Glycine Species Research Articles

Phylogenetic and genomic relationships in the genus Glycine Willd. based on sequences from the ITS region of nuclear rDNA.

Phylogenetic relationships among all 18 species of the genus Glycine were inferred from nucleotide sequence variation in the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. Pairwise sequence divergence values ranged from 0.2% (a single nucleotide) between Glycine max and Glycine soja to 8.6% between Glycine hirticaulis and Glycine falcata. The length of the ITS1 and ITS2 sequences ranged from 215 to 238 nucleotides and from 205 to 222 nucleotides, respectively, and that of 5.8S was 168 nucleotides across all the species. Phylogenetic analyses of the ITS region clearly resolved all the genomic groups that were established previously based on cytogenetic and biochemical studies. Based on this study, we assign new genome symbols: HH to Glycine arenaria, H1H1 to Glycine hirticaulis, H2H2 to Glycine pindanica, II to Glycine albicans, and I1I1 to Glycine lactovirens. Parsimony analysis of the entire ITS region, using subgenus Soja as outgroup, resulted in a trichotomy consisting of the clades: G. falcata (F genome), Glycine cyrtoloba and Glycine curvata (C genome), and all other species (A, B, D, E, H, and I genomes) of the subgenus Glycine.

The occurrence and gene expression of vegetative storage proteins and a Rubisco Complex Protein in several perennial soybean species

Vegetative storage proteins (VSPs) have been extensively studied in Glycine max, but not in perennial relatives of the cultivated soybean. The occurrence and gene expression of VSPs and a Rubisco Complex Protein (RCP) in several Glycine species was investigated by mRNA blot hybridization and protein immunoblotting. RCP had a developmental pattern of gene expression that closely paralleled that of VSP. The RCP gene was also induced by depodding, methyljasmonate treatment, wounding, and to a lesser extent by nitrogen fertilization, as was previously found for the VSPs. VSP in leaves of 13 perennial soybeans was heterogeneous in apparent size and number of bands detected by immunoblotting following SDS-PAGE. In contrast, RCP was detected as a single band of nearly identical mobility in all species. Both proteins were most abundant in young leaves of the perennials, and methyljasmonate and wounding induced both VSP and RCP gene expression in perennial soybeans. These results suggest that the VSPs in perennial soybeans function as storage reserves, as they do in G. max.

Genomic relationships in the genusGlycine (Fabaceae: Phaseoleae): use of a monoclonal antibody to the soybean Bowman‐Blrk inhibitor as a genome marker

We investigated the use of a monoclonal antibody (MAb 238) to the soybean Bowman‐Birk inhibitor (BBI) to verify and understand the intergenomic relationships among the wild perennial Glycine species. Competitive enzyme linked immunosorbent assay and western blot screening studies revealed that the accessions of B‐genome (G. latifolia, G. microphylla, and G. tabacina, 2n = 40) and C‐genome (G. curvata and G. cyrtoloba) species did not contain the MAb 238 crossreactive proteins (BBI‐nulls). By contrast, all the A‐genome (G. argyrea, G. canescens, G. clandestina, and G. latrobeana), E‐genome (G. tomentella, 2n = 38), and F‐genome (G. falcata) species, G. arenaria (genome unknown), and the polyploid (2n = 78,80) G. tomentella accessions were BBI‐positive. The D‐genome G. tomentella (2n = 40) and tetraploid G. tabacina (2n = 80) contained both BBI‐null and BBI‐positive type accessions. Among the recently described species, G. hirticaulis (2n = 40), G. lactovirens, and G. pindanica contained the MAb 238 crossreactive proteins while G. albicans did not. Glycine hirticaulis, G. pindanica, and G. tomentella (2n = 38) displayed highly similar MAb 238 crossreactive isoelectric focusing banding patterns, indicating that they are genomically close to each other. Glycine hirticaulis was found to have both diploid (2n = 40) and tetraploid (2n = 80) cytotypes. We demonstrated that the MAb 238 was specific to the trypsin inhibitor domain of the BBI. The MAb 238 clearly reflected all the previously established relationships in the genus Glycine, validating its use as a genome marker.

Genomic Relationships in the Genus Glycine (Fabaceae: Phaseoleae): Use of a Monoclonal Antibody to the Soybean Bowman-Birk Inhibitor as a Genome Marker

We investigated the use of a monoclonal antibody (MAb 238) to the soybean Bowman-Birk inhibitor (BBI) to verify and understand the intergenomic relationships among the wild perennial Glycine species. Competitive enzyme linked immunosorbent assay and western blot screening studies revealed that the accessions of B-genome (G. latifolia, G. microphylla, and G. tabacina, 2n = 40) and C-genome (G. curvata and G. cyrtoloba) species did not contain the MAb 238 crossreactive proteins (BBI-nulls). By contrast, all the A-genome (G. argyrea, G. canescens, G. clandestina, and G. latrobeana), E-genome (G. tomentella, 2n = 38), and F-genome (G. falcata) species, G. arenaria (genome unknown), and the polyploid (2n = 78,80) G. tomentella accessions were BBI-positive. The D-genome G. tomentella (2n = 40) and tetraploid G. tabacina (2n = 80) contained both BBI-null and BBI-positive type accessions. Among the recently described species, G. hirticaulis (2n = 40), G. lactovirens, and G. pindanica contained the MAb 238 crossreactive proteins while G. albicans did not. Glycine hirticaulis, G. pindanica, and G. tomentella (2n = 38) displayed highly similar MAb 238 crossreactive isoelectric focusing banding patterns, indicating that they are genomically close to each other. Glycine hirticaulis was found to have both diploid (2n = 40) and tetraploid (2n = 80) cytotypes. We demonstrated that the MAb 238 was specific to the trypsin inhibitor domain of the BBI. The MAb 238 clearly reflected all the previously established relationships in the genus Glycine, validating its use as a genome marker.

Natural glycine unidentate on copper(II): a novel mode of bonding

The compound of composition copper(II)–glycinate–picric acid–water (1/2/2/6) has been prepared and its crystal structure determined. It contains a centrosymmetric copper(II) ion, co-ordinated in a distorted octahedron by two water molecules in trans positions, two unidentate glycine moieties, linked to the metal in a trans arrangement via single carboxylate oxygens, and two more distant water molecules. The unidentate glycine species, probably zwitterions, represent an unexpected non-chelated form of the ligand. New similar compounds of (S)-alanine and (S)-lysine have also been prepared. The nature of the complexed ions in solution (‘speciation’) is considered.

The identification and distribution of Glycine latrobeana (Meissn.) Benth. in Tasmania

Glycine latrobeana has commonly been confused in Tasmania with other Glycine species and also with Desmodium gunnii.This paper details the known distribution and habitat characteristics of G. latrobeana and presents a key to the species of Glycine in Tasmania.

Backcross (BC2‐BC4)‐Derived Fertile Plants from Glycine max and G. tomentella Intersubgeneric Hybrids

Wild perennial relatives of the soybean [Glycine max (L.) Merr.] have not been utilized to broaden the genetic base of the crop. This study provides information on production, morphology, cytology, and breeding behavior of backcross (BC2 to BC4)‐derived fertile plants from soybean (2n = 40, genome GG) and G. tomentella Hayata (2n = 78, genome DDEE). The main hurdle was to obtain BC~ plants (G. max cv. Altona) × (G. tomentella, Pl 483218)] → F1, 2n = 59, genome GDE → colchicine treatment (CT) → 2n = 118, GGDDEE × soybean cv. Clark 63 → BC1, 2,n = 76 (expected 2n = 79). Three sterile BC2 plants (2n = 58, 2n = 56, 2n = 55) were produced. Procedures for immature seed‐rescue were utilized to obtain F, amphiploid, BC1 and BC2 plants. Soybean cv. Clark 63 was used as the recurrent parent in the backcrossing programs. The range of chromosome number among BC3 plants was 2n = 41 to 2n = 52, and among BC4 plants was 2n = 40 to 64. A hypertriploid (2n = 64) in BC4 plant originated after fertilization of an unreduced egg (n = 44) by a normal haploid (n = 20) male gamete. Phenotypes of BC3 and BC4 plants were dissimilar but generally resembled closely to the soybean cv. Clark 63. We expect to isolate a series of aneuploid lines from BC3 to BC6 generations that may open up the path to introgress desired traits from wild perennial Glycine species to the cultivated soybean.

Genomic diversity in aneudiploid (2n = 38) and diploid (2n = 40) Glycine tomentella revealed by cytogenetic and biochemical methods

Of the 15 perennial species of the subgenus Glycine Willd., G. tomentella Hayata is unique in that it has four cytotypes (2n = 38, 40, 78, and 80) and a wide range of geographical distributions. The objective of this study was to uncover the genomic diversity among accessions of aneudiploid (2n = 38) and diploid (2n = 40) G. tomentella based on crossability rate, hybrid seed and seedling viability, meiotic chromosome pairing of F1 hybrids, and seed protein and protease inhibitor profiles. Aneudiploid and diploid G. tomentella accessions were divided into two (D1 and D2) and three [D3(A,B,C), D4, and D5] groups, respectively, based on previous isozyme studies. Crossability rate, intergenomic hybrid viability, degree of chromosome pairing, total seed protein profiles, and trypsin and chymotrypsin inhibitor banding patterns confirmed the isozyme grouping with minor disagreements. A consistent variation was not observed among the aneudiploid accessions in any method of analysis used in this study. Similarly, cytogenetic analysis and the total seed protein profiles did not show dissimilarity among the accessions from Papua New Guinea (PNG; the D3 group) and north of Mitchell River in Northern Queensland [N.Qld(n)]. However, trypsin and chymotrypsin inhibitor analysis revealed that the PNG accessions were distinctly different from N.Qld(n) accessions. The D4 and D5 group accessions were clearly distinguishable by both cytogenetic and biochemical methods. Thus, this study indicates the presence of four genomic groups among G. tomentella (2n = 38, 40) accessions, including the aneudiploids D1 and D2 in one group and diploids in three groups (D3, D4, and D5). These findings will be useful in further genome analysis and add to our present understanding of the biosystematics of the genus Glycine.

Glycine pindanica (Fabaceae, Phaseolae), a New Species from West Kimberley, Western Australia

A new species, Glycine pindanica, is described from the western side of Dampier Peninsula, Western Australia. Its distribution is mapped. A key is provided to the species of Glycine in north-western Australia. G. pindanica and the closely allied G. hirticaulis Tindale & Craven form a small group of non-rhizomatous, non-stoloniferous species with digitately trifoliate leaves, linear leaflets and hairs having persistent, slightly enlarged bases. Opportunist amphicarpy occurs in both species. An illustration of G. pindanica is provided.

日本におけるダイズアブラムシの生活環

1) 日本でのダイズアブラムシの生活環は野外観察と飼育実験から,クロウメモドキ属のクロウメモドキ,クロツバラを一次寄主とすることを明らかにした。2) ダイズアブラムシの生活環と一次寄主の落葉期,萌芽期との同調性について考察した。3) これまで記載のなかった,ダイズアブラムシの幹母,幹雌,産卵雌虫と雄ならびに簡単な記載だけだった産雌虫を図示し,ワタアブラムシと比較して記載した。

Characterization of trypsin and chymotrypsin inhibitors in the wild perennial Glycine species

Seven wild perennial species were used for characterizing the seed protease inhibitors in the genus Glycine Willd. subgenus Glycine to determine the presence and examine the variability and expression patterns. Seeds of all the species contained trypsin and chymotrypsin inhibitors. There were highly significant variations among the wild perennial species in the electrophoretic profiles of trypsin and chymotrypsin inhibitors, migration patterns of anti-KTI and anti-BBI (mAB 238)immunocrossreactive proteins, and trypsin and chymotrypsin inhibitor activities of seeds

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

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.

Shoot formation from somatic hybrid callus between soybean and a perennial wild relative

Protoplasts isolated from hypocotyl tissue of dark-grown soybean ( Glycine max L.) seedlings, were labelled with fluorescein, using fluorescein diacetate, and fused electrically with green mesophyll protoplasts from seedling cotyledons of a wild perennial relative. The resulting heterokaryons were bifluorescent owing to the green fluorescent label of the soybean parent and the red chlorophyll autofluorescence from the cotyledon protoplasts of the wild Glycine sp. G1171. Post-fusion protoplast populations comprised between 1.61% and 7.40% heterokaryons with a mean of 3.77 ± 2.13%. The heterokaryons were isolated from unfused protoplasts and homokaryons by flow cytometry. They were cultured in agarose-solidified medium, where they developed into cell colonies. The latter produced friable callus if transferred to a rich culture medium, or green, nodular tissue and occasionally abnormal shoots, if grown on regeneration media previously used for Glycine species. Phenotypically normal shoots were obtained when callus on SC2 medium was transferred to SC2 supplemented with kinetin, zeatin and gibberellic acid (GA 3). However, these shoots did not develop further. Polyacrylamide gel electrophoresis of total protein revealed that heterokaryon-derived tissues grown on both K8 and SC2 media produced more isoenzymes of aspartate aminotransferase than an exact summation of the parental zymograms.

Nodulation and Nodulin Gene Expression in an Interspecific Hybrid Between Glycine max and Glycine tomentella

When the genome of a wild species, Glycine tomentella, is combined with that of the cultivated soybean, Glycine max in a hybrid, nodulation specificity is altered. Two strains of Rhizobium fredii that nodulate the G. rnax parent fail to nodulate the hybrid, indicating the presence of a restriction gene, while three Bradyrhizobium japonicum strains that nodulate G. max are highly effective on the hybrid. Some strains of (Brady)rhizobium, originally isolated from wild Australian Glycine species, nodulate only one of the two parental species. One strain ineffective on G. max is effective on the hybrid. Two other strains, while effective on one parent, appear more effective on the hybrid. In three fully examined cases in which effective nodules are produced on the hybrid, the late nodulins leg-hemoglobin, glutamine synthetase, and xanthine dehydrogenase are expressed from both genomes. Thus, although a rhizobial strain may nodulate only one or the other of the parental types, if it successfully nodulates the hybrid, the nodulation process provides signals or internal conditions that lead to expression of late nodulins from both genomes.

Sources of Resistance to Soybean Rust in PerennialGlycineSpecies

Accessions of 12 perennial Glycine species were evaluated for resistance to Phakopsora pachyrhizi, the causal agent of soybean rust. A total of 23% of the accessions were resistant, 18% were moderately resistant, and 58% were susceptible. In two experiments, 59 and 40% of the accessions of G. tabacina (2n=80) were resistant. Resistance to P. pachyrhizi was identified in accessions of G. argyrea, G. canescens, G. clandestina, G. latifolia, G. microphylla, and G. tomentella, but not in accessions of G. arenaria, G. cyrtoloba, G. curvata, and G. falcata

WHOLE- AND PART-FLOWER SELF-POLLINATION IN GLYCINE CLANDESTINA AND G. ARGYREA AND THE EVOLUTION OF AUTOGAMY.

The overall rate of self-fertilization can be viewed as the sum of two distinct processes: 1) self-pollination of all ovules in a flower (whole-flower self-pollination); and 2) self-pollination of some of the ovules in a flower, occurring together with outcrossing of the remaining ovules (part-flower self-pollination). In some situations these processes may be equated with different modes of self-pollination. A model of the mating system in which the progeny of separate fruits serve as the unit of observation is presented. The model partitions the overall rate of self-pollination into components attributable to whole- and part-flower selfing. When the mating system is estimated using information on marker genotypes from chasmogamous fruits in two species of Glycine together with the whole- and part-flower selfing model, the results indicate that the chasmogamous flowers in a subalpine population of G. clandestina underwent a significant level of whole-flower selfing, whereas in another, lower elevation population of G. clandestina and in a subtropical population of G. argyrea, they did not. This difference is thought to be related to the contrast in the variability of environmental conditions for insect-mediated pollination between the habitats sampled. In particular, the large component of whole-flower selfing observed in the subalpine population of G. clandestina may be due to self-pollination that is induced during periods unfavorable to insect-mediated pollination. It can be demonstrated that such induced selfing will be selected whenever environmental conditions are such that pollinator activity limits seed set, and moreover that induced selfing can result in the selection of overall levels of self-pollination that are intermediate between 0 and 1. Monte Carlo simulation is employed to show that ignoring the correlation of self-fertilization events that result from whole- and part-flower selfing may lead to biased estimates of mating system parameters.

Variation in the DNA Content of Glycine Species

Nuclei were isolated from cotyledons of a range of accessions from 14 species of Glycine. These were stained with ethidium bromide and the relative fluorescence for each genotype was measured by flow cytometry. The DNA content was estimated by comparison of relative fluorescence with that from nuclei from seedling leaves of Allium cepa, whose DNA content has been calculated previously by chemical assay. The 4C amounts for diploid Glycine ranged from 3-80 to 6-59 pg. Two groups of diploid species appeared from the analysis. The first consisted of species with amounts ranging from 3-80 to 516 pg and included G. canescens (AA), G. argyrea (AlA1), G. clandestina (A2A2), G. microphylla (BB), G. latifolia (B, B, ), G. tabacina 2n = 40 (B2B2), G. tomentella In = 38 (EE) and In = 40 (DD), G. max and G. soja (GG), G. arenaria and G. latrobeana. A second group had higher DNA contents ranging from 5-27 to 6-59 pg, and consisted of G. curvata, G. cyrtoloba (CC), and G.falcata (FF). The polyploid species, G. tabacina 2n = 80 (AABB, BBB^J, G. tomentella In = 78 and 2n = 80 (AAEE and DDEE, respectively) contained amounts approximating to the sums of the respective parental diploid species thought to have given rise to these allotetraploids. Intraspecific variation was detected in the DNA content of G. canescens. Within the overall distribution of DNA amounts found in A genome species, each genome contained a range of DNA contents specific to that species. This phenomenon was also detected amongst B genome species.

5024944 Transformation, somatic embryogenesis and whole plant regeneration method for glycine species: Glenn B Collins, David F Hildebrand, Paul Lazzeri, Thomas Adams, Wayne A Parrott, Lynn M Hartweck assigned to Lubrizol Genetics Inc; The University of Kentucky Research Foundation Inc

5024944 Transformation, somatic embryogenesis and whole plant regeneration method for glycine species: Glenn B Collins, David F Hildebrand, Paul Lazzeri, Thomas Adams, Wayne A Parrott, Lynn M Hartweck assigned to Lubrizol Genetics Inc; The University of Kentucky Research Foundation Inc

Nutrient culture medium components affecting plant recovery from immature embryos of three Glycine genotypes and an interspecific hybrid grown in vitro

Immature embryos of Glycine max (L.) Merr. cv. ‘Bay’, G. tabacina (Labill.) Benth. and G. tomentella Hayata were cultured on 72 media combinations to identify a nutrient medium which would allow a greater percentage of interspecific plants to be recovered from cultured embryos of G. max × various perennial Glycine species. The highest mean plant recovery rate of 79% was from a medium containing ‘B5’ nutrient salts as reported by Gamborg et al. (1968), vitamin components according to Williams (1978) and 30 g/l sucrose. This is as compared with 25% from a medium used previously. In an additional test, 67% of hybrid embryos of G. max × G. tomentella were recovered from the same medium, from which G. tomentella was most effectively recovered in all testing.

Examination of 2,4-D tolerance in perennial Glycine species

High levels of tolerance to 2,4-D [(2,4-dichlorophenoxy)acetic acid] were expressed at the cell and whole plant level by several accessions of wild perennial Glycine species. To determine the mechanism(s) responsible for tolerance, heterotrophic cell suspension cultures of soybean [ Glycine max (L.) Merr.] and four perennial Glycine accessions were used to investigate[ 14C]2,4-D uptake and metabolism. Uptake of 2,4-D was similar between soybean and the perennials. However, soybean metabolized only 7% of the absorbed 2,4-D at 2 and 10 μ M concentrations. The four perennial accessions metabolized 2,4-D at least 79% at 10 μ M and 64% at 50 μ M 2,4-D. The primary metabolite produced by the tolerant perennial lines was the glycoside conjugate of 4-OH-2,5-D, as determined by high-performance liquid chromatography cochromatography and a gas chromatography-mass spectrometer. 2,4-D-sensitive soybean and the least tolerant perennial accession produced high quantities of amino acid conjugates of 2,4-D. Varying levels of 2,4-D tolerance among the perennial Glycine accessions may also be influenced by differential sensitivity of each accession to 2,4-D and/or metabolites.