Ultrastructural Study of Cytoplasmic Inclusions in Plants Infected with Potyviruses

The ultrastructure of cell inclusions of different ornamental plants infected with several filamentous viruses of the potyviruses was studied at the transmission electron microscope. The plants examined were: Crocus sativus, Hyacinthus sp., Gladiolus sp., rhizomatous Iris, Muscari sp., Nerine bowdenii and Sparaxis sp. Differences in morphology of cylindrical inclusions have been used to separte the potyviruses in several subdivisions.     

Bioactive Compounds with Added Value Prepared from Terpenes Contained in Solid Wastes from the Olive Oil Industry

Starting from solid wastes from two‐phase olive‐oil extraction, the pentacyclic triterpenes oleanolic acid and maslinic acid were isolated. These natural compounds were transformed into methyl olean‐12‐en‐28‐oate ( 5 ), which then was transformed into several seco‐C‐ring triterpene compounds by chemical and photolytic modifications. The triene seco‐products were fragmented through several oxidative procedures to produce, simultaneously, cis‐ and trans‐decalin derivatives, both potential synthons for bioactive compounds. The chemical behavior of the isolated fragments was investigated, and a suitable approach to several low‐molecular‐weight terpenes was performed. These are interesting processes for the value‐addition to solid waste from the olive‐oil industry.     

Plasmid Encoded AcrAB–TolC Tripartite Multidrug-Efflux System in Acidiphilium symbioticum H8

Acidophilic bacterium, Acidiphilium symbioticum H8, is resistant to high levels of several heavy metals, hydrophobic agents, and organic solvents. The ~9.6 kb plasmid pASH8, was purified, digested with HindIII, and sub-cloned in pUC19 at the respective site. Three different fragment size clones were achieved. The clones were completely sequenced and analyzed. The first clone encodes for a single putative open reading frame (ORF), which showed significant homology to several rusticyaninA1 proteins. The second clone encodes for a 43-kDa protein, which has conserved domain homology with several outer envelop TolC proteins. The clone with pASH8 tolC gene can functionally complement an Escherichia coli tolC mutant strain, making it resistant to several toxic hydrophobic agents, earlier for which it was sensitive. The tolC gene was found to be essential for imparting resistance to the clone toward these toxic hydrophobic agents. The third clone encodes for a putative 318-aa AcrA (acriflavine resistance protein A) protein and the clone was resistance to plasmid curing dye acriflavine. The clone also has a truncated ORF, which showed significant homology to cation-efflux pump AcrB. This study is the first to report a multi-drug efflux system to be encoded on a plasmid of any Acidiphilium strain.     

巴西橡胶Pto类抗病同源序列的克隆与系统发育重建

番茄Pto基因是一类可以编码丝氨酸/苏氨酸激酶(STK)序列的广谱抗性候选基因,其序列克隆与鉴定为深入了解番茄的抗病机制奠定了基础.在该研究中,一对依据Pto基因的保守序列设计的简并引物被用来扩增巴西橡胶中Pto基因抗病同源序列,扩增得到了一个约550 bp的基因片段,其随后被克隆并测序.序列分析发现,其中的7个抗病同源序列与Pto基因高度同源(BLASTX E value <3e-53),所以其被认为是Pto基因抗病同源序列(Pto-RGCs).通过巴西橡胶的Pto-RGCs多序列比对表明,这些序列包含了多个STKs保守的次级结构域.此外,系统发育分析也表明,巴西橡胶的Pto-RGCs属于Pto基因同源的R基因.该研究结果中Pto-RGCs可为巴西橡胶抗病的发展提供一个有效的基因资源.     

Identification of Rhizobium strains on antibiotic concentration gradients

Antibiotic concentration gradients were used to characterise several cultures of R. phaseoli and Rhizobium spp. (isolated from Cicer arietinum) by differences in intrinsic antibiotic resistance (IAR). Differentiation between cultures was facilitated by use of cluster analyses. The method permitted 15/16 cultures of R. phaseoli to be distinguished on 14 antibiotics. Two cultures which exhibited similar IAR patterns were shown to be the same strain obtained from different collections. The validity of the technique for strain identification was demonstrated by fluorescent antibody tests which gave corresponding identity for 50 nodule isolates from plants inoculated with a mixture of three strains of R. phaseoli. The method was less suitable for characterising cultures of the slow-growing Rhizobium spp. because several antibiotics produced growth lacking a clearly defined boundary between resistance and susceptibility. Although 15/16 cultures of Rhizobium spp. could be differentiated, several isolates were distinguishable only by a difference on a single antibiotic. Similarity between stock cultures and derivative nodule isolates indicated that IAR on gradient plates was a stable property unaffected by plant passage.     

Molecular characterization and cytogenetic analysis of highly repeated DNAs of lake trout,Salvelinus namaycush

The chromosomes of lake trout (Salvelinus namaycush) contain a considerable amount of heterochromatin located at the centromeres and/or telomeres of several chromosomes, including a sex-specific block located distally on the X chromosome. In order to investigate further the repetitive DNAs of lake trout, genomic DNA from a female was size fractionated (<600 bp) with the restriction endonuclease AluI and fragments were cloned into the bacteriophage M13. A total of 42 clones were isolated. Relative copy number of individual inserts within the lake trout genome was estimated by Southern analysis. Twelve clones were determined to be highly repetitive and were chosen for further investigation. Inserts of these clones contained sequences similar to the AluI/RsaI, EcoRI/DraI, DraI/BstEII, and MboI/BglII families reported from Arctic char (Salvelinus alpinus). The chromosomal location of several of these fragments was determined in lake trout by fluorescence in situ hybridization (FISH). Two related AluI/RsaI sequences (Type A, 140 bp, and Type B, 120 bp) showed differential hybridization. Type A hybridized to the centromeres of all metacentric as well as several acrocentric chromosomes. Type B hybridized to the centromeres of most acrocentric chromosomes. A sequence with homology to the EcoRI/DraI family hybridized to the centromeres of several acrocentric chromosomes. Sequences with partial similarity to the DraI/BstEII family hybridized to the major rDNA sites (nucleolar organizer regions, NORs) and several minor telomeric sites. The interstitial and telomeric heterochromatin of lake trout, including that of the X chromosome, appears to comprise sequences belonging to the MboI/BglII family.     

Chloroplast-located <Emphasis Type="Italic">BjFer1</Emphasis> together with anti-oxidative genes alleviate hydrogen peroxide and hydroxyl radical injury in cytoplasmic male-sterile <Emphasis Type="Italic">Brassica juncea</Emphasis>

We studied how plant cell modulated redox homeostasis in cytoplasmic male-sterility (CMS) Brassica juncea. The CMS Brassica juncea was identified to be mutated in several mitochondrial genes that suggested the changes of cell redox homeostasis. We observed that it was not associated with increased oxidative stress as shown by decreased H₂O₂ and ^∙OH contents in this type of CMS. The expressions of several anti-oxidative genes were up-regulated in 5-day-old seedlings of CMS than MF lines under light and dark conditions. The mitochondrial alternative oxidase pathway was not activated, as indicated by no increased expression of AOX1a gene in CMS. Interestingly, the expression of Ferritin1 gene was markedly activated in 5-day-old seedlings of CMS than MF line under light and dark conditions. Consequently, we detected increased content of total iron in 30-day-old leaves in CMS than MF line. We isolated Ferritin1 orthologous gene from Brassica juncea, which was targeted to the chloroplast and induced by Fe-citrate and H₂O₂, not ABA. Taken together, we proposed that increased expressions of BjFer1 and several antioxidant genes protected cell from oxidative stress in CMS Brassica juncea.     

Response to potyvirus infection and genetic mapping of resistance loci to potyvirus infection in Lactuca

 We have investigated the interaction between two different potyviruses and resistant cultivars of Lactuca sativa. Turnip mosaic virus (TuMV) and lettuce mosaic virus (LMV) were used to inoculate several cultivars under different temperature regimes to characterize the resistance reaction. Resistance conferred by the recessive mo locus against LMV infection did not provide immunity. Virus accumulated in plant tissues to different levels depending on the genetic background of the cultivar, suggesting that several genes were involved in the resistance phenotype. Under temperature regimes that enhanced the hypersensitive reaction, resistant cultivars produced necrotic reactions. In contrast, resistance to TuMV infection conferred by the dominant Tu locus resulted in complete immunity in the plant. No virus accumulated in inoculated leaves nor was any necrotic reaction observed. The resistance loci were characterized at the genetic level by mapping them relative to molecular markers. Only weak linkages could be identified to mo, again supporting the hypothesis that several genes are involved. The Tu locus was mapped in two different crosses relative to several markers, the closest two linked at less than 1 cM. A high-resolution genetic map of the Tu locus was constructed by screening 500 F₂ individuals for recombinants around that locus. Received: 4 June 1996/Accepted: 15 November 1996     

Trypanosomatidae Isolated from Zelus leucogrammus (Perty, 1834) (Hemiptera,Reduviidae), with a Discussion on Flagellates of Insectivorous Bugs*

The insectivorous bug Zelus leucogrammus (Perty, 1834) (Hemiptera, Reduviidae) was found to harbor several species of trypanosomatids of the genera Leptomonas, Crithidia, Herpetomonas and Blastocrithidia. However, laboratory bred nymphs given sterile food were infected only by Blastocrithidia, the other flagellates appearing in their intestinal tract when they were fed on other insects. The Blastocrithidia was thought to belong to a single species and to be a parasite proper of Z. leucogrammus (and possibly other Hemiptera), producing luxuriant infections in the intestinal tract of the bugs captured in nature and spreading easily among isolated populations through cannibalism and ingestion of cysts; it was also abundant in the genital tract of gravid females but transovarian transmission was not proved. This Blastocrithidia experimentally infected Triatoma infestans and is very close to B. triatomae Cerisola, del Prado, Rohwedder and Bozzini, 1971. The other trypanosomatids, never numerous in naturally infected Z. leucogrammus, are acquired by the bug from many different insects, chiefly Diptera, on which it was seen to feed. Herpetomonas muscarum and C. luciliae, the common trypanosomatids among several muscoid Diptera, were isolated both from flies and from the bugs fed on them.     

A potato hypersensitive resistance gene against potato virus X maps to a resistance gene cluster on chromosome 5

 The dominant Nb gene of potato confers strain-specific hypersensitive resistance against potato virus X (PVX). A population segregating for Nb was screened for resistance by inoculating with PVX strain CP2, which is sensitive to Nb. Through a combination of bulked segregant analysis and selective restriction fragment amplification, several amplified fragment length polymorphism (AFLP) markers linked to Nb were identified. These were cloned and converted into dominant cleaved amplified polymorphic sequence (CAPS) markers. The segregation of these markers in a Lycopersicon esculentum×L. pennellii mapping population suggested that Nb is located on chromosome 5. This was confirmed by examining resistant and susceptible potato individuals with several tomato and potato chromosome-5-specific markers. Nb maps to a region of chromosome 5 where several other resistance genes– including R1, a resistance gene against Phytophthora infestans, Gpa, a locus that confers resistance against Globodera pallida, and Rx2, a gene that confers extreme resistance against PVX–have previously been identified. Received: 2 January 1997/Accepted: 7 February 1997     

Occurrence of the non-phosphorylative pathway for gluconate degradation in different fungi

The presence of the non-phosphorylative pathway for gluconate degradation in several different fungi has been determined. The pathway was found to be present to varying extents in Penicillium and Mucor species, but was not detected in Fusaria and some Aspergillus. Most of the organisms tested were able to grow on gluconate as the sole source of carbon.     

6-Pentyl-α-pyrone from Trichoderma harzianum: Its Plant Growth Inhibitory and Antimicrobial Properties

Gluconobacter suboxydans has a highly active respiratory chain which oxidizes several sugars and sugar alcohols. The results of this study indicate that the sugar-oxidizing respiratory chain consists of ubiquinone, several cytochromes c and a cytochrome o. The respiratory chain was shown to contain at least five cytochromes, including two cytochromes c associated with alcohol or aldehyde dehydrogenase, two cytochromes c that react with carbon monoxide and a single cytochrome o, some of which were characterized in this study. Furthermore, several lines of evidence suggest that the respiratory chain branchs at the site of ubiquinone with KCN-sensitive and -insensitive terminal oxidases, which may correspond to cytochrome o oxidase and an alternative oxidase consisting of a possible cytochrome c, respectively.     

A REAPPRAISAL OF THE SUBGENUS GLYCINE

The genus Glycine Willd. is divided into three subgenera, Glycine Willd., Soja (Moench) F. J. Herm., and Bracteata Verdc. Six species are currently recognized in the subgenus Glycine: G. canescens F. J. Herm., G. clandestina Willd., G. falcata Benth., G. latrobeana (Meissn.) Benth., G. tabacina (Labill.) Benth., and G. tomentella Hayata. Distribution of the subgenus extends from south China to Tasmania and includes several Pacific islands. A collection of these species was examined cytologically and morphologically in an attempt to evaluate existing variability between and within taxa. Chromosome counts confirmed G. canescens, G. clandestina and G. falcata to be diploid with 2n = 40. Both tetraploids (2n = 80) and diploids were found in G. tabacina, the latter restricted to Australia. Glycine tomentella accessions were primarily tetraploid, but several collections from New South Wales, Australia, were found to be aneuploid with 78 chromosomes. One collection was aneuploid at the diploid level with 38 chromosomes. Meiosis appeared normal in the aneuploids with regular bivalent formation. Several accessions previously identified as G. tomentella were diploid. Seed of G. latrobeana was not available for analysis. Numerical techniques in the form of cluster analysis and principal components analysis were applied to morphological data on vegetative and inflorescence characters obtained from each collection. Numerical analysis grouped the accessions essentially according to current species delimitations with some exceptions. Glycine tabacina specimens from Taiwan approached G. clandestina in several characteristics. The diploid G. tomentella specimens formed a separate cluster and appeared morphologically distinct from the remaining taxa.     

Early regeneration of Calluna heathland under various fertilization treatments

Summary A sod-cutting and fertilization experiment was performed on a Calluna-dominated heathland in The Netherlands to determine appropriate management regimes for Calluna regeneration, and to further understand the nutrient responses of heathland species. Replicated permanent plots were analysed by multivariate techniques. Sod-cutting alone caused Calluna regeneration from its soil seed bank. A single fertilization at the start of the experiment caused initial vegetation differences which disappeared after a few years as the nutrients were lost from the system, except that one application of nitrogen enhanced the rate of Calluna regeneration. Repeated fertilization caused large differences in the vegetation: repeated nitrogen enhanced several bryophyte species while greatly inhibiting Calluna, repeated phosphate partly inhibited Calluna while greatly favouring several lichen species, and the most striking result of repeated calcium was also an increase in bryophytes, but the species were different from those favoured by nitrogen. Treatments which inhibited Calluna tended to increase species diversity because of the lessened Calluna dominance.     

Morphological variation in Bosmina longirostris (O.F. Müller, 1785) (Crustacea: Cladocera): consequence of cyclomorphosis or indication of cryptic species?

Samples of the polymorphic cladoceran Bosmina longirostris were collected in several lakes and ponds in Western Germany between 1993 and 1999. The shape of the first antennule was used as a qualitative criterion to classify 1241 individuals to three morphs (`longirostris', `cornuta', `pellucida') according to Lieder (Crustacea: Cladocera: Bosminidae. Stuttgart, Germany: Gustav. Fischer Verlag, 1996). Eight morphometric variables were subsequently measured in each individual to quantify size‐ and shape‐related features. Principal component and discriminant analyses were used to describe the morphological variation within and among the three morphs. Seasonal environmental changes modified several traits and thus, cyclomorphosis accounted for a high proportion of within‐morph variation. However, there was no gradual change from one morph to another. As well among‐morph variation was considerably greater than within‐morph variation. Consequently, the disagreement between the qualitative pre‐classification and the assignment basing on morphometric discriminant functions was low (4.6–12.0%). Considering that each morph is morphologically well‐defined, and that the different morphs coexisted over several generations in the same lake we conclude that they represent different species rather than variants of one polymorphic species. Therefore, we recognize beside the nominal taxon Bosmina longirostris ( 26 ) two more species: Bosmina cornuta ( 10 ) and Bosmina pellucida 30 .     

CHLAINOMONAS KOLII(HARDY ET CURL) COMB. NOV. (CHLOROPHYTA,VOLVOCALES), A REVISION OF THE SNOW ALGA,TRACHELOMONAS KOLII HARDY ET CURL (EUGLENOPHYTA,EUGLENALES)1,2

The snow alga Trachelomonas kolii is transferred from the Euglenophyta (Euglenales) to the Chlorophyta (Volvocales) as Chlainomonas kolii comb. nov. As a result of critical examination of both living and type material, this species was found to have 4 flagella per vegetative cell, true starch, and 1 axial plastid per cell with several peripheral lobes. Vegetative cells of Trachelomonas kolii were described originally as having 1 flagellum, lacking true starch but having paramylum, and as having several parietal plastids per cell. The reticulate markings on the outer envelope of vegetative cells were found to be different from those in the original illustrations. Vegetative cells and resting spores of Chlainomonas kolii and Chlainomonas rubra are compared. The similarities of resting spores of Chlainomonas kolii and Chlamydomonas nivalis are discussed. These are the first records of Chlainomonas kolii from snow in Washington State.     

Long-term survival of isolates of various Cladosporium and Fusicladium species under mineral oil

Lima bean agar cultures of several Cladosporium and Fusicladium species were placed under sterile mineral oil and stored in a refrigerator in 1956. Isolates of C. herbarum, C. fulvum, C. cucumerinum, and F. dendriticum were found to be non-viable in 1983 (after 27 years), but an isolate of C. carpophilum was found to be still viable. Earlier, in 1980, several isolates of C. caryigenum (syn. F. effusum and C. effusum) were found to be viable and capable of sporulating on various agar media (after 24 years). In 1983, re-oiled cultures of surviving C. caryigenum isolates were tested for pathogenicity to its host (Carya illinoensis — the pecan tree). One isolate was found to be still pathogenic.     

EVOLUTION OF THE LATICIFER IN EUPHORBIA AS INTERPRETED FROM STARCH GRAIN MORPHOLOGY

The morphology of plastid starch grains from several succulent and nonsucculent species of Euphorbia was examined in parenchyma and in non-articulated laticifiers. Several classes of grains were identified: small oblong-round, rod, somewhat osteoid, osteoid, lobed osteoid, discoid, and round grains. Parenchyma possessed only small oblong-round grains, whereas grains of different morphology were present in laticifers. Different species of this genus can be characterized by the morphology of the large, or mature, starch grains present in the laticifer. The rod-shaped grain, which was somewhat wider at the midregion than at the ends, was present in several nonsucculent forms, E. terracina, E. pulcherrima and E. heterophylla. The somewhat osteoid grain was represented by the succulent species E. viguieri, E. milii, and E. mauritanica, where the mature grains developed somewhat enlarged ends. Grains with much enlarged ends were represented in the succulent species of E. abyssinica, E. pseudocactus, and E. tirucalli. Alteration of the pattern of starch deposition in which several lobes were formed at the ends of the grain has given rise to a lobed osteoid class (E. inconstantia). Lobes also may be formed with greater frequency along the midregion of the grain in this than in other species. Euphorbia lactea had the most complex grain in which lobing was frequent at the ends as well as along the midregion, resulting in a large discoid grain. Subclasses in which grains differed significantly in length between species were evident in all classes containing several species. The average length of grains in any subclass was similar for subclasses between the classes. The study suggests that the elongated grain of the laticifer was derived from the round or oblong grain present in the more primitive parenchymatous cell. Progressive changes in the pattern of starch deposition have given rise to osteoid and discoid grains of increasing morphological complexity which is interpreted to represent trends in laticifer evolution between different species of Euphorbia as reflected by subtle changes for starch deposition within the plastids of this cell.     

An experimental demonstration of trophic interactions affecting water quality of Rice Lake, Ontario (Canada)

Eight cylindrical enclosures (3 m diameter, 2.7 m long, V = 20m³) were installed in eutrophic Rice Lake (Ontario, Canada) in late spring of 1987. Fish (yearling yellow perch (Perca flavescens) and macrophytes (Potamogeton crispus) presence and absence were set at the beginning of the experiment to yield four combinations of duplicate treatments. The purpose of the experiment was to determine if the phytoplankton, zooplankton, macrophytes and fish species resident in the lake interact to influence water quality (major ions, phosphorus, algal densities and water clarity).The presence of fish was associated with: (1) decreased biomass of total zooplankton, (2) decreased number of species in the zooplankton, (3) decreased average size of several zooplankton taxa, (4) higher total phosphorus concentrations, (5) higher phytoplankton and chlorophyll a concentrations, (6) lower water clarity, (7) lower potassium levels during macrophyte die-back, (8) lower pH and higher conductivity in the presence of macrophytes. Biomass of large Daphnia species (but not total zooplankton) was highly correlated with the algal response (r ² = 0.995) and was associated with reduced biomass of several algal taxa including some large forms (Mougeotia, Oedogonium) and several colonial blue-green algae. However, no significant control of late summer growth of the bloom-forming blue-green alga Anabaena planctonica Brun. was achieved by the Daphnia presence-fish absence treatment. Release of phosphorus to the water column during the die-back of P. crispus was not an important phenomenon.