Species-specific ability ofChlorella strains (Chlorophyceae) to form stable symbioses withHydra viridis

46 strains ofChlorella, identified by physiological and biochemical characters, were examined for their ability to form stable symbioses with aposymbioticHydra viridis. It was found to be a species-specific characteristic. Among the 15 taxa studied, onlyC. saccharophila var.ellipsoidea, C. saccharophila var.saccharophila, C. fusca var.vacuolata, C. kessleri, C. luteoviridis, andC. protothecoides formed stable symbioses withHydra viridis. Among the 11 known physiological and biochemical characters of theseChlorella species, only acid tolerance seems to be correlated with symbiosis: All symbiotic species are capable of growing at or below pH 4.0.     

Deoxyribonucleic acid reassociation in the taxonomy of the genus Chlorella

The genetic relationships of nine strains of Chlorella saccharophila were determined by DNA hybridization techniques. Four strains are closely related to the type strain 211-9a and one strain seems to be moderately related, whereas the taxonomic position of the remaining three strains is not clear. C. saccharophila, like C. sorokiniana, is another species of Chlorella containing strains which are heterogeneous in their overall DNA base sequence and partly also in morphological, biochemical and physiological characters.     

Deoxyribonucleic acid reassociation in the taxonomy of the genus Chlorella

DNA hybridization techniques showed Chlorella fusca var. vacuolata and C. kessleri to be homogeneous species with DNA homologies of 90–100% C. fusca var. fusca and var. rubescens, however, have only about 15% DNA homology with C. fusca var. vacuolata and should no longer be regarded as varieties. A good correlation was found so far between biochemical and physiological characters used in the taxonomy of Chlorella and DNA relatedness. Mutant strains of Chlorella were tested for DNA homologies to prove the reliability of the taxonomical interpretation.     

Physiological and biochemical contributions to the taxonomy of the genus prototheca

Five physiological and biochemical characters, which had proved to be valuable for the taxonomy of the genus Chlorella, were studied in the genus Prototheca. There is no hydrogenase activity and no liquefaction of gelatin. Most strains are very acidtolerant (limit of growth at pH 2.0 or 2.5) and very salt-tolerant (limit of growth at 4 or 5% NaCl). Two strains grow well at 38°C. The 16 strains, which were previously assigned to seven taxa, fall into four different groups. Our results tend to support the assumption that Prototheca might be related to Chlorella protothecoides.     

Biochemical variation of Cryptococcus neoformans

Ninety-seven strains of Cryptococcus neoformans and C. bacillisporus were examined for 44 biochemical characters and the results were analyzed numerically. One phenon emerged at the 86% level of similarity when strains were clustered according to their M-similarity values. All strains grew in ten carbon sources (D-glucose, D-galactose, arbutin, maltose, sucrose, D-melezitose, D-xylose, D-mannitol, D-glucitol, and meso-inositol), and also grew at 37 ° C and produced urease and phenoloxidase. None of them grew in melibiose, lactose, nor valine, and none reduced nitrate to nitrite. Comparison of selected biochemical characters, creatinine utilization, and serotypes of 49 aberrant strains is presented. Forty-eight of the 97 strains produced the Filobasidiella state either alone or when paired with a strain of compatible mating-type. Filobasidiella neoformans serotypes A and D were interfertile with compatible mating-types of F. bacillispora serotypes B and C. The 44 biochemical characters and 4 serotypes did not predict barriers to mating competence. The present study further substantiates that Filobasidiella neoformans and F. bacillispora are one species.     

Physiological and biochemical contributions to the taxonomy of the genus Chlorella

Starch hydrolysis, i.e., the production of extracellular amylolytic enzymes, was found to be a specific character for most species of the genus Chlorella. C. fusca var. vacuolata, C. spec. 211-30, and C. spec. 211-11r hydrolyse starch, whereas C. vulgaris, C. fusca var. rubescens, C. zofingiensis, C. fusca var. fusca, C. minutissima, C. homosphaera, C. kessleri, C. luteoviridis, and C. protothecoides are unable to hydrolyse starch. Only C. sorokiniana and C. saccharophila appear heterogenous; within C. sorokiniana, 7 strains hydrolyse starch and 9 do not; and within C. saccharophila, 6 strains exhibit amylolytic activity and 2 do not. — A key for the identification, according to 9 easily determined physiological and biochemical characters, of the Chlorella species is presented.     

Using multiple sources of characters to delimit species in the genus Crataegus (Rosaceae): the case of the Crataegus rosei complex

The aim of this study was to delimit the taxa of the Crataegus rosei complex using an integrative approach that incorporates a suite of molecular (cpDNA and nuclear microsatellite markers), morphological, and geometric morphometric characters. One hundred and ten plants from 19 populations that encompass the entire distribution range of the species complex were collected and examined along with herbarium specimens. Parsimony and Bayesian inference analyses were run using morphological, molecular, and both the morphological and molecular data sets combined. Analyses to determine genetic structure based on microsatellite data and multivariate analyses incorporating geometric morphometrics were also done to identify differences in leaf shape. The results supported the recognition of two taxa: C. rosei with high levels of gene flow among its populations, remarkable morphological variation and a wide distribution range and C. rosei var. amoena, composed of a few isolated populations in the high elevation location of Cerro Potosí; a new specific epithet will be decided for the latter in accordance with the International Code of Nomenclature for algae, fungi, and plants.     

Analysing diversity among Indian isolates of Anabaena (Nostocales, Cyanophyta) using morphological, physiological and biochemical characters

A set of 24 strains belonging to the genus Anabaena (Phylum Cyanobacteria), isolated from diverse geographic locations in India, were evaluated along with three International type strains of Anabaena (ATCC 29414, ATCC 29208 and ATCC 27899) for their morphological, physiological and biochemical diversity. The morphological dataset, consisting of 58 variants for 15 characters, and SDS-PAGE protein profiles comprising 17 polymorphic bands were utilized to differentiate the selected Anabaena strains and explore the patterns of diversity through cluster analysis. Physiological and biochemical characterization with respect to nitrogen fixation and accumulation of chlorophyll and phycobiliproteins led to the identification of some highly promising Anabaena strains for use as biofertilizers and source of pigments. The study highlighted the tremendous inter and intraspecific diversity within the Anabaena isolates and indicated the potential as well as constraints of the morphological and protein profiling datasets for unambiguous differentiation and analyses of diversity among the Anabaena strains.     

A MORPHOLOGICAL AND TAXONOMIC CONSIDERATION OF TABELLARIA (BACILLARIOPHYCEAE) FROM THE NORTHCENTRAL UNITED STATES1,2

The taxonomy, distribution, and ecology of the diatom genus Tabellaria Ehr. from a portion of the northcentral United States were studied from collections made from August 1968 to October 1971. Using the transmission electron microscope with carbon replicas of the frustule surfaces, taxonomically significant characters were identified and related to those visible with the light microscope, four morphologically distinguishable taxa were recognized. The frustular morphology of T. fenestrata (Lyngbye) kütz. em. Knudson var. fenestrata was confirmed by both light and electron microscopy. T. flocculosa (Roth) Kütz. var. flocculosa is composed of 3 morphologically distinguishable strains. One of which is sufficiently different from the nominate variety to be described as T. flocculosa var. linearis var. nov. Also, a planktonic ecotype of one of the strains was identified.     

Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress

Fatty acid composition and degree of fatty acid saturation during temperature stress in thermo-intolerant (Phaeodactylum tricornutum) and thermo-tolerant (Chaetoceros muelleri) marine diatoms were investigated. A greater number of fatty acids were observed in C. muelleri than in P. tricornutum regardless of treatment. The major fatty acids detected were 14:0, 16:0, 16:1, 16:2, 16:3, 18:0, 18:1(n-9)c, 18:2(n-6) and 20:5(n-3) with additional fatty acids 18:1(n-9)t and 20:4(n-6) detected in C. muelleri. Short duration (2 h) temperature increase above optimal growth temperature had a greater effect on fatty acid composition in C. muelleri than in P. tricornutum and the degree of fatty acid saturation was affected more by temperature in C. muelleri than in P. tricornutum during both short and long duration (24 h) treatments. Total protein assay results suggest that P. tricornutum, but not C. muelleri, was undergoing stress under our growing conditions although lipids in both diatoms were affected by increased temperature. Immunodetection of proteins with anti-rubisco indicates that the rubisco large subunit was undergoing greater turnover in C. muelleri than in P. tricornutum. However, the integrity of rubisco as a suitable indicator of lipid status needs further study. This work supports the hypothesis that a particular temperature, and not treatment duration, has the greater effect on changes in fatty acid composition. Furthermore, changes in fatty acid composition and degree of fatty acid saturation occurred more quickly in the diatoms in response to increased temperature than previously observed in nutrient starvation studies. Since diatom lipids represent an important resource for growth and reproduction of marine animals, the rapid alteration of their lipid composition under temperatures normally encountered in marine environments warrants further study.     

A Note on the Limitations of Identifying Soft Rot Erwinias by Temperature Tolerances and Sensitivity to Erythromycin on a Pectate Medium

Forty-five erwinia strains from 14 different hosts have been tested lor growth on a crystal violet pectate medium with or without 70 μ/ml erythromycin at 27, 33.5 and 37°C. On the basis of positive data in the literature it was tried to identify the strains as Erwnia carotovora subsp. atroseptica, E. carotovora subsp. carotovora or E. chrysanthemi using this method. It appeared that relatively few strains reacted differentially, apparently due to strain variation (all strains were identified also by biochemical and physiological tests). The conclusion is that for a reliable identification of field isolates of soft rot crwinias other (biochemical, physiological and serological) methods have to be applied.     

Factors Affecting the Mixotrophic Flagellate Poterioochromonas malhamensis Grazing on Chlorella Cells

Grazing behaviour between protozoa and phytoplankton exists widely in planktonic ecosystems. Poterioochromonas malhamensis is a well‐known and widespread mixotrophic flagellate, which is recognized to play an important role within marine and freshwater planktonic ecosystems and regarded as the greatest contamination threat for mass algal cultures of Chlorella. In this study, a comprehensive range of factors, including morphological characters, biochemical compositions, and specific growth rate of ten species or strains of Chlorella, were evaluated for their effect on the feeding ability of P. malhamensis, which was assessed by two parameters: the clearance rate of P. malhamensis on Chlorella spp. and the specific growth rate of P. malhamensis. The results showed that the clearance rate of P. malhamensis was negatively correlated with cell wall thickness and specific growth rate of Chlorella spp., while the specific growth rate of P. malhamensis was positively correlated with carbohydrate percentage and C/N ratio and negatively correlated with protein, lipid percentage, and nitrogen mass. In conclusion, the factors influencing feeding selectivity include not only the morphological character and chemical composition of Chlorella, but also its population dynamics. Our study provides useful insights into the key factors that affect the feeding selectivity of P. malhamensis and provides basic and constructive data to help in screening for grazing‐resistant microalgae.     

THE MOLECULAR PHYLOGENETIC AND GEOMETRIC MORPHOMETRIC EVALUATION OF MICRASTERIAS CRUX‐MELITENSIS/M. RADIANS SPECIES COMPLEX1

We investigated nine strains of the Micrasterias crux‐melitensis (Ehrenb.) Hassall ex Ralfs and M. radians W. B. Turner species complex. A combination of molecular, morphological, and geometric morphometric data was used to reveal the patterns of their phenotypic and phylogenetic differentiation. The molecular data based on internal transcribed spacer (ITS) rDNA, glycine transfer RNA (trnG^uuc) intron, and SSU rDNA sequences revealed three phylogenetic lineages. One of them comprised the six European and North American strains that were morphologically identified as M. crux‐melitensis. Phenotypic data illustrated high morphological variability of strains within this genetically homogenous lineage that spanned several traditional infraspecific taxa, including strains corresponding to M. crux‐melitensis var. janeira (Racib.) Grönblad and M. crux‐melitensis var. superflua W. B. Turner, whose morphometric characteristics profoundly differed. Three strains of M. radians formed two separate phylogenetic lineages corresponding to traditional varieties M. radians var. evoluta (W. B. Turner) Willi Krieg. and M. radians var. bogoriensis (C. J. Bernard) G. S. West. The morphological types corresponding to the former variety have, so far, only been reported from Africa. Therefore, we cannot preclude that geographic isolation may play a role in species differentiation of relatively large freshwater protists, such as Micrasterias.     

Xanthomonas campestris pv.parthenii pathovar nov. incitant of leaf blight of parthenium

A new bacterial leaf blight disease of parthenium (Parthenium hysterophorus L.) is described for the first time. The disease-causing bacterium was isolated and its morphological, physiological and biochemical characters were determined. The pathogenicity of bacterium is apparently limited only to parthenium. The pathogen was identified asXanthomonas campestris pv.parthenii pathovar nov. on the basis of morphological, physiological, biochemical and pathogenic characteristics.     

Molecular and morphological analyses of solitary forms of brackish Thalassiosiroid diatoms (Coscinodiscophyceae), with emphasis on their phenotypic plasticity

Blooms of centric diatoms are a common feature in the Bilbao estuary during summer when river flow is at its lowest and water temperature is above 20ºC. To gain insight into the specific composition of these diatom blooms, net samples and cultures of estuarine isolates were analysed under the scanning electron microscope (SEM) and by molecular analyses of the Internal Transcribed Spacers 1 and 2 plus the coding region 5.8S (ITS region) and the 28S rRNA gene. Seven species of solitary centric diatoms belonging to four genera were found in the estuary including: Conticribra weissflogii, Cyclotella atomus var. atomus, Cyclotella cryptica, Cyclotella marina, Cyclotella meneghiniana, Discostella pseudostelligera and Thalassiosira pseudonana. Dominant species during blooms were C. meneghiniana and Co. weissflogii in the upper estuary and D. pseudostelligera and T. pseudonana in the middle estuary. The morphological traits used to differentiate between species pairs of similar morphology (C. meneghiniana/C. cryptica or D. pseudostelligera/D. woltereckii) were observed to vary with environmental conditions, denoting a great deal of phenotypic plasticity which would hinder accurate identification of the species when using morphological approaches alone.     

A multivariate statistical approach to Centaurea classification using essential oil composition data of some species from Turkey

The composition of the essential oils of ten Centaurea species from Turkey, Centaurea aladaghensis, C. antiochia var. prealta, C. antitauri, C. babylonica, C. balsamita, C. cheirolepidoides, C. deflexa, C. iconiensis, C. lanigera, C. ptosimopappoides have been studied. Multivariate statistical analyses (Principal Component Analysis, Multidimensional Scaling, Hierarchical Cluster Analysis) applied to GC-MS data, seem to be very useful to investigate and establish the natural taxonomic delimitation of this very difficult genus. The groupings resulted independent from the ecological similarities (i.e. plants that live in the same habitats or share similar morphological characteristics), so it seems that the environment has no influence on the biosynthesis of the volatiles of these plants.     

Numerical taxonomy of some charophytes in Iran

Morphological characters of 10 species of Chara from different localities in northern, western and central Iran have been analyzed, with Nitella hyalina and Tolypella glomerata as outgroups. Phenetic and cladistics analyses of populations belonging to the 10 Chara species were undertaken to reveal their inter‐population morphological variation and inter‐specific relationships. Clustering was based on 54 characters (41 qualitative and 13 quantitative). This produced two major clusters, grouping Chara crassicauliss, C. vulgaris var. longibracteata, C. vulgaris var. vulgaris and C. contraria separate from C. grovesii, C. gymnophylla var. gymnophylla, C. gymnophylla var. rohlenae, C. kirghisorum and C. tomentosa. A smaller cluster of C. socotrensioides, C. fibrosa and Chara sp. was separate from these two clusters. A factor analysis identified the most variable morphological characters as type of cortification, prescence and length of the spine cells, stipulode length, type of branchlet cortication, bract cells length, placement of the gametangia, character of end segment, length of branchlet and internode, development of posterior bract cells, oogonia length and width, oospore length, width and length/width ratio, fossa breath, and number of striae.     

Phylogeny,morphology, and physiology of Micractinium strains isolated from shallow ephemeral freshwater in Antarctica

Cryotolerant eukaryotic microalgae were isolated from meltwater streams on Ardley Island and King George Island in Antarctica, and their morphological, molecular, and physiological characteristics were investigated. Owing to their simple morphology, distinctive characters were not observed with neither light microscopy nor transmission electron microscopy. However, molecular phylogenetic inferences drawn from the concatenated small subunit rRNA and internal transcribed spacer sequence data indicated that these microalgal strains belonged to the genus Micractinium. All the Micractinium strains showed cryotolerant properties, while their optimum growth temperature was around 20°C. Similar to other cryotolerant organisms, these Antarctic microalgae also contained a higher ratio of polyunsaturated fatty acids to saturated fatty acids. In this study, new Antarctic Micractinium spp. were discovered and added to the culture collection. These cryotolerant strains may serve as a promising source of nutritionally important linoleic (C_18:2 ω6) and α‐linolenic (C_18:3 ω3) acids.     

Biosystematic study ofSedum L. SubgenusAizoon (Crassulaceae)

Variation in chromosome number and morphological characters ofSedum aizoon L. var.floribundum Nakai were investigated to analyze, correlations between them. Geographical variation in chromosome number was also examined. Chromosome numbers of 189 individuals from 55 localities were counted as 2n=32, 33, 34, 48, 61, 64, 78, 80, 84, 85, 88, 93, 94, 95, 96, 97 and 102. InSedum subgenusAizoon, which has the basic number of X=8, var.floribundum of the speciesaizoon showed a polyploid series from tetraploid to dodecaploid. Tetraploids were found most frequently in this variety. More than two chromosome numbers were found in all the populations with 2n=61 or more. Tetraploids were mainly distributed in the eastern part of Japan, and higher polyploids (higher than hexaploid level except octaploids) were distributed in the western part of Japan. Thirteen morphological characters were examined in 119 individuals belonging to 30 populations. The principal component analysis and the cluster analysis of these characters indicated that populations with similar chromosome numbers were not always morphologically similar. Ten morphotypes can be recognized based on the combination of chromosome number and morphological characters. ThusS. aizoon var.floribundum can be regarded as a polyploid-aneuploid complex.