Allozyme variation and genetic integrity of Dactylorhiza incarnata (Orchidaceae)

Eleven Danish populations of the diploid Dactylorhiza incarnata s.lat. have been examined for allozyme variation in DIA, PGD, PGI, PGM, SKD, TP1, and UGPP. The results reveal a deficiency of heterozygotes in several populations. Possible reasons for this deficiency are considered. The results also indicate that no genetic barrier seems to prevent gene flow between D. incarnata var. incarnata , var. dunensis , and var. ochrantha — and some indication exists that a considerable amount of gene exchange takes place between sympatric populations of var. incarnata and var. ochrantha . On the other hand, there is no sign of recent introgression between D. incarnata s.lat. and any of the sympatric tetraploid species ( D. maculata, D. majalis, D. purpurella s.lat.). It is suggested that knowledge on the various degrees of genetic integrity of morphologically recognizable entities should be elaborated and subsequently utilized for a biosystematic approach to Dactylorhiza .     

AFLP analysis of Cynodon dactylon (L.) Pers. var. dactylon genetic variation.

Cynodon dactylon (L.) Pers. var. dactylon (common bermudagrass) is geographically widely distributed between about lat 45 degrees N and lat 45 degrees S, penetrating to about lat 53 degrees N in Europe. The extensive variation of morphological and adaptive characteristics of the taxon is substantially documented, but information is lacking on DNA molecular variation in geographically disparate forms. Accordingly, this study was conducted to assess molecular genetic variation and genetic relatedness among 28 C. dactylon var. dactylon accessions originating from 11 countries on 4 continents (Africa, Asia, Australia, and Europe). A fluorescence-labeled amplified fragment length polymorphism (AFLP) DNA profiling method was used to detect the genetic diversity and relatedness. On the basis of 443 polymorphic AFLP fragments from 8 primer combinations, the accessions were grouped into clusters and subclusters associating with their geographic origins. Genetic similarity coefficients (SC) for the 28 accessions ranged from 0.53 to 0.98. Accessions originating from Africa, Australia, Asia, and Europe formed major groupings as indicated by cluster and principal coordinate analysis. Accessions from Australia and Asia, though separately clustered, were relatively closely related and most distantly related to accessions of European origin. African accessions formed two distant clusters and had the greatest variation in genetic relatedness relative to accessions from other geographic regions. Sampling the full extent of genetic variation in C. dactylon var. dactylon would require extensive germplasm collection in the major geographic regions of its distributional range.     

Status of Carex bergrothii (Cyperaceae) on Gotland, SE Sweden

Material of Carex oederi s.lat. was studied at four localities on the Baltic island of Gotland. Each locality included material referable to C. bergrothii , i.e., tall sedges with long leaves and with comparatively large and broad utricles, as well as material of C. oederi s.str., i.e., short sedges with smaller utricles. Univariate and multivariate statistical analyses show continuous variation in C. oederi s. lat. on Gotland and it is concluded that C. bergrothii cannot be recognised as a separate species. However, a large proportion of the phenotypic variance in C. oederi s. lat. on Gotland is due to genotypic differentiation, and C. bergrothii is considered as an extreme ecorype of C. oederi . This ecotype is apparently adapted to grow in deep water on calcareous clay among loose stands of Cladium mariscus .     

Evaluation of Morphological Variation in the Lichen Diploschistes ocellatus (Ascomycota, Ostropales): Evidence from Nuclear rDNA ITS Sequence Data

Abstract: A morphologically distinct population (var. almeriensis ) of the lichen species Diploschistes ocellatus from southern Spain is examined using morphological and molecular methods. While the morphological examination revealed some unique features of var. almeriensis, the ITS sequence data did not support a distinction from D. ocellatus s. str. The Kishino-Hasegawa test did not rule out trees distinguishing between the two morphotypes, although spectral analytical data support an inclusion of var. almeriensis in D. ocellatus. The influence of potentially erronously aligned ambiguous sites is evaluated by analysis of different alignments. However, all alignments agree in placing all specimens of D. ocellatus together and, consequently, in the consensus tree D. ocellatus s. lat. forms a monophyletic clade with high bootstrap support.     

Variation in Galium palustre s. lat. in the Baltic land-uplift area

Morphological variation, chromosome numbers, reproductive system and ecology were studied in Galium palustre s. lat. in some areas in the Baltic land-uplift area. Two cytotypes were found; diploids with 2n = 24 and octoploids with 2n = 96. Each cytotype comprises a large amount of habitat-correlated variation. Despite this, it is fairly easy to destinguish morphologically between the two cytotypes. It is therefore suitable to treat the diploid cytotype as one species, Galium palustre s. str., and the octoploid cytotype as another species, Galium elongatum . Both species grow in wet places, such as ditches, lake-shores and wet woodland. There are practically no differences in habitat ecology between them, but a slight tendency can be discerned for octoploids to grow in localities which never dry out, while diploids can grow in drier localities. Only G. palustre s. str. grows on fairly exposed sea-shores. A morphologically distinct from can also be distinguished, including diploids from sea-shores and coastal areas below 5 m. a. s. 1. This ecotype is described as Galium palustre var. balticum var. nov. It is characterised by smooth or nearly smooth stems and short, narrow leaves. It has always a very rich flowering and fruit set, which may be regarded as an adaptation to coastal conditions.     

Taxonomy of the Genus Triticum L.

In the present paper, an brief historical account and the comments on the modern taxonomic systems of the genus Triticum L. are made. The author suggests that principles to determine a species be (1) a special type of genome, and (2) reproductive isolation. The principles to determine a subspecies are the special co-type of genome and incomplete reproductive isolation. There are no differences on the level of genome constitution and no reproductive isolation between the varieties or concultivars. According to these principles, the author schemes a taxonomic system of the genus Triticum L. based on biosystematics as follows: Triticum monococcum L. sensu lat. subsp. boeoticum (Boiss.) Yen, st. nov. var. thaoudar (Reut ) Flaksb. concv. Einkhorn subsp. urartu (Tum.)Vap. T. timopheevi Zhuk. sensu lat. var. araraticum (Jakubz.) Yen, st. nov. T. zhukovskyi Men. et Er. T. turgidum L. sensu lat. var. dicoccoides (Körn. in litt. in Schweinf.) Bowden concv. (1) Emmer, (2) Durum wheat, (3) Rivet wheat, (4) Polish wheat, (5) Persian wheat. T. aestivum L. sensu lat. concv. (1) Tibetian weed wheat, (2) Spelt, (3) Vavilov wheat, (4) Macha wheat, (5) Yunnan hulled wheat, (6) Winter common wheat, (7) Spring common wheat, (8) Branch-eared wheat, (9) Club wheat, (I0) Indian dwarf wheat, (11) Xinjiang ricewheat.     

Comparison of the distribution of the repetitive DNA sequences in three variants of Cucumis sativus reveals their phylogenetic relationships

Repetitive DNA sequences with variability in copy number or/and sequence polymorphism can be employed as useful molecular markers to study phylogenetics and identify species/chromosomes when combined with fluorescence in situ hybridization (FISH). Cucumis sativus has three variants, Cucumis sativus L. var. sativus, Cucumis sativus L. var. hardwickii and Cucumis sativus L. var. xishuangbannesis. The phylogenetics among these three variants has not been well explored using cytological landmarks. Here, we concentrate on the organization and distribution of highly repetitive DNA sequences in cucumbers, with emphasis on the differences between cultivar and wild cucumber. The diversity of chromosomal karyotypes in cucumber and its relatives was detected in our study. Thereby, sequential FISH with three sets of multi-probe cocktails (combined repetitive DNA with chromosome-specific fosmid clones as probes) were conducted on the same metaphase cell, which helped us to simultaneously identify each of the 7 metaphase chromosomes of wild cucumber C. sativus var. hardwickii. A standardized karyotype of somatic metaphase chromosomes was constructed. Our data also indicated that the relationship between cultivar cucumber and C. s.var. xishuangbannesis was closer than that of C. s. var. xishuangbannesis and C. s. var. hardwickii.     

In vitro susceptibility characteristics of Cryptococcus neoformans varieties from AIDS patients in Goiânia, Brazil

Sixty clinical isolates of Cryptococcus neoformans from AIDS from Goiania, state of Goiás, Brazil, were characterized according to varieties, serotypes and tested for antifungal susceptibility. To differentiate the two varieties was used L-canavanine-glycine-bromothymol blue medium and to separate the serotypes was used slide agglutination test with Crypto Check Iatron. The Minimal Inhibitory Concentration (MIC) of fluconazole, itraconazole, and amphotericin B were determined by the National Committee for Clinical Laboratory Standards macrodilution method. Our results identified 56 isolates as C. neoformans var. neoformans serotype A and 4 isolates as C. neoformans var. gattii serotype B. MIC values for C. neoformans var. gattii were higher than C. neoformans var. neoformans. We verified that none isolate was resistant to itraconazole and to amphotericin B, but one C. neoformans var. neoformans and three C. neoformans var. gattii isolates were resistant to fluconazole. The presence of C. neoformans var. gattii fluconazole resistant indicates the importance of determining not only the variety of C. neoformans infecting the patients but also measuring the MIC of the isolate in order to properly orient treatment.     

Detection of Colletotrichum spp. Resistant to Benomyl by Using Molecular Techniques

Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 μg/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.     

A gene encoding lysine 6-aminotransferase, which forms the beta-lactam precursor alpha-aminoadipic acid, is located in the cluster of cephamycin biosynthetic genes in Nocardia lactamdurans.

A gene (lat) encoding lysine 6-aminotransferase was found upstream of the pcbAB (encoding alpha-aminoadipylcysteinyl-valine synthetase) and pcbC (encoding isopenicillin N synthase) genes in the cluster of early cephamycin biosynthetic genes in Nocardia lactamdurans. The lat gene was separated by a small intergenic region of 64 bp from the 5' end of the pcbAB gene. The lat gene contained an open reading frame of 1,353 nucleotides (71.4% G + C) encoding a protein of 450 amino acids with a deduced molecular mass of 48,811 Da. Expression of DNA fragments carrying the lat gene in Streptomyces lividans led to a high lysine 6-aminotransferase activity which was absent from untransformed S. lividans. The enzyme was partially purified from S. lividans(pULBS8) and showed a molecular mass of 52,800 Da as calculated by Sephadex gel filtration and polyacrylamide gel electrophoresis. DNA sequences which hybridized strongly with the lat gene of N. lactamdurans were found in four cephamycin-producing Streptomyces species but not in four other actinomycetes which are not known to produce beta-lactams, suggesting that the gene is specific for beta-lactam biosynthesis and is not involved in general lysine catabolism. The protein encoded by the lat gene showed similarity to ornithine-5-aminotransferases and N-acetylornithine-5-aminotransferases and contained a pyridoxal phosphate-binding consensus amino acid sequence around Lys-300 of the protein. The evolutionary implications of the lat gene as a true beta-lactam biosynthetic gene are discussed.     

Germinazione E Accrescimento Dell'Abete Bianco (Abies Alba Mill.) In Diverse Condizioni Termiche E Fotoperiodiche

Abstract

Germination and growth of ABIES ALBA mill. in different light and temperature conditions. — The results of a research on white fir from different localities are summarized as follows:

a) Germination of seeds from three italian localities (Tuscany, lat. 43°, 44'; Abruzzo, lat. 41°, 52'; Calabria, lat. 38°, 33') has been tested in several ecological conditions (temperature, light and chilling); it has been found that 22 °C, 8 hrs. of light and a 3 weeks prechilling at 5 °C, are stimulating conditions for all the seeds. The Cala-brian seeds seem the most reactive.

b) Growth in different photoperiodic conditions has been tested on seedlings from the Appennine, from France (lat. 49°, 10') and Poland (lat. 49°, 39'). Growth is evidently stimulated by long-day conditions in all the seeds experimented and growth acceleration touches its maximum in the Calabrian seedlings, provided they germinated at 12°C in the ligth.

It has been concluded that Calabrian Abies behaves as a Biotype, evidently distinguished from all the others, and an ecological explanation of this behaviour has been attemped.     

Environmental response of Galium palustre s. lat. in a coastal area in Central Sweden

Galium palustre s. lat. is a polymorphic taxon. The differences in morphology are correlated with the level of ploidy, but there is also a certain amount of phenotypic plasticity. The plasticity probably contributes to this taxon's successful colonization of a wide variety of habitats. The amount of plasticity was studied for plants from different kinds of habitats in Sweden, both for diploids (G. palustre s. str.) and octoploids (G. elongatum ), which were cloned. This cloned material was then transplanted to three different localities in Sweden: a sunny exposed sea-shore, a ditch, and a shady alder swamp. The general pattern of morphology was similar for all material, independent of its origin and level of ploidy, with smaller plants (smaller in several vegetative characters) and a more abundant flowering on the exposed seashore than in the ditch, and the largest plants with a fairly sparse flowering in the alder swamp. The octoploid plants originating from the wooded wetland are the least plastic ones but are very variable within the localities. G. palustre var. balneum shows a similar variation pattern as the others, but there are also genetically fixed differences in some characters between it and G. palustre var. palustre . The changes in morphology of the plants in the different localities occur fairly rapidly (in one month), although the differences are more spectacular one year later.     

Chemical taxonomy of Torrubiella s. lat.: zeorin as a marker of Conoideocrella

The insect pathogens in the genus Torrubiella s. lat. were recently divided into new genera based on molecular phylogenetic characters. Isolates collected at various locations in Thailand, were tested for their productivity of a hopane-type triterpene, zeorin (6α,22-dihydroxyhopane), when cultured in potato dextrose broth under static conditions. Among the 49 strains of Torrubiella s. lat. species, Conoideocrella luteorostrata (ten strains) and C. tenuis (seven strains), all collected on scale insects (Hemiptera), produced zeorin, whereas another six strains of Orbiocrella petchii (which was recently removed from Torrubiella) failed in the detection of this secondary metabolite. All other Torrubiella s. lat. (26 strains), collected on other insect hosts including leafhoppers (eight strains), Lepidoptera (one strain), and spiders (17 strains), produced no detectable zeorin. Paecilomyces cinnamomeus (nine strains), the anamorph of C. luteorostrata, also produced zeorin. These results correspond with the recent taxonomic reclassification based on multigene phylogeny.     

A numerical study of the common narrow-leaved taxa of chenopodium occurring in the Western United States

A numerical taxonomic study was conducted on the common narrow-leaved members of the genusChenopodium that occur in the western United States. As many as six and as few as two species have been recognized. Thirty-five individual popu lations were used as OTU’s, and 35 characters were considered. The data were subjected to cluster (both the weighted-pair group method and the within-group dispersion method) and principal component analysis. The results of both analyses are in general agreement and suggest phenetic relationships that differ from current interpretations in the literature. It is concluded thatC. desiccatum var.leptophylloides is much more closely related phenetically toC. atrovirens (and may not be distinct from it) than it is toC. desiccatum var.desiccatum. Chenopodium hians andC. leptophyllum appear to be phenetically distinct despite the fact that they have sometimes been viewed as conspecific.Cheno podium subglabrum is not very similar toC. leptophyllum, and should probably be treated as a separate species rather than as a variety of the latter. Popu lations referable toC. incognitum form a rather loose cluster that appears some what intermediate between populations ofC. desiccatum var.leptophylloides,C. atrovirens, C. hians, andC. leptophyllum.     

Genetic diversity of expressed Plasmodium falciparum var genes from Tanzanian children with severe malaria

ABSTRACT: BACKGROUND: Severe malaria has been attributed to the expression of a restricted subset of the var multigene family, which encodes for Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1). PfEMP1 mediates cytoadherence and sequestration of infected erythrocytes into the post-capillary venules of the vital organs such as the brain, lung or placenta. Var genes are highly diverse and can be classified in three major groups (ups A, B and C) and two intermediate groups (B/A and B/C) based on the genomic location, gene orientation and upstream sequences. The genetic diversity of expressed var genes in relation to severity of disease in Tanzanian children was analysed. METHODS: Children with defined severe (SM) and asymptomatic malaria (AM) were recruited. Fulllength var mRNA was isolated and reversed transcribed into var cDNA. Subsequently, the DBL and N-terminal domains, and up-stream sequences were PCR amplified, cloned and sequenced. Sequences derived from SM and AM isolates were compared and analysed. RESULTS: The analysis confirmed that the var family is highly diverse in natural Plasmodium falciparum populations. Sequence diversity of amplified var DBL-1alpha and upstream regions showed minimal overlap among isolates, implying that the var gene repertoire is vast and most probably indefinite in endemic areas. var DBL-1alpha sequences from AM isolates were more diverse with more singletons found (p<0.05) than those from SM infections. Furthermore, few var DBL-1alpha sequences from SM patients were rare and restricted suggesting that certain PfEMP1 variants might induce severe disease. CONCLUSIONS: The genetic sequence diversity of var genes of P. falciparum isolates from Tanzanian children is large and its relationship to disease severity has been studied. Observed differences suggest that different var genes might have fundamentally different roles in the host-parasite interaction. Further research is required to examine clear disease-associations of var gene subsets in different geographical settings. The importance of very strict clinical definitions and appropriate large control groups needs to be emphasized for future studies on disease associations of PfEMP1.     

夏蜡梅属的细胞地理学研究

本文首次报道西美蜡梅(Calycanthus occidentalis)的核型为K(2n)=22=20m(2SAT)+2sm,与美国蜡梅的变种光叶红(C.floridus var.oblongifolius)同属“2A”类型但较为原始,它们都比“1A”的夏蜡梅(C.chinensis)进化,三者由原始到进步的顺序可能为夏蜡梅—→西美蜡梅—→美国蜡梅(光叶红)。夏蜡梅属可能以较原始的夏蜡梅起源于东亚(中国),再东向迁移到北美洲(美国)形成西美蜡梅和美国蜡梅(光叶红)。     

Conservation genetics and taxonomic status of the rare Kentucky lady's slipper: Cypripedium kentuckiense (Orchidaceae)

Cypripedium kentuckiense is a recently described rare orchid found in Arkansas (predominantly) and in eight other states. Much debate has focused on whether this taxon should be recognized as a distinct species or considered to be an extreme manifestation of the variability present in the widespread taxon Cypripedium parviflorum var. pubescens. In this study, 12 isozyme loci were analyzed for 14 populations of C. parviflorum var. pubescens and eight populations of C. kentuckiense. These data were used to examine the genetic similarity of these taxa, assess whether isozyme data support the continued recognition of C. kentuckiense as a distinct species, and assess whether a newly discovered disjunct Virginia population of C. kentuckiense is genetically isolated from other C. kentuckiense populations. The isozyme data revealed that the two taxa are very closely related with a high interspecific genetic identity. However, C. kentuckiense populations contain a subset of the variation present in C. parviflorum var. pubescens, and they have expected levels of heterozygosity that are one-quarter that of C. parviflorum var. pubescens populations. Cypripedium kentuckiense also possesses one widespread unique allele and a unique multilocus genotype. These data suggest that C. kentuckiense should be recognized as a distinct species, possibly of recent origin from C. parviflorum. Lastly, the isozyme data support the hypothesis that gene flow between the Virginia population and other populations of C. kentuckiense has been restricted.     

Variation in morphological characters and habitat requirements in Carex capitata and Carex arctogena (Cyperaceae) in Norway and Sweden

Norwegian and Swedish populations of Carex capitata s. lat. were studied for variation in morphological characters. Canonical variates analysis (CVA) arranges populations in two discrete groups, corresponding to C. capitata s. str. and C. arctogena , respectively. In principal components analysis (PCA) of the total material a small overlap could be seen. However, separate PCA: s for the two regions in Scandinavia where the two taxa are sympatric show perfect separation. Student's t-test revealed significant differences in all morphological characters studied. Studies of companion plants, analysed with detrended correspondence analysis (DCA), indicate that the taxa have rather similar habitat demands in Oppland, Central Norway, although C. capitata s. str. seems to prefer wetter, and C. arctogena drier sites, respectively. Since the two taxa are distinct in morphology, they are best treated as separate species. The name Carex capitata is lectotypified.