 2679), which appeared to be the most promising isolate among the tested fungi, by spraying five concentrations of its conidia (1.0 × 10⁶, 1.5 × 10⁶, 7.5 × 10⁶, 1.0 × 10⁷ and 1.5 × 10⁷ conidia ml⁻¹). The estimation of the LC₅₀ (95% confidence limits) was 3.4 × 10⁶ (9.3 × 10⁵ − 6.9 × 10⁶) conidia ml⁻¹. This study showed that P. fumosoroseus (isolate no: 2679) could be a good candidate as a microbial control agent against T. pityocampa in forest environment.     

Effect of inundation duration on <Emphasis Type="Italic">Salix gracilistyla</Emphasis> density and size on a gravel bar

To clarify the effects of inundation on the establishment of Salix gracilistyla trees, the relationship between total inundation duration and the distribution and size of trees was examined on a gravel bar along the Miya River in central Japan. The density of S. gracilistyla was low at both low and high elevations on the gravel bar, but high at middle elevations. The density increased with up to 165 days of inundation and declined gradually with more than 165 days of inundation. Density had a negative effect on crown area, whereas inundation duration had a positive effect. At lower elevations, growth limitation of S. gracilistyla is mainly caused by inundation stress, whereas at higher elevations it seems to be primarily due to drought stress, although it depends on the total duration of inundation in a year (TDIY). Consequently, the density is highest at middle elevations where trees do not become too large. These results suggest that inundation duration is one factor determining the density of S. gracilistyla trees, which in turn affects the size of each tree.     

忍冬和华南忍冬叶表皮扫描电镜观察及其生态适应性研究

利用扫描电镜观察了忍冬（Lonicera jàponica Thunb）和华南忍冬（Lonicera confus）叶表皮形态特征,观察指标包括：气孔器、表皮毛、表皮细胞等.结果表明：气孔仅分布在下表皮,气孔器呈无规则型散生;上表皮细胞的垂周壁呈沟槽状下陷;下表皮密被表皮毛（包括腺状毛和非腺毛）,同时还分布有瘤状的草酸钙簇晶.为进一步证明这种瘤状结构是否为草酸钙簇晶,作者发现利用Fluo-3/AM（钙离子荧光指示剂）处理的活体叶片在激光共聚焦显微镜下能够发出强烈的荧光.此外,通过电子探针对瘤状物进行分析,发现其元素组成为C、O、Ca.上述叶表皮构造特征,有利于减少植物体水分散失,还有助于植物适应富钙的石灰土,从而适应我国西南岩溶区特殊的干旱、富钙的环境.电镜观察结果表明,在忍冬叶上表皮中脉处有呈线状分布的腺状表皮毛,而华南忍冬叶上表皮则无表皮毛分布,这一微形态特征可以作为二者种间特异性差别,从而为二者的合理区分提供一定的依据.     

Nutrient element contents of cutting seedlings of hybrid species （ Liriodendron chinense x tulipifera）

The cutting seedlings ofLiriodendron chinense xtulipifera were treated with the different concentrations of auxin (treatment₁: IBA of 50 g·kg⁻¹+NAA of 300 g·kg⁻¹; treatment₂; IBA of 100 g·kg⁻¹+NAA of 300 g·kg⁻¹). The biomass nutrient element contents for different organs (root, stem, leaf) of cutting seedling ofLiriodendron chinense xtulipifera were measured by the dry method, Kjeldahl method and Atomic Absorption Spectroscopy method. The result showed that the biomass of root, stem, and leaf of the cutting seedling treated with auxin was all remarkably increased. The contents of element C in root, stem and leaf had no significant difference between the control and auxin treatments, while the contents of N, P, K and Ca in stem were much lower than that in leaf and root. Variance analysis showed that for the same organ with different concentration treatment of auxin, the four nutrient elements (N, P, K, and Ca) had no significant difference in contents, while there existed significant or very significant difference in contents of the four nutrient elements in different organs with the same concentration auxin treatment. The N, P, K and Ca contents were very low in cutting seedlings; as a result, additional fertilizer should be applied to the seedlings when they were planted in the field. Foundation item: This paper was supported by Jiangsu Province Science Foundation (BE96350). Biography: ZHANG Xiao-ping, (1972-), female, Ph. Doctor in Nanjing Forestry University. Nanjing 210037, P. R. China. Responsible editor: Zhu Hong     

Antioxidative and antiradical activities of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> leaf oils from Thailand

The leaf essential oils (six samples) from three clones of Eucalyptus camaldulensis Dehnh. were characterized by gas chromatography-mass spectrometry. Radical scavenging and antioxidant properties were investigated by the 1,1-diphenyl-2-picylhydrazyl (DPPH) assay and the β-carotene bleaching test. It was found that the whole essential oil and its fractions had significant antioxidant effects when they were tested by each method. In the DPPH assay, the E. camaldulensis leaf oils showed IC₅₀ inhibitory concentrations in the range of 1.75–12.62 mg/ml. In the β-carotene bleaching test, the IC₅₀ valued were in the range of 14.30–118.55 μg/ml. Part of this report was presented at the 18th meeting of the Chugoku-Shikoku Branch of the Japan Wood Research Society, November 2006     

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.     

Vitality and storage condition of Syringa pollen

The fresh pollen vitality,the effect of different storage conditions on the pollen vitality,and the difference of vitality among the species of seven species of Syringa were determined in Shenyang,China.The results indicated that the pollen vitality (81.5%) of Syringa villosa was the highest among the seven tested species,followed by S.microphylla and S.meyeri,and that of S.oblata var.affinis was the lowest.The low temperature was the best condition for storage of pollen of Syringa,and the most proper temperature for the storage was 0-2℃.The storability of S.microphylla was the best of all,and it could be stored over 60 days at the temperature of 0-2℃,next was S.villosa and S.meyeri.     

Host instar suitability of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> (Genn.) (Hom.: Aleyrodidae) for the parasitoid <Emphasis Type="Italic">Eretmocerus mundus</Emphasis> (Hym.: Aphelinidae)

Efficient use of parasitoids in pest control depend on the knowledge of the biological relationships between host and parasitoid. Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) is one the most important natural enemies of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) that is found in fields and greenhouses in Adana, Turkey. Although more studies have been done to determine the relationships between E. mundus and B. tabaci, the differences among strains of E. mundus could have important biological concequences. In this study, biological characteristics of native population of E. mundus parasitization of B. tabaci (Q biotype) were determined using bean plants Phaseolus vulgaris (L.) at a constant temperature of 25 ± 1°C, relative humidity 70 ± 10% and 16:8 (L:D) photoperiod regimen. All B. tabaci nymphal instars were parasitized by E. mundus, but the mean number of daily-parasitized nymphs was highest in the second instars (15.3). In addition, second instar females had the shortest mean development time (15.2 days). Mean development times of parasitoids obtained from first, second, third and fourth B. tabaci instars varied from 15.2 to 16.2 days and 15.2 to 15.9 days for female and male, respectively. Proportion of female (♀/♂+♀) varied from first (0.53) to second (0.34) host instars. Impact of results on potential biological control of B. tabaci was discussed through conservation native natural enemies.     

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.     

Temperature-dependent development of <Emphasis Type="Italic">Diglyphus isaea</Emphasis> (Hymenoptera: Eulophidae) on <Emphasis Type="Italic">Liriomyza sativae</Emphasis> (Diptera: Agromyzidae) on cucumber

The development of Diglyphus isaea (Walker), a parasitoid of leafminers, was studied under laboratory conditions at seven constant temperatures (10, 15, 20, 25, 30, 35, and 40°C) on Liriomyza sativae Blanchard reared on cucumber (Cucumis sativus L.). The total development period (oviposition to adult emergence) decreased with increasing temperature between 15 and 35°C. In 10 and 40°C no development rate was found in D. isaea, so that it may be claimed that these temperatures fall outside the temperature range for development. Linear regression was used to describe the relationship between development rate and temperature. For egg to adult development, males of D. isaea required 153.8 degree-days (DD) above the theoretical threshold of 9.2°C and females required 161.3 DD above 9.4°C. Data were fitted to four nonlinear temperature-dependent models. Evaluation of the models took place based on the following criteria: fit to data, number and biological value of the fitted coefficient, and accuracy on the estimation of the thresholds. It could be concluded that the Briere-1 and Briere-2 models are suitable for estimating the minimum, maximum and optimal temperature thresholds of D. isaea. Thermal requirements and temperature thresholds can be used to predict the occurrence, number of generations and population dynamics of D. isaea.     

Controlling the sugar beet beetle <Emphasis Type="Italic">Cassida vittata</Emphasis> with entomopathogenic nematodes

Sugar beet, Beta vulgaris L. is a strategic crop of sugar industry in Egypt. It is threatened by several insect pests among most important of them is the beet beetle Cassida vittata. This work deals with the biological control of this insect using four Egyptian and imported entomopathogenic nematodes (EPNs). The nematodes included Steinernema carpocapsae S2, Heterorhabditis bacteriophora S1, S. feltiae and H. bacteriophora (1-3). Daily mortality of larvae, pupae and adults of C. vittata were recorded after treatment with serial concentrations (from 500 to 4,000 infective juveniles/ml) of each of four studied EPNs. Development of nematodes in insect bodies was followed up. S. carpocapsae S2 was chosen for the application against different stages of the pest in a sugar beet field. In the field, single application of S. carpocapsae S2 killed 65% of the larvae, 92% of the pupae and 57.3% of the adults of C. vittata within a week. This work is the first report on using the EPNs to control sugar beet beetle.     

Inheritance and cross-resistance of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> insecticidal crystal protein Cry1Ac resistance in cotton bollworm <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> Hubner population from Tamil Nadu,India

Cotton bollworm, Helicoverpa armigera Hubner selected for five generations with Bacillus thuringiensis insecticidal crystal protein Cry1Ac in the laboratory developed 12.98-fold resistance. Resistance and susceptible populations were mass crossed to study the dominance of resistance. The Cry1Ac—selected (BCR) population showed 5.8-fold resistance to Cry1Aa and 5.04-fold resistance to Cry1Ab. The degree of dominance (D) was 0.34 and 0.40 for the R × S and S × R hybrids, respectively, which indicates incomplete recessive character of Cry1Ac resistance in the population. The estimated realized heritability (h ²) and response quotient (Q) of resistance for Cry1Ac were 0.52 and 0.15, respectively. This indicated the lower phenotypic variation in the selected population. The resistance risk assessment based on h ² indicated that the resistance would increased tenfold after <9 generations for Cry1Ac in the resistant population. The results show the ability of H. armigera to develop resistance against Cry1Ac and cross-resistance to Cry1Aa and Cry1Ab.