Why species delimitation matters for fungal ecology: Colletotrichum diversity on wild and cultivated cashew in Brazil

Anthracnose is one of the most important plant diseases globally, occurring on a wide range of cultivated and wild host species. This study aimed to identify the Colletotrichum species associated with cashew anthracnose in Brazil, determine their phylogenetic relationships and geographical distribution, and provide some insight into the factors that may be influencing community composition. Colletotrichum isolates collected from symptomatic leaves, stems, inflorescences, and fruit of cultivated and wild cashew, across four Brazilian biomes, were identified as Colletotrichum chrysophilum, Colletotrichum fragariae, Colletotrichum fructicola, Colletotrichum gloeosporioides sensu stricto, Colletotrichum queenslandicum, Colletotrichum siamense and Colletotrichum tropicale. Colletotrichum siamense was the most dominant species. The greatest species richness was associated with cultivated cashew; leaves harbored more species than the other organs; the Atlantic Forest encompassed more species than the other biomes; and Pernambuco was the most species-rich location. However, accounting for the relative abundance of Colletotrichum species and differences in sample size across strata, the interpretation of which community is most diverse depends on how species are delimited. The present study provides valuable information about the Colletotrichum/cashew pathosystem, sheds light on the causal agents identification,and highlights the impact that species delimitation can have on ecological studies of fungi.     

Effects of varying pollen load on fruit set,seed set and seedling performance in apple and pear

Two apple crosses and one pear cross were carried out using three different pollen densities. In one of the apple crosses, both fruit and seed set decreased when a mixture of pollen and powder (diluted pollen treatment) was used for pollination. Also, seedlings from the undiluted treatment were more vigorous than those originating from pollination with diluted pollen. In the other apple cross, however, no effect on fruit set, seed set or growth in the first year was found. Mean diameter after the second year differed among treatments, but did not seem to be related to pollen density. For pear, the number of good seeds decreased and the number of deaf, probably unfertilized, seeds increased with decreasing pollen density. However, there was no apparent significant effect of pollen density on seedling growth. With respect to scab resistance and mildew attack in both apple crosses, no differences among treatments were found. The results indicate that gametophytic competition may occur in apple crosses, but that it is probably weaker in pear.     

A rapid lateral-flow immunoassay for phytosanitary detection of Erwinia amylovora and on-site fire blight diagnosis

Fire blight is an invasive disease caused by Erwinia amylovora that threatens pome fruit production globally. Effective implementation of phytosanitary control measures depends upon rapid, reliable pathogen detection and disease diagnosis. We developed a lateral-flow immunoassay specific for E. amylovora with a detection limit of log 5.7 CFU/ml, typical of pathogen concentrations in symptomatic plant material. The simple assay had comparable sensitivity to standard culture plating, serum agglutination and nested PCR when validated for application in a phytosanitary laboratory as a confirmatory test of cultured isolates and for first-line diagnosis of phytosanitary samples that represent the full range of commercial, ornamental and forestry host species. On-site validation in ring-trials with local plant inspectors demonstrated robust and reliable detection (compared to subsequent plating and PCR analysis). The simplicity, inspector acceptance and facilitation of expedited diagnosis (from 2 days for laboratory submitted samples to 15 min with the immunoassay), offers a valuable tool for improved phytosanitary control of fire blight.     

Molecular systematics in the genus Mucor with special regards to species encountered in cheese

The genus Mucor, a member of the order Mucorales, comprises different species encountered in cheeses. Although fungi play a fundamental role in cheese manufacturing and ripening, the taxonomy of many fungal species found in cheese is poorly defined; indeed, this is the case for Mucor spp. In the present study, we assessed the phylogenetic relationships among 70 Mucor strains, including 36 cheese isolates, by using a five gene phylogenetic approach combined with morphological analyses. Overall, at least six species of Mucor were identified among the cheese isolates including a possible new taxon. The present study also suggests that the genus Mucor comprises undescribed taxa and needs to be properly defined.     

Correlation in the tolerance to ozone between sporophytes and male gametophytes of several fruit and nut tree species (Rosaceae)

We analyzed relative sensitivities to ozone partial pressure for male gametophytes (pollen and pollen tubes) of five tree-crop species in the family Rosaceae (almond, apple, apricot, nectarine/peach, pear) and of four cultivars of almond. Relative sensitivities to ozone partial pressure were previously described using field-based, whole-plant, physiological measurements for the sporophytes of each of the species and cultivars used in this study. Our objective was to determine the extent of correlation between sporophyte and gametophyte responses to ozone among and within these tree species. If the relative sensitivities prove to be similar for the two generations, two research strategies become possible for breeders interested in developing new cultivars with improved tolerance to atmospheric ozone: First, because screening large numbers of sporophytes for their responses to ozone partial pressure is a formidable task, exploiting the much more easily manipulated male gametophyte generation for large-scale screening of individuals for ozone tolerance could greatly facilitate identification and selection of tolerant genotypes. Second, it opens the possibility of applying selection pressure to the gametophyte generation (as during pollen germination or pollen tube growth) to favor production of ozonetolerant progeny. Results indicate that, among and within species, relative responses of pollen tube growth to ozone partial pressure are highly correlated with that of the sporophytes. Pollen germination was inhibited by ozone treatments for all but one species (pear), but differences among and within species were neither significant nor (with the exception of pear) did they correlate with relative sporophyte sensitivities.     

Mycobacterium tuberculosis Phylogeny Reconstruction Based on Combined Numerical Analysis with IS1081, IS6110, VNTR, and DR-Based Spoligotyping Suggests the Existence of Two New Phylogeographical Clades

This paper deals with phylogenetic relationships among a set of 90 clinical strains representative of the worldwide diversity of the Mycobacterium tuberculosis complex (Kremer et al. 1999) using eight independent genetic markers: IS6110, IS1081, the direct repeat (DR) locus, and five variable number of tandem DNA repeat loci (VNTR). In a preliminary experiment, phylogenetic trees based on single markers were constructed that led to the detection of some similarities between the VNTR-based and the spoligotyping-based phylogenetic trees. In the second step, a more global phenetic approach based on pairwise comparison of strains within each typing system was used, followed by calculations of mean genetic distances based on all the eight loci and the use of the neighbor-joining algorithm for tree reconstruction. This analysis confirmed our preliminary observations and suggested the existence of at least two new phylogeographical clades of M. tuberculosis, one defined as the ``East African–Indian family' (EA-I), which may find its origin on the African or Asian continents, and the other as the ``Latin American and Mediterranean' (LA-M) family. The existence of these two families was also validated by an independent phylogenetic analysis of spoligotyping on a larger set of shared types (n= 252) and further corroborated by VNTR and katG–gyrA results. The potential origin of these families of bacilli is discussed based on cattle domestication and human migration history. In conclusion, the information contained in insertion sequence and repetitive DNAs may serve as a model for the phylogenetic reconstruction of the M. tuberculosis complex.     

Trypanosoma rangeli is phylogenetically closer to Old World trypanosomes than to Trypanosoma cruzi

Trypanosoma rangeli and Trypanosoma cruzi are generalist trypanosomes sharing a wide range of mammalian hosts; they are transmitted by triatomine bugs, and are the only trypanosomes infecting humans in the Neotropics. Their origins, phylogenetic relationships, and emergence as human parasites have long been subjects of interest. In the present study, taxon-rich analyses (20 trypanosome species from bats and terrestrial mammals) using ssrRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH), heat shock protein-70 (HSP70) and Spliced Leader RNA sequences, and multilocus phylogenetic analyses using 11 single copy genes from 15 selected trypanosomes, provide increased resolution of relationships between species and clades, strongly supporting two main sister lineages: lineage Schizotrypanum, comprising T. cruzi and bat-restricted trypanosomes, and Tra[Tve-Tco] formed by T. rangeli, Trypanosoma vespertilionis and Trypanosoma conorhini clades. Tve comprises European T. vespertilionis and African T. vespertilionis-like of bats and bat cimicids characterised in the present study and Trypanosoma sp. Hoch reported in monkeys and herein detected in bats. Tco included the triatomine-transmitted tropicopolitan T. conorhini from rats and the African NanDoum1 trypanosome of civet (carnivore). Consistent with their very close relationships, Tra[Tve-Tco] species shared highly similar Spliced Leader RNA structures that were highly divergent from those of Schizotrypanum. In a plausible evolutionary scenario, a bat trypanosome transmitted by cimicids gave origin to the deeply rooted Tra[Tve-Tco] and Schizotrypanum lineages, and bat trypanosomes of diverse genetic backgrounds jumped to new hosts. A long and independent evolutionary history of T. rangeli more related to Old World trypanosomes from bats, rats, monkeys and civets than to Schizotrypanum spp., and the adaptation of these distantly related trypanosomes to different niches of shared mammals and vectors, is consistent with the marked differences in transmission routes, life-cycles and host-parasite interactions, resulting in T. cruzi (but not T. rangeli) being pathogenic to humans.     

In vitro susceptibilities of Neoscytalidium spp. sequence types to antifungal agents and antimicrobial photodynamic treatment with phenothiazinium photosensitizers

Neoscytalidium spp. are ascomycetous fungi consisting of pigmented and hyaline varieties both able to cause skin and nail infection. Their color-based identification is inaccurate and may compromise the outcome of the studies with these fungi. The aim of this study was to genotype 32 isolates morphologically identified as Neoscytalidiumdimidiatum or N. dimidiatum var. hyalinum by multilocus sequence typing (MLST), differentiate the two varieties by their sequence types, evaluate their susceptibility to seven commercial antifungal drugs [amphotericin B (AMB), voriconazole (VOR), terbinafine (TER), 5-flucytosine (5FC), ketoconazole (KET), fluconazole (FLU), and caspofungin (CAS)], and also to the antimicrobial photodynamic treatment (APDT) with the phenothiazinium photosensitizers (PS) methylene blue (MB), new methylene blue (NMBN), toluidine blue O (TBO) and the pentacyclic derivative S137. The efficacy of each PS was determined, initially, based on its minimal inhibitory concentration (MIC). Additionally, the APDT effects with each PS on the survival of ungerminated and germinated arthroconidia of both varieties were evaluated. Seven loci of Neoscytalidium spp. were sequenced on MLST revealing eight polymorphic sites and six sequence types (ST). All N. dimidiatum var. hyalinum isolates were clustered in a single ST. AMB, VOR and TER were the most effective antifungal agents against both varieties. The hyaline variety isolates were much less tolerant to the azoles than the isolates of the pigmented variety. APDT with S137 showed the lowest MIC for all the isolates of both varieties. APDT with all the PS killed both ungerminated and germinated arthroconidia of both varieties reducing the survival up to 5 logs. Isolates of the hyaline variety were also less tolerant to APDT. APDT with the four PS also increased the plasma membrane permeability of arthroconidia of both varieties but only NMBN and S137 caused peroxidation of the membrane lipids.     

金秋梨和新高梨的分子遗传学对比分析

金秋梨和新高梨的形态性状、生物学性状以及酯酶同工酶、过氧化物酶同工酶、细胞色素氧化酶同工酶酶谱均产生了变化。利用异源DNA探针—小麦rDNA克隆pTA71,与四种核酸内切酶PstⅠ、BamHⅠ、EcoRⅠ及BglⅡ酶切消化的金秋梨和新高梨叶片总DNA杂交,结果表明,pTA71-PstⅠ和pTA71-BamHⅠ二种探针—酶组合可在金秋梨和新高梨中检测到RFLP差异。     

Diallelic multilocus model of neutral genes

The evolution of a completely linked diallelic multilocus system of neutral genes in a finite population is studied. A diffusion model incorporating random genetic drift and mutation is used. We neglect the recombination. To begin with, the spectral analysis of the Kolmogorov backward equation for this model is investigated. We apply this to two extreme situations when the number of sites approaches to infinity. One is a DeMoivre-Laplace type approximation and the other is a Poisson type approximation. The former is applied to the study of the simultaneous distribution and evolution of a large number of neutral genes. It is applicable to the distribution of a polygenic character controlled by clustered loci on a chromosome, and we show that it differs from the normal distribution on account of random genetic drift and linkage disequilibrium. The latter is applied to the distribution of the number of segregating sites in DNA nucleotide sequences, and the rate of evolution is obtained.     

梨锈病病菌生态位研究

安徽主栽梨树品种对锈病菌的抗性差异显著,而对感病品种以病情指数和病叶数为依据的锈菌寄主生态位进行研究,结果生态位宽度依次是０．７５５５和０．７９５１;各品种水平空间和垂直空间生态位宽度均接近于１。此研究表明梨锈病菌是以随机扩散传播侵染的。     

辽宁省生态学会成立大会暨首届生态学术讨论会在沈阳召开

辽宁省生态学会成立大会暨首届生态学术讨论会于1985年6月26日至28日在沈阳召开。来自全省科研、教学、生产以及政府部门和新闻出版单位的117名代表出席了会议。辽宁省副省长彭祥松、辽宁省人大常委会副主任冯友松出席了大会并讲了话。辽宁省科协副主席涂长晟同志、中国科学院沈阳分院副院长盛士骏同志以及中国土壤学会、黑龙江省生态学会、辽宁省动物学会等兄弟学会的代表到会指导并祝贺。大会还收到了中国生态学会理事长马世骏先生、中共中央书记处农村发展中心顾问石山同志、中国社会科学院顾问于光远同志、植物生态学家侯学煜先生、中国《人与生物圈》国家委员会副主席阳含熙先生的贺信。     

梨锈病病菌生态位研究

安徽主栽梨树品种对锈病菌的抗性差异显著,而对感病品种以病情指数和病叶数为依据的锈菌寄主生态位进行研究,结果生态位宽度依次是0.7555和0.7951;各品种水平空间和垂直空间生态位宽度均接近于1.此研究表明梨锈病菌是以随机扩散传播侵染的.     

Basal relationships in the Drosophila melanogaster species group

The Drosophila melanogaster species group is a popular model for evolutionary studies due to its morphological and ecological diversity and its inclusion of the model species D. melanogaster. However, phylogenetic relationships among major lineages within this species group remain controversial. In this report, the phylogeny of 10 species representing each of the well-supported monophyletic clades in the melanogaster group was studied using the sequences of 14 loci that together comprise 9493 nucleotide positions. Combined Bayesian analysis using gene-specific substitution models produced a 100% credible set of two trees. In the strict consensus of these trees, the ananassae subgroup branches first in the melanogaster species group, followed by the montium subgroup. The remaining lineages form a monophyletic clade in which D. ficusphila and D. elegans branch first, followed by D. biarmipes, D. eugracilis, and the melanogaster subgroup. This strongly supported phylogeny resolves most basal relationships in the melanogaster species group, and provides a framework that can be extended in the future to encompass more species.     

Multilocus autosomal sex determination

The two basic one locus sex determination models, diploid individual sex determination and parental sex determination, are generalized to the multilocus framework. As in the single locus case, it is established that there are two classes of polymorphic equilibria, equilibria with even sex ratio and equilibria with equal allele frequencies in the two sexes. The condition for external stability of this second class equilibria to invasion by a new mutant allele is that a new appropriately averaged sex ratio near the equilibrium be moved closer to the even sex ratio. However, stable polymorphisms with noneven sex ratio are not those that have a sex ratio as close as possible to 1/2, in contrast to the single locus case.Research supported in part by NIH grants GM 28016 and GM 10452 and a grant from the U.S.-Israel Binational Science Foundation     

梨不同DNA提取方法的效果研究

以7个梨品种为实验材料,比较分析了SDS法、CTAB法、SDSCTAB法、改良的CTAB法、高盐低pH值法、分步离心法对梨总DNA提取的效果。结果表明:利用以上6种方法提取的梨总DNA在纯度和量上有很大的差别。所得到的平均DNA量从大到小依次为:分步离心法、SDS法、SDSCTAB法、改良的CTAB法、CTAB法、高盐低pH值法。DNA提取纯度依次为分步离心法、SDSCTAB法、改良的CTAB法、高盐低pH值法、CTAB法、SDS法。RAPD和自交不亲和基因(S基因)特异性引物扩增实验结果都比较理想,但分步离心法和SDSCTAB法提取的DNA双酶切效果较好。分步离心法提取的梨总DNA更适用于后续的分子生物学实验操作。