The use of transposons for the mutagenesis of R. phaseoli and R. japonicum

Plasmid pSUp2011 has been used to transfer transposon Tn5 into the cells of R. japonicum 110 and R. phaseoli 693. Transposition of Tn5 into the chromosomes of R. japonicum and R. phaseoli has been demonstrated,resulting in isolation of auxotrophic and symbiotic mutants of both strains. The frequencies of selected auxotrophic mutations have reached 4% in R. japonicum 110 and 0.6% in R. phaseoli 693. Streptomycin resistance gene locating on Tn5 has been found to be phenotypically expressed in R. japonicum 110 and R. phaseoli 693 cells.     

大黄属3种大黄植物不同部分蒽醌含量的测定与比较

采用C18反相柱高效液相色谱方法分离、外标法定量对大黄属掌叶组唐古特大黄、波叶组波叶大黄、穗序组穗序大黄的根（及根茎）、叶片、叶柄、主茎四部分的芦荟大黄素、大黄酸、大黄素、大黄酚4种游离和结合蒽醌的含量进行了测定和比较。结果表明：唐古特大黄中,叶片中的游离蒽醌含量高于其它部分,游离蒽醌总量地上部分远高于地下部分;结合蒽醌则根中最高,蒽醌总量地下部分远高于地上部分。波叶大黄和穗序大黄中,游离和结合蒽醌均为根中最高,穗序大黄蒽醌总量地下部分远高于地上部分,而波叶大黄中,游离蒽醌总量地上部分高于地下部分,结合蒽醌总量地上部分与地下部分相差不大,地上部分略高于地下部分。     

Comparison of in vitro susceptibilities of Rickettsia prowazekii, R. rickettsii, R. sibirica and R. tsutsugamushi to antimicrobial agents

The in vitro activities of 16 antimicrobial agents against Rickettsia prowazekii (Breinl strain), R. rickettsii (Bitterroot strain), R. sibirica (ATCC No. VR151) and R. tsutsugamushi (Gilliam, Karp, Kato, Shimokoshi, Kawasaki and Kuroki strains) were determined by the cell culture method. Tetracycline, demethylchlortetracycline, doxycycline, minocycline, chloramphenicol, kitasamycin and rifampicin were generally effective (MIC, 0.005-0.78 micrograms/ml) to all strains tested. Quinolones such as norfloxacin, ciprofloxacin and ofloxacin were moderately active, but they were less active against R. tsutsugamushi than other rickettsial species. Penicillins and cephems showed low activity against most of the strains tested, but high concentrations of benzylpenicillin (MIC, 25-50 micrograms/ml) inhibited R. prowazekii, R. rickettsii and R. sibirica. These findings may be applicable for differentiation of species of genus Rickettsia.     

Gates, R.R. Early historico-botanical records of theOenotheras

Ohne Zusammenfassung

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Gates, R.R. Mutation inOenothera

     

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.     

Experimental infection of domestic animals with R. prowazeki and R. canada]

The authors infected lambs with R. prowazeki and R. canada to ascertain their possible role in the natural infection of the animals. The lambs were infected subcutaneously with increasing doses; rickettsiemia was recorded with the aid of tests on guinea pigs and Ixodidae and Argasidae ticks fed on the lambs. Dynamics of antibody formation was ascertained in the infected animals in the agglutination reaction and in the complement fixation test. The antigenic affinity of R. canada and rickettsia of the typhoid group and the presence of common antigenic determinants with the Proteus OX19 was confirmed. The absence of any clinical manifestations, the character of antibody formation, impossibility of inducing the generalized infection and of the isolation of the causative agent from the blood pointed to the low susceptibility of lambs to R. prowazeki and R. canada; thus a possibility of circulation of the causative agents of typhius among the domestic animals scarcely probable.     

A cytogenetic study of R. constantinopolitanus and R. sericeus (Ranunculaceae) in Iran

With 600 species Ranunculus is the largest genus in Ranunculaceae, and has a broad global distribution. We studied the karyotypes of R. constantinopolitanus and R. sericeus species of Ranunculaceae and identified their symmetry level. New chromosome numbers of 2n = 21 (Nodeh woods population) and 2n ca. 63 (Javaherdeh population) are reported for R. constantinopolitanus. Two different populations of R. sericeus had two different chromosome results. We investigated morphological and karyological studies along with pollens micromorphology. Different populations of R. constantinopolitanus and R. sericeus, with different chromosome numbers showed morphological and micromorphological differences. Therefore, we considered the two populations of R. sericeus as cytotypes. There was a correlation between the studied morphological characters and pollen size with ploidy levels in the two R. constantinopolitanus populations.     

Polymorphism of the ITS-2 region of the ribosomal DNA of the Triatominae Rhodnius domesticus, R. pictipes, R. prolixus and R. stali

Abstract The polymerase chain reaction?restriction fragment length polymorphism technique (PCR?RFLP) was used to compare Rhodnius domesticus (Neiva & Pinto), R. pictipes (Stal), R. prolixus (Stal) and R. stali (Lent, Jurberg & Galvão) (Hemiptera: Reduviidae). The enzyme BstUI differentiated R. domesticus, R. pictipes and R. prolixus, and HhaI differentiated R. domesticus, R. pictipes and R. stali. With the fingerprinting analysis generated by these two enzymes, it was possible to clearly identify all four species in the study.     

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.     

Cytological Studies of Hybrids of Roegneria ciliaris with R.alashanica and R.magnicaespes(Poaceae:Triticeae)

In order to investigate the relationships between Roegneria.ciliaris(Trin.)Nevski and R.alashanica Keng,and between R.ciliaris and R.magnicaespes D.F.Cui.R.ciliaris(Trin.)Nevski was crossed with the other two species,through the aid of embryo rescue. Hybrids were obtained from these two combinations.Chromosome pairing behavior of parents an dhybrids was observed at metaphase I of pollen mother cells. Meiotic configuratons were 10.62Ⅰ＋8.17Ⅱ＋0.32Ⅲ＋0.02Ⅳ for R.ciliaris×R.magnicaespes and 18.00Ⅰ＋4.76Ⅱ＋0.16Ⅲ for R.ciliaris ×R.magnicaespes;c-value of the two hybrids were 0.44 and 0.21,respectively.     

Genome constitutions of Roegneriaalashanica,R. elytrigioides,R. magnicaespes and R. grandis (Poaceae: Triticeae) via genomic in‐situ hybridization

The genomic constitutions of Roegneria alashanica, R. elytrigioides, R. magnicaespes and R. grandis were studied using GISH. DNA of Pseudoroegneria spicata (St), P. libanotica (St), P. stipifolia (St), R. ciliaris (StY), Lopophyllum elongatum (E^e), Agropyron cristatum (P) and Hordeum bogdanii (H) were used for probing, respectively. The results indicated that: 1) R. alashanica and R. magnicaespes contained one St genome, the other genome was unidentified, however, it was not an E, P, H or Y genome; 2) R. elytrigioides contained two St genomes and should on this basis be included in Pseudoroegneria; 3) R. grandis contained an St and an St^g genome. The St^g genome is suggested to be a modified form of the St genome of Pseudoroegneria and to be homoeologous with the Y genome. It might be an intermediate type between the St and Y genomes. Therefore, R. elytrigioides should be treated as Pseudoroegneriaelytrigioides.Roegneria alashanica, R. magnicaespes and R. grandis does apparently not belong in the genus Roegneria but further studies are needed to establish their correct taxonomic position.     

Comparative Ecology of Rubus Species (Rosaceae) II. Reproductive Characteristics of Three Rubus Species, R. palmatus var. coptophyllus, R. microphyllus and R. crataegifolius

Abstract The reproductive characteristics of three Rubus species ( R. palmatus Thunberg var. coptophyllus (A. Gray) O. Kuntze, R. microphyllus L.f., and R. crataegifolius Bunge) were critically investigated using materials collected from the forest floor of an 85-year-old Hinoki ( Chamaecyparis obtusa ) plantation in central Honshu, Japan. The underground branching patterns of these species were also investigated at the nursery. R. microphyllus produced the largest number of fruits in the same cane size among the three species (164 fruits per cane of 300 cm³ in D₀2H). However, the largest number of seeds produced per cane was in R. crataegifolius , where the maximum seed number attained 74,500 per cane, which was twice as many as those of the other two species. There was no apparent difference in dry matter allocation to fruits in these species. The three species produced numerous suckers by underground branching, forming large clones. In the open site at the nursery, the number of suckers produced by underground branching increased exponentially. On the other hand, in natural populations occurring in closed sites, the number of suckers produced by underground branching in the three species was by contrast much smaller. Three Rubus species proved to possess similar perennation as well as reproductive strategies, although R. palmatus var. coptophyllus shows much lower seed production and slower sucker production than the other two species.     

Genetic determinant of pyocin R2 in Pseudomonas aeruginosa PAO. II. Physical characterization of pyocin R2 genes using R-prime plasmids constructed from R68.45

The chromosome segment which contains the genes responsible for production of pyocin R2 in P. aeruginosa PAO was defined physically using R-prime plasmids constructed in vivo from R68.45. The previous conclusion from genetic mapping that the cluster of pyocin R2 genes is located in between trpC and trpE genes was confirmed by deletion mapping of various R prime plasmids bearing the trpC gene. The pyocin R2 gene cluster was further localized on two contiguous HindIII fragments of 16 kb and 8.0 kb. PML14 strain, in which R-type pyocin genes were completely deleted, had only one 11 kb HindIII fragment instead. Heteroduplexes between this 11 kb fragment with the two HindIII fragments of PAO revealed that the cluster of pyocin R2 genes was an insertion 13 kb long.     

Role of arginine 177 in the MnII binding site of manganese peroxidase. Studies with R177D, R177E, R177N, and R177Q mutants.

Previously, we reported that Arg177 is involved in MnII binding at the MnII binding site of manganese peroxidase isozyme 1 (MnP1) of Phanerochaete chrysosporium by examining two mutants: R177A and R177K. We now report on additional mutants: R177D, R177E, R177N, and R177Q. These new mutant enzymes were produced by homologous expression in P. chrysosporium and were purified to homogeneity. The molecular mass and the UV/visible spectra of the ferric and oxidized intermediates of the mutant enzymes were similar to those of the wild-type enzyme, suggesting proper folding, heme insertion, and preservation of the heme environment. However, steady-state and transient-state kinetic analyses demonstrate significantly altered characteristics of MnII oxidation by these new mutant enzymes. Increased dissociation constants (Kd) and apparent Km values for MnII suggest that these mutations at Arg177 decrease binding of MnII to the enzyme. These lowered binding efficiencies, as observed with the R177A and R177K mutants, suggest that the salt-bridge between Arg177 and the MnII binding ligand Glu35 is disrupted in these new mutants. Decreased kcat values for MnII oxidation, decreased second-order rate constants for compound I reduction (k2app), and decreased first-order rate constants for compound II reduction (k3) indicate that these new mutations also decrease the electron-transfer rate. This decrease in rate constants for compounds I and II reduction was not observed in our previous study on the R177A and R177K mutations. The lower rate constants suggest that, even with high MnII concentrations, the MnII binding geometries may be altered in the MnII binding site of these new mutants. These new results, combined with the results from our previous study, clearly indicate a role for Arg177 in promoting efficient MnII binding and oxidation by MnP.     

Combined action of penicillin, tetracycline and rifampicin on R. sibirica and R. prowazekii in experiments on chick embryos

The effect of combinations of penicillin, tetracycline and rifampicin on R. prowazekii (the causative agent of typhus) and R. sibirica (the causative agent of tick-borne rickettsiosis of the North Asia) was studied. It was shown that tetracycline and penicillin used in combination had a summation effect on both R. sibirica and R. prowazekii. The dose of each antibiotic was 2 times lower than the doses of the antibiotics used alone. However, R. sibirica was less sensitive to this combination than R. prowazekii: the minimum rickettsiocidic doses of the combination were 0.5 mg of tetracycline + 10000 units of penicillin per embryo with respect to R. sibirica and 0.1 mg of tetracycline + 10000 units of penicillin per embryo with respect to R. prowazekii. The combinations of rifampicin with penicillin or tetracycline in the concentrations used had no rickettsiocidic effect on either R. sibirica or R. prowazekii. However, it should be noted that these combinations had a synergistic action and provided a rickettsiostatic effect on R. prowazekii: the dose of rifampicin in its combination with penicillin was decreased 10 times and in the combination of rifampicin with tetracycline the doses of both rifampicin and tetracycline were decreased 10 times. Still, penicillin even in a dose of 20000 units per embryo had only a rickettsiostatic effect on R. sibirica and R. prowazekii.     

Historical and recent evidence for close relationships among Rickettsia parkeri,R. conorii,R. africae,and R. sibirica: Implications for rickettsial taxonomy

Rickettsia parkeri, a member of the spotted fever group rickettsias, was first described in 1939 and was thought to be non‐pathogenic until recently, when it was found to cause a spotted fever‐like illness in humans and areas of necrosis (eschars) at the sites of tick bites. Accordingly, there is currently much interest in this emerging pathogen. In this study, all published articles concerning R. parkeri were reviewed and analyzed for evidence of relatedness among this agent and other spotted fever group (SFG) rickettsiae which also produce similar clinical syndromes and/or eschars, including R. conorii, R. africae, and R. sibirica. A synthesis of the historical (antigenic) and recent (molecular) data supporting a phylogenetic sub‐grouping of these SFG organisms is presented and comments are offered about the taxonomy of rickettsial organisms in general, and R. parkeri in particular.     

Effect of divalent cations on succinate transport in Rhizobium tropici,R. leguminosarum bv phaseoli and R. loti

Rhizobium tropici, R. leguminosarum bv phaseoli and R. loti each have an active C₄-dicarboxylic acid transport system dependent on an energized membrane. Free thiol groups are probably involved at the active site. Since EDTA inhibited succinate transport in R. leguminosarum bv phaseoli and R. loti, divalent cations may participate in the process; the activity was reconstituted by the addition of Ca²⁺ or Mg²⁺. However, EDTA had no effect on succinate transport in R. tropici, R. meliloti or R. trifolii strains. Ca²⁺ or Mg²⁺ had a similar effect on the growth rates of R. tropici and R. leguminosarum bv phaseoli; R. tropici did not require Ca²⁺ to grow on minimal medium supplemented with succinate but R. leguminosarum bv phaseoli required either or both of the divalent cations Ca²⁺ and Mg²⁺. A R. tropici Mu-dI (lacZ) mutant defective in dicarboxylic acid transport, was isolated and found unable to form effective bean nodules.The authors are with the Division of Biochemistry, Instituto de Investigaciones Biológicas Clemente Estable, Avda, Italia 3318, 11.600 Montevideo, Uruguay     

Comparative Ecology of Rubus Species (Rosaceae) I. Ecological Distribution and Life History Characteristics of Three Species, R. palmatus var. coptophyllus, R. microphyllus and R. crataegifolius

Abstract The ecological distribution and life history characteristics of three Rubus species, i.e., R. palmatus Thunb. var. coptophyllus (A. Gray) O. Kuntze, R. microphyllus L.f. and R. crataegifolius Bunge were investigated, with particular emphasis on the seasonal growth cycles, modes of the alternation of aerial organs, and patterns of dry matter allocation, and these features were critically compared with one another. It was found that seasonal changes of aboveground biomass are strongly related to the modes of cane alternation, to underground branching producing stolons, and to their dry matter allocation patterns. The modes of cane alternation, underground branching and dry matter allocation patterns were very similar in R. microphyllus and R. crataegifolius , taking about five months from May to September. But R. palmatus var. coptophyllus showed much more gradual replacement of aerial organs, extending over seven months from May to December. These three Rubus species are often predominant in the early successional stages of pioneer communities which develop after cutting of forests in central Japan, and differentiations of the above-mentioned life history traits found in Rubus species seem to be related to the constraints inherent in their somewhat labile, changing environments, e.g., at the forest margin or clearings produced temporarily.