7种木层孔菌属真菌的培养特性

描述了7种木层孔蓖属（Phellinus)真菌的培养特性,它们是淡黄木层孔菌（P.gilvus),火木层孔菌（P.igniarius),裂蹄木层孔菌（P.linteus),松木层孔菌（P.pini),冷杉木层孔菌（P.pinivar.abietis) ,苹果木层孔菌（P.pomaceus)和稀硬木层孔菌（P.robustus)。     

新组合欧氏拟木层孔菌——“西方拟木层孔菌”的合法名称

拟木层孔菌属Phellinopsis是最近从木层孔菌属Phellinus中独立出来的一个属。拟木层孔菌属的全部4个种中,只有西方拟木层孔菌P. occidentalis在中国没有分布。简要回顾了西方拟木层孔菌这一名称的历史,其基元异名西方层孔菌Fomes occidentalis属于无效发表,因此将西方拟木层孔菌处理为无效名称,并将该名称所代表种的合法名称欧氏木层孔菌Phellinus overholtsii组合为欧氏拟木层孔菌Phellinopsis overholtsii。     

中国木层孔菌属三个新记录种

报道了木层孔菌属Phellinus 3个中国新记录种。赤杨木层孔菌Phellinus alni典型特征为菌盖具有较宽的同心环带和清晰的密集环纹菌核;黑木层孔菌P. nigricans具有较大的担孢子;东方木层孔菌P. orienticus具有平伏的子实体,担孢子相对较小。对这3个种进行了详细的描述和显微结构绘图。     

三种木层孔菌子实体不同溶剂提取物抗氧化活性的研究

以抗坏血酸为阳性对照,分别对桑木层孔菌Phellinus mori、鲍姆木层孔菌Phellinus baumii和瓦宁木层孔菌Phellinus vaninii野生子实体甲醇提取物(ME)、氯仿提取物(CE)、乙酸乙酯提取物(EAE)、正丁醇提取物(BE)和热水提取物(WE)的体外清除DPPH(1,1-二苯基-2-三硝基苯肼)自由基、羟自由基和总抗氧化能力进行了检测。结果表明,3种木层孔菌的不同溶剂提取物都具有体外抗氧化活性。3种菌EAE清除DPPH自由基能力较强,其中瓦宁木层孔菌EAE清除DPPH自由基能力最强,IC50为10.65μg/mL。3种木层孔菌热水浸提物清除羟自由基的能力较强,鲍姆木层孔菌WE的清除率最高,1mg/mL时达到73.52%。另外,3种木层孔菌不同提取物总抗氧化能力和清除DPPH自由基能力基本相同,瓦宁木层孔菌BE的总抗氧化能力最强,1mg/mL时达到了167.7U/mL。3种木层孔菌相比,瓦宁木层孔菌不同提取物的体外抗氧化能力最强,其次是鲍姆木层孔菌,桑木层孔菌的体外抗氧化能力最弱。     

忍冬木层孔菌液体培养物的化学成分研究

忍冬木层孔菌(Phellinus lonicerinus(Bond.)Bond.et Sing)为湖北等地习用的桑黄品种,也是土家族常用药.对忍冬木层孔菌液体培养物的化学成分及其体外抗肿瘤活性进行研究.通过硅胶柱色谱、SephadexLH-20柱色谱及半制备型HPLC等色谱分离技术从忍冬木层孔菌发酵液及菌丝体中分离得到...     

三种白腐菌生物学特性与木质纤维素酶基因遗传多样性

以采自东北林业大学帽儿山实验林场的3种白腐真菌——木蹄层孔菌(Fomes fomentarius)、鲍姆鲍姆木层孔菌(Phellinus baumiibaumii)和火木层孔菌(Phellinus igniarius)为材料,用菌落直径测量法比较3种白腐菌在马铃署葡萄糖固体培养基上的生长速度,采用菌丝体干重法比较其在马铃署葡萄糖液体培养基中的生物量变化。结果显示:马铃薯葡萄糖固体培养基上3种白腐菌均为快速生长类型,其生长速度木蹄层孔菌火木层孔菌鲍姆鲍姆木层孔菌;马铃署葡萄糖液体培养基中生物量增长速度木蹄层孔菌鲍姆鲍姆木层孔菌火木层孔菌。用比色法测量其木质纤维素酶活性,结果显示:木蹄层孔菌产锰过氧化物酶和漆酶量较高,鲍姆鲍姆木层孔菌和火木层孔菌产木质素过氧化物酶量较高;木蹄层孔菌、鲍姆鲍姆木层孔菌和火木层孔菌3种白腐菌的3种主要木质素酶(锰过氧化物酶、漆酶和木质素过氧化物酶)的表达量,种间差异显著(F=3.75*、5.20**、3.01*),白桦木屑诱导处理与对照间差异显著(F=3.84*、4.19*、5.28*);两种主要纤维素酶(葡聚糖内切酶、葡聚糖外切酶)的表达量,种间差异不显著,受碳源影响作用显著(F=3.99*、4.04*)。筛选29对引物组合,对3种白腐菌几种主要木质纤维素酶基因进行TRAP-PCR分子标记检测,比较3菌种间遗传差异,扩增总条带数为357条,多态性条带数为255条,多态性条带的比例为71.43%,其中木质素降解酶基因总多态位点比率为73.77%,纤维素降解酶基因总多态位点比率为68.97%。3种白腐菌的木质纤维素降解酶基因在种间均存在较高的遗传差异。因此,特定基因的TRAP分子标记可以用于木腐菌的遗传变异分析。     

6种“桑黄”石油醚提取物的体内抗肿瘤活性

对6种＂桑黄＂的石油醚提取物进行了抗肿瘤体内试验,测定其对H22荷瘤小鼠抑瘤率、免疫器官指数、免疫因子含量和生存率的影响。试验结果表明：粗毛纤孔菌、鲍姆木层孔菌、火木层孔菌和瓦宁木层孔菌的石油醚提取物高、低剂量（100,50 mg/kg）以及黑壳目层孔菌石油醚提取物高剂量（100 mg/kg）对肿瘤均具有一定抑制作用,抑瘤率均〉40%;瓦宁木层孔菌石油醚提取物低剂量组（50 mg/kg）的抑瘤效果最佳,为76.82%,其脾指数、胸腺指数均明显高于阳性组（P〈0.01）,IL 2的含量也明显高于对照组和阳性组,能延长小鼠的生存期;椭圆嗜蓝孢孔菌石油醚提取物也具有一定的抑瘤效果,高、低剂量（100,50 mg/kg）抑瘤率分别为39.30%和36.17%。     

四种多孔菌子实体粗多糖抗肿瘤活性的比较研究

对粗毛纤孔菌、椭圆嗜蓝孢孔菌、火木层孔菌、木蹄层孔菌4种多孔菌子实体粗多糖成分的含量及其体内抗肿瘤活性进行了比较研究。结果表明粗毛纤孔菌子实体中的粗多糖含量为4.1%,高于其他3种多孔菌;同时,4种多孔菌子实体粗多糖对H22荷瘤小鼠均显示出一定的抗肿瘤活性,除木蹄层孔菌外,其他3种多孔菌给药剂量为500mg/mL和1 000mg/mL时抑瘤率均大于40%,其中粗毛纤孔菌子实体粗多糖抑瘤率最高,低剂量组（500mg/kg）为58.12%,高剂量组（1 000mg/kg）为47.75%。     

忍冬木层孔菌液体培养过程中多酚含量及抗氧化活性研究

忍冬木层孔菌是木层孔菌属的一种药用真菌,目前对它的研究少有报道。以液体培养为基础,研究忍冬木层孔菌培养过程中生物量、胞内外多酚含量的变化,以及胞内外提取物抗氧化活性的变化。结果显示,菌丝体生物量在培养168–216h时达到最大值（8.33g/L）;胞内多酚和胞外多酚含量分别在216h和168h时达到最大值（15.60mg GAE/g和106.76mg GAE/L）;液体培养胞内外提取物的抗氧化活性与多酚含量正相关,多酚含量高时提取物对DPPH[1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl]自由基的抑制率也高。     

火木层孔菌发酵菌粉及其组分对小鼠移植性肝癌H22的体内生长作用

采用肝癌H22荷瘤小鼠的肿瘤模型,对火木层孔菌(桑黄)Phellinus igniarius发酵菌粉及其各提取物组分的体内抗肿瘤活性进行评价。结果表明,火木层孔菌发酵菌粉及其各提取物组分对荷瘤小鼠肝癌H22都具有一定的抗肿瘤作用并能延长荷瘤小鼠生存期,其中,火木层孔菌发酵菌粉(1,000mg/kg/d)及其组分I(I多糖组分)(360mg/kg/d)具有较为显著的抗肿瘤作用,抑瘤率分别为33.5%和40.3%。组织病理学研究结果表明在菌粉及其多糖组分作用后,肿瘤细胞坏死细胞明显增多,免疫组化检测表明火木层孔菌发酵菌粉及其多糖组分能明显的降低瘤组织中Bcl-2基因蛋白的表达,提高小鼠瘤组织中的Bax基因蛋白的表达。火木层孔菌发酵菌粉及其多糖组分具有较好的体内抗肿瘤活性。     

Functional expression of a bark beetle cytochrome P450 that hydroxylates myrcene to ipsdienol

The final steps in the pheromone-biosynthetic pathway of the pine engraver beetle, Ips pini (Say) (Coleoptera: Scolytidae) are unknown, but likely involve myrcene (7-methyl-3-methylene-1,6-octadiene) hydroxylation to produce the aggregation pheromone component, ipsdienol (2-methyl-6-methylene-2,7-octadien-4-ol). We have isolated a full-length I. pini cDNA encoding a cytochrome P450, CYP9T2. The recovered cDNA is 1.83kb and the open reading frame encodes a 532 amino acid protein. CYP9T2 is regulated by the same physiological factors that induce pheromone production. Quantitative real-time PCR experiments showed that feeding on host phloem induced CYP9T2 expression in males, but not females, and that basal expression levels are highest in male midguts, similar to other I. pini pheromone-biosynthetic genes. Microsomes prepared from Sf9 cells co-expressing baculoviral-mediated recombinant CYP9T2 and housefly (Musca domestica) NADPH-cytochrome P450 reductase converted myrcene to ipsdienol. The product identified by coupled GC-MS was mostly (4R)-(-)-ipsdienol, an important aggregation pheromone component for western North American I. pini. These results are consistent with CYP9T2 encoding a myrcene hydroxylase that functions near the end of the pheromone-biosynthetic pathway.     

Reciprocal interactions between the bark beetle-associated yeast Ogataea pini and host plant phytochemistry

Here we report the first experiments testing reciprocal effects between the bark beetle-associated yeast, Ogataea pini, and phytochemicals present in tree tissues (Pinus ponderosa). We tested two hypotheses: (i) tree phytochemicals mediate O. pini growth and (ii) O. pini affects chemical composition of plant tissues. We tested six monoterpenes on O. pini biomass growth in vitro and found that most monoterpenes inhibited O. pini growth; however mean O. pini biomass increased 21.5% when treated with myrcene and 75.5% when treated with terpinolene, relative to control. Ogataea pini was grown on phloem tissue ex vivo to determine whether O. pini affected phloem chemistry. Monoterpene concentrations declined in phloem over time, but phloem colonized by O. pini had significantly different concentrations of monoterpenes at two periods than phloem with no yeast. After 7 d, when O. pini was present, concentrations of the monoterpene Δ-3-carene was 42.9% lower than uncolonized phloem and concentrations of the monoterpene terpinolene was 345.0% higher than uncolonized phloem. After 15 d phloem colonized by O. pini had 505.4% higher concentrations of α-pinene than uncolonized phloem. These experiments suggest that O. pini responds to phytochemicals present in host tissues and the presence of O. pini might alter the chemical environment of phloem tissues during the early stages of beetle development. The interactions between O. pini and phytochemicals in pine vascular tissues might have consequences for the bark beetle that vectors O. pini, Dendroctonus brevicomis.     

New genetic markers for identifying Cronartium flaccidum and Peridermium pini and examining genetic variation within and between lesions of Scots pine blister rust in Sweden

Microsatellite markers were developed as an identification tool and for analysis of the genetic variation in the pathogens Cronartium flaccidum and Peridermium pini, causing Scots pine blister rust in Pinus spp. Six reference aeciospore samples from Finland were used to examine genetic differences between the two pathogens. Genetic variation within and between 27 lesions on Scots pines from seven locations in Sweden was also investigated. Aeciospores were collected from single aecia within the lesions. Reference samples from P. pini were homozygous for all seven microsatellite loci investigated, while the three C.?flaccidum samples contained heterozygous loci. These results confirm previous studies, where homozygous aeciospores were indicated to be characteristic for P. pini. The majority of aeciospores had two nuclei in both heterozygotic and homozygotic samples. Five of the Swedish lesions contained only homozygotic aecia, while the aecia in the remaining 22 lesions were heterozygotic. All lesions with homozygotic aecia contained only one single multilocus genotype, while many of the lesions with heterozygotic aecia contained several genotypes. The latter indicates the occurrence of multiple matings within a lesion between the resident spermogonia and alien fertilizing spermatia.     

Ips pini (Curculionidae: Scolytinae) is a vector of the fungal pathogen, Sphaeropsis sapinea (Coelomycetes), to Austrian pines, Pinus nigra (Pinaceae)

Sphaeropsis sapinea (Fr.:Fr.) Dyko and Sutton, is among the most common and widely distributed pathogens of conifers worldwide. S. sapinea is disseminated over short distances by rain splash and moist wind, but significant knowledge gaps regarding long-range dispersal remain. Our objective was to determine whether or not the pine engraver beetle, Ips pini Say, is a vector of the pathogen onto Austrian pines (Pinus nigra Arnold). In 2004 and 2005, individuals of I. pini were collected with pheromone traps at two locations in central Ohio (197 and 1,017 individuals for 2004 and 2005, respectively) and screened for the presence of S. sapinea. In the field, fresh logs of Austrian pine were baited with pheromone lures, mechanically wounded, or left undisturbed. After 2 mo, logs were evaluated for insect feeding and the presence of S. sapinea along beetle galleries. Fresh logs were also inoculated in the greenhouse with adult I. pini that were either artificially infested or uninfested with S. sapinea spores to determine vectoring potential. Phoresy rates for individual collections ranged from 0 to 4.1%; average rates were 1.5 and 2.0% for 2004 and 2005, respectively. Isolation frequencies of S. sapinea from baited (15 +/- 5%) and unbaited logs (3 +/- 1%) differed significantly (P=0.009). I. pini was also capable of transmitting the pathogen under controlled conditions. Based on phoresy rates, association, and artificial inoculation studies, we conclude that I. pini is able to transmit S. sapinea to Austrian pine stems.     

Multispecies trapping of six pests of scots pine in Sweden and Poland

Abstract:  Multispecies sex pheromone trapping (trapping of more than one species in the same trap) for the pine shoot moth Rhyacionia buoliana , the pine moth, Dendrolimus pini , the nun moth, Lymantria monacha , the pine beauty moth, Panolis flammea and the pine sawflies Diprion pini and Neodiprion sertifer was evaluated. The catch from traps baited with the pheromone of a single species was compared with the catch from traps baited with pheromones for several species. The catch in the multispecies traps was significantly reduced in comparison with the single species traps for Dendrolimus pini , L. monacha and N. sertifer . Neodiprion sertifer was most likely inhibited by the Diprion pini pheromone. A follow-up study of Dendrolimus pini and L. monacha showed no pheromonal interference between them. Further studies are needed to evaluate the feasibility of multispecies monitoring. However, for practical applications a slight decrease in catch, due to pheromonal interference, could probably be tolerated as long as the catch reflects total population density.     

Analysis of volatiles from black pine (Pinus nigra): significance of wounding and egg deposition by a herbivorous sawfly

The composition of headspace volatiles of black pine (Pinus nigra) was analysed by coupled gas chromatography-mass spectrometry (GC-MS). It has been shown in a previous study that egg deposition of the sawfly Diprion pini on P. sylvestris induces a quantitative change of the pine volatile blend . Chrysonotomyia ruforum, an egg parasitoid of D. pini, is known to be attracted by volatiles from egg-carrying P. sylvestris, but not by odour from egg-laden P. nigra. Therefore, the present study focused on the question whether also P. nigra as another host plant of this sawfly responds to egg deposition by change of its volatile blend. The headspace of untreated, egg-carrying, and artificially wounded P. nigra twigs were compared. The artificial damage inflicted to the twigs mimicked the damage by the sawfly female prior to egg deposition. Thirty five mainly terpenoid compounds that were identified in more than 50% of the egg-carrying P. nigra twigs could also be detected in the headspace of untreated and artificially wounded twigs. Quantitative differences of the blends of volatiles from differently treated P. nigra twigs were compared by multivariate data analyses. PLS-DA (projection to latent structures-discriminant analysis) revealed that blends of volatiles from differently treated P. nigra differed significantly. When comparing volatiles from egg-carrying and artificially wounded P. nigra with respective P. sylvestris samples, qualitative and quantitative differences were detected. The differences in volatile composition of P. nigra and P. sylvestris are discussed with special respect to the egg parasitoid's response to odours of egg-carrying pine twigs.     

Different defense strategies of Dendrolimus pini, Galleria mellonella, and Calliphora vicina against fungal infection

The resistance of Galleria mellonella, Dendrolimus pini, and Calliphora vicina larvae against infection by the enthomopathogen Conidiobolus coronatus was shown to vary among the studied species. Exposure of both G. mellonella and D. pini larvae to the fungus resulted in rapid insect death, while all the C. vicina larvae remained unharmed. Microscopic studies revealed diverse responses of the three species to the fungal pathogen: (1) the body cavities of D. pini larvae were completely overgrown by fungal hyphae, with no signs of hemocyte response, (2) infected G. mellonella larvae formed melanotic capsules surrounding the fungal pathogen, and (3) the conidia of C. coronatus did not germinate on the cuticle of C. vicina larvae. The in vitro study on the degradation of the insect cuticle by proteases secreted by C. coronatus revealed that the G. mellonella cuticle degraded at the highest rate. The antiproteolytic capacities of insect hemolymph against fungal proteases correlated well with the insects' susceptibility to fungal infection. The antiproteolytic capacities of insect hemolymph against fungal proteases correlated well with the insects' susceptibility to fungal infection. Of all the tested species, only plasmatocytes exhibited phagocytic potential. Exposure to the fungal pathogen resulted in elevated phagocytic activity, found to be the highest in the infected G. mellonella. The incubation of insect hemolymph with fungal conidia and hyphae revealed diverse reactions of hemocytes of the studied insect species. The encapsulation potential of D. pini hemocytes was low. Hemocytes of G. mellonella showed a high ability to attach and encapsulate fungal structures. Incubation of C. vicina hemolymph with C. coronatus did not result in any hemocytic response. Phenoloxidase (PO) activity was found to be highest in D. pini hemolymph, moderate in G. mellonella, and lowest in the hemolymph of C. vicina. Fungal infection resulted in a significant decrease of PO activity in G. mellonela larvae, while that in the larvae of D. pini remained unchanged. PO activity in C. vicina exposed to fungus slightly increased. The lysozyme-like activity increased in the plasma of all three insect species after contact with the fungal pathogen. Anti E. coli activity was detected neither in control nor in infected D. pini larvae. No detectable anti E. coli activity was found in the control larvae of G. mellonella; however, its exposure to C. coronatus resulted in an increase in the activity to detectable level. In the case of C. vicina exposure to the fungus, the anti E. coli activity was significantly higher than in control larvae. The defense mechanisms of D. pini (species of economic importance in Europe) are presented for the first time.     

Effect of host tree seasonal phenology on substrate suitability for the pine engraver (Coleoptera: Scolytidae): implications for population dynamics and enemy free space

This study evaluated the effects of seasonal phenology on the substrate quality of susceptible hosts to the pine engraver, Ips pini (Say). We also determined the effects of the duration and method of storage on host quality for purposes of laboratory rearing. Live red pine trees were felled at various times during the season, and I. pini adults from a laboratory colony were established on the logs. Subsamples of logs were stored for various intervals, and then provided to beetles. Subsamples of stored logs were waxed at both ends to prevent water loss before being submitted to the same assays. Suitability of red pine phloem tissue in susceptible hosts declined for I. pini throughout the growing season. As the season progressed, the number of beetle progeny that emerged from colonized hosts dropped substantially. This decline was associated with simultaneous reductions in phloem moisture content. Reduction in host suitability may partially offset any advantage I. pini may gain from colonizing trees after the major predators have become less abundant. Bark beetle brood production decreased significantly with length of storage, regardless of the month of tree felling or the method of storing. Implications for bark beetle population dynamics and laboratory rearing systems are discussed.