CONTRIBUTION TOWARD A MONOGRAPH OF RAMARIA III. R. SANGUINEA,R. FORMOSA. AND TWO NEW SPECIES FROM EUROPE

A neotype specimen is designated for R. formosa and a representative specimen for R. sanguined, thus securing taxonomic concepts for these names. Two very similar taxa are proposed as new: R. eosanguinea, which differs from R. sanguinea chiefly in exhibiting clamp connections, and R. neoformosa which differs from R. formosa chiefly in the absence of clamps. The subject of mimic taxa is briefly discussed.     

Inositol 1,4,5-trisphosphate receptors. Localization in epithelial tissue.

Using a polyclonal antiserum raised against the inositol 1,4,5-trisphosphate receptor (IP3R) purified from rat cerebellum, we examined the subcellular distribution of IP3R in canine pancreatic homogenates. IP3R was present primarily in a smooth microsomal fraction (low density), a (high density) rough microsomal (RM) fraction previously shown to consist of highly purified rough endoplasmic reticulum (RER) vesicles, and, to a much lesser extent, in an intermediate density microsomal fraction which did not contain markers for RER or plasma membrane. When the RM fraction was subjected to isopycnic centrifugation on sucrose gradients, IP3R equilibrated at high sucrose densities. When ribosomes were extracted from the RM fraction by treatment with puromycin/high salt, IP3R equilibrated at considerably lighter sucrose densities. This shift in density indicated that IP3R which was present in the RM fraction is associated with the RER. Because of a significant amount of IP3R fractionating into the smooth microsomal fraction (which contains plasma membrane, among other "smooth" membranes) and a considerable amount of IP3R present in the nuclear pellet which is also enriched in plasma membrane, we examined the possibility that IP3R may be present in plasma membrane. Further subfractionation of a crude plasma membrane pellet from rat liver revealed that IP3R coenriched with a plasma membrane marker and strongly suggested an association of IP3R with plasma membrane. The issue of why the same receptor is found in multiple biochemically and morphologically distinct membrane fractions is discussed in terms of the possibility of RER subcompartmentalization and IP3R subtypes. The fractionation pattern of IP3R in pancreas is significantly different from that previously reported for calcium (Ca2+)-binding proteins and an intracellular Ca-ATPase (Nigam, S. K. and Towers, T. (1990) J. Cell Biol. 111, 197-200), raising questions as to links between these latter proteins and IP3 sensitive Ca2+ pools. Nevertheless, although the fractionation patterns are different, all of these proteins are clearly associated with the RER.     

Ionization of adenine derivatives: EPR and ENDOR studies of X-irradiated adenine.HCl.1/2H2O and adenosine.HCl.

Following X irradiation of adenine.HCl.H2O at 10 K, evidence for five distinct radical products was present in the EPR/ENDOR. (In both adenine.HCl.1/2H2O and adenosine.HCl, the adenine base is present in a cationic form as it is protonated at N1). From ENDOR data, radical R1, stable at temperatures up to 250 K, was identified as the product of net hydrogen loss from N1. This product, evidently formed by electron loss followed by proton loss, is equivalent to the radical cation of the neutral adenine base. Radical R2, unstable at temperatures above 60 K, was identified as the product of net hydrogen addition to N3, and evidently formed by electron addition followed by proton addition. Radicals R3-R5 could not be identified with certainty. Similar treatment of adenosine.HCl provided evidence for six identifiable radical products. Radical R6, stable to ca. 150 K, was identified as the result of net hydrogen loss from the amino group, and evidently was the product of electron loss followed by proton loss. Radical R7 was tentatively identified as the product of net hydrogen addition to C4 of the adenine base. Radical R8 was found to be the product of net hydrogen addition to C2 of the adenine base, and R9 was the product of net hydrogen addition to C8. Radical R10 was identified as the product of net hydrogen abstraction from C1' of the ribose, and R11 was an alkoxy radical formed from the ribose. With the exception of R11, all products were also found following irradiation at 65 K. Only radical R8 and R9 were stable at room temperature. Most notable is the different deprotonation behavior of the primary electron-loss products (radical R1 vs. R6) and the different protonation behavior of the primary electron-gain products (radical R2 vs. no similar product in adenosine.HCl). The major structural difference in the two crystals is the electrostatic environment of the adenine base. Therefore, this study provides further evidence that environmental influences are important in determining proton transfer processes.     

Fingerprinting bacterial chromosomal DNA with restriction endonuclease EcoRI: comparison of Rhizobium spp. and identification of mutants.

Total cellular DNA from Rhizobium trifolii, R. melitoti, and R. japonicum strains 110 and 117 were prepared. DNA fragments generated with restriction endonuclease EcoRI from these DNA samples were compared in agarose gels after electrophoresis. DNA cleavage patterns generated from R. japonicum strain 110, R. trifolii, and R. meliloti were clearly distinguishable from each other. Restriction endonuclease cleavage patterns of DNA from R. japonicum strain 110 and presumptive R. trifolii mutant strains that nodulate soybean were found to be similar. Rhizobium trifolii mutant strains were also lysed by a phage specific for R. japonicum strain 110. These results show that "R. trifolii mutant strains" are indeed derivatives of R. japonicum strain 110 and not R. trifolii.     

Chimeric twins. T.S. and M.R. reexamined

A pair of chimeric twins, T.S. (male) and M.R. (female), were examined. The amounts of 'foreign' blood cells in each twin found on three occasions were compared. The percentages of M.R. cells found in the blood of T.S. in 1977 and in 1982 were similar and about 1/5 of that found in 1970. The amount of T.S.-blood cells found in M.R. was declining slowly from about 31% in 1970 to about 25% in 1982.     

Phylogeny of Rosellinia capetribulensis sp. nov. and its allies (Xylariaceae)

A new Rosellinia species, R. capetribulensis isolated from Calamus sp. in Australia is described. R. capetribulensis is characterized by perithecia immersed within a carbonaceous stroma surrounded by subiculum-like hyphae, asci with large, barrel-shaped amyloid apical apparatus and large dark brown spores. Morphologically, R. capetribulensis appears to be similar to R. bunodes, R. markhamiae and R. megalospora. To gain further insights into the phylogeny of this new taxon we analyzed the ITS-5.8S rDNA using maximum parsimony and likelihood methods. In addition, a morphological dataset also was analyzed phylogenetically to investigate possible affinities. ITS rDNA based phylogenies reveal that R. capetribulensis is closely related to other Rosellinia species showing closest affinity to R. arcuata, RL necatrix and R. pepo. However, analysis of R. capetribulensis forms an unsupported branch sister to these taxa. Results from the morphological matrix indicate a close morphological affinity to members of Rosellinia subgenus Rosellinia. Despite that ITS rDNA and morphological analyses present difficulties in constructing a proper phylogenetic framework among Rosellinia and allied genera, there is sufficient evidence to support the establishment of the new taxon in the genus Rosellinia. The morphological similarities and differences between R. capetribulensis and allied genera such as Astrocystis and Entoleuca are also briefly discussed.     

Chromosome mapping in Pseudomonas aeruginosa PAT.

A linkage map of Pseudomonas aeruginosa PAT has been derived from the results of conjugation experiments using the plasmids FP2-2, R68, R91-5, and R68.45. FP2-2 and R68 each mobilize the chromosome from single, distinct transfer origins. R91-5 appears to mobilize the chromosome from two such origins, and R68.45 utilizes a number of transfer origins. R68 and R91-5 have both been shown to mobilize the chromosome with a polarity opposite to that by FP2-2. The locations of the transfer origins of these plasmids are such that it has not been possible to demonstrate chromosomal circularity by means of interrupted mating experiments. However, the available time-of-entry data combined with linkage data from plate mating experiments support the conclusion that the chromosome of P. aeruginosa is circular.     

Pararhinebothroides hobergi n. gen. n. sp. (Eucestoda: Tetraphyllidea) in Urobatis tumbesensis (Chondrichthyes: Myliobatiformes) from coastal Ecuador.

A new species of tetraphyllidean eucestode inhabiting Urobatis tumbesensis from inshore waters of southeastern Ecuador shares 3 synapomorphies with Rhinebothroides spp.: apical bothridial suckers poorly differentiated from the marginal loculi, internal seminal vesicles, and insertion of the vas deferens dorsally closer to the poral than the aporal end of the cirrus sac. The new species differs from Rhinebothroides spp. by lacking medial bothridial septa and loculi and having symmetrical ovarian arms, and possesses an apparent autapomorphic trait by having the vas deferens tapering to a narrow tube before entering the cirrus sac, extending posteriorly to the posterior end of the cirrus sac where it expands into an external seminal vesicle running ventral to the cirrus sac anteriorly to anterior to the vagina. In Rhinebothroides spp., the vas deferens is expanded into an external seminal vesicle near the insertion into the cirrus sac As the sister group of Rhinebothroides, we propose a new genus to accommodate the new species. Phylogenetic evaluation of phyllobothriids recently assigned to Anthocephalum shows that they represent a paraphyletic assemblage of species of varying degrees of relatedness to Rhinebothroides spp. and the new species. Uncovering the relationships of the new species and the various species assigned to Anthocephalum permitted reevaluation of character transformations used in previous phylogenetic analysis of Rhinebothroides. Transformation series for 3 characters, previously based on functional outgroup comparisons, changed and a new character, length of cirrus sac, was added. The new phylogenetic analysis differs from the previous hypothesis only in placing R. scorzai as the sister species of R. circularisi + R. venezuelae + R. moralarai rather than of R. freitasi + R. glandularis + R. mclennanae. The occurrence of the sister species of Rhinebothroides in a Pacific Ocean stingray adds additional support to the hypothesis of Pacific origins of South American freshwater stingrays.     

Studies on bucephalid digeneans parasitising molluscs and fishes in Finland. II. The description of Rhipidocotyle fennica n. sp. and its discrimination by principal components analysis

Rhipidocotyle fennica n. sp. (= Rhipidocotyle Type A of Taskinen et al., 1991) from the intestine of Esox lucius in central Finland is described and compared by means of a principal components analysis (PCA) with R. campanula (= Rhipidocotyle Type B of Taskinen et al., 1991). Its cercaria develops in the bivalve Anodonta anatina and the metacercaria occurs in the skin and fins of Rutilus rutilus. The metacercaria is discriminated from that of R. campanula by PCA and is described along with aspects of the chaetotaxy of the cercaria. The new species is distinguished from R. campanula, R. kovalae, R. papillosa and R. septpapillata.     

Species delimitation of three species within the Russula subgenus Compacta in Korea: R. eccentrica,R. nigricans,and R. subnigricans

Distinguishing individual Russula species can be very difficult due to extensive phenotypic plasticity and obscure morphological and anatomical discontinuities. In this study, we use the internal transcribed spacer (ITS) and 28S nuclear ribosomal large subunit (LSU) markers to identify and study the genetic diversity of species in the Russula subgenus Compacta in Korea. We focus on two morphologically similar species that are often misidentified for each other: R. nigricans and R. subnigricans. Based on molecular phylogenetic analyses, we identify three subgroups of R. nigricans, with two from Asia and one from Europe/North America. Surprisingly, we find Korean R. subnigricans are more closely related to R. eccentrica from North America than the type specimen of R. subnigricans from Japan. These molecular data, along with habitat data, reveal that Korean R. subnigricans had previously been misclassified and should now be recognized as R. eccentrica. Both ITS and LSU exhibit high interspecific and low intraspecific variation for R. eccentrica, R. nigricans, and R. subnigricans. These markers provide enough resolutional power to differentiate these species and uncover phylogeographic structure, and will be powerful tools for future ecological studies of Russula.     

Studies of non-disjunction of the major autosomes in Drosophila melanogaster. II. Effects of dose-fractionation, low radiation doses, EMS, and ageing.

To test whether the induction of two-hit events plays a role in the induction of disomic gametes by exposure to X-rays of the oocytes of females carrying compound second chromosomes, exposures of 1000 R, 2000 R or 3000 R were given as an acute dose or as two equal fractions separated by a 3-h interval. The dose-effect relationship for acute exposure is linear over this exposure range (1000–3000 R) and the yields obtained are remarkebly similar to those recorded in an earlier study by Clark and Sobels. At 3000 R a significant reduction in yield was observed after exposure fractionation. At exposure levels of 2000 and 1000 R, however exposure fractionation tends to enhance the yield od disomics.The dose-effect relationship study was then extended to 125, 250 and 500 R. After an exposure of 125 R the induced frequency of disomics was significantly higher than control, but not different from the frequencies induced by 250, 500 and 1000 R. There is no obvious explanation for this plateau which, together with the linear increase from 1000–3000 R, provide an indication that there may be several mechanisms involved in the induction of disomic gemetes.Treatment with ethyl methanesulfonate (EMS) and ageing of the females for a week did not affect the yield of disomic progeny.     

Aspects of the Comparative Physiology of Ranunculus bulbosus L. and Ranunculus repens L.: I. Response to Nitrogen

Ranunculus bulbosus and R. repens are closely related taxonomicallyand coexist in the same general habitat but differ in that R.bulbosus only exceptionally shows vegetative multiplicationcontrasting with R. repens which reproduces extensively by meansof long branched stolons which root and form new daughter plantsat the nodes. The response of these two species to nitrogen was similar inthat an increase from 10 to 50 ppm nitrogen gave rise to greatlyincreased total dry weight and higher shoot to root ratios,whereas further nitrogen increase from 50 to 200 ppm had littleeffect. However, they differed in that at low nitrogen levelsthe perennating organ (corm) took priority in R. bulbosus withsexual reproduction assuming greater importance at the highernitrogen levels, whereas in R. repens the main storage organ(the stock) was not especially favoured at low nitrogen levelsreflecting perhaps the differences between this structure andthe corm in R. bulbosus. In R. repens the greatest responseto nitrogen was shown by the stolons although this was due toincreased branching and a higher dry weight per unit lengthat the higher levels of nitrogen (50 and 200 ppm). Primary stolonlength was not greatly affected by nitrogen level. Dry weightsof flowers and fruits in R. repens were very much lower thanin R. bulbosus. It is suggested that in R. bulbosus the emphasis lies firstin replacement of the parent at approximately the same siteby a daughter from the new corm and secondly in sexual reproductivespread. In R. repens lateral spread by vegetative propagationtakes precedence over all else and under low nitrogen conditionsbranching is sacrificed and energy is diverted to maintaininglong primary stolons. These, in turn, have the potential forproducing daughter plants at some distance from the parent andthis may be of considerable importance in field situations sincethe daughter may come to occupy more favourable sites than theparent.     

Characterization of the umu-complementing operon from R391.

In addition to conferring resistances to antibiotics and heavy metals, certain R factors carry genes involved in mutagenic DNA repair. These plasmid-encoded genes are structurally and functionally related to the chromosomally encoded umuDC genes of Escherichia coli and Salmonella typhimurium. Three such plasmid operons, mucAB, impCAB, and samAB, have been characterized at the molecular level. Recently, we have identified three additional umu-complementing operons from IncJ plasmid R391 and IncL/M plasmids R446b and R471a. We report here the molecular characterization of the R391 umu-complementing operon. The nucleotide sequence of the minimal R plasmid umu-complementing (rum) region revealed an operon of two genes, rumA(R391) and rumB(R391), with an upstream regulatory signal strongly resembling LexA-binding sites. Phylogenetic analysis revealed that the RumAB(R391) proteins are approximately equally diverged in sequence from the chromosomal UmuDC proteins and the other plasmid-encoded Umu-like proteins and represent a new subfamily. Genetic characterization of the rumAB(R391) operon revealed that in recA+ and recA1730 backgrounds, the rumAB(R391) operon was phenotypically indistinguishable from mucAB. In contrast, however, the rumAB(R391) operon gave levels of mutagenesis that were intermediate between those given by mucAB and umuDC in a recA430 strain. The latter phenotype was shown to correlate with the reduced posttranslational processing of the RumA(R391) protein to its mutagenically active form, RumA'(R391). Thus, the rumAB(R391) operon appears to possess characteristics that are reminiscent of both chromosome and plasmid-encoded umu-like operons.     

Interleukin-6 receptor signaling. II. Bio-availability of interleukin-6 in serum.

Interleukin-6 (IL-6) is used as a growth factor by various tumor cells. It binds to a gp80 specific receptor (IL-6R) and then to a gp130 transducing chain. Both receptor chains are released as soluble functional proteins which circulate in biological fluids. With a view to studying the physiological role of these soluble receptors, both proteins were purified from human plasma. Surface plasmon resonance was used to measure the kinetic constants of equilibria between IL-6 and natural sIL-6R, and between the IL-6/sIL-6R complex and soluble gp130. Kd values were found to be 0. 9 and 2.3 nM respectively. Soluble natural IL-6R and gp130 were also found to interact with a Kd of 2.8 nM in the absence of IL-6. By using these Kd values, a mathematical simulation predicted that 1) within a large range of IL-6, sIL-6R and sgp130 concentrations, free IL-6 represents 30% of the total circulating cytokine, 2) sIL-6R overconcentrations lead to dramatic changes of the concentration of free IL-6, 3) increased concentrations of sgp130 should produce an efficient buffering effect on the IL-6/sIL-6R complex without incidence on the level of free IL-6. According to this model, the IL-6/sIL-6R complex appears to be an important support of IL-6 signaling in the most commonly encountered in vivo situations. The concentration of this complex is directly under the control of the concentration of sIL-6R; its bio-availability should be efficiently buffered by increased sgp130 concentrations.     

Polynucleotide fragments from the 28S ribosomal RNA of insects.

If RNA is extracted from the ribosomes which had been isolated from frozen-thawed tissue of Galleria mellonella, the 28 S RNA, when heated or treated with urea, dissociates into seven different species of polynucleotide fragments. They were designated as R1, R2, R3, R4, R5, R6, and R7, whose molecular weights were estimated to be 1.15x10-6, 0.75x10-6, 0.55x10-6, 0.40x10-6, 0.30x10-6, 0.25x10-6, 0.20x10-6 daltons, respectively. It is likely that R1 and R5 arise from a single nick in original 38 S rRNA. Experiments with isolated R1 suggest that it is made up of a hydrogen-bonded complex of R2 and R4. R5 is a complex of R6 and an unidentified species, X. It is suggested that these fragments result from nicks which are introduced, secondarily, in the phosphodiester bonds by an endogenous endonuclease(s). Since the secondary nicks are limited in number and located in specific points of the molecule, it appears that the reaction is quite specific. It was also shown that the 28 S aphid RNA, which apparently lacks the primary nick, is susceptible to nicking.     

Overproduction and crystallization of FokI restriction endonuclease.

To overproduce FokI endonuclease (R.FokI) in an Escherichia coli system, the coding region of R.FokI predicted from the nucleotide sequence was generated from the FokI operon and joined to the tac promoter of an expression vector, pKK223-3. By introduction of the plasmid into E. coli UT481 cells expressing the FokI methylase gene, the R.FokI activity was overproduced about 30-fold, from which R.FokI was purified in amounts sufficient for crystallization. The removal of a stem-loop structure immediately upstream of the R.FokI coding region was essential for overproduction.     

On the Difference Between Elachanthemum Y. Ling et Y. R. Ling and Stilpnolepis Krasch.

It is quite unreasonable reducing Elachanthemum Y. Ling et Y. R. Ling into Stilprolepis Krasch. and it is so wrong idea attributing the achenes and cupuliform corolla (as a matter of fact, the cupuliform corolla is originally from the Stilpnolepis Krasch., not from Elachanthemum Y. Ling et Y. R. Ling) of Elachanthemum Y. Ling et Y. R. Ling to “earlier Development” in the paper published in Act. Phytotax. Sin. 23(6): 470-472. 1985. In Elachanthemum Y. Ling et Y. R. Ling, bracts of capitula herbaceous, obviously floccose on the abaxial surface and membranaceous only on the margin, corolla of bisexual florets tubular, achenes oblique, obovoid, and the exine of pollen grains minutespinulate, but in Stilpnolepis Krasch., on the contrary, whole bracts membranaceous, glabrous, corolla of hermaphrodite florets cupuliform or campanulate, achenes long-clavate or fuciform and the exine of pollen grains remarkably spiny.     

Three ascomycetes on leaves of evergreen Ilex trees from Japan: Rhytisma ilicis-integrae sp. nov., R. ilicis-latifoliae, and R. ilicis-pedunculosae sp. nov.

Three species belonging to the genus Rhytisma causing tar spot were collected on leaves in evergreen trees of Ilex species from Japan. Rhytisma ilicis-latifoliae, the known species, is found on Ilex latifolia, and R. ilicis-integrae sp. nov. and R. ilicis-pedunculosae sp. nov. are found on I. integra and I. pedunculosa, respectively. Ascomata are formed on the abaxial part of the stromata in all the Rhytisma species studied, and spermogonia are formed on the amphigenous parts in R. ilicis-latifoliae and on the adaxial part in R. ilicis-integrae and R. ilicis-pedunculosae. Shape and size of asci, ascospores, and spermatia are distinctly different among the three species. The morphology of germination tubes from ascospores and appressoria is unique for each Rhytisma species. Yellowish spots arise on the newly developing leaves in mid-May, then abundant spermatia are produced in spermogonia in the three Rhytisma species. In the next year, ascospores are produced in ascomata from early April to late May in R. ilicis-integrae and from early April to early June in R. ilicis-latifoliae and R. ilicis-pedunculosae, and they are considered to be the inocula of disease infection.     

Genetic differentiation of U.S.S.R. house mice: electrophoretic study of proteins

Protein electrophoresis at 24 loci was used to characterize house mice from 56 localities in the U.S.S.R., concentrating on samples from Moldavia to Primorye (extreme south-east of the U.S.S.R.). Mus -2A is the most widespread form, extending over the European part of the U.S.S.R., Middle Asia and Siberia as far east as the Pacific Ocean. In Moldavia the group is sympatric with Mus-iB . It is found with Mus -4A in Transcaucasus, where it may hybridize with Mus -1. In Primorye Mus -2A and M. raddei have a wide zone of hybridization with Mus -2C.     

A comparison of the unfolding and dissociation of the large ribosome subunits from Rhodopseudomonas·spheroides N.C.I.B. 8253 and Escherichia coli M.R.E. 600

1. The behaviour of the large ribosomal subunit from Rhodopseudomonas spheroides (45S) has been compared with the 50S ribosome from Escherichia coli M.R.E. 600 (and E. coli M.R.E. 162) during unfolding by removal of Mg(2+) and detachment of ribosomal proteins by high univalent cation concentrations. The extent to which these processes are reversible with these ribosomes has also been examined. 2. The R. spheroides 45S ribosome unfolds relatively slowly but then gives rise directly to two ribonucleoprotein particles (16.6S and 13.7S); the former contains the intact primary structure of the 16.25S rRNA species and the latter the 15.00S rRNA species of the original ribosome. No detectable protein loss occurs during unfolding. The E. coli ribosome unfolds via a series of discrete intermediates to a single, unfolded ribonucleoprotein unit (19.1S) containing the 23S rRNA and all the protein of the original ribosome. 3. The two unfolded R. spheroides ribonucleoproteins did not recombine when the original conditions were restored but each simply assumed a more compact configuration. Similar treatments reversed the unfolding of the E. coli 50S ribosomes; replacement of Mg(2+) caused the refolding of the initial products of unfolding and in the presence of Ni(2+) the completely unfolded species (19.1S) again sedimented at the same rate as the original ribosomes (44S). 4. Ribosomal proteins (25%) were dissociated from R. spheroides 45S ribosomes by dialysis against a solution with a Na(+)/Mg(2+) ratio of 250:1. During this process two core particles were formed (21.2S and 14.2S) and the primary structures of the two original rRNA species were conserved. This dissociation was not reversed. With E. coli 50S approximately 15% of the original ribosomal protein was dissociated, a single 37.6S core particle was formed, the 23S rRNA remained intact and the ribosomal proteins would reassociate with the core particle to give a 50S ribosome. 5. The ribonuclease activities in R. spheroides 45S and E. coli M.R.E. 600 and E. coli M.R.E. 162 50S ribosomes are compared. 6. The observations concerning unfolding and dissociation are consistent with previous reports showing the unusual rRNA complement of the mature R. spheroides 45S ribosome and show the dependence of these events upon the rRNA and the importance of protein-protein interactions in the structure of the R. spheroides ribosome.