DNA fingerprinting in soybean (Glycine max (L.) Merrill) with oligonucleotide probes for simple repetitive sequences

Summary Soybean DNA fingerprints were analyzed by digoxigenin-labeled oligonucleotide probes complementary to simple repetitive sequences. The clearest and most polymorphic patterns were obtained with (AAT)₆ as a probe, with which all 47 soybean cultivars tested could be distinguished. However, DNA fingerprints of individuals within cultivars showed the same pattern. Using (CT)₈, (GAA)₅ or (AAGG)₄ as probes, clear polymorphic patterns among cultivars and accessions in the subgenus Soja (Glycine max and Glycine soja) were not observed, while quite different patterns were found in accessions in the subgenus Glycine. The results suggest that G. max and G. soja are closer in their genome structure. DNA fingerprints of reciprocal crosses between cultivars and accessions in the subgenus Soja were similar, and contained bands of both parents. In an F₂ population from these crosses, such bands segregated in a Mendelian fashion.     

A cluster analysis of wild and domesticated soybean phenotypes

Summary Cluster analysis techniques were used to examine phenotypic variation within the USDA Soybean Germplasm Collection. This study included ten Plant Introductions each of the domesticated soybean, Glycine max (L). Merr., the wild soybean, G. soja Sieb. & Zucc., and a disputed species, G. gracilis Skvortz. G. max and G. soja were found to be morphologically distinct entities and G. gracilis was found to be conspecific with G. max. Because G. gracilis probably represents weedy races associated with the cultivated soybean and because gracilis phenotypes can be distinguished from G. max, the designation, G. max forma gracilis, is recommended.Joint contribution: North Central Region, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, and Journal Paper No. J-9389 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011; Project 2107.     

Characterizing morphological traits and estimating genetic relationship for intermediate soybean collected from South Korea

Twenty‐two wild, 16 intermediate and 20 cultivated soybean varieties were used to analyse relationships among the species of subgenus Soja, genus Glycine using 11 morphological traits and simple sequence repeat (SSR) markers. These genotypes using eleven agronomic characteristics were divided into three clusters: cluster I included 20 G. max, cluster II included 12 intermediate, and cluster III included 22 G. soja and four intermediate lines. Genetic relationships among the species of subgenus Soja showed three distinct clusters. Cluster I consisted of 20 G. max cultivars and two intermediate type lines, and cluster II consisted of 13 intermediate type and two G. soja lines; however, cluster III consisted of 20 G. soja and one intermediate type lines. These phenotypic and genetic data suggest that the intermediate type lines could be distinguished between G. max and G. soja lines. However, the intermediate type could not be classified as a new species.     

Differences in Ion Accumulation and Salt Tolerance among Glycine Accessions

Salinity reduces crop yield by limiting water uptake and causing ion‐specific stress. Soybean [Glycine max (L.) Merr.] is sensitive to soil salinity. However, there is variability among soybean genotypes and wild relatives for salt tolerance, suggesting that genetic improvement may be possible. The objective of this study was to identify differences in salt tolerance based on ion accumulation in leaves, stems and roots among accessions of four Glycine species. Four NaCl treatments, 0, 50, 75 and 100 mm , were imposed on G. max, G. soja, G. tomentella and G. argyrea accessions with different levels of salinity tolerance. Tolerant genotypes had less leaf scorch and a greater capacity to prevent Na⁺ and Cl^? transport from soil solution to stems and leaves than sensitive genotypes. Magnitude of leaf injury per unit increase in leaf Na⁺ or Cl^? concentrations was lower in tolerant than in susceptible accessions. Also, plant injury was associated more with Na⁺ rather than with Cl^? concentration in leaves. Salt‐tolerant accessions had greater leaf chlorophyll‐meter readings than sensitive genotypes at all NaCl concentrations. Glycine argyrea and G. tomentella accessions possessed higher salt tolerance than G. soja and G. max genotypes.     

Glycine max and Glycine soja are capable of cold acclimation

Soybean has been considered a cold intolerant species; based largely upon seed germination and soil emergent evaluations. This study reports a distinct acquisition of cold tolerance, in seedlings, following short acclimation periods. Diversity in cold responses was assessed in eight cultivars of Glycine max and six accessions of G. soja. All varieties of soybean significantly increased in freezing tolerance following acclimation. This study indicates soybean seedlings are indeed capable of sensing cold and acquiring cold tolerance. Germination rates after cold imbibition were negatively correlated with maturity group, but positively correlated with cold acclimation potential in G. soja. Seed fatty acid composition was varied between the species, with Glycine soja accessions containing about 2‐times more linolenic acid (18:3) than G. max. Furthermore, high levels of linoleic acid (18:2) in seeds were positively correlated with germination rates following cold imbibition in G. soja only. We suggest that domestication has not impacted the overall ability of soybean to cold acclimate at the seedling stage and that there is little variation within the domesticated species for ability to cold acclimate. Thus, this brief comparative study reduces the enthusiasm for the “wild” species as an additional source of genetic diversity for cold tolerance.     

Intersubgeneric Crossability in the Genus Glycine Willd.

In intersubgeneric crosses between wild perennial Glycine species of submenus Glycine and soybeans, Glycine max (L.) Merr., of subgenus Soja, all the putative hybrid pods aborted from 9 to 30 days after pollination (DAP). Spraying of 70 ppm gibberellic acid on hybridized gynoecia facilitated pod retention. Fluorescence microscopy revealed normal pollen germination and pollen tube growth in all the intersubgeneric crosses except in the G. cyrtoloba × G. max cross.     

Heterosis of interspecific soybean hybrids for traits related to N2 fixation

Summary Three greenhouse experiments were conducted to compare the performance of soybean (Glycine max (L.) Merr.), wild soybean (G. soja Sieb. et Zucc.), and soybean x wild soybean hybrids for traits relating to N₂ fixation including nodulation, acetylene reduction, nodule leghemoglobin concentration, and nitrogen (N) accumulation and dry matter (DM) accumulation. In all three experiments G. max generally exceeded G. soja in nodulation, acetylene reduction, and N and DM accumulation while the soybean possessed higher nodule leghemoglobin concentration. In Experiment I, the mean of the hybrids did not differ significantly from the G. max parent in nodule mass, acetylene reduction activity, nodule leghemoglobin concentration, or DM accumulation. The hybrids did exceed the soybean parent in N accumulation, thus demonstrating high parent heterosis. In Experiments IIA and IIB with a more carefully chosen set of G. soja parents, high parent heterosis of individual crosses was common. Across the three experiments average high parent heterosis was 34, 28, and 28%, respectively, for nodule mass, N accumulation, and DM accumulation. If one accepts the assumption that hybrid vigor results from the accumulation of dominant alleles, then these alleles could theoretically be accumulated via selection in a homozygous genotype. If this is true than the results of the experiments reported here suggest that interspecific soybean x wild soybean crosses could serve as sources of homozygous lines which would exceed currently available soybean cultivars in nodule mass, and N and DM accumulation.This work was supported in part by the USDA Competitive Grants Program, Grant 82-CRCR-1-1039 and Cooperative Agreement 58-32U4-2-370 between the U.S. Department of Agriculture and the Agronomy Department, University of Maryland, Scientific Article No. A-4648, Contribution No. 7644 of the Maryland Agricultural Experiment Station.     

Association of isozyme genotypes with agronomic and seed composition traits in soybean

Summary Two crosses between Glycine max (L.) Merr. and G. soja Sieb. & Zucc. parents were used to study the association between isozyme marker loci and agronomic and seed composition traits in soybean. The parents possessed different alleles at six isozyme loci for Cross 1 (A80-244036 × PI 326581) and at eight isozyme loci for Cross 2 (A81-157007 × PI 342618A). A total of 480 BC₂F_4:6 lines from the two crosses was evaluated for 13 traits in two environments. Lines were grouped in locus classes from 0 to 5 according to the number of loci homozygous for the G. soja alleles that they possessed. Within each locus class, each isozyme genotype was represented by five random lines.Selection for G. max alleles at the isozyme loci was not effective in recovering the recurrent parent phenotype in either cross. In cross 1, however, BC₂F₄-derived lines in the 0- or 1- locus class more closely resembled the G. max parent than lines in the 4- or 5- locus classes for most of the agronomic and seed composition traits evaluated. Significant associations were found between particular isozyme genotypes and every trait analyzed. The estimated effect of genes linked to the Pgm1 locus was a delay in maturity of 6.0±3.4 days. In cross 1, the Idh2 locus was associated with a significant effect on linolenic acid content. The percentage of variation accounted for by the models of estimation varied according to the heritability of the trait. The R² was high (up to 78%) for maturity, lodging, and vining, and low (up to 21%) for seed yield. Most of the variation was associated with the BC₂F₁ family from which the lines were derived. There was little evidence that digenic epistasis was an important source of variation.Journal Paper No. J-13505 of the Iowa Agric. Home Econ. Exp. Stn., Ames, IA, Project 2475.     

The Importance of the Bolivian Wild Potato Species in Breeding for Globodera pallida Resistance

Screening for resistance to the potato cyst nematode, Globodera pallida, in potatoes from. Bolivia, was carried out in 1983 and 1984, using a mixture of four nematode populations representing pathotypes Pa₁, Pa₂ and Pa₃ From the 66 accessions of 17 species and subspecies evaluated, highly resistant genotypes were identified in 21 accessions from seven species. All had Pf/Pi values of 2 or less, whereas the susceptible control, Solanum tuberosum cv. ‘Disiree’ had Pf/Pi values of more than 2G in both tests. Two diploid wild species, S. brevicaule and S. leptophyes, showed the best resistant. The geographical distributional of resistant populations and the evolution of resistance in wild potato populations are discussed.     

The effectiveness of evaluating wild species: searching for sources of resistance to bruchid beetles in the genus Vigna subgenus Ceratotropis

A species level germplasm collection representing 76% of known taxa in the genus Vigna subgenus Ceratotropis was evaluated for resistance to two species of bruchid beetles, Callosobruchus chinensis and C. maculatus. Seven taxa consisting of 29 accessions were found to be resistant to C. chinensis and 4 taxa consisting of 24 accessions were found to be resistant to C. maculatus. This compared with no resistant accessions being found in several hundred landrace accessions of mungbean, V. radiata var. radiata, in the same subgenus. Sometimes resistance was found in all accessions of a particular taxon, such as complete resistance to both C. chinensis and C. macualtus in V. umbellata. Other taxa showed intra taxon variation for resistance such as V. reflexo-pilosa andV. minima. The levels and patterns of resistance among taxa were diverse. The results suggest that various factors cause resistance to bruchid in the subgenus Ceratotropis. While the number of eggs laid on seeds generally reflected seed size, one small seeded cultivar of V. mungo var. mungo, black gram, had an unusually high number of eggs laid per seed. No correlation was found between seed size and levels of resistance. The species level germplasm collection, which reflects the core collection concept in trying to maximize genetic diversity in a limited number of accessions, has enabled a large number of potentially useful sources of resistance to bruchid beetles to be found efficiently. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sources of resistance to Mayetiola Destructor in breadwheat and durum wheat in Morocco

Summary Grain yield reductions of both breadwheat (Triticum aestivum L.) and durum wheat (Triticum turgidum L. var. durum) caused by attacks of Hessian fly (Mayetiola destructor Say) are second perhaps only to those caused by inadequate soil moisture in Morocco. To identify effective sources of resistance, 817 entries of common wheat and durum wheat reported to be resistant to Hessian fly were evaluated under natural infestations in Morocco. A large number of genes conferring virulence are present in populations of Moroccan Mayetiola. The genes H₁, H₂, H₃, h₄, H₆, H₇, H₈, H₉, H₁₀, H₁₁, H₁₄, H₁₅, and H₁₆ as well as the Marquillo, Kawvale and PI 94587 resistance sources are not useful for cereal improvement in North Africa. Luso, which has the gene H₁₂, also appeared susceptible in limited testing. Genotypes having the genes H₅ and H₁₃ were identified as significantly reducing larval survial in natural populations of Mayetiola. Of 11 resistant breadwheats identified with unknown genes, seven were from Portugal and three were from the Soviet Union. Although none of the durums tested had high levels of reistance, the two most promising durums were from Portugal. It is proposed that initially H₅ be deployed in durum wheats and H₁₃ be used in common wheat improvement. Leaf pubescence appears of little use in reducing the larval survival of Mayetiola.     

Mapping and use of QTLs controlling pod dehiscence in soybean

While the cultivated soybean, Glycine max (L.) Merr., is more recalcitrant to pod dehiscence (shattering-resistant) than wild soybean, Glycine soja Sieb. & Zucc., there is also significant genetic variation in shattering resistance among cultivated soybean cultivars. To reveal the genetic basis and develop DNA markers for pod dehiscence, several research groups have conducted quantitative trait locus (QTL) analysis using segregated populations derived from crosses between G. max accessions or between a G. max and G. soja accession. In the populations of G. max, a major QTL was repeatedly identified near SSR marker Sat_366 on linkage group J (chromosome 16). Minor QTLs were also detected in several studies, although less commonality was found for the magnitudes of effect and location. In G. max × G. soja populations, only QTLs with a relatively small effect were detected. The major QTL found in G. max was further fine-mapped, leading to the development of specific markers for the shattering resistance allele at this locus. The markers were used in a breeding program, resulting in the production of near-isogenic lines with shattering resistance and genetic backgrounds of Japanese elite cultivars. The markers and lines developed will hopefully contribute to the rapid production of a variety of shattering-resistant soybean cultivars.     

A partially male-sterile mutant line of soybeans,Glycine max (L.) Merr.: inheritance

Summary A partial male sterility system in the soybean (Glycine max (L.) Merr.) germplasm population AP6(SI)CI was found to be controlled monogenically by a recessive allele, msp. Observations of msp msp plants in different environments suggested that environmental conditions significantly affect expressivity of the msp allele with respect to male sterility. We obtained no experimental evidence of cytoplasmic effects on msp expression. Background genotypes, however, seem to affect msp expressivity through their determination of flowering dates and resultant interactions with varying environmental conditions.Homogeneous populations of partially male-sterile plants can be generated by increasing families of msp msp plants in fertility-inducing environments, if measures are implemented to prevent the introduction and/or build-up of fertile genotypes that arise from natural cross-pollinations.Joint contribution: North Central Region, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, and Journal Paper No. J-9596 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011; Project 2107.     

Characterization of flowering time and SSR marker analysis of spring and winter type Brassica napus L. germplasm

Flowering dates and life forms of all available Brassica napus accessions conserved at the North Central Regional Plant Introduction Station (NCRPIS) were characterized, and a survey of molecular variation was conducted by using simple sequence repeats (SSR) in order to support better management of accessions with diverse life forms. To characterize flowering phenology, 598 B. napus accessions from the NCRPIS collection were planted in Iowa and Kansas field sites together with a current commercial cultivar and observed for days to flowering (first, 50% and 100% flowering) in 2003. Days from planting to 50% flowering ranged from 34 to 83 in Iowa and from 53 to 89 in Kansas. The mean accumulated growing degree days (GDD) to 50% flowering were 1,997 in Iowa, and 2,106 in Kansas. Between locations, the correlation in flowering time (r = 0.42) and the correlation in computed GDD (r = 0.40) were both significant. Differences in flowering-time rank were observed for several accessions. Accessions that failed to flower in Iowa in a single growing season comprised 28.5% of the accessions; of the flowering accessions, 100% plant flowering was not always achieved. Accessions were grouped according to flowering time. A stratified sample of 50 accessions was selected from these groups, including 10 non-flowering and 40 flowering accessions of diverse geographic origins and phenological variation. The flowering time observed in the sampled accessions when grown in the greenhouse were found to be significantly correlated to the flowering time observed in the field locations in Iowa (r = 0.79) and Kansas (r = 0.49). Thirty SSR markers, selected across 18 Brassica linkage groups from BrassicaDB, and 3 derived from Brassica expressed sequence tags (ESTs) were scored in the stratified sample. An average of three bands per SSR primer pair was observed. Associations of SSR marker fragments with the life forms were determined. Analysis of molecular variation by using cluster analysis and ordination resulted in recognizable, distinct groups of annual and biennial life-form types, which may have direct applications for planning and management of future seed regenerations. Mention of commercial brand names in this paper does not constitute an endorsement of any product by the U.S. Department of Agriculture or cooperating agencies.     

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

Summary 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.Contribution No. 89-276-J of the Kansas Agric. Exp. Stn., Manhattan.     

Heritability and gene effects for incorporating pubescence into cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L) Walp.) from <Emphasis Type="Italic">V. rhomboidea</Emphasis> Burtt. Davy

Cowpea, an important subsistence pulse crop in the tropics and subtropics is susceptible to several insect pests that seriously limit its yield potential. Pubescence (hairiness) on the parts of some of its wild relatives has been reported to reduce the damage caused by some of the insect pests. A generation mean analysis was conducted between a cowpea variety IT82D-716 and two accessions of Vigna rhomboidea to investigate the gene effects and heritability for incorporating pubescence into cultivated cowpea from V. rhomboidea. The additive-dominance model that was adopted in the analysis was observed to sufficiently explain the mode of inheritance of leaf and stem pubescence with the additive effect being more important than the dominance effect. A six-parameter model with epistatic gene interactions was adequate for explaining the inheritance of pod pubescence. Heritability estimates, in the narrow sense were high for pubescence density and pubescence length. Inheritance of pubescence in crosses between cowpea and V. rhomboidea was governed by one and two genes. Significant and higher additive gene effects and high-heritability suggest that backcross selection schemes should be responsive in the development of pubescent cowpea lines.     

A linkage group in rice (Oryza sativa L.) involving anti-inhibitory genes

Summary Inheritance studies of purple pigmentation in T-160 x AC-177 revealed that purple colour in the coleoptile is based on four genes (Pc ₁ , Pc₂, I-Pc, Ai-Pc), in the internode on two genes (Pin _a1 , Pin _a2 ), in the junctura on three genes (Pja, Pjb ₁ , Pjb ₂ ) and in the apiculus on three genes (Pa, I-P, Ai-P). For panicle density monogenic segregation (Lx) was obtained. Linkage was established between the genes for the apiculus (Pa and Ai-P), panicle density (Lx), the internode (Pin _a ), the coleoptile (Ai-Pc) and the junctura (Pja), in that sequence. This linkage group forms part of the IV mp group of Misro et al. (1966) which corresponds to the g group of japonica.The two anti-inhibitory genes Ai-Pg and Ai-Pau of the linkage group Ai-Pg=Ai-Pc=Ai-Pau reported earlier (Dhullappanavar et al., 1975) are also allotted to the IV linkage group of Misro et al. (1966) since Ai-Pc is placed in that group in the present studies.     

Morphological and biochemical variation in Sardinian populations ofMedicago polymorpha L. suitable for rainfed Mediterranean conditions

Summary Forty five accessions of Sardinian germplasm ofMedicago polymorpha L., derived from a wide range of climatic and edaphic conditions, were evaluated to identify useful genetic resources for the development of cultivars for rainfed Mediterranean conditions. The accessions were assessed for variation in morphological, agronomic and biochemical traits. Considerable variation was found between populations indicating the potential for selection.Work supported by the Italian Ministry of Agriculture and Forestry (MAF), Special Project Foraggicoltura Prativa, subproject Miglioramento Genetico.     

Pollen production in soybeans with respect to genotype,environment, and stamen position

Summary Our objective was to determine the average numbers of pollen grains from fertile plants (Ms ₁) and the average numbers of coenocytic microspores from genetic male sterile plants (ms ₁ ms ₁) in soybeans, Glycine max L. Merr. Comparisons were made between the average numbers of pollen grains and the average numbers of coenocytic microspores with respect to environment where plants were grown and to stamen position in the flower. Five male sterile lines were used. They included the North Carolina ms ₁ mutant, the cultivar Hark with the ms ₁ gene, and mutants identified as the Urbana, Tonica, and Ames male steriles. Three environments used were the Agronomy Farm, University of Georgia, Athens, Georgia; the Agronomy and Agricultural Engineering Research Center, Iowa State University, Ames, Iowa; and the Agronomy Greenhouse, Iowa State University, Ames, Iowa.Pollen production from fertile plants varied from 374 to 760 pollen grains per anther among genetic lines and environments. This variation may be an important consideration in selecting a male parent to use as a pollinator for hybrid seed production.Among fertile plants, the average numbers of pollen grains per anther of the separate stamen and and of the lower whorl of stamens were significantly different only in greenhouse-grown plants. Among male sterile plants, the average numbers of coenocytic microspores per anther of the separate stamen and of the lower whorl of stamens were significantly different in three genotype x environment combinations. These three exceptions did not conform to any genetic or environmental pattern. Deviations from the expected ratio of 4 pollen grains from fertile plants: 1 coenocytic microspore from sterile plants were attributed to initial differences in the average number of microspore mother cells between the two genotypes.Joint contribution: Agricultural Research Service, USDA, North Central Region, and Journal Paper No. J-8910 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011; Project 2107.     

Screening wild Lycopersicon species for resistance to powdery mildew (Oidium lycoperiscum)

Since the late 1980s powdery mildew, designated Oidium lycopersicum, frequently invaded the tomato crop in Western Europe. All commercial cultivars are susceptible. To screen for resistance in wild species a reliable and efficient disease test was developed. Young plants with two to three true leaves are inoculated at high relative humidity by spraying with a freshly prepared suspension of 2×10⁴ conidia, ml^–1. Symptoms are periodically evaluated according to a scale based on the percentage of leaf area with mycelium.One hundred and twenty seven accessions, representing eight wild Lycopersicon species, were screened for resistance to O. lycopersicum. A large variation in resistance was found between species. L. hirsutum was the most resistant species; L. pennellii was moderately resistant; species of the subgeneric group of L. esculentum and of the peruvianum-complex were all susceptible. L. parviflorum was classified separately due to a large variation between accessions. Except for this species, a low variation was found between accessions within species. High levels of resistance were observed in four accessions of L. hirsutum, in one of L. parviflorum and in one of L. peruvianum. This resistance is characterized by a very low disease incidence and a strongly restricted mycelium growth and lack of sporulation.