锁阳及其寄主白刺内生真菌的生态分布及遗传关系

首次研究了寄生植物锁阳及其寄主白刺内生真菌的分布特征及遗传关系.采用组织块法分离天然白刺、寄生体中锁阳和白刺的内生真菌,利用ITS-rDNA分子序列并结合形态学方法鉴定菌种,研究内生真菌的分离率、定殖率、分离频率、多样性指数、均匀度指数及相似性系数等的差异,以及寄生关系中内生真菌的多样性、遗传关系及分布特征等.结果表明: 本次获得的49株内生真菌隶属于18个分类单元,95.9%为子囊菌,4.1%为担子菌;内生真菌总分离率为15.3%,总定殖率为25.0%;天然白刺中内生真菌Shannon多样性指数最大,为2.13;锁阳花序与锁阳茎的内生真菌相似性系数最大,为0.50;镰孢菌属为白刺的优势菌属,青霉属为锁阳的优势菌属.锁阳与白刺寄生体中真菌类群的差异性分布表明寄生关系对内生真菌群落存在一定影响.     

Quinolizidine alkaloid status of Styphnolobium and Cladrastis (Leguminosae)

Reports of quinolizidine alkaloids in Styphnolobium Schott and Cladrastis Raf. (Leguminosae) conflict with their position in recent molecular phylogenies because they are not members of a major clade of quinolizidine alkaloid-accumulating taxa. The alkaloid status of these two genera was therefore re-investigated using gas chromatography–mass spectrometry. Quinolizidine alkaloids could not be detected in extracts of leaves, flowers or seeds of S. japonicum (L.) Schott, nor in leaves of S. affine (Torrey & A. Gray) Walp., C. delavayi (Franch.) Prain, C. kentukea (Dum.-Cours.) Rudd or C. platycarpa Mak. In contrast, Calia secundiflora (Ortega) Yakovlev, also currently placed outside the major clade of quinolizidine alkaloid-producing genera in molecular phylogenies, was confirmed to accumulate a range of quinolizidine alkaloids.     

Potential of fungal antagonists for biocontrol of Fusarium spp. in wheat and maize through competition in crop debris

Pathogenic Fusarium spp. cause head blight in wheat or ear rot in maize leading to yield losses and also a reduction in quality due to mycotoxin contamination of the grain. Infected crop residues are the main inoculum source for epidemics. Saprophytic fungi, obtained from cereal tissues or necrotic tissues of other crops, were screened for their ability to colonise wheat straw and maize stalks and to suppress sporulation of pathogenic Fusarium spp. Results of bio-assays conducted under controlled conditions were variable among Fusarium spp. and host substrates for most antagonists tested, such as yeasts, Trichoderma spp. and non-pathogenic Fusarium spp. Isolates of Clonostachys rosea consistently suppressed sporulation of F. culmorum and F. graminearum on wheat straw, and of F. culmorum, F. graminearum, F. proliferatum and F. verticillioides on maize stalks. Isolates of C. rosea, C. cladosporioides and F. equiseti were applied to pieces of maize stalks or flowering ears in preliminary experiments conducted under field conditions. The colonisation of stalk pieces by pathogenic Fusarium spp. was assessed after 9 months. Colonisation of stalk pieces by pathogenic Fusarium spp. was significantly reduced at several sampling dates. However, results obtained with the antagonists were not consistent for all sampling dates and between experiments.     

A yeast clade near Candida kruisii uncovered: nine novel Candida species associated with basidioma-feeding beetles

Yeasts similar to Candida kruisii were isolated repeatedly from the digestive tracts of basidioma-feeding beetles, especially nitidulids inhabiting and feeding on a variety of agarics in the southeastern USA and Barro Colorado Island, Panama. Based on the identical sequences of the D1/D2 domains of the LSU rRNA gene (rDNA) and host beetle information, the isolates were grouped into 19 genotypes which varied from C. kruisii by up to 38 nucleotide differences in the D1/D2 region. Phylogenetic analysis of rDNA sequences and phenotypic traits placed the isolates in C. kruisii and in nine undescribed taxa. The new species and type strains are designated as Candida pallodes (NRRL Y-27653^T), C. tritomae (NRRL Y-27650^T), C. panamensis (NRRL Y-27657^T), C. lycoperdinae (NRRL Y-27658^T), C. atbi (NRRL Y-27651^T), C. barrocoloradensis (NRRL Y-27934^T), C. aglyptinia (NRRL Y-27935^T), C. stri (NRRL Y-48063^T), and C. gatunensis (NRRL Y-48064^T). A phylogeny based on analysis of a combined database of sequences of SSU and LSU rDNA and the ITS region showed that the nine new species formed a novel sister clade to C. kruisii that was strongly supported by bootstrap analysis. Candida pallodes, C. tritomae, C. panamensis, and C. lycoperdinae formed one subclade, while C. atbi, C. barrocoloradensis, C. aglyptinia, C. stri, and C. gatunensis formed a second distinct subclade within the larger clade. Candida pallodes and C. atbi showed a strong host specificity to beetle species in the genus Pallodes (Coleoptera: Nitidulidae) collected from a variety of agarics. On the other hand, C. panamensis, C. tritomae, and C. lycoperdinae were associated with several unrelated beetles in Erotylidae, Scarabaeidae, Tenebrionidae, and Curculionidae as well as Lycoperdina ferruginea (Nitidulidae). Candida pallodes, C. tritomae, C. lycoperdinae, and C. atbi have been isolated repeatedly in the USA, while the other five new species have been found only at Barro Colorado Island, Panama.     

A novel group I intron in Candida dubliniensis is homologous to a Candida albicans intron

In the present study, we determined the sequence of group I self-splicing introns found in the large ribosomal RNA subunit of Candida albicans, Candida stellatoidea and the recently-described species Candida dubliniensis. It was found that both the intron and ribosomal RNA nucleotide sequences are almost perfectly identical between different C. albicans strains as well as between C. albicans and C. stellatoidea strains. Comparisons of ribosomal RNA sequences suggest that local isolates of atypical C. albicans from individuals infected with human immunodeficiency virus can be assigned to the C. dubliniensis species. C. dubliniensis strains also harbor a group I intron in their ribosomal RNA, as observed in about 40% of C. albicans strains and all C. stellatoidea strains. This novel C. dubliniensis group I intron is identical to the C. albicans and C. stellatoidea intron, except for two widely divergent stem-loop regions. Despite these differences, the C. dubliniensis intron possesses self-splicing ability in an in vitro assay. Taken together, these data support the idea that C. albicans and C. stellatoidea should be joined together as variants of the same species while C. dubliniensis is a distinct but closely related microorganism. To our knowledge, the C. albicans and C. dubliniensis introns are the first example of a pair of homologous group I introns differing only by the presence of apparently facultative sequences in some stem-loops suspected to be involved in stabilization of tertiary structure.     

Isolation and characterization of some mycelia inhabiting Tuber ascomata

Tuber spp. are ectomycorrhizal ascomycetes that produce subterranean ascomata known as truffles. Truffles can be regarded as complex microhabitats hosting bacteria and yeasts. In this paper we show that guest filamentous fungi are also associated to truffle ascomata, regardless of the Tuber spp., and report the morpho-molecular characterization of seven truffle-hosted mycelia isolated from healthy and intact Tuber ascomata. Some of these isolates were shown to be related to the fungal endophytes of plants. Interestingly, the truffle-hosted mycelia grew stuck to the hyphal wall of their partner when co-cultivated with the Tuber borchii mycelium, but not when co-cultivated with the test species Agaricus macrosporus. The present data suggest that guest filamentous fungi can be added to the list of truffle-interacting microorganisms.     

凋落叶多样性对杉木幼苗生长及吸收15N标记硫铵的影响

利用¹⁵N硫铵研究了凋落叶多样性对杉木幼苗生长及养分吸收的影响.结果表明,凋落叶多样性的增加有利于盆栽杉木幼苗的生长.杉木、火力楠、红栲和刺楸4种凋落叶混合处理后,杉木幼苗的生长量最大;杉木、火力楠、刺楸3种凋落叶混合处理后的杉木幼苗生长量次之,其它依次为杉木、火力楠、红栲3种凋落叶混合处理>杉木和刺楸凋落叶处理>杉木和红栲凋落叶处理>对照>杉木和火力楠2种凋落叶混合处理>杉木凋落叶处理.就杉木幼苗对硫铵氮的吸收率而言,不作任何处理的杉木幼苗吸收率最高,其次为杉木、火力楠、红栲和刺楸4种凋落叶混合处理,其它依次为杉木、火力楠、刺楸3种凋落叶混合处理和杉木、火力楠、红栲3种凋落叶混合处理>杉木和刺楸凋落叶处理>杉木和红栲凋落叶处理>杉木和火力楠2种凋落叶混合处理>杉木凋落叶处理.另外,用凋落叶处理后,土壤中硫铵氮的残留量比不作凋落叶处理的土壤多,硫铵氮的总回收量也比不作凋落叶处理的土壤大幅增加,而且凋落叶多样性的增加也会增加硫铵氮的残留量.     

Species of Colletotrichum on Agavaceae

Species of Colletotrichum cause diseases on a wide range of hosts, frequently infecting plants in the Agavaceae (monocotyledons: Liliales). Three species of Colletotrichum restricted to the Agavaceae were detected through morphological studies of specimens and molecular sequence analyses of the LSU of the nu-rDNA and the ITS region of the nu-rDNA from cultures. Colletotrichum agaves on Agave is fully described and illustrated. Colletotrichum dracaenophilum is described as a new species for isolates having long conidia and occurring on Dracaena sanderiana from China. Colletotrichum phormii and Glomerella phormii are determined to be the correct scientific names for the asexual and sexual states, respectively, of a species commonly referred to as C. rhodocyclum and G. phacidiomorpha occurring mainly on Phormium. In addition, C. gloeosporioides and C. boninense were isolated from plants in the Agavaceae. All species of Colletotrichum described on Agavaceae were evaluated based on type specimens. A key to the five species of Colletotrichum on Agavaceae is included. This paper includes one new species, Colletotrichum dracaenophilum, and three new combinations, Colletotrichum phormii, Glomerella phormii, and Phaeosphaeriopsis phacidiomorpha.     

Fungal root endophytes of the carnivorous plant Drosera rotundifolia

As carnivorous plants acquire substantial amounts of nutrients from the digestion of their prey, mycorrhizal associations are considered to be redundant; however, fungal root endophytes have rarely been examined. As endophytic fungi can have profound impacts on plant communities, we aim to determine the extent of fungal root colonisation of the carnivorous plant Drosera rotundifolia at two points in the growing season (spring and summer). We have used a culture-dependent method to isolate fungal endophytes and diagnostic polymerase chain reaction methods to determine arbuscular mycorrhizal fungi colonisation. All of the roots sampled contained culturable fungal root endophytes; additionally, we have provided molecular evidence that they also host arbuscular mycorrhizal fungi. Colonisation showed seasonal differences: Roots in the spring were colonised by Articulospora tetracladia, two isolates of uncultured ectomycorrhizal fungi, an unidentified species of fungal endophyte and Trichoderma viride, which was present in every plant sampled. In contrast, roots in the summer were colonised by Alatospora acuminata, an uncultured ectomycorrhizal fungus, Penicillium pinophilum and an uncultured fungal clone. Although the functional roles of fungal endophytes of D. rotundifolia are unknown, colonisation may (a) confer abiotic stress tolerance, (b) facilitate the acquisition of scarce nutrients particularly at the beginning of the growing season or (c) play a role in nutrient signalling between root and shoot.     

少花蒺藜草入侵对沙质草地氮库的影响

以我国北方大面积发生的入侵植物少花蒺藜草为对象,研究了其对科尔沁沙质草地土壤氮库和地上植物氮库的影响,并在室内条件下,以自然生境中少花蒺藜草伴生牧草披碱草和扁穗冰草为对照,利用¹⁵N同位素技术分析了少花蒺藜草的生物固氮能力.结果表明:与对照裸地和本地植物鹅观草样地相比,少花蒺藜草入侵地土壤总氮库分别显著增加47.5%和20.8%,土壤铵态氮库分别显著降低25.6%和25.2%.与鹅观草相比,少花蒺藜草地上部分植物氮库显著降低18.7%.少花蒺藜草的¹⁵N原子丰度、¹⁵N原子百分超、¹⁵N原子加权百分超均显著低于披碱草和扁穗冰草.少花蒺藜草和披碱草的氮肥利用率分别为48.5%和47.0%,差异不显著.与披碱草相比,少花蒺藜草生物固氮率为60.2%Ndfa.少花蒺藜草对氮素高效利用的特征,揭示了其适应沙质草地生态系统并蔓延的一种生态适应机制.     

A fourth gene from the Candida albicans CDR family of ABC transporters

Using primers derived from a region of the Candida albicans CDR1 (Candida drug resistance) gene that is conserved in other ABC (ATP-binding cassette) transporters, a DNA fragment from a previously unknown CDR gene was obtained by polymerase chain reaction (PCR). After screening a C. albicans genomic library with this fragment as a probe, the complete CDR4 gene was isolated and sequenced. CDR4 codes for a putative ABC transporter of 1490 amino acids with a high degree of homology to Cdr1p, Cdr2p and Cdr3p from C. albicans (62, 59 and 57% amino acid sequence identity, respectively). Cdr4p has a predicted structure typical for cluster I.1 of yeast ABC transporters, characterized by two homologous halves, each comprising an N-terminal hydrophilic domain with consensus sequences for ATP binding and a C-terminal hydrophobic domain with six transmembrane helices. In contrast to the CDR1/CDR2 genes, the genetic structure of the CDR4 gene was conserved in 59 C. albicans isolates from six different patients. Northern hybridization analysis showed that the CDR4 gene was expressed in most isolates, but no correlation between CDR4 mRNA levels and the degree of fluconazole resistance of the isolates was found. In addition, a C. albicans mutant in which both copies of the CDR4 gene were disrupted by insertional mutagenesis was not hypersusceptible to fluconazole as compared to the parent strain. Unlike CDR1 and CDR2, CDR4 does not, therefore, seem to be involved in fluconazole resistance in C. albicans.     

Lignin degradation by white rot fungi on spruce wood shavings during short-time solid-state fermentations monitored by near infrared spectroscopy

Due to their outstanding capability of degrading the recalcitrant biomacromolecule lignin, white rot fungi have been attracting interest for several technological applications in mechanical pulping and wood surface modification. However, little is known about the time course of delignification in early stages of colonisation of wood by these fungi. Using a Fourier transform near infrared (FT-NIR) spectroscopic technique, lignin loss of sterilised spruce wood shavings (0.4–2.0 mm particle size) that had been degraded by various species of white rot fungi could be monitored already during the first 2 weeks. The delignification kinetics of Dichomitus squalens, three Phlebia species (Phlebia brevispora, Phlebia radiata and Phlebia tremellosa), three strains of Ceriporiopsis subvermispora as well as the white rot ascomycete Hypoxylon fragiforme and the basidiomycete Oxyporus latemarginatus were determined. Each of the fungi tested was able to reduce the lignin content of spruce wood significantly during the first week. The amount of delignification achieved by the selected white rot fungi after 2 weeks ranged from 7.2% for C. subvermispora (FPL 105.752) to 2.5% for P. radiata. Delignification was significant (P = 95%) already after 3 days treatment with C. subvermispora and P. tremellosa. Activities of extracellular ligninolytic enzymes (laccase, manganese peroxidase and/or lignin peroxidase), expressed by each of the tested fungi, were determined. Lignin was degraded when peroxidase activity was detected in the fungal cultures, but only a low level of correlation between enzyme activities and the extent of delignification was found.     

马尾松人工林林窗边缘效应对樟和红椿凋落叶难降解物质分解的影响

采用凋落物分解袋, 以四川低山丘陵区马尾松人工林人工砍伐形成的7个不同面积的林窗边缘(100、225、400、625、900、1225、1600 m²)为研究对象, 以林下为对照, 研究了2种乡土树种——樟和红椿凋落叶难降解物质(木质素、纤维素、总酚、缩合单宁)在不同大小林窗边缘的降解动态特征.结果表明:马尾松人工林林下与不同大小林窗边缘相比较,红椿凋落叶中除纤维素外,其余难降解物质的降解率以及樟凋落叶木质素降解率均显著高于林下.在全年分解过程中, 2种凋落叶4种难降解物质的降解率总体均呈现持续上升的趋势.其中,缩合单宁降解最快,其次是总酚和纤维素,而木质素降解最慢.随林窗面积的增大, 红椿凋落叶除纤维素外,其余难降解物质在中型林窗边缘(400、625 m²)具有相对较高的降解率,而樟凋落叶的木质素在625 m²林窗边缘时也表现出较高的降解率.在凋落叶分解过程中,难降解物质降解率与凋落叶袋内温度和凋落物质量均呈显著相关.中型林窗(400～625 m²)对凋落物分解过程中难降解物质的降解具有更显著的边缘效应, 而这种边缘效应与物种有一定关联.     

Plant and fungal responses to elevated atmospheric carbon dioxide in mycorrhizal seedlings of Pinus sylvestris

The effects of elevated CO₂ concentration upon the mycorrhizal relationships of Scots pine (Pinus sylvestris) seedlings were investigated. Plants were grown for 4 months with their shoots exposed to ambient (C_AMB=360 μl l⁻¹) or elevated (C_ELEV=700 μl l⁻¹) CO₂ environments while their root systems, either colonised by the mycorrhizal fungi Paxillus involutus or Suillus bovinus, or left in the non-mycorrhizal condition, were maintained in sealed dishes. In one series of these plants the effects of C_ELEV upon the extent of mycorrhizal development and upon their growth and nutrition were determined, while another series were transferred from the dishes after 1 month, to transparent observation chambers before being returned to the two CO₂ environments. In these chambers, the effects of C_ELEV upon development of the external mycelial systems of the two mycorrhizal fungi was determined by measuring the advance of the hyphal fronts of the mycorrhizal fungi across non-sterile peat from the colonised plants. The dry mass and number of mycorrhizal tips were significantly higher in C_ELEV than in the C_AMB condition in plants colonised by both fungi in the dishes. Yields of whole plants and of shoots were higher in the C_ELEV treatment whether or not they were grown in the mycorrhizal condition, but the greater yields were not associated in these sealed systems with enhanced nutrient gain. The dry mass of non-mycorrhizal plants was greater than that of those colonised by mycorrhizal fungi under elevated CO₂. This is thought to be attributable to the energetic cost of production of the larger mycorrhizal systems in this treatment. The extent of development of the mycorrhizal mycelial systems of both fungi was greatly increased in C_ELEV relative to that in C_AMB environments. It is hypothesised that increased allocation of carbon to mycorrhizal root systems and their associated mycelia would provide the potential for enhancement of nutrient acquisition in open systems of greater fertility.     

Allozyme differentiation in four species of the Crocus cartwrightianus group and in cultivated saffron (Crocus sativus)

The genetic variability and divergence of four species of the genus Crocus L., related to Crocus cartwrightianus group, namely Crocus thomasii Ten., Crocus hadriaticus Herbert, Crocus oreocreticus B.L. Burtt, C. cartwrightianus Herbert, has been studied by means of starch gel isozyme electrophoresis. For each population the following enzymatic loci were analyzed: PGI-1, PGI-2, G6PDH-1, G6PDH-2, IDH-1, IDH-2, 6PGDH-1, 6PGDH-2, SKDH-1, SKDH-2, AK-1 and AK-2. The genetic variability was estimated through the parameters A (mean number of alleles per locus), P (percent of polymorphic loci), H_o (mean observed heterozygosity), and H_e (mean expected heterozygosity). The genetic differentiation has been assayed by Wrigth's F-statistics, and the genetic divergence by Nei's index. Our data confirmed that the taxa are distinct species, in spite of their similar morphology and karyology. C. thomasii is more genetically similar to C. cartwrightianus and C. oreocreticus than to C. hadriaticus. We hypothesized an autopolyploid origin of saffron, probably from C. cartwrightianus, considering the genotypic classes of Crocus sativus and the other related species of the C. cartwrightianus group studied here.     

西藏东南部柏木属群落类型及物种多样性

西藏东南部柏木属植物群落是青藏高原天然林资源的重要组分,在维持高原生物多样性与生态安全方面作用显著.为明确其群落类型及物种多样性,本研究通过在米林县、朗县和波密县设置典型样地和划分群落类型,分析了不同群落物种组成、物种多样性指数及建群种径级结构.结果表明: 柏木属植物群落可划分为4个群落类型:西藏柏木-柳叶忍冬+绒毛山梅花-凤尾蕨+紫马唐群落(群落Ⅰ)、巨柏-圆锥山蚂蝗+砂生槐-线形草沙蚕+芸香草+固沙草群落(群落Ⅱ)、巨柏-甘肃野丁香-杂配藜+固沙草群落(群落Ⅲ)、巨柏-小蓝雪花-藏沙蒿+固沙草+蒺藜群落(群落Ⅳ).各群落Margalef丰富度指数、Shannon多样性指数、Pielou均匀度指数及种间相遇机率指数均表现为:群落Ⅰ＞群落Ⅱ＞群落Ⅲ＞群落Ⅳ.4个群落的建群种幼龄级个体多而中老龄级少,植株由幼龄级向中龄级过渡阶段死亡率较高,柏木属植物种群呈衰退趋势.     

Susceptibility assays of Candida tropicalis to miconazole

In vitro evaluation of antifungal activity of imidazole derivatives is made difficult by the inhibitory effects of several factors such as inoculum size, growth form of the fungus, incubation temperature, the presence of complex substances, including divalent ions, which strongly influence final results. This is particularly evident when testing clinical isolates of C. tropicalis strains resistant to imidazole drugs. Our data based on assays of miconazole nitrate and miconazole sulfosalicylate against C. tropicalis show that it is possible to abolish various interference activities on the antimicrobial activity by suitable modifications of some cultural conditions. Thus, a study has been carried out to assess miconazole sulfosalicylate activity on C. tropicalis throughout experiments performed by contact test and agar diffusion test. The use of these techniques made it possible to display some activity of the imidazoles even against strains of C. tropicalis, which were defined as resistant using usual susceptibility assay conditions. Experimental conditions which cause the increase of susceptibility of C. tropicalis are related to factors that modify the barrier function and cellular permeability as demonstrated mainly by the effect of electrical conductivity (E.C.), pH of the medium and pretreatment of fungal inoculum with sodium dioctylsulfosuccinate (SDSS). Our results suggest that the correlation between drug dosage and inhibitory activity in vitro can be improved by the modifications proposed in this paper.     

Variations in chromosome size and organization in Candida albicans and Candida stellatoidea

Candida albicans and the closely related species Candida stellatoidea are medically important diploid asexual yeasts. Clinical isolates frequently show variant electrophoretic karyotypes, apparently due largely to chromosomal translocations. These translocations seem to occur at hot spots characterized by the repeated DNA sequence RSP1. A programmed karyotypic rearrangement occurs in C. stellatoidea. Karyotypic rearrangement may serve as a source of genetic variation in these asexual yeasts.     

Review on safety of the entomopathogenic fungi Beauveria bassiana and Beauveria brongniartii

The commercial use of entomopathogenic fungi and their products as mycoinsecticides necessitates their registration. Worldwide, several registration guidelines are available, however, most of them focus on similar or even the same safety issues. With respect to the two entomopathogenic fungi, Beauveria bassiana (Bals.-Criv.) Vuill. and Beauveria brongniartii (Sacc.) Petch, many commercial products have been developed, and numerous papers on different biological, environmental, toxicological and other safety aspects have been published during the past 30-40 years. The aim of the present review is to summarise these data. The following safety issues are presented: (1) identity of Beauveria spp.; (2) biological properties of Beauveria spp. (history, natural occurrence and geographical distribution, host range, mode of action, production of metabolites/toxins, effect of environmental factors); (3) analytical methods to determine and quantify residues; (4) fate and behaviour in the environment (mobility and persistence in air, water and soil); (5) effects on non-target organisms (non-target microorganisms, plants, soil organisms, aquatic organisms, predators, parasitoids, honey bees, earth worms and nontarget arthropods); (6) effects on vertebrates (fish, amphibia, reptiles and birds); and (7) effects on mammals and human health. Based on the present knowledge it is concluded that both Beauveria species are considered to be safe.