Phylogenetic relationships of the chloroplast genomes in the genus Glycine inferred from four intergenic spacer sequences

The nucleotide sequences of four intergenic spacer regions of chloroplast DNA, atpB-rbcL, trnS-trnG, rps11-rpl36, and rps3-rpl16, were analyzed in the genus Glycine. Phylogenetic analysis based on the sequence data using Neonotonia wightii as the outgroup generated trees supporting the classification of two subgenera, Soja and Glycine, and three plastome groups in the subgenus Glycine. The results were consistent with the presence of diversified chloroplast genomes within tetraploid plants of G. tabacina and G. tomentella, as well as with a close relationship between G. tomentella and G. dolichocarpa that had been suggested based on morphological analyses. Little sequence variation was found in the subgenus Soja, suggesting that G. soja rapidly expanded its distribution in East Asia. The analysis also showed that the differentiation into three plastome groups in the subgenus Glycine occurred in the early stages of its evolution, after the two subgenera diverged.     

Genomic relationships among diploid wild perennial species of the genus Glycine Willd. subgenus Glycine revealed by crossability,meiotic chromosome pairing and seed protein electrophoresis

Summary The nomenclature of species beased on classical taxonomy can be verified from cytogenetic, biochemical and molecular studies. The objective of the study presented here was to provide further information on genomic affinities among species of the genus Glycine Willd. based on crossability, meiotic chromosome pairing of F₁ hybrids and seed-protein profiles. Meiotic chromosome pairing data revealed no genomic similarity between G. microphylla (BB) and G. falcata (FF), nor between G. tomentella (2n = 38; EE) and G. microphylla (BB). Despite morphological similarity between G. cyrtoloba (CC) and G. curvata no F₁ hybrid was obtained, although 748 flowers were pollinated. The seed-protein banding patterns showed G. latrobeana to be closer to the A-genome species than to others. Based on these results we assign genome symbol A₃A₃ to G. latrobeana. Likewise, G. curvata was allotted the designation C₁C₁ because the seed-protein banding patterns of G. curvata and G. cyrtoloba are similar. The genome designations of Glycine species based on cytogenetic investigations may be further extended by results obtained from biochemical and molecular approaches.Research supported in part by the Illinois Agricultural Experiment Station and US Department of Agriculture Competitive Research Grant 88-37231-4100     

Differentiation of nodules of Glycine max

Plants of Glycine max var. Caloria, infected as 14 d old seedlings with a defined titre of Rhizobium japonicum 3Il b85 in a 10 min inoculation test, develop a sharp maximum of nitrogenase activity between 17 and 25 d after infection. This maximum (14±3 nmol C₂H₄ h^-1 mg nodule fresh weight^-1), expressed as per mg nodule or per plant is followed by a 15 d period of reduced nitrogen fixation (20–30% of peak activity). 11 d after infection the first bacteroids develop as single cells inside infection vacuoles in the plant cells, close to the cell wall and infection threads. As a cytological marker for peak multiplication of bacteroids and for peak N₂-fixation a few days later the association of a special type of nodule mitochondria with amyloplasts is described. 20 d after inoculation, more than 80% of the volume of infected plant cells is occupied by infection vacuoles, mostly containing only one bacteroid. The storage of poly--hydroxybutyrate starts to accumulate at both ends of the bacteroids. Non infected plant cells are squeezed between infected cells (25d), with infection vacuoles containing now more than two (up to five) bacteroids per section. Bacteroid development including a membrane envelope is also observed in the intercellular space between plant cells. 35 d after infection, more than 50% of the bacteroid volume is occupied by poly--hydroxybutyrate. The ultrastructural differentiation is discussed in relation to some enzymatic data in bacteroids and plant cell cytoplasm during nodule development.     

Variability in rDNA loci in the genus Oryza detected through fluorescence in situ hybridization

The 17s-5.8s-25s ribosomal RNA gene (rDNA) loci in Oryza spp. were identified by the fluorescence in-situ hybridization (FISH) method. The rDNA loci were located on one-to-three chromosomes (two-to-six sites) within the eight diploid Oryza spp. One of the rDNA loci gave the weakest hybridization signal. This locus is reported for the first time in the genus Oryza. The chromosomes containing the rDNA loci were determined to be numbers 9, 10 and 11 in descending order of the copy number of rDNA. The application of image analysis methods, after slide preparation treatments (post-treatments), and the use of a thermal cycler, greatly improved the reproducibility of the results. The evolutionary significance of the variability of rDNA loci among the Oryza spp. is discussed.     

The genomic relationships among six wild perennial species of the genus Glycine subgenus Glycine Willd.

Summary Based on meiotic chromosome behavior in intra- and interspecific hybrids, genome symbols were assigned to the following diploid (2n=40) Glycine species: G. canescens = AA; G. clandestina- Intermediate pod (Ip)=A ₁ A ₁; G. clandestina-Short pod (Sp)=BB; G. latifolia = B ₁ B ₁; G. tabacina = B ₂ B ₂; G. cyrtoloba = CC; and G. tomentella = DD. Genome symbol GG was reserved for the soybean, G. max. At metaphaseI, loose chromosome associations were observed in completely sterile interspecific hybrids whose parents differed in their genomes, suggesting some chromosome homologies among species. Although G. clandestina-Sp, G. latifolia and G. tabacina are morphologically distinct species, they differ only by a paracentric inversion. Similar observations were recorded for G. canescens and G. clandestina-Ip. Evidence is presented that demonstrates that G. tabacina (2n=80) and G. tomentella (2n=78, 80) are allotetraploid species complexes. Hybrid weakness, sterility, seedling lethality and seed inviability were found in intra- and interspecific hybrids.This research was supported in part by the Illinois Agricultural Experiment Station and the U.S. Department of Agriculture (Special grant 82-CRSR-2-2007). Travel grants to collect Glycine germplasm were received from the Rockefeller Foundation, the Illinois Soybean Program Operating Board, the National Science Foundation (INT76-14753) and the International Board for Plant Genetic Resources     

Glycine max leaflets lack a base-tip gradient in growth rate

The paradigm of dicotyledonous leaf expansion describes high relative growth rates at the leaf base, dampening towards the leaf tip. Soybean leaflet expansion was measured under different growth conditions with both conventional and time-lapse video techniques. In contrast to findings from other species, maximum growth rates occurred at ∼2 a.m. and the basipetal growth pattern was absent, suggesting the factors controlling soybean expansion are distinct from other species.     

Ribosomal gene variation in soybean (Glycine) and its relatives

Summary The genes encoding the 18S25S ribosomal RNA gene repeat in soybean (Glycine max) and its relatives in the genus Glycine are surveyed for variation in repeat length and restriction enzyme site locations. Within the wild species of subgenus Glycine, considerable differences in repeat size occur, with a maximum observed in G. falcata. Repeat length and site polymorphisms occur in several species, but within individual plants only single repeat types are observed. The rDNA of the cultivated soybean and its wild progenitor, G. soja are identical at the level of this study, and no variation is found in over 40 accessions of the two species. Data from rDNA mapping studies are congruent with those of previous biosystematic studies, and in some instances give evidence of divergences not seen with other approaches.     

Plant regeneration from seedling explants of perennial Glycine species

More than 5000 cultures, from 30 accessions of six Glycine species, were established to assess the rôle of plant genotype in the response to an agar-solidified culture medium containing B5 salts and vitamins, 3% w/v sucrose, 1.1 mg 1^–1 BAP and 0.005 mg 1^–1 IBA, already known to induce shoot regeneration in callus of G. clandestina. Shoot initiation was obtained in a variety of explants from G. canescens, G. falcata, G. latrobeana and G. tomentella. With the exception of G. latrobeana, development of buds into shoots followed transfer to B5-based medium with 0.2 mg^–1 BAP and 0.005 mg 1^–1 IBA. Shoots readily produced roots in hormone-free half-strength B5 medium. In G. latrobeana, both extension and rooting occurred on this medium. Shoot regeneration was obtained in 12 of 30 accessions evaluated, but one accession of G. canescens, G1171, produced shoots and plantlets at a consistently higher frequency than other accessions, with plantlet recovery in more than 70% of the cultures. Bud formation in callus of G. canescens G1171 also occurred if BAP was replaced by 1.0 mg 1^–1 kinetin, 2i-p or zeatin, albeit at a lower frequency.     

Chromosome numbers in the genus Lippia (Verbenaceae)

The genus Lippia (Verbenaceae) comprises about 200 taxa mainly distributed in Brazil, Mexico, Central America, Africa, Argentina and Paraguay. Some problems involving the number and delimitation of species have been reported. In order to contribute to the solving of these problems, the chromosome numbers of 14 Lippia species are documented. The following species were collected at Espinhaço Range, Southeast Brazil: Section Zapania (L. corymbosa, L. diamantinensis, L. hermannioides, L. lacunosa, L. rotundifolia, L. rubella), section Rhodolippia (L. florida, L. lupulina, L. pseudothea, L. rosella), section Goniostlachyum (L. glandulosa, L. pohliana, L. sidoides) and section Dioicolippia (L.filifolia). Immature inflorescences were collected and the ideal size for chromosome observation was determined. The majority of species have a haploid chromosome number from 10 to 14. Few species have a higher chromosome number, which suggests the occurrence of polyploidy. The relationships between chromosome numbers and the taxonomic sections are also discussed.     

Genomic analysis in the genus Aegilops.

Summary Hybrids between different Aegilops species and Secale cereale were studied at metaphase I by means of a C-banding technique. On the basis of differences in the C-banding patterns of some of the chromosomes of these hybrids it was possible to carry out an accurate analysis of several types of Aegilops-Aegilops and Aegilops-Secale chromosome associations and, consequently, to establish intraspecific and intergeneric genome relationships. Genomes present in the majority of polyploid Aegilops species are shown to maintain similar patterns of evolutionary affinity to those reported for their proposed diploid parents although in some species there are differences indicating either that differentiations occurred during the evolution of the polyploid species or, on the contrary, that the diploid donors proposed are not the correct ones. On the other hand, differences in the relationships not only between the R genome and different Aegilops genomes but also among different homoeologous groups have been found.     

Subculture-induced protein synthesis in tissue cultures of Glycine max and Phaseolus vulgaris

The effects of subculture of tissue cultures on the levels of certain mRNAs have been investigated, and the action of cytokinins on the disposition of certain mRNAs between possible non-translating and translating pools has been determined. mRNA preparations were assayed by cell free translation with message-dependent reticulocyte lysate and the in vitro products resolved by polyacrylamide gel electrophoresis. Subculture of the cells caused a rapid stimulation of polysome formation. It also increased the translatable levels of a small group of mRNAs, one of which was present in both bean and soybean cultures. Cytokinins caused a slight increase in polysome levels after subculture, but had no effect on the levels of particular mRNAs, nor on the distribution of mRNAs between a non-translating and translating pool, nor on polysome levels in the absence of subculture.Abbreviations EDTA ethylenediaminetetra acetic acid - EGTA 1,2-di-(2-aminoethoxy)ethane-NNNN-tetra acetic acid - IEF isoelectric focusing - SDS sodium dodecyl sulphate     

A guide to the homoeology of chromosomes within the Triticeae

Summary A method of assessing chromosome homoeologies within the genomes of the Triticeae which does not require the ultimate test of substitution of the chromosomes into wheat is presented. This takes the form of a table listing key characters that are associated with specific homoeologous groups. These characters include marker genes, chromosome morphology, and plant morphologies of wheat-alien chromosome addition and wheat tetrasomic lines.     

Cytokinin inhibition of respiration by cells and mitochondria of soybean,Glycine max (L.) Merrill

Cells of a soybean tissue strain, suspended in an aerated liquid medium, caused disappearance of p-coumaric acid from the medium and oxidation of guaiacol, benzidine, pyrogallol, L-dihydroxyphenylalanine and L-epinephrine. Both the disappearance and the oxidations were inhibited by 6-benzylaminopurine (BAP) at a concentration of 0.5 mM. BAP at other concentrations either promoted or inhibited oxidation of epinephrine in precisely the pattern reported earlier for the disappearance of coumarate; therefore, the disappearance of coumarate probably involves its oxidation. The effectiveness of other cytokinins in inhibiting the oxidation was studied.At 0.5 mM, and perhaps even at 0.5 M, some of the several cytokinins tested inhibited oxygen consumption by the soybean cells. This inhibition, which did not require any of the above metabolizable compounds, was especially marked in the presence of cyanide, azide or Antimycin A, and was detectable in 10 min or less. Either Antimycin A or salicylhydroxamic acid alone promoted O₂ consumption but together they were quite inhibitory. The soybean cells apparently have an alternate respiratory pathway and cytokinins may influence its operation.Several cytokinins at 0.5 mM, and perhaps at 0.5 M, also inhibited oxygen consumption by mitochondrial preparations from the soybean cells, the inhibition being evident in about 20 s. The consumption required a substrate such as malate, succinate or NADH. Cytokinins and related compounds varied in effectiveness as follows: BAP and 6-isopentenyla-minopurine 9-tetrahydropyranyl-BAP > kinetin, ribosyl-isopentenylaminopurine, 9-methyl-BAP and 9-methoxymethyl-BAP > 6,6-dimethylaminopurine and zeatin (slight activity) > 6-methylaminopurine, nicotinamide and adenine (ineffective). To a great extent this order parallels the order of effectiveness of the compounds in causing cell division. Mitochondria, therefore, may contain a site for an important cytokinin action.Abbreviations BAP 6-benzylaminopurine - IPA 6-(²-isopentenyl)aminopurine     

Rate of synthesis of spermine and spermidine in germinating seeds of Glycine,Helianthus and Triticum

The spermine, spermidine, and the total protein content of embryos or embryonic axes from Triticum durum, Helianthus annuus, and Glycine max seeds at different times of early germination was evaluated. Mitotic activity of root-tip meristems from germinating seeds was also determined. The hypothesis is suggested that differences in polyamine and protein pattern during early germination could be correlated with the onset of mitotic activity and with the different characteristics of the seeds assayed.Abbreviations SPM spermine - SPD spermidine     

Cytogenetic relationship ofAegilops longissima chromosomes with common wheat chromosomes

The relationships of three wheat-Aegilops longissima chromosome addition lines A, C, and D with homoeologous wheat chromosomes were studied in PMC meiosis. Substitutions of alien chromosome A for wheat chromosome 6 B, chromosome C for 1 B and chromosome D for 4 B were obtained. The production of 4 BS/C and 7 BS/D chromosome translocations indicated cytogenetic relationships of C partially to homoeologous wheat chromosomes of group 1 and 4, and D partially to homoeologous wheat chromosomes of group 4 and 7.     

An electrophoretic karyotype for Candida albicans reveals large chromosomes in multiples

Summary Using field-inversion gel electrophoresis we defined an electrophoretic karyotype for the yeast, Candida albicans. The karyotype is distinct from other species of Candida and is species specific. A total of five distinct chromosomal mobility groups were observed, at least four of which are composed of a minimum of two fragments each. From the apparent sizes of these fragments relative to the large chromosomes of the morphologically related yeast Saccharomyces cerevisiae, together with the known genome size of this organism, we conclude that the karyotype is the result of the migration of intact chromosomes.     

Adventitious roots inGlycine subg.Glycine (Leguminosae): Morphological and taxonomic indicators of the B genome

Adventitious roots were observed on 3 wild perennialGlycine species, in short-poddedG. clandestina, G. latifolia and mostG. tabacina (2x, 4x, 6x), while other species lacked adventitious roots. This suggests that the adventitious roots trait is associated with B genome species. Intra- and interspecific F₁ hybrids reveal that adventitious roots apparently are inherited recessively. The presence of adventitious roots or short-poddedG. clandestina, coupled with infertility with longer-poddedG. clandestina, and enzymatic and leaflet morphology differences between the twoG. clandestina subgroups, supports segregation of the short-podded form as a separate taxon.     

Biosynthesis of pectic galactan by membrane-bound galactosyltransferase from soybean (<Emphasis Type="Italic">Glycine max</Emphasis> Merr.) seedlings

We investigated the properties of a galactosyltransferase (GalT) that is involved in the synthesis of -(14)-galactan side chains of pectins. A membrane preparation of etiolated 6-day-old soybean (Glycine max Merr.) hypocotyls transferred [¹⁴C]Gal from UDP-[¹⁴C]Gal into intact and partially hydrolyzed lupin -(14)-galactans of various chain lengths as exogenous acceptors, while activity to endogenous acceptors was negligible. Maximal activity occurred at pH 6.5 and 20–25°C in the presence of 25 mM Mn²⁺ and 0.75% Triton X-100. The transfer reaction onto the unmodified commercial pectic galactan (M _r>150,000) from lupin we used was very low but increased when the M _r of the galactan was reduced by partial acid hydrolysis. Among the partially hydrolyzed galactans, high-M _r (average M _r 60,000) -(14)-galactan was a more efficient acceptor [specific activity 2,000–3,000 pmol min^–1 (mg protein)^–1] than low-M _r (average M _r 10,000 and 5,000) polymers. Digestion of the radiolabeled product from high-M _r galactan with endo--(14)-galactanase released mainly radioactive -(14)-galactobiose and Gal, indicating that the transfer of [¹⁴C]Gal occurred through -(14)-linkages. HPLC analysis showed that the enzyme also catalyzes incorporation of Gal into pyridylaminated (PA) -(14)-galactooligomers with degree of polymerization at least 5. Gal₇-PA chains were elongated by attachment of one, two, or three Gal residues leading to the formation of Gal_8–10-PA.Abbreviations AGP Arabinogalactan-protein - Ara Arabinose - DP Degree of polymerization - GalA Galacturonic acid - Gal _n -PA Pyridylaminated -(14)-galactooligosaccharides - GalT Galactosyltransferase - MALDI–TOF–MS Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry - Rha Rhamnose Sugars described in this paper belong to the d-series unless otherwise noted     

Synchronization of protoplasts from Glycine max (L.) Merr. and Brassica napus (L.)

Cells of Glycine max originating in a suspension culture and cells of Brassica napus prepared from hypocotyls were synchronized. Synchronization was achieved by preparing protoplasts in the usual way and subsequently letting the protoplasts regenerate into cells by removing the cell-wall-digesting enzymes. More than 70% of the cells had divided synchronously at the end of the first cycle as determined by the mitotic index. The high frequency of mitosis critically depended on the osmolality of the medium. The duration of the S-phase was estimated by measuring the activity of thymidylate kinase as well as incorporation of [³H]deoxythymidine into acid-insoluble material. The data indicate that synchronization is induced by resetting the cell cycle.Abbreviations dTMP deoxythymidine 5-monophosphate - TCA trichloroacetic acid