Micromorphological characteristics of decayed wood and laccase produced by the brown-rot fungus <Emphasis Type="Italic">Coniophora puteana</Emphasis>

Microscopic examination showed the cell wall decay pattern produced by the brown-rot fungus Coniophora puteana to be different from the degradation pattern known to be typical for brown-rot fungi. Erosion and thinning of cell walls in patterns considered to be characteristic of white-rot decay were observed. In particular, the fungal strain COP 20242 degraded secondary cell wall layers extensively, and also degraded lignin-rich middle lamellae. Some strains of C. puteana produced soft-rot type cavities in the S2 layer. All strains of C. puteana employed in the present work showed a positive reaction to tannic acid in the Bavendamm test, indicating the production of laccase. Microscopic and enzymatic studies provided evidence to suggest that the wood decay by C. puteana is unique both in terms of micromorphological and enzymatic patterns of cell wall degradation. This is because brown-rot fungi are not generally known to form cavities in the cell walls or to produce lignin-degrading enzymes. These observations suggest that lignin degradation capacity of brown-rot fungi may be greater than previously considered.     

兰花致病真菌拮抗细菌的筛选与鉴定

[目的]为获得高效拮抗3种兰花病原真菌胶孢炭疽菌、尖孢镰孢菌和腐皮镰孢菌的菌株。[方法]使用平板对峙法筛选拮抗菌株,结合形态特征观察、生理生化试验和16S rRNA基因序列分析的方法对目的菌株进行鉴定。[结果]筛选获得5株同时拮抗三种病原菌的菌株,其中菌株GT312拮抗效果明显。16S rRNA基因序列分析显示,该菌株与Bacillus amyloliquefaciens(模式菌株FZB42T)相似性最高,为99.93%。菌株形态特征观察、生理生化试验结果与B.amyloliquefaciens描述一致。[结论]菌株GT312可同时拮抗3种兰花病原真菌,鉴定为解淀粉芽孢杆菌,为兰花病害的生物防治提供了菌株资源。     

Effects of high temperature kiln drying on the practical performances of Japanese cedar wood (Cryptomeria japonica) I: changes in hygroscopicity due to heating

The effect of heating on the hygroscopicity of Japanese cedar wood was investigated as a simple evaluation of thermal degradation in large-dimension timber being kiln-dried at high temperatures (>100°C). Small wood pieces were heated at 120°C in the absence of moisture (dry heating) and steamed at 60°, 90°, and 120°C with saturated water vapor over 2 weeks, and their equilibrium moisture contents (M) at 20°C and 60% relative humidity (RH) were compared with those of unheated samples. No significant change was induced by steaming at 60°C, while heating above 90°C caused loss in weight (WL) and reduction in M of wood. The effects of steaming were greater than those of dry heating at the same heating temperature. After extraction in water, the steamed wood showed additional WL and slight increase in M because of the loss of water-soluble decomposition residue. The M of heated wood decreased with increasing WL, and such a correlation became clearer after the extraction in water. On the basis of experimental correlation, the WL of local parts in large-dimension kiln-dried timber was evaluated from their M values. The results indicated that the thermal degradation of inner parts was greater than that of outer parts.     

Screening for condensed tannin-degrading fungi with a synthetic <Superscript>14</Superscript>C-labeled compound

To find fungi that are potent for degradation of condensed tannin, a two-step screening was used. This involved measurement of fungal growth rate on Japanese cedar (Cryptomeria japonica) bark, followed by determination of [¹⁴C]-labeled CO₂ generated from fungal degradation of synthetic [¹⁴C]-labeled condensed tannin model. In the first screening, 75 strains of wood rot fungi were tested, and 19 strains effectively decreased bark weight and/or the weight of the methanol-soluble fraction. For the second screening, [¹⁴C]-labeled condensed tannin model compound was synthesized in 11.8% yield based on radioactivity measurements. Over the incubation period, Coriolus hirsutus K-2671, Lentinus edodes Is, and Lampteromyces japonicus Nn showed higher cumulative [¹⁴C]-labeled CO₂ emissions than the other strains and mineralized the [¹⁴C]-labeled condensed tannin model compound by 3.7%, 3.0%, and 3.0%, respectively. Fractionation of the methanol extracts from the medium by gel permeation chromatography after fungal treatment suggested that fungi that can induce the emission of significant levels of [¹⁴C]-labeled CO₂ can extensively depolymerize condensed tannins.     

A new belamcandaquinone from the seeds of Iris bungei Maxim

A novel dimeric 1,4-benzoquinone and resorcinol derivative, Belamcandaquinone N (1), and two known compounds, 3-hydroxyirisquinone (2) and 5-[(Z)-10-heptadecenyl] resorcinol (3), were isolated from the seeds of Iris bungei Maxim. Their structures were elucidated by spectroscopic methods and comparing with literature data of known compounds. These compounds showed remarkable cytotoxic activity against RM-1 cell lines.     

Efficacy of modified diatomaceous earth on different cockroach species (Orthoptera, Blattellidae) and silverfish (Thysanura, Lepismatidae)

Defined populations of American (Periplaneta americana), German (Blattella germanica), and Oriental (Blatta orientalis) cockroaches, and silverfish (Lepisma saccharina) were observed after exposure to deposits (25 g/m²) of a new 1,1,1–trimethyl-N-trimethylsilane-modified, highly hydrophobic diatomaceous earth (DE) formulation by using a computer-aided device measuring motility, circadian rhythm, and mortality under defined environmental and climatic field-simulating and exposure-enforced conditions. In a humid climate (85% relative humidity) with water and food offered ad libitum, complete population eradication could be achieved on the sixth day against B. germanica, on the eighth day against P. americana, and on the ninth day against L. saccharina, respectively. No population eradication occurred within 10 days of exposure when testing B. orientalis, showing a mean survival rate of 29.4 ± 6.7 % of the populations. When comparing the species-specific mortality rates with the results obtained from corresponding reference control groups, significantly higher mortality rates could be observed in B. germanica (F = 66; df = 52; P < 0.00001), P. americana (F = 344; df =66; P < 0.00001), L. saccharina (F = 253; df = 24; P < 0.00001), and B. orientalis (F = 422; df = 11; P < 0.00001). Overall, the efficacy of the hydrophobised DE examined ranked as follows: B. germanica > P. americana (F = 51; df = 24; P < 0.00001) > L. saccharina (F = 43; df = 24; P < 0.00001) >> B. orientalis (F = 9; df = 15; P < 0.000001). DE exposure resulted in complete disruption of the circadian activity in B. germanica and P. americana, but not when tested against B. orientalis, where the species-specific circadian motility peak was still preserved at lower levels after 10 days of exposure. In contrast to the cockroach species examined, no specific circadian rhythm could be measured in the L. saccharina control and treatment groups. Results indicate that hydrophobised DE originating from freshwater diatoms modified with 1,1,1-trimethyl-N-trimethylsilane can be successfully used for the control of infestations with German and American cockroaches as well as silverfish, but not against Oriental cockroaches. It is concluded that species-specific morphological, physiological and behavioural characteristics of insects influencing DE efficacy as well as the toxicological risk of modified DE to humans deserve further investigation.     

Mutualistic symbiosis between Bursaphelenchus xylophilus and bacteria of the genus Pseudomonas

Interactions between the pine wood nematode (PWN), Bursaphelenchus xylophilus, and bacteria of the genus Pseudomonas were examined by cultivating axenic PWN and bacterial strains using callus of Pinus thunbergii. Ten (Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas cepacia and Pseudomonas spp.) of the 29 bacterial strains tested, significantly increased the reproduction of PWN. The rest of the bacteria (19 strains of 10 species) inhibited the reproduction of PWN completely. The growth of 18 of the 29 bacterial strains tested, including the 10 strains promoting PWN reproduction, was significantly increased by the presence of PWN. It indicated a mutualistic symbiotic relationship between PWN and the 10 bacterial strains in the genus Pseudomonas. The bacterial mutualistic symbionts are organisms, which may have co‐evolved with PWN rather than being accidentally associated. The finding provides further evidence for our hypothesis that pine wilt disease is complex, induced by both PWN and associated phytotoxin‐producing bacteria.     

Infection of red palm weevil,<Emphasis Type="Italic"> Rhynchophorus ferrugineus</Emphasis>, by spore-forming bacilli indigenous to its natural habitat in Egypt

A search for means of biologically controlling the red palm weevil (RPW) Rhynchophorus ferrugineus resulted in the isolation of several microbial cultures belonging to bacteria, actinomycetes and fungi. Among the obtained cultures, three potent spore-forming bacilli were further isolated from natural habitats associated with insect-damaged date palms in Egypt. The isolated bacterial cultures, (strains 73, 15 and 27) were identified as variants of Bacillus sphaericus, B. megaterium and B. laterosporus, respectively. Under standard bioassay conditions, the mortalities of the second-instar larvae of the target insect induced by the three isolated bacilli variants ranged between 40 and 60%. The most active culture (strain no. 73) was identified as B. sphaericus. This isolate formed spherical endospores and crystalline endotoxin comparable to those of B. sphaericus standard strain 2362, which is pathogenic to mosquito larvae, as shown by electron microscopy. However, the amino acid composition of the spore-endotoxin complexes was markedly different.     

Degradation of lipophilic wood extractive constituents in Pinus sylvestris by the white-rot fungi Bjerkandera sp. and Trametes versicolor

The white-rot fungi Trametes versicolor and Bjerkandera spp. are among the most frequent decomposers of angiosperm wood in forest ecosystems and in wood products in service. Wood extractives have a major impact on wood properties and wood utilization. This work evaluated the ability of two white-rot fungal strains (Bjerkandera sp. strain BOS55 and T. versicolor strain LaVec94-6) to degrade the main lipophilic extractive constituents in Scots pine (Pinus sylvestris L.). The time course of wood decay and wood extractive degradation was monitored in stationary batch assays incubated for eight weeks. The strains tested eliminated high levels of total resin, 34 to 51% in two weeks. Wood triglycerides were the most readily degraded extractive components (over 93% elimination in only two weeks). Free fatty acids and resin acids, which are potential fungal inhibitors, were also rapidly decomposed by the fungal strains. Sterols were used more slowly, nonetheless, the fungal degradation of this extractive fraction ranged from 50 to 88% after four weeks. Received 19 March 1999     

Structural features of a new heteropolysaccharide from the fruit bodies of Melia azedarach and its effect on cytotoxic activity

Water-soluble polysaccharides were extracted from the fruit pulps of Melia azedarach and some columns were undertaken to isolate the major polysaccharide (MPS-III). Its structural features were elucidated by IR analysis, carbohydrate analysis, periodate oxidation, Smith degradation, methylation analysis and NMR spectroscopy. Then it was evaluated for the cytotoxic activity in vitro against four human cell lines, using the sulphorhodamine B assay. The data obtained indicated that MPS-III contains a α(1→4) main chain backbone composed of arabinose, mannose in a molar ratio of 1.31:1.0 and has α(1→6) branch structure. And MPS-III showed a strong cytotoxic activity in the BGC-823 cell line.     

Analysis of synonymous codon usage in chloroplast genome of Populus alba

The pattern of codon usage in the chloroplast genome of Populus alba was investigated. Correspondence analysis （a commonly used multivariate statistical approach） and method of effective number of codons （ENc）-plot were conducted to analyze synonymous codon usage. The results of correspondence analysis showed that the distribution of genes on the major axis was significantly correlated with the frequency of use of G＋C in synonymously variable third position of sense codon （GC3S）, （r=0.349）, and the positions of genes on the axis 2 and axis 3 were significantly correlated with CAI （r=-0.348, p〈0.01 and r=0.602, p〈0.01）. The ENc for most genes was similar to that for the expected ENc based on the GC3s, but several genes with low ENc values were lying below the expected curve. All of these data indicated that codon usage was dominated by a mutational bias in chloroplast gcnome ofP. alba. The selection in nature for translational efficiency only played a minor role in shaping codon usage in the chloroplast genome ofP alba.     

西南桦和尾巨桉凋落叶分解及其与土壤性质的相关性

[目的]探讨南亚热带西南桦和尾巨桉人工纯林的凋落叶分解动态及其与土壤化学性质之间的相关关系.[方法]采用原位分解袋法研究凋落叶的分解过程.[结果]表明:西南桦、尾巨桉人工林凋落叶分解系数分别为0.96 a^-1和0.88 a^-1.在为期12个月的分解试验中,2种凋落叶有机C含量在整个分解过程中呈逐渐下降趋势;全K含量和C/N比在分解前期迅速下降,之后趋于平缓;全N含量和全P含量在整个分解过程中呈逐渐上升趋势;2种凋落叶N/P比则呈先升高后下降的趋势.无论是分解前期还是分解后期,凋落叶质量损失与N含量均呈显著正相关(前期R=0.877;后期R=0.855),与C/N均呈显著负相关(前期R=-0.735;后期R=-0.697).与尾巨桉林地土壤性质相比,西南桦凋落叶分解提高了林地0~10、10~20 cm土壤的有机C、全N、全P、全K、N/P,对2030 cm土壤有机C、全K、pH值、C/N、N/P则未产生显著影响.相关分析表明:凋落叶初始有机C含量与土壤有机C、全N、全P、全K、N/P显著相关;凋落叶初始全N含量与土壤全N、pH值、C/N显著相关.[结论]凋落叶的养分含量与土壤养分的关系紧密;与尾巨桉相比,西南桦凋落叶的养分含量明显较高,分解速率更快,释放到土壤中的养分也更多.     

薄壳山核桃叶及青皮水浸液对3种植物的化感作用

采用蒸馏水浸提法收集薄壳山核桃叶及青皮化感物质,运用室内生物测定法检测其不同浓度(0.005、0.01、0.1 g·mL^-1)浸提液对小麦、油菜和绿豆种子萌发和幼苗生长的影响。结果表明:薄壳山核桃叶及青皮水浸液处理对3种受体植物的种子萌发和幼苗的苗高基本上表现为"低(0.005 g·mL^-1)促高(0.1 g·mL^-1)抑"的双重浓度效应,个别受体植物的化感作用虽然表现不同于"低促高抑"的整体趋势,但随着浓度的增加也表现为促进作用降低或抑制作用增强的现象;对3种受体植物根长的抑制作用大于对苗高的抑制作用,而对幼苗鲜质量、干质量的影响则因受体种类的不同而不同。薄壳山核桃叶水浸液处理对3种受体植物的综合化感效应依次为:油菜> 小麦> 绿豆;青皮水浸液处理对3种受体植物的综合化感效应依次为:小麦> 油菜> 绿豆。综合3种受体植物的化感效应认为,在相同浓度下,薄壳山核桃叶水浸液处理对3种受体植物的促进作用最强,而青皮水浸液处理对3种受体植物的抑制作用最强。     

松材线虫Bxpel2基因的克隆及RNA干扰载体构建

[目的]为了构建松材线虫(Bursaphelenchus xylophilus)果胶酶Bxpel2基因干扰载体.[方法]通过Trizol 法提取松材线虫总RNA,反转录合成cDNA,设计带T₇启动子的果胶酶Bxpel2基因引物,以cDNA为模板扩增出果胶酶Bxpel2基因片段,连接到RNA干扰载体,再以干扰载体为模板,PCR扩增出目的片段后进行测序鉴定,合成果胶酶Bxpel2基因双链RNA(dsRNA),采用RT-PCR检测松材线虫Bxpel2基因干扰后的表达情况.[结果]表明:1)提取的松材线虫总RNA完整性好,无降解;2)成功克隆出松材线虫果胶酶Bxpel2基因片段(790 bp)并将其连接至pMD19-T载体;3)以RNA干扰载体为模板合成dsRNA,浓度分别为1.313 mg·mL^-1和1.152 mg·mL^-1;4)RT-PCR结果显示,松材线虫经过dsRNA干扰后,Bxpel2基因表达基本受到抑制.[结论]成功构建松材线虫果胶酶Bxpel2基因干扰载体,为进一步研究Bxpel2基因在松材线虫致病过程中的作用和功能奠定了良好的基础.     

Microbial Inoculants for Sustainable Forests

Abstract

Fungal root pathogens are widespread and may cause substantial seedling losses in conifer nurseries. Furthermore, poor seedling survival and growth on reforestation sites results in reduced forest regeneration. Use of microbial inoculants for disease control and plant growth promotion has become an important endeavour. A microbial culture collection of 500 strains was assessed for biological control of fungal root pathogens and/or plant growth promotion of conifer seedlings. Seven of these strains showed significant suppres-sive effects on various soil-bome fungal pathogens. On Douglas fir, two strains, RAL3 and 64-3, reduced disease caused by Fusariumby. 7-42% in repeated growth room assays. The same strains significantly increased healthy stand of white spruce seedlings inoculated with Fusariumand Pythiumin a conifer nursery, and increased the survival of bare-root white spruce seedlings planted on a reforestation site by 19-23%. Both strains also significantly increased new root and total plant dry weights. Strain RAL3 in commercial formulation maintained a viable population of about log 8-9 cfu/ml for over a year when stored at 5°C. Strain survival on seed varied with conifer species. No decreases in bacterial populations were observed on seeds of jack pine or Douglas fir after 37 to 44 days storage at 5°C, but decreases were observed on seeds of white spruce and Scots pine. This study has provided candidate beneficial microbial inocu-lants which offer promise for development of commercial inoculants for the forestry industry.     

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.     

Genetic variation and mtCOI phylogeny for Bemisia tabaci (Hemiptera, Aleyrodidae) indicate that the ‘B’ biotype predominates in Iran

Despite a large number of investigations on the molecular genetics and population structure of the whitefly Bemisia tabaci (Gennadius) complex, no such study had been conducted in Iran. The genetic variation of B. tabaci was examined using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) for 18 field collections from cucumber, eggplant, and tomato in four provinces of Iran. PCR amplification and restriction digestion with two enzymes detected 388 RFLP fragments, of which 16 fragments showed polymorphisms. Cluster analysis of these data placed all B. tabaci individuals within a single group, and there was no evidence for between- or within-population genetic variation. Phylogenetic (Clustal W) analysis of 42 B. tabaci mtCOI sequences (n = 21 field collections) from Iran, and a comparison with well-studied haplotype or biotype reference sequences available in public sequence databases, revealed that the Iranian B. tabaci populations were most closely related to the B biotype at 0–1.2% nucleotide identity. The B biotype is a well-known member of a sister clade from the Middle East–North African region of the world, owing to its nearly worldwide distribution and invasive characteristics. This report indicates that a single major haplotype of B biotype is prevalent in Iran and that its closest relative is the B biotype. Also, given the extent of known variation in the Middle East and African continent, data indicate somewhat surprisingly that the B. tabaci collections sampled in Iran had limited genetic variation and population substructure. Knowledge that the B biotype of B. tabaci predominates in Iran is important for designing effective pest management strategies given that biotypes of B. tabaci are known to differ greatly with respect to insecticide resistance, host range, virus–vector interactions, and other key biological characteristics.     

Bacillus pumilus and Stenotrophomonas maltophilia as two potentially causative agents involved in Persian oak decline in Zagros forests (Iran)

The oak decline is known as one of the most destructive complex diseases causing high economic losses around the world, especially in Iran. The main objective of the present study was to investigate the possible role of bacteria as causative agents of oak decline in the Zagros forests of Iran. To do this, stem, root and leaf samples were taken from symptomatic Persian oak trees (Quercus brantii) in different zones of Zagros forests (Ilam Province, Iran). From 150 bacterial isolates, 20 showed pathogenicity against Geranium seedlings. Among 20 hypersensitivity test positive strains, four strains showed pathogenicity against oak saplings. Based on morphological and 16S rRNA gene sequence analysis, three strains were identified as Bacillus pumilus and one strain as non‐sporulating Gram‐negative Stenotrophomonas maltophilia. Pathogenicity studies of different B. pumilus and S. maltophilia strains revealed that they have potential to cause the disease in oak saplings and symptoms of disorder in Persian oak trees. To our knowledge, there are no previous records of B. pumilus and S. maltophilia causing decline on Fagaceous trees like Q. brantii. More detailed field and molecular studies are required to confirm the absolute role of such bacteria in occurrence of oak decline in Zagros forests.     

Gas exchange,water relations and morphology of yellow-cedar seedlings and stecklings before planting and during field establishment

Information is lacking on how yellow-cedar (Chamaecyparis nootkatensis (D. Don.) Spach) stecklings compare with seedlings in their response to potentially limiting field site environmental conditions. Before planting, yellow-cedar seedlings and stecklings were characterized at 5 and 22°C root temperatures for gas exchange parameters (n=6), root growth capacity (n=12) and plant water movement (n=6) using a fully randomized design in a controlled environment growth room. Seedlings, compared with stecklings (t-test), showed a faster (p=0.01) recovery of net photosynthesis (P_n) and stomatal conductance (g_wv) after root temperature increased from 5 to 22°C. Seedlings had greater (p=0.05) root growth capacity (number of new roots 0.5 cm in length) than stecklings after 14 days at 22°C and after 14 days at 5°C followed by 15 days at 22°C. Seedlings had a lower (p=0.01) resistance to plant water movement measured after 29 days (14 days at 5°C followed by 15 days at 22°C root temperature).Seedlings and stecklings were planted on a coastal British Columbia field site and monitored for 1) morphological development and physiological response to ambient site conditions throughout the first growing season, and 2) physiological response to drought conditions during late summer. Both trials used a fully randomized design and stock type means were compared using t-tests (p=0.10, 0.05, and 0.01). Initially, seedlings were taller (p=0.01) than stecklings but otherwise morphologically similar (n=20). Over a 35 week period after planting, stecklings had greater incremental height growth, while seedlings produced a greater (p=0.01) number of roots extending out of the root plug. Seedlings and stecklings (n=6) had similar P_n and g_wv patterns under late spring environmental conditions. However, seedlings had greater P_n and g_wv under cold temperature (p=0.05 and p=0.1, respectively) and drought conditions (p=0.05 and p=0.01, respectively), but during conditions of high evaporative demand and adequate soil moisture, stecklings had greater (p=0.01) P_n and g_wv. Seedlings had higher (p=0.01) daily integrated shoot water potential values (D) at different times during the growing season, and maintained higher (p=0.05) predawn (_pd) shoot water-potential values, higher (D) (p=0.01) and lower (p=0.01) resistance to plant water movement (R_SPAC) during drought conditions.