Requirements for in vitro propagation of seven nitrogen-fixing Alnus species

Studies on the in vitro propagation of Alnus crispa, A. glutinosa, A. incana, A. japonica, A. rubra, A. sinuata and A. viridis indicated interspecific as well as intraspecific variations in their requirements for in vitro culture. The WPM and Blaydes media supported, respectively, growth of A. glutinosa and A. crispa but not that of both species, while the MS medium induced equal or significantly better growth than WPM and Blaydes media for both species. The optimum type and concentration of sugar to be used in the multiplication medium varied with species. Only A. glutinosa showed good growth on sucrose while glucose was optimum for all other species but at different concentrations. All species rooted in 3 weeks on half-strength MS medium including 1 M IBA. All clones of A. glutinosa and A. rubra rooted 100%, whereas easy-to-root and difficult-to-root clones were observed in the other species. In the rooting medium, glucose promoted rooting of the difficult-to-root clones better than sucrose. Survival following transfer to an artificial substrate was 100% for all species. Nodulation tests using pure cultures of two Frankia strains showed 100% nodulation on all Alnus clones.     

Quantitative analyses of C-banded karyotypes,and systematics in the cultivated species of theScilla siberica group (Liliaceae)

Quantified C-band karyograms are presented for the related speciesScilla siberica, S. mordakiae, S. ingridae, S. amoena, andS. mischtschenkoana. Chromosome structure, banding style, and heterochromatin characters suggest a systematic grouping of two more closely related species pairs:S. siberica andS. mordakiae, S. ingridae andS. amoena; they are part of a larger aggregate, well separated fromS. mischtschenkoana. Four different heterochromatin fractions can be recognized inS. siberica and its relatives, but only two inS. mischtschenkoana. C-bands do not replace, but they are added to euchromatin. The particular origin and history of the cultivatedS. amoena and the triploidS. siberica cv. Spring Beauty appear to be responsible for their karyotype constancy, but chromosome conservatism obviously is genuine inS. mischtschenkoana. The banding data support the systematic grouping proposed on a morphological basis, and provide additional evolutionary evidence.Evolution ofScilla and Related Genera, IV.     

β-D-Glycan synthases and the CesA gene family: lessons to be learned from the mixed-linkage (1→3),(1→4)β-D-glucan synthase

Cellulose synthase genes (CesAs) encode a broad range of processive glycosyltransferases that synthesize (14)-D-glycosyl units. The proteins predicted to be encoded by these genes contain up to eight membrane-spanning domains and four `U-motifs' with conserved aspartate residues and a QxxRW motif that are essential for substrate binding and catalysis. In higher plants, the domain structure includes two plant-specific regions, one that is relatively conserved and a second, so-called `hypervariable region' (HVR). Analysis of the phylogenetic relationships among members of the CesA multi-gene families from two grass species,Oryza sativa and Zea mays, with Arabidopsis thaliana and other dicotyledonous species reveals that the CesA genes cluster into several distinct sub-classes. Whereas some sub-classes are populated by CesAs from all species, two sub-classes are populated solely by CesAs from grass species. The sub-class identity is primarily defined by the HVR, and the sequence in this region does not vary substantially among members of the same sub-class. Hence, we suggest that the region is more aptly termed a `class-specific region' (CSR). Several motifs containing cysteine, basic, acidic and aromatic residues indicate that the CSR may function in substrate binding specificity and catalysis. Similar motifs are conserved in bacterial cellulose synthases, the Dictyostelium discoideum cellulose synthase, and other processive glycosyltransferases involved in the synthesis of non-cellulosic polymers with (14)-linked backbones, including chitin, heparan, and hyaluronan. These analyses re-open the question whether all the CesA genes encode cellulose synthases or whether some of the sub-class members may encode other non-cellulosic (14)-glycan synthases in plants. For example, the mixed-linkage (13)(14)-D-glucan synthase is found specifically in grasses and possesses many features more similar to those of cellulose synthase than to those of other -linked cross-linking glycans. In this respect, the enzymatic properties of the mixed-linkage -glucan synthases not only provide special insight into the mechanisms of (14)-glycan synthesis but may also uncover the genes that encode the synthases themselves.     

Chemotaxonomical investigations of the generaBlackstonia andCentaurium (Gentianaceae)

Methanolic extracts from aerial parts and capsules of plants of 5 populations ofBlackstonia perfoliata and 99 populations of nine European and two AmericanCentaurium species (Gentianaceae) have been screened by means of TLC for the secoiridoid glucosides: sweroside, swertiamarin, gentiopicroside and the m-hydroxybenzoyl esters of sweroside, namely centapicrin, desacetylcentapicrin, decentapicrin A and B as well as for the xanthones: 1,8-dihydroxy-3,5-dimethoxyxanthone, 1,8-dihydroxy-3,7-dimethoxyxanthone, 1,8-dihydroxy-3,5,6,7-tetramethoxyxanthone and xanthone--mono-glucosides. The taxonomical significance of the results is discussed. On the basis of chemotaxonomical evidence twoCentaurium species,C. pulchellum andC. tenuiflorum, are placed in sect.Parviflora instead of sect.Centaurium subsect.Parviflora. Part 8 in the series Secoiridoids and Xanthones in the genusCentaurium. For part 7 see:Van der Sluis & Labadie (1985). — Parts of this study were presented at the 10th annual congress of Farmacognosie en Natuurstofchemie in Utrecht, Nederland, Nov. 11, 1983. For summary see:van der Sluis, W. G., & Labadie, R. P. (1984), Pharm. Weekbl.119, 905–906.     

Soil-eating byAlouatta andAteles

Among 12 species of New World monkeys studied in La Macarena Region and the River Caquetá basin of Colombia, onlyAlouatta seniculus andAteles belzebuth were frequently observed to eat soil. They do this at particular sites on the ground called salados by local people. They also eat termite nests found on tree trunks. OnlyAteles drink the water of salado sites. The chemical properties of 17 soil samples and 5 water samples were analyzed. The results are discussed in relation to the question of whyAlouatta andAteles eat soil.     

Ultrastructure of the parasitic interface ofPucciniastrum,Thekopsora, Naohidemyces,andCalyptospora (Uredinales,Pucciniastraceae) in the dikaryotic stage

The ultrastructure of dikaryotic haustoria of sevenPucciniastrum species,Thekopsora galii, Naohidemyces vaccinii, andCalyptospora goeppertiana was investigated.Pucciniastrum actinidiae, P. agrimoniae, P. pyrolae, andCalyptospora goeppertiana revealed haustoria whose necks were wrapped by a fold of the extrahaustorial matrix. The matrix-fold ofCalyptospora goeppertiana was characteristically shaped.Pucciniastrum circaeae, P. epilobii, P. hikosanense, P. styracinum, Thekopsora galii, andNaohidemyces vaccinii showed typical haustorial necks which were not sheathed by a matrix-fold. Haustorial necks which were wrapped by a fold of the extrahaustorial matrix were designated velopedunculate, and those which were naked gymnopedunculate. The application of haustorial ultrastructure as a character for use in systematics is discussed.Part 112 of the series Studies in Heterobasidiomycetes.     

Meiotic studies ofElymus nutans andE. jacquemontii (Poaceae,Triticeae) and their hybrids withPseudoroegneria spicata and seventeenElymus species

Hybridizations ofElymus nutans andE. jacquemontii were carried out with one species ofPseudoroegneria (S genome), and 20Elymus species, each containing either of the SH, SY, SYH, or SYW genomes. Chromosome configurations were analysed at metaphase I of the two target taxa and their interspecific hybrids. It is concluded that (i)E. nutans is an allohexaploid containing the SYH genomes, andE. jacquemontii is an allotetraploid having the SY genomes; (ii) the genomic affinity is associated with the geographic distance between the species studied; (iii) minor genomic structural rearrangements have occurred within the hexaploid taxon ofE. nutans.     

A Cytogenetic Study of the Leaf Beetle Genus Cyrtonus (Coleoptera, Chrysomelidae)

The chromosomes of ten species of Cyrtonus and the genome sizes of six are surveyed. Among the total of 15 chromosomally studied species, 11 have 2n=28 chromosomes and a 13+Xy_p male meioformula, three have 2n=40 and 19+Xy_p and one 2n=46 and 22+Xy_p. All but one species with 28 chromosomes show only metacentric or submetacentric chromosomes, whereas the species with 40 and 46 chromosomes display some telocentrics or subtelocentrics, that are probably derived from the former by centric fissions. However, since the number of major chromosome arms is strikingly higher in these latter species (NF=70 and 78) than in the 28-chromosome species (mostly NF=56), other chromosomal rearrangements such as pericentric inversions or heterochromatin accretions could also be involved. The genome sizes display a narrow range, from 1C=0.6–1.22pg, and they are not significantly correlated with the chromosome numbers. Some possible factors implied in the rough chromosomal evolution of Cyrtonus are discussed in relation to a few other genera of the subfamily Chrysomelinae.     

Hair morphology and the relationships of species inSolanum sect.Basarthrum

The usefulness of features of leaf hairs in distinguishing subgenera and sections is well documented in bothRhododendron andSolanum. In this analysis of the taxa ofSolanum sect.Basarthrum (23 species), and of a sample of closely related taxa from sect.Petota (22 species), it is shown that such features serve to delineate subsectional groups such as series and some species as well. SectionBasarthrum has an unexpected diversity of hair types. Although this group has been characterized by 2-celled bayonet hairs, more than one half of the taxa in the section bear multicellular finger hairs, and 3 species also possess branched hairs. Thus, major rearrangements of the species previously assigned to sect.Basarthrum are indicated or supported by pubescence features. The taxa studied from seriesEtuberosa andJuglandifolia (both of sect.Petota) show hair types that a) are relatively primitive for the section, and b) show linkage between sects.Petota andBasarthrum.     

Genetics of the sex ratio anomaly in Drosophila hybrids of the Virilis group

Summary A study was made of the effect of genotype and temperature (25 and 17°C) on sex ratio in the hybrids D. virilis Sturt. X D. littoralis Sokolov. A genetic system has been found controlling sex-differential viability. In the F₁ of the reciprocal hybrids D. virilis X D. littoralis the sex ratio is normal, though at 17°C females are slightly excessive. The abnormal sex ratio is observed only in the progeny of test crosses.The major gene causing the death of female progeny of the cross [ (, D. virilis x , D. littoralis) x D. virilis] x D. littoralis is located on chromosome 2 of D. virilis. It is expressed as a lethal if chromosome 5 is heterogeneous virilis-littoralis. Chromosome 3 of D. virilis bears a modifier-enhancer and chromosome 5, a suppressor, of this lethal found in chromosome 2. This genetic system has a maternal effect and functions at 25°C, interacting with the X-chromosome of D. littoralis. If the maintainance temperature is lowered to 17°C, the progeny of the cross hybrid FB₁ x D. littoralis is predominantly female. Partial death of males is accounted for by a disturbance in the interaction between the genes of X-chromosome in certain combinations with the D. virilis autosomes and the Y-chromosome of the paternal species D. littoralis.Sex-differential mortality in the hybrids D. virilis x D. littoralis is one of the isolating factors between these species which does not appear to act until the second and subsequent F₁ generations due to the formation of the recombination load.     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

Intra- and interspecific genetic differentiation in closely related pines fromPinus subsectionSylvestres (Pinaceae) in the former Soviet Union

Thirty-seven natural populations of four closely related species ofPinus subsect.Sylvestres, P. mugo, P. funebris, P. pallasiana, andP. sylvestris, occurring in the former Soviet Union were investigated by starch-gel electrophoresis. In the populations assayed 127 allelic variants at 25 loci were revealed.Nei's distance coefficient (Dn) was used to estimate the level of genetic differentiation amongP. sylvestris races and among closely related species. A dendrogram constructed using Dn values shows that of the fiveP. sylvestris races analyzed only the geographically isolated var.hamata exhibited sufficient differences at theDia-2 locus (a mean Dn value relative to the other four races is 0.025) to recognize it as a distinct taxon. The remaining races, sylvestris, cretacea, lapponica, and sibirica, have a similar gene pool (Dn values are not greater than 0.010), and they should be regarded as a single taxon,P. sylvestris var.sylvestris. Interspecific comparisons revealed thatP. sylvestris andP. mugo have the closest genetic affinities to each other withNei's genetic distance of 0.108. The dendrogram demonstrates thatP. funebris is closer toP. sylvestris andP. mugo thanP. pallasiana. The available paleontologic data allowed us to conclude thatNei's (1975) time scale estimate for the time of divergence of the taxa was more accurate thanNei's (1971) time scale estimate.     

Epimerization of Hydroxyl Group in Lupan Series Triterpenoids

Two methods of obtaining 3-betulinic acid and related compounds from their 3-epimers were studied: the reaction of bimolecular substitution and the stereoselective reduction of 3-ketoderivatives. The substitution of acyloxy by formyloxy group in 3--tosyllupeol or of the belulin hydroxyl by benzoyloxy group resulted only in ^{2, 3}-elimination products, with none of the expected products of bimolecular substitution being found. The catalytic hydrogenation of betulonic acid over Raney nickel resulted only in reduction of the isopropenyl double bond, whereas the use of 5% Ru/C gave a 60 : 40 mixture of epimers of dihydrobetulinic acid. Practically the same mixture of betulinic acid epimers was obtained when reducing betulonic acid with L-Selectride. The cytotoxic activity of 3-betulinic acid increased toward the Bro melanoma cells and decreased toward the MS melanoma cells.     

Developmental aspects of long-term callus culture ofVicia faba L.

Summary Vicia faba callus line (VFS 1), isolated from expiants of immature embryo, grew satisfactorily onMurashige andSkoog complete medium with 1.38 M 2,4-D, or with 0.92 M 2,4-D to which 1.0 M kinetin was added. It also grew well on the B 5 modified medium containing 2.3 M 2,4-D and 25.0 M kinetin. On the last of these media the cultures grew more uniformly and without necrosis. They also showed diminishing variation in polyploidy in favour of diploids and corresponding aneuploids (hypodiploids).After being cultured for nearly three years on MS containing 1.38 M 2,4-D, 8–33% of cultures of VFS 1 were able to regenerate roots when transferred to either MS half strength with 5.37 M NAA, or to a medium without 2,4-D, or else to media with the addition of kinetin only (in various concentrations).     

Evidence of β-carotene 7,8(7′,8′) oxygenase (β-cyclocitral,crocetindial generating) in Microcystis

A -carotene oxygenase is described which occurs in the Cyanobacterium Microcystis. It cleaves -carotene and zeaxanthin specifically at the positions 7,8 and 7,8, while echinenone and myxoxanthophyll are not affected. The oxidative cleavage of -carotene leads to the formation of -cyclocitral and crocetindial and that of zeaxanthin to hydroxy--cyclocitral and crocetindial in nearly stoichiometric amounts. Oxidant is dioxygen as has been demonstrated by high incroporation (86%) of ¹⁸O₂ into -cyclocitral. -Carotene oxygenase is membrane bound, sensitive to sulfhydryl reagents, antioxidants and chelating agents. Iron seems to be an essential part of the enzyme activity. Cofactors necessary for the reaction could not be detected.Abbreviations TLC thin layer-chromatography - PIPES piperazine-N,N-bis-(2-ethanesulfonate) Na - TES 2{[tris-(hydroxymethyl)-methyl]-amino} ethanesulfonic acid Dedicated to Professor G. Drews on occasion of his 60th birthday     

Steroid Polyols from the Far Eastern Starfish Henricia sanguinolenta and H. leviuscula leviuscula

Two new asterosaponins, (20R)-3-O--D-(2-O-methylxylopyranosyl)-24-propylcholest-4-ene-3,6,8,15,16,29-hexaol (sanguinoside A) and (20R,24S)-3-O--D-(2,3,4-tri-O-methylxylopyranosyl)-5-cholestane-3,4,6,8,15,24-hexaol (sanguinoside B), were isolated from two species of Pacific Far Eastern Starfish Henricia sanguinolenta and H. leviuscula leviuscula, collected in the Sea of Okhotsk. Both glycosides contain aglycones with pentahydroxysteroid nuclei of similar structures, which are substituted at the 3-hydroxy group with differently methylated -D-xylosyl residues. Sanguinoside A has an unusual structure of its aglycone side chain, whereas sanguinoside B has a unique permethylated carbohydrate chain. In addition, laevisculoside G, a known glycoside, was identified in the H. leviuscula starfish. The structures of the isolated glycosides were established by interpreting their spectral data and by comparing their spectral characteristics with those of known compounds.     

Theβ-subunit of human chorionic gonadotropin containsN-glycosidic trisialo tri- and tri′-antennary carbohydrate chains

TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz¹H-NMR spectroscopy to beAbbreviations hCG human chorionic gonadotropin - hCG- -subunit - hCG- -subunit - PNGase-F peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52) - endo-F endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96) - SDS sodium dodecyl sulphate - PAGE polyacrylamide gel electrophoresis - CBB coomassie brilliant blue R 250 - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose     

Hexagonally ordered “rosettes” of particles in the plasma membrane ofMicrasterias denticulata Bréb. and their significance for microfibril formation and orientation

Summary Cells ofMicrasterias denticulata Bréb. at the stage of secondary wall formation have been studied by freeze-etching. It was found that the plasma membrane exhibits oval areas in which arrays of membrane particles occur. These particles form rosettes which are arranged in a hexagonally ordered lattice with a center to center spacing of 25 nm. Nearly the same periodicities can be found between microfibrils. It is concluded that the rosettes probably together with the thickened area of the plasma membrane below them represent the apparatus for the production and orientation of microfibrils. The hypothesis suggesting the incorporation of membrane templates functional in microfibril formation, originally advanced byKiermayer andDobberstein (1973) has received further support.     

DNA base composition within the genusScenedesmus (Chlorophyta)

The DNA base compositions of 60 strains ofScenedesmus were determined and found to be a valuable indicator for the differentiation within this genus (except for the very closely related species of the subsect.Desmodesmus). The range for allScenedesmus species was 49.9–69.3 mol% GC for nuclear DNA and 36.8–39.9 mol% GC for chloroplast DNA. The separation of the genusTetradesmus cannot be verified by GC values, becauseS. obliquus andS. (Tetradesmus)wisconsinensis have a similar GC content.S. (Chlorella)ultrasquamata, S. costato-granulatus (sect.Costato-granulati) andS. lunatus are separated clearly from all other species of the genus because of their high GC content.     

Meiotic analysis ofElymus caucasicus,E. longearistatus,and their interspecific hybrids with twenty-threeElymus species (Triticeae,Poaceae)

Interspecific hybridizations were carried out between the two tetraploidsElymus caucasicus andE. longearistatus, and 23 tetraploids and hexaploids ofElymus containing SH, SY, SYH, and SYW genomes and representing various geographical regions. Meiotic pairing was studied in the two target species and their hybrids. It is concluded from this study that (i) interspecific hybridization is fairly easy to perform although strong reproductive barriers exist between the species; (ii)Elymus caucasicus andE. longearistatus are allotetraploids, and share the diverged SY genomes; (iii) the divergence of SY genomes is correlated with the geographic distance between theElymus spp. studied.