食用牛肝菌不同部位紫外指纹图谱鉴别分析

采用紫外光谱技术结合主成分分析法,建立快速鉴别牛肝菌的不同部位及不同种类、产地的方法。通过单因素实验确定提取牛肝菌特征成分的最佳条件,制备测试液并进行紫外光谱测定。结果表明,牛肝菌样品的重现性、精密度和10h内稳定性的RSD（%）分别在0.09–1.81、0.11–1.92、0.06–2.33之间;牛肝菌不同部位紫外指纹图谱具有明显的指纹特性;主成分分析（PCA）表明菌盖和菌柄前3个主成分累积贡献率分别为94.797%和92.961%,能够反映样品的主要信息;SIMCA软件分析显示牛肝菌菌盖和菌柄化学成分积累不同。根据牛肝菌不同部位紫外光谱信息和主成分分析能区分同一牛肝菌的不同部位,鉴别不同种类、不同产地的食用牛肝菌。     

Analysis of the genes encoding the largest subunit of RNA polymerase II in Arabidopsis and soybean

We have cloned and sequenced the gene encoding the largest subunit of RNA polymerase II (RPB1) from Arabidopsis thaliana and partially sequenced genes from soybean (Glycine max). We have also determined the nucleotide sequence for a number of cDNA clones which encode the carboxyl terminal domains (CTDs) of RNA polymerase II from both soybean and Arabidopsis. The Arabidopsis RPB1 gene encodes a polypeptide of approximately 205 kDa, consists of 12 exons, and encompasses more than 8 kb. Predicted amino acid sequence shows eight regions of similarity with the largest subunit of other prokaryotic and eukaryotic RNA polymerases, as well as a highly conserved CTD unique to RNA polymerase II.The CTDs in plants, like those in most other eukaryotes, consist of tandem heptapeptide repeats with the consensus amino acid sequence PTSPSYS. The portion of RPB1 which encodes the CTD in plants differs from that of RPB1 of animals and lower eukaryotes. All the plant genes examined contain 2–3 introns within the CTD encoding regions, and at least two plant genes contain an alternatively spliced intron in the 3 untranslated region. Several clustered amino acid substitutions in the CTD are conserved in the two plant species examined, but are not found in other eukaryotes. RPB1 is encoded by a multigene family in soybean, but a single gene encodes this subunit in Arabidopsis and most other eukaryotes.     

Multi-gene phylogenies indicate ascomal wall morphology is a better predictor of phylogenetic relationships than ascospore morphology in the Sordariales (Ascomycota, Fungi)

Ascospore characters have commonly been used for distinguishing ascomycete taxa, while ascomal wall characters have received little attention. Although taxa in the Sordariales possess a wide range of variation in their ascomal walls and ascospores, genera have traditionally been delimited based on differences in their ascospore morphology. Phylogenetic relationships of multiple representatives from each of several genera representing the range in ascomal wall and ascospore morphologies in the Sordariales were estimated using partial nuclear DNA sequences from the 28S ribosomal large subunit (LSU), beta-tubulin, and ribosomal polymerase II subunit 2 (RPB2) genes. These genes also were compared for their utility in predicting phylogenetic relationships in this group of fungi. Maximum parsimony and Bayesian analyses conducted on separate and combined data sets indicate that ascospore morphology is extremely homoplastic and not useful for delimiting genera. Genera represented by more than one species were paraphyletic or polyphyletic in nearly all analyses; 17 species of Cercophora segregated into at least nine different clades, while six species of Podospora occurred in five clades in the LSU tree. However, taxa with similar ascomal wall morphologies clustered in five well-supported clades suggesting that ascomal wall morphology is a better indicator of generic relationships in certain clades in the Sordariales. The RPB2 gene possessed over twice the number of parsimony-informative characters than either the LSU or beta-tubulin gene and consequently, provided the most support for the greatest number of clades.     

The mismatch in distributions of vertebrates and the plants that they disperse

Little is known about how mutualistic interactions affect the distribution of species richness on broad geographic scales. Because mutualism positively affects the fitness of all species involved in the interaction, one hypothesis is that the richness of species involved should be positively correlated across their range, especially for obligate relationships. Alternatively, if mutualisms involve multiple mutualistic partners, the distribution of mutualists should not necessarily be related, and patterns in species distributions might be more strongly correlated with environmental factors. In this study, we compared the distributions of plants and vertebrate animals involved in seed‐dispersal mutualisms across the United States and Canada. We compiled geographic distributions of plants dispersed by frugivores and scatter‐hoarding animals, and compared their distribution of richness to the distribution in disperser richness. We found that the distribution of animal dispersers shows a negative relationship to the distribution of the plants that they disperse, and this is true whether the plants dispersed by frugivores or scatter‐hoarders are considered separately or combined. In fact, the mismatch in species richness between plants and the animals that disperse their seeds is dramatic, with plants species richness greatest in the in the eastern United States and the animal species richness greatest in the southwest United States. Environmental factors were corelated with the difference in the distribution of plants and their animal mutualists and likely are more important in the distribution of both plants and animals. This study is the first to describe the broad‐scale distribution of seed‐dispersing vertebrates and compare the distributions to the plants they disperse. With these data, we can now identify locations that warrant further study to understand the factors that influence the distribution of the plants and animals involved in these mutualisms.     

First report of the ectomycorrhizal status of boletes on the Northern Yucatan Peninsula, Mexico determined using isotopic methods

Despite their prominent role for tree growth, few studies have examined the occurrence of ectomycorrhizal fungi in lowland, seasonally dry tropical forests (SDTF). Although fruiting bodies of boletes have been observed in a dry tropical forest on the Northern Yucatan Peninsula, Mexico, their occurrence is rare and their mycorrhizal status is uncertain. To determine the trophic status (mycorrhizal vs. saprotrophic) of these boletes, fruiting bodies were collected and isotopically compared to known saprotrophic fungi, foliage, and soil from the same site. Mean δ¹⁵N and δ¹³C values differed significantly between boletes and saprotrophic fungi, with boletes 8.0‰ enriched and 2.5‰ depleted in ¹⁵N and ¹³C, respectively relative to saprotrophic fungi. Foliage was depleted in ¹³C relative to both boletes and saprotrophic fungi. Foliar δ¹⁵N values, on the other hand, were similar to saprotrophic fungi, yet were considerably lower relative to bolete fruiting bodies. Results from this study provide the first isotopic evidence of ectomycorrhizal fungi in lowland SDTF and emphasize the need for further research to better understand the diversity and ecological importance of ectomycorrhizal fungi in these forested ecosystems.     

Evolutionary relationships of the cup-fungus genus Peziza and Pezizaceae inferred from multiple nuclear genes: RPB2, beta-tubulin, and LSU rDNA

To provide a robust phylogeny of Pezizaceae, partial sequences from two nuclear protein-coding genes, RPB2 (encoding the second largest subunit of RNA polymerase II) and beta-tubulin, were obtained from 69 and 72 specimens, respectively, to analyze with nuclear ribosomal large subunit RNA gene sequences (LSU). The three-gene data set includes 32 species of Peziza, and 27 species from nine additional epigeous and six hypogeous (truffle) pezizaceous genera. Analyses of the combined LSU, RPB2, and beta-tubulin data set using parsimony, maximum likelihood, and Bayesian approaches identify 14 fine-scale lineages within Pezizaceae. Species of Peziza occur in eight of the lineages, spread among other genera of the family, confirming the non-monophyly of the genus. Although parsimony analyses of the three-gene data set produced a nearly completely resolved strict consensus tree, with increased confidence, relationships between the lineages are still resolved with mostly weak bootstrap support. Bayesian analyses of the three-gene data, however, show support for several more inclusive clades, mostly congruent with Bayesian analyses of RPB2. No strongly supported incongruence was found among phylogenies derived from the separate LSU, RPB2, and beta-tubulin data sets. The RPB2 region appeared to be the most informative single gene region based on resolution and clade support, and accounts for the greatest number of potentially parsimony informative characters within the combined data set, followed by the LSU and the beta-tubulin region. The results indicate that third codon positions in beta-tubulin are saturated, especially for sites that provide information about the deeper relationships. Nevertheless, almost all phylogenetic signal in beta-tubulin is due to third positions changes, with almost no signal in first and second codons, and contribute phylogenetic information at the "fine-scale" level within the Pezizaceae. The Pezizaceae is supported as monophyletic in analyses of the three-gene data set, but its sister-group relationships is not resolved with support. The results advocate the use of RPB2 as a marker for ascomycete phylogenetics at the inter-generic level, whereas the beta-tubulin gene appears less useful.     

One large subunit of ribulose 1,5-bisphosphate carboxylase oxygenase in Medicago,Spinacia and Nicotiana

Summary Isoelectric focusing of subunits of ribulose 1,5-bisphosphate carboxylase oxygenase of Medicago, Spinacia and Nicotiana were investigated, using a rapid isolation technique, without S-carboxymethylation. RuBPC-ase and its subunits were isolated by gel electrophoresis. Isoelectric focusing of RuBPC-ase of M. sativa and M. falcata showed that this enzyme consists of one large subunit (LSU) polypeptide and two or three small subunits (SSU), depending on the genotype. The pl of the LSU's was identical, but the pl of SSU's of the two genotypes was different. Amino acid composition and tryptic peptide maps further supported the concept of a conserved nature of LSU and heterogeneity of SSU polypeptides in Medicago. It was also found that S. oleracea, N. tabacum, N. glutinosa and N. excelsior have a single LSU polypeptide, but they differ in respect of pl values. The SSU polypeptides appeared to be variable. S-carboxymethylation affected the number as well as the pl values of LSU and SSU polypeptides. It is suggested that one LSU polypeptide is probably the general rule in higher plants, rather than the three LSU polypeptides demonstrated by Chen et al. (1977) and Wildman (1979).     

Diversity of ant-plant interactions: protective efficacy in Macaranga species with different degrees of ant association

The pioneer tree Macaranga in SE Asia has developed manyfold associations with ants. The genus comprises all stages of interaction with ants, from facultative relationships to obligate myrmecophytes. Only myrmecophytic Macaranga offer nesting space for ants and are associated with a specific ant partner. The nonmyrmecophytic species are visited by a variety of different ant species which are attracted by extrafloral nectaries (EFN) and food bodies. Transitional Macaranga species like M. hosei are colonized later in their development due to their stem structure. Before the colonization by their specific Crematogaster partner the young plants are visited by different ant species attracted by EFN. These nectaries are reduced and food body production starts as soon as colonization becomes possible. We demonstrated earlier that obligate ant partners can protect their Macaranga plants against herbivore damage and vine cover. In this study we focused on nonspecific interactions and studied M. tanarius and M. hosei, representing a non-myrmecophyte and a transitional species respectively. In ant exclusion experiments both M. tanarius and M. hosei suffered significantly higher mean leaf damage than controls, 37% versus 6% in M. hosei, 16% versus 7% in M. tanarius. M. tanarius offers both EFN and food bodies so that tests for different effects of these two food rewards could be conducted. Plants with food bodies removed but with EFN remaining had the lowest mean increase of herbivore damage of all experimental groups. Main herbivores on M. hosei were mites and caterpillars. Many M. tanarius plants were infested by a shootborer. Both Macaranga species were visited by various ant species, Crematogaster spp. being the most abundant. We found no evidence for any specific relationships. The results of this study strongly support the hypothesis that non-specific, facultative associations with ants can be advantageous for Macaranga plants. Food bodies appear to have lower attractive value for opportunistic ants than EFN and may require a specific dietary adaptation. This is also indicated by the fact that food body production in the transitional M. hosei does not start before stem structure allows a colonization by the obligate Crematogaster species. M. hosei thus benefits from facultative association with a variety of ants until it produces its first domatia and can be colonized by its obligate mutualist.     

Leaf morphology of a facultative rheophyte,Farfugium japonicum var.luchuense (Compositae)

The gross morphology and anatomy of leaves in a facultative rheophyte,Farfugium japonicum var.luchuense, were examined in order to clarify how the two characters are correlated with one another and how the facultative rheophyte differs from obligate rheophytes in leaf morphology and anatomy. Most rheophytes of the variety have narrowly cuneate leaf base, while most inland plants usually have truncate to cordate one, although the habitat-morphology correlation is not so clear. The leaf shape or the divergence angle of leaf base is strongly correlated with the number of primary veins and intervein-distance dependent on the number of cells intervening between veins. This makes a marked contrast to many reported cases of obligate rheophytes in which the leaf shape is strongly correlated with cell size. There is a rough tendency that narrower leaves of rheophytes have a thicker cuticular layer. However, the cell size and the volume (area) of intercellular space differ only slightly with the leaf shape.     

不同气候条件下沙冬青属植物在我国的潜在分布——基于生态位模型预测

沙冬青属（Ammopiptanthus）植物是古地中海第三纪孑遗濒危物种,包括沙冬青（Ammopiptanthus mgolicus）和矮沙冬青（Ammopiptanthus nanus）,主要分布在我国西北干旱、半干旱地区,其不仅具有较高的研究价值,同时对我国西北干旱地区生态环境具有十分重要的作用。近年来由于全球气候变化及人为干扰等因素,沙冬青属植物天然分布面积骤缩,濒临灭绝。本研究利用MaxEnt模型、Bioclim模型和Domain模型对沙冬青属植物在我国末次间冰期（Last Interglacial）、末次冰盛期（Last Glacial Maximum）、当代和2050年（RCP4.5和RCP8.5）4个时期气候情景下的潜在适生区进行预测。结果表明：MaxEnt模型对沙冬青属植物潜在分布区的预测具有极高的准确度,所有模型的平均受试者工作特征曲线下面积（AUC测试值）均高于0.80。当代沙冬青最佳及高适生区占全国总面积的2.78%,主要集中在内蒙古中部、宁夏北部和甘肃北部等地;未来沙冬青最佳及高适生区在现有分布范围呈现向外扩张的趋势,主要分布在内蒙古鄂托克旗、鄂尔多斯、阿拉善左旗、宁夏吴忠和甘肃民勒县等地。当代矮沙冬青最佳及高适生区占全国总面积的2.23%,主要集中在新疆南部;未来矮沙冬青最佳及高适生区向新疆乌恰县南部、乌鲁木齐北部移动和扩大,主要分布在新疆乌恰县、乌苏市、吐鲁番市和乌鲁木齐市。未来2050年（RCP4.5和RCP8.5）两种气候情景下沙冬青和矮沙冬青的潜在分布总面积均有所增加,与当代相比变化不明显,但不同适生等级的潜在分布面积变化较大,在更高的CO₂排放量（RCP8.5）情景下沙冬青和矮冬青的最佳及高适生区范围的预测结果都将减少。从气候因素角度考虑,研究表明未来气候情景下沙冬青属植物的适生区变化过程中,年均温（Bio1）、最湿月降水量（Bio13）和温度季节性变化（Bio4）是影响沙冬青属植物分布的关键因子,并为我国西北干旱半干旱地区具有重要的经济价值并将持续其生态服务功能。     

Molecular evolution and genome divergence at <Emphasis Type="Italic">RPB2</Emphasis> gene of the St and H genome in <Emphasis Type="Italic">Elymus</Emphasis> species

Molecular evolution of the second largest subunit of low copy nuclear RNA polymerase II (RPB2) in allotetrploid StH genomic species of Elymus is characterized here. Our study first reported a 39-bp MITE stowaway element insertion in the genic region of RPB2 gene for all tetraploid Elymus St genome and diploid Pseudoroegneria spicata and P. stipifolia St genome. The sequences on 3′-end are highly conserved, with AGTA in all sequences but H10339 (E. fibrosis), in which the AGTA was replaced with AGAA. All 12 Stowaway-containing sequences encompassed a 9 bp conserved TIRs (GAGGGAGTA). Interestingly, the 5′-end sequence of GGTA which was changed to AGTA or deleted resulted in Stowaway excision in the H genome of Elymus sepcies, in which Stowaway excision did not leave footprint. Another two large insertions in all St genome sequences are also transposable-like elements detected in the genic region of RPB2 gene. Our results indicated that these three transposable element indels have occurred prior to polyploidization, and shaped the homoeologous RPB2 loci in St and H genome of Eymus species. Nucleotide diversity analysis suggested that the RPB2 sequence may evolve faster in the polyploid species than in the diploids. Higher level of polymorphism and genome-specific amplicons generated by this gene indicated that RPB2 is an excellent tool for investigating the phylogeny and evolutionary dynamics of speciation, and the mode of polyploidy formation in Elymus species.     

Implications of complete nuclear large subunit ribosomal RNA molecules from the harmful unarmored dinoflagellate Cochlodinium polykrikoides (Dinophyceae) and relatives

The D1/D2 domains of large subunit (LSU) rDNA have commonly been used for phylogenetic analyses of dinoflagellates; however, their properties have not been evaluated in relation to other D domains due to a deficiency of complete sequences. This study reports the complete LSU rRNA gene sequence in the causative unarmored dinoflagellate Cochlodinium polykrikoides, a member of the order Gymnodiniales, and evaluated the segmented domains and secondary structures when compared with its relatives. Putative LSU rRNA coding regions were recorded to be 3433 bp in length (49.0% GC content). A secondary structure predicted from the LSU and 5.8S rRNAs and parsimony analyses showed that most variation in the LSU rDNA was found in the 12 divergent (D) domains. In particular, the D2 domain was the most informative in terms of recent evolutional and taxonomic aspects, when compared with both the phylogenetic tree topologies and molecular distance (approximately 10 times higher) of the core LSU. Phylogenetic analysis was performed with a matrix of LSU DNA sequences selected from domains D2 to D4 and their flanking core sequences, which showed that C. polykrikoides was placed on the same branch with Akashiwo sanguinea in the “GPP” complex, which is referred to the gymnodinioid, peridinioid and prorocentroid groups. A broad phylogeny showed that armored and unarmored dinoflagellates were never clustered together; instead, they were clearly divided into two groups: the GPP complex and Gonyaulacales. The members of Gymnodiniales were always interspersed with peridinioid, prorocentroid and dinophysoid forms. This supports previous findings showing that the Gymnodiniales are polyphyletic. This study highlights the proper selection of LSU rDNA molecules for molecular phylogeny and signatures.     

Chorioactidaceae: a new family in the Pezizales (Ascomycota) with four genera

Molecular phylogenetic and comparative morphological studies provide evidence for the recognition of a new family, Chorioactidaceae, in the Pezizales. Four genera are placed in the family: Chorioactis, Desmazierella, Neournula, and Wolfina. Based on parsimony, likelihood, and Bayesian analyses of LSU, SSU, and RPB2 sequence data, Chorioactidaceae represents a sister clade to the Sarcosomataceae, to which some of these taxa were previously referred. Morphologically these genera are similar in pigmentation, excipular construction, and asci, which mostly have terminal opercula and rounded, sometimes forked, bases without croziers. Ascospores have cyanophilic walls or cyanophilic surface ornamentation in the form of ridges or warts. So far as is known the ascospores and the cells of the paraphyses of all species are multinucleate. The six species recognized in these four genera all have limited geographical distributions in the northern hemisphere.     

Comparison of primary and secondary 26S rRNA structures in twoTetrahymena species: Evidence for a strong evolutionary and structural constraint in expansion segments

Summary We have determined the nucleotide sequence of the 26S large subunit (LSU) rRNA genes for twoTetrahymena species,T. thermophila andT. pyriformis. The inferred rRNA sequences are presented in their most probable secondary structures based on compensatory mutations, energy, and conservation criteria. The majority of the nucleotide changes between the twoTetrahymena LSU rRNAs and the positions of a relatively large deletion and of the processing cleavage sites resulting in the generation of the hidden break are all located within the so-called divergent domains or expansion segments. These are regions within the common core of secondary structure where expansions have taken place during the evolution of the rRNA of higher eukaryotes.The dispensable nature of some of the expansion segments has been taken as evidence of their non-functionality. However, our data show that a considerable selective constraint has operated to presesrve the secondary structure of these segments. Especially in the case of the D2 and D8 segments, the presence of a considerable number of compensatory base changes suggests that the secondary structure of these regions is of functional importance. Alternatively, these expansion segments may have maintained characteristic folding patterns because only such structures are being tolerated within otherwise functionally important regions.     

<i>Natarajania thailandica</i> sp. nov. (Stilbosporaceae,Diaporthales) from Thailand

A fungus similar to the monotypic genus Natarajania, isolated from dead wood and collected in Thailand, is reported. Analysis of partial ribosomal LSU and a protein coding gene (RPB2) demonstrated that the new isolate belonged to Stilbosporaceae, Diaporthales and genetically different from N. indica. It is unique in producing synnematous conidiophores, smooth-walled conidiogenous cells and a flared collarette but lacks an elongated collar-canal which is distinct in the type species. Therefore, sequence data and morphological traits are used to introduce the new species, Natarajania thailandica.     

Migration between continents: geographical structure and long-distance gene flow in Porpidia flavicunda (lichen-forming Ascomycota)

Historical and contemporary geographical distribution ranges with their associated gene flow patterns interact to produce the genetic diversity observed today. Often it is not possible to separate out the impacts of historical events, e.g. past fragmentation, and contemporary gene flow, e.g. long-distance dispersal. Porpidia flavicunda is a lichen-forming ascomycete occurring circumpolar in the boreal to arctic zones for which vegetation history suggests that its distribution pattern has stayed broadly the same over the past millennia. DNA-sequence diversity in P. flavicunda can, thus, be expected to predominantly represent geographical population differentiation and its contemporary migration rates. The population sample consists of 110 specimens collected in Northern Québec, Baffin Island, Western Greenland and Northern Scandinavia. DNA-sequence data sets of three nuclear gene fragments (LSU, RPB2 and beta-tubulin) were analysed for genetic diversity within, and differentiation between, geographical regions. Tests of population subdivision employing analyses of molecular variance and exact tests of haplotype frequency distributions showed significant structure between the geographical regions. However, the lack of fixed nucleotide polymorphisms and the wide sharing of identical haplotypes between geographical regions suggest recurrent long-distance gene flow of propagules. Still, the means by which propagules are dispersed remain to be discovered. Inference of migration rates shows that in many cases a sufficiently high amount of migrants is exchanged between geographical regions to prevent drastic population differentiation through genetic drift. The observed haplotype distributions and migration rates point to a gene flow model of isolation by distance.     

<Emphasis Type="Italic">Cantharellus pseudoformosus</Emphasis>, a new species associated with <Emphasis Type="Italic">Cedrus deodara</Emphasis> from India

The genus Cantharellus is known to have a global distribution and to form ectomycorrhizal associations with a diverse set of host plants. Here we describe Cantharellus pseudoformosus as a species new to science with a possible association with Cedrus deodara. ITS and LSU data demonstrated that the material from India is distinct from Cantharellus formosus and other closely related species.     

Senescence in wheat leaves: is a cysteine endopeptidase involved in the degradation of the large subunit of Rubisco?

In wheat (Triticum aestivum L.), leaf senescence can be initiated by different factors. Depending on the plant system (intact plants or detached leaves) or the environmental conditions (light, nutrient availability), the symptoms of senescence differ. The aim of this work was to elucidate the catabolism of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC. 4.1.1.39) under various senescence-inducing conditions. Leaf senescence was initiated in intact plants by darkness or by N-deprivation and in leaf segments by exposure to light or darkness. Depending on the treatment, a 50 kDa fragment of Rubisco was observed. The formation of this fragment was enhanced by leaf detachment and low light. In segments exposed to high light and in intact plants induced to senesce by N-deprivation, the fragment was essentially absent. Since an antibody against the N-terminus of a large subunit of Rubisco (LSU) did not cross-react with the fragment, it appears likely that a smaller fragment was removed from the N-terminus of LSU. Inhibitor studies suggest that a cysteine endopeptidase was involved in the formation of the 50 kDa fragment. Non-denaturing-PAGE followed by SDS-PAGE revealed that the fragment was produced while LSU was integrated in the holoenzyme complex, and that it remained there after being produced. It remains open how the putative endopeptidase reaches the stromal protein Rubisco. The results indicate that depending on the senescence-inducing conditions, different proteolytic enzymes may be involved. The involvement of vacuolar proteases must be considered as occurring during LSU degradation, which takes place in darkness, low light or under carbon limitation.     

The statistical analysis of potato anther culture data

Methods for the production and utilisation of haploids have been developed for a range of crop plants. Genetical and environmental factors are known to influence both the rate of haploid induction and the mode of regeneration. Investigations designed to examine the parameters that influence haploid production from anthers are usually based on the overall percentage of responding anthers. Previous studies have rarely taken into account the frequency or distribution of anther culture response within a potato flower. In this paper the binomial, poisson and multiplicative binomial distributions are used for the first time to describe the distribution of anther culture response within a Solanum tuberosum flower.