Inheritance and linkage analysis of ten enzyme and blood protein loci in the Japanese brown frogRana japonica

In order to clarify the genetic linkage groups of the Japanese brown frogRana japonica, and compare them with those of other vertebrates, the inheritance of 10 enzyme and blood protein loci was examined in 267 offspring derived from 18 crosses using 10 males heterozygous at these loci. Most of the segregation tests exhibited no significant deviations from the expected normal Mendelian ratios. Of 32 pairs of loci tested for linkage, 29 pairs showed independent assortment in all crosses examined. In the three other locus pairs, betweenADH-1 andAlb, IDH-1 andHb-1, andLDH-B andMPI, all offspring analyzed were parental, and there were no recombinants. These three linkage groups comprising six loci were thus established inR. japonica, whereas no linkage between the other four loci,AAT-1, ADA, GPI, andPEP-A, was observed.     

Joint Segregation of Biochemical Loci in Salmonidae. II. Linkage Associations from a Hybridized Salvelinus Genome (S. NAMAYCUSHxS. FONTINALIS)

The results of more than 300 pairwise examinations of biochemical loci for joint segregation in brook trout (Salvelinus fontinalis) and in the hybridized genome of lake trout (S. namaycush) x brook trout are summarized. Nineteen loci have been assigned to the following eight linkage groupings on the basis of nonrandom assortment, including cases of both classical linkage and pseudolinkage: ODH with PMI with PGI-3, PGI-2 with SDH, ADA-1 with AGP-2, AAT-(1,2) with AGP-1 with MDH-1, MDH-3 with MDH-4, LDH-3 with LDH-4, IDH-3 with ME-2 and GUS with CPK-1. Pseudolinkage (an excess of nonparental progeny types) was observed only for male testcross parents. The results suggest that this phenomenon involves homeologous chromosome arms as evidenced by the de novo association of presumed duplicate loci in each case. Classical linkage has not been found for the five pairs of duplicate loci examined in Salvelinus, suggesting that not all of the eight metacentrics in the haploid complement involve fusions of homeologous chromosomes. Females consistently showed a greater degree of recombination.     

Linkage relationships among 12 enzyme loci in cattle fever ticks of the genus Boophilus

Electrophoretic variation at 12 enzyme loci in cattle fever ticks were examined for mode of inheritance and linkage relationships. All variants exhibited codominant Mendelian inheritance, and PGI and GOT were sex-linked. Four pairs of enzyme loci were linked (PGI and GOT, ACON-A and alpha GPD, MDH-1 and LDH, and EST-3 and EST-4). Four additional enzyme loci (ACON-C, MDH-2, PEP, and PP) did not show linkage to any other locus tested, giving eight presumptive linkage groups that have enzyme markers in the genus Boophilus.     

Biochemical Polymorphism in Yellow Catfish, Mystus nemurus (C&V), from Thailand

Yellow catfish, Mystus nemurus (Cuv. & Val.), is becoming one of the major freshwater species farmed by aquaculturists in Southeast Asia. It was of interest to examine levels of genetic subpopulation differentiation among samples of this species obtained from parts of its range, as well as to compare the genetics of wild and hatchery-bred fish. Horizontal starch gel electrophoresis and histochemical staining techniques were used to examine genetic variation within and among eight wild and one hatchery populations of M. nemurus from northern, northeastern, central and southern Thailand. Four tissues (heart, liver, kidney, and muscle) from individual specimens were used to analyze variations at 23 protein-coding loci. Fifteen of the 23 loci examined (65.22%), namely, ACP*, AAT-1*, EST-1*, EST-2*, GPI*, IDH-1*, IDH-2*, MDH-1*, MDH-2*, MDH-3*, ME*, PGM*, 6PGD*, SOD*, and HB*,were polymorphic at the 0.95 level. Observed heterozygosities ranged from 0.041 to 0.111, with an average of 0.068 ± 0.028. Genetic distances ranged from 0.005 to 0.164. The greatest genetic distance was found between the Chainat and the Suratthani populations (0.164), a level indicative of subspecific differentiation in M. nemurus from within Thailand.     

Allozyme Polymorphisms and Biochemical-Genetic Taxon Markers in Lampreys (Lampetra, Eudontomyzon, Petromyzon)

Variation at 23 putative enzyme-coding loci was scored in 424 lampreys, including 321 European brook lampreys (Lampetra planeri), 83 European river lampreys (L. fluviatilis), 11 Ukrainian brook lampreys (Eudontomyzon mariae), and nine sea lampreys (Petromyzon marinus). Twelve polymorphisms are described for Lampetra species (LDH*, SOD-2*, PNP*, AAT-1*, AK-1*, ES-2*, LGL*, MPI*, GPI-1*, GPI-2*, PGM*, IDHP-2*), and two each for E. mariae (GPI-1*, ME-2*) and P. marinus (MDH-1*, ME-2*). Diagnostic allozymes are presented for the discrimination of lamprey taxa, some of which are difficult to recognize from the morphology of ammocoetes larvae, the life stage usually encountered when collecting cyclostomes. The allelic markers described permit the clear allocation to a genus, except for the species L. fluviatilis and L. planeri, which are not differentiated by qualitative allozyme analysis.     

马尾松GOT、LDH和MDH同工酶的遗传方式和连锁关系

本文采用聚丙烯酰胺凝胶电泳研究了马尾松三种同工酶的遗传方式和连锁关系。结果表明,GOT、LDH和MDH三种同工酶的遗传表达稳定,分别受4个、2个和4个位点控制。只有一对位点(MDH-3和MDH-4)间存在着紧密的连锁关系。     

Variations de la malate déshydrogénase dans quatre espèces de Jaera albifrons: Etude génétique et variation géographique

Malate dehydrogenase has been detected in four species of the super-species Jaera albifrons. Two polymorphic enzymatic systems occur showing a one banded pattern for homozygotes and a three banded pattern for heterozygotes. Two independant autosomal loci control the variation as is shown by intra- and interspecific crosses. In European populations of the three species studied, MDH-1 is polymorphic and MDH-2 is monomorphic. In the American species, J. posthirsuta, the locus MDH-2 is polymorphic and MDH-1 is monomorphic.     

Temporal genetic variation of brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis> L. from Vorobiev creek (White sea) based on allozyme analysis

We performed an analysis of allozyme variation in brown trout from Vorobiev creek. Seventeen allozyme loci encoding glycerol-3-phosphate dehydrogenase (G3PDH), aspartate aminotransferase (AAT), malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase D (EST-D) were studied. We found statistically significant differences in allele frequencies for the AAT-1,2*, G3PDH-2,3*, LDH-5*, and MDH-2* loci between brown trout samples collected in 1981–1982 and/or 1992–1995. We suggest that temporal changes of allele frequencies in brown trout from Vorobiev Creek are associated with gene drift.     

Four independent electrophoretic markers in spadefoot toads

Four enzyme-encoding genes (Idh-1, Idh-2, Ldh-1, and Mdh-1), with alleles diagnostic for the spadefoot toads Scaphiopus multiplicatus and Scaphiopus bombifrons, were investigated in order to characterize their inheritance and linkage relationships. Electrophoretic phenotypes in the offspring of natural crosses exhibit Mendelian segregation, behaving as genotypes produced by alternative alleles at four independently assorting loci. These phenotypes are useful markers of genetic identity and the degree of genetic admixture in the analyses of hybrid zone dynamics for these two species. Concurrent use of a morphological index verified the diagnostic value of the markers. The morphological index is useful for the identification of parental species and many F1 hybrids, however offspring of backcrosses usually express parental species characteristics. Electrophoretic typing allows the identification of all hybrids and 87 percent of the offspring from backcrosses. It also facilitates the identification of tadpoles, which are extremely difficult to distinguish morphologically. The technique is useful for the assessment of introgression and the evaluation of reproductive interaction in these species, even though it slightly underestimates backcross offspring.     

EVIDENCE FOR A MOSAIC HYBRID ZONE IN THE GRASS SHRIMP PALAEMONETES KADIAKENSIS (PALAEMONIDAE) AS REVEALED BY MULTIPLE GENETIC MARKERS

Molecular techniques provide powerful tools for studying the geographic structure of hybrid zones and the dynamics of gene exchange between incipient species. We examined allozyme variation at five loci (PGM, GPI, MDH-1, MDH-2, and LDH) for 27 populations of Palaemonetes kadiakensis from the central, coastal, and eastern regions of Texas. Central Texas populations of P. kadiakensis exhibited highly significant linkage disequilibrium and departures from Hardy-Weinberg genotype proportions. In populations with linkage disequilibrium, allelic differences at GPI defined two types of P. kadiakensis, designated A and B. Both types existed in central Texas with little or no evidence of interbreeding, whereas the populations from all other localities showed complete introgression of type B alleles into the type A gene pool. We also examined ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) variation in a subset of populations, chosen to cover a range of geographic locations and levels of linkage disequilibrium. Two groups of mtDNA haplotypes and two restriction fragment patterns for the rDNA corresponded to allozyme type A and B individuals in populations exhibiting linkage disequilibrium. In populations with ongoing hybridization, all hybrid animals (N= 15) exhibited type A mtDNA. Exhibition of type A mtDNA indicated that type A females had mated successfully with type B males, but type B females had not mated successfully with type A males. Genotype distributions suggest reduced reproduction by hybrid offspring in central Texas populations. These patterns are consistent with a mosaic model of hybrid zone dynamics.     

Mapping genetic loci that regulate lipid levels in a NZB/B1NJxRF/J intercross and a combined intercross involving NZB/B1NJ, RF/J, MRL/MpJ, and SJL/J mouse strains

The NZB/B1NJ (NZB) mouse strain exhibits high cholesterol and HDL levels in blood compared with several other strains of mice. To study the genetic regulation of blood lipid levels, we performed a genome-wide linkage analysis in 542 chow-fed F2 female mice from an NZBxRF/J (RF) intercross and in a combined data set that included NZBxRF and MRL/MpJxSJL/J intercrosses. In the NZBxRF F2 mice, the cholesterol and HDL concentrations were influenced by quantitative trait loci (QTL) on chromosome (Chr) 5 [logarithm of odds (LOD) 17-19; D5Mit10] that was in the region identified earlier in crosses involving NZB mice, but two QTLs on Chr 12 (LOD 4.7; D12Mit182) and Chr 19 (LOD 5.7; D19Mit1) were specific to the NZBxRF intercross. Triglyceride levels were affected by two novel QTLs at D12Mit182 (LOD 8.7) and D15Mit13 (LOD 3.5). The combined-cross linkage analysis (1,054 mice, 231 markers) 1) identified four shared QTLs (Chrs 5, 7, 14, and 17) that were not detected in one of the parental crosses and 2) improved the resolution of two shared QTLs. In summary, we report additional loci regulating lipid levels in NZB mice that had not been identified earlier in crosses involving the NZB strain of mice. The identification of shared loci from multiple crosses increases confidence toward finding the QTL gene.     

Genetic Mapping in Xenopus Laevis: Eight Linkage Groups Established

Inheritance of alleles at 29 electrophoretically detected protein loci and one pigment locus (albinism) was analyzed in Xenopus laevis by backcrossing multiply heterozygous individuals generated by intersubspecies hybridization. Pairwise linkage tests revealed eight classical linkage groups. These groups have been provisionally numbered from 1 to 8 in an arbitrarily chosen order. Linkage group 1 includes ALB-2 (albumin), ADH-1 (alcohol dehydrogenase), NP (nucleoside phosphorylase), and ap (periodic albinism). Linkage group 2 contains ALB-1 and ADH-2, and probably is homeologous to group 1. Linkage group 3 comprises PEP-B (peptidase B), MPI-1 (mannosephosphate isomerase), SORD (sorbitol dehydrogenase), and mIDH-2 (mitochondrial isocitrate dehydrogenase). Linkage group 4 contains GPI-1 (glucosephosphate isomerase) and EST-4 (esterase 4). Linkage group 5 contains GPI-2 and PEP-D (peptidase D). Linkage group 6 comprises ACP-3 (acid phosphatase), sME (cytosolic malic enzyme), and GLO-2 (glyoxalase). Linkage group 7 consists of sSOD-1 (cytosolic superoxide dismutase), GPD-2 (glycerol-3-phosphate dehydrogenase), mME (mitochondrial malic enzyme), and the sex determining locus. Linkage group 8 includes FH (fumarate hydratase) and TRF (transferrin). Recombination frequencies between linked loci showed differences related to the genomic constitution (parental subspecies) and to the sex of the heterozygous parent. Independent assortment was observed between the duplicate ALB loci. This is true for the duplicate ADH, GLO, and MPI loci as well, supporting the view that these genes have been duplicated as part of a genome duplication that occurred in the evolutionary history of X. laevis. Comparative analysis of genetic maps reveals a possible conservation of several linkages from the Xenopus genome to the human genome.     

GENETIC EVIDENCE FOR DIPLOIDY IN EQUISETUM

Sporophytes and gametophytes of Equisetum arvense, E. laevigatum, and E. telmateia were analyzed using enzyme electrophoresis to estimate isozyme number. Despite their uniformly high chromosome numbers (2n = 216), these three species exhibited isozyme numbers typical of diploid seed plants for the enzymes AAT, ADH, ALD, GDH, [NADP]IDH, LAP, MDH, [NADP]ME, PGI, PGM, SkDH, and 6PGDH. All three species exhibited an additional isozyme for TPI. There is, therefore, no genetic evidence for low base numbers such as x = 9 and x = 12 suggested for Equisetum. Intact chloroplasts were isolated from E. arvense and the chloroplast extract compared electrophoretically to whole plant extracts. The single enzymes observed for LAP, GDH, [NADP]IDH, and [NADP]ME were absent from the chloroplast extract. Isozymes AAT-1, ALD-2, MDH-3, PGI-1, PGM-2, SkDH-2, 6PGDH-2, TPI-2, and TPI-3 were active in the chloroplast fraction; 6PGDH-1, PGI-2, PGM-1, and TPI-1 were lacking from the chloroplast fraction and were considered cytosolic. Isozymes AAT-2, MDH-1, MDH-2, MDH-4, and SkDH-1 were also lacking from the chloroplast fraction but because AAT, MDH, and SkDH have been reported from several subcellular compartments, their localization is unknown. These findings indicate that isozymes in Equisetum species are subcellularly compartmentalized as has also been demonstrated for homosporous ferns, gymnosperms, and angiosperms.     

Inheritance of malate dehydrogenase nulls in soybean

Three chlorophyll-deficient mutants (CD-1, CD-2, and CD-3), derived from the progeny of independent germinal revertants from the w4-mutable soybean line [Glycine max (L.) Merrill], were characterized genetically. Electrophoretic analyses indicated that these lines lacked two of three mitochondrial malate dehydrogenase isozymes (MDH-). The absence of two MDH bands was conditioned by a recessive allele at a locus designated Mdh1. All three CDs were allelic to each other and to T253, a Harosoy isoline y20-k2 MDH- from the Genetic Type Collection. The MDH- phenotype and the yellow-green plant phenotype were each inherited as single recessive alleles. No recombination between the two traits was found in nine F2 populations from crosses of the CDs by wild-type soybean lines. Complete linkage of the Mdh1 and y20 loci suggested that the mutations in the chlorophyll-deficient lines were deletions. Phenotypic differences among the CDs suggested that the deletions may have different endpoints. The chromosomal aberrations were not large enough to affect transmission of y20 and Mdh1 mutant alleles through the pollen or ovule. CD-1, CD-2, and CD-3 were added to the Soybean Genetic Type Collection as T323, T324, and T325, respectively.     

Genetics of isozymes in grasspea

We studied the inheritance and linkage of ACO-1, ACO-2, AAT-1, AAT-2, EST-3, EST-6, FDH, LAP-1, PGD-2, SKDH, and TPI-1 in four F2 populations of grasspea (Lathyrus sativus L.) using horizontal starch-gel electrophoresis. Mendelian inheritance was observed for all of the isozymes studied. All isozymes showed codominant gene expression except for EST-3, which was expressed in a dominant fashion due to the presence of a null allele. Monomeric quaternary structure was observed for ACO-1, ACO-2, EST-6, LAP-1, and SKDH. Dimeric quaternary structure was observed for AAT-1, AAT-2, FDH, PGD-2, and TPI-1. The isozyme loci Aat-2 and Skdh were linked with a map distance of 28 cM.     

Genic heterozygosity, chromosomal interchanges and fitness in rye: any relationship?

Relationship between heterozygosity at allozyme loci, chromosomal interchanges and fitness was analyzed in a rye cultivar showing a polymorphism for such rearrangements. Nine allozyme systems (ACO, ACPH, GOT, GPI, LAP, MDH, PER, PGD and PGM) and five components of fitness (number of fertile tillers, total offspring, egg cell fertility, flowers/ear and seeds/ear) were studied. The estimated selection coefficients against interchange heterozygotes ranged from s = 0.12 to s = 0.34. A significant effect of the genic heterozygosity on some fitness components was observed in interchange heterozygotes (tillering and total offspring), in their standard homozygous sibs (flowers/ear and seeds/ear) and in the descendants of the crosses between standard karyotypes (flowers/ear, seeds/ear and egg cell fertility). However, the main effect was linked to genetic background associated to different crosses. Significant differences for Acph-1, Gpi-1, Lap-1, Mdh-1, Mdh-4, Pgd-2 and Pgm-1 loci were also found in some of these crosses although these differences were inconsistent. This suggests that probably the allozyme loci analyzed were not directly contributing to the fitness and that they are linked, in some cases, to different deleterious alleles depending on both cross and locus. This fact could support the local effect hypothesis as explanation although we do not discard the existence of some inbreeding level (general effect hypothesis) since all crosses and loci studied show a overall consistent trend of increased fitness with increased heterozygosity.     

Inheritance of isozyme variation and heterozygosity in Pinus ponderosa.

Techniques are presented to detect 23 isozyme loci in the long-lived perennial plant, ponderosa pine. Meiotically derived megagametophyte from seeds is used to examine directly the segregation of allelic variants. Approximately seven seeds were initially examined for 12 enzymes from each of 47 trees from ten stands throughout the northern Rocky Mountain region. Additional seeds were also examined from selected families to confirm the inheritance of observed electrophoretic variants at 13 polymorphic loci and to estimate linkage relationship. Significant norandom segregation was consistently detected for three pairs of loci: ADH-1:AAT-2, ADH-1:PGI-1, and LAP-2:6PG-1. Preliminary estimates of population parameters reveal a relatively high average heterozygosity (H = 0.123). This is partitioned into a high amont of genetic variation within local stands, with only approximately 12% of the total heterozygosity resulting from genic difference between stands.     

Molecular and whole-plant responses to selection for enzyme activity in alfalfa root nodules: evidence for molecular compensation of aspartate aminotransferase expression

Summary The enzyme aspartate aminotransferase (AAT) plays a key role in the assimilation of fixed-N in alfalfa (Medicago sativa L.) root nodules. AAT activity in alfalfa nodules is due to the activity of two dimeric isozymes, AAT-1 and AAT-2, that are products of two distinct genes. Three forms of AAT-2 (AAT-2a, -2b, and-2c) have been identified. It was hypothesized that two alleles occur at the AAT-2 locus, giving rise to the three AAT-2 enzymes. In a prior study bidirectional selection for root nodule AAT and asparagine synthetase (AS) activities on a nodule fresh weight basis in two diverse alfalfa germ plasms resulted in high nodule enzyme activity subpopulations with about 20% more nodule AAT activity than low enzyme activity subpopulations. The objectives of the study presented here were to determine the inheritance of nodule AAT-2 production and to evaluate the effect of bidirectional selection for AAT and AS on AAT-2 allelic frequencies, the relative contributions of AAT-1 and AAT-2 to total nodule activity, nodule enzyme concentration, and correlated traits. Two alleles at the AAT-2 locus were verified by evaluating segregation of isozyme phenotypes among F₁ and S₁ progeny of crosses or selfs. Characterization of subpopulations for responses associated with selection was conducted using immunoprecipitation of in vitro nodule AAT activity, quantification of AAT enzyme protein by ELISA, and AAT activity staining of native isozymes on PAGE. Results indicate that selection for total AAT activity specifically altered the expression of the nodule AAT-2 isozyme. AAT-2 activity was significantly greater in high compared to low activity subpopulations, and high AAT subpopulations from both germ plasms had about 18% more AAT-2 enzyme (on a nodule fresh weight basis). No significant or consistent changes in AAT-2 genotypic frequencies in subpopulations were caused by selection for AAT activity. Since changes in AAT activity were not associated with changes in AAT-2 genotype, selection must have affected a change(s) at another locus (or loci), which indirectly effects the expression of nodule AAT.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture, and does not imply its approval to the exclusion of other products or vendors that might also be suitable     

Inheritance of isozyme variation and heterozygosity in Pinus ponderosa

Techniques are presented to detect 23 isozyme loci in the long-lived perennial plant, ponderosa pine. Meiotically derived megagametophyte from seeds is used to examine directly the segregation of allelic variants. Approximately seven seeds were initially examined for 12 enzymes from each of 47 trees from ten stands throughout the northern Rocky Mountain region. Additional seeds were also examined from selected families to confirm the inheritance of observed electrophoretic variants at 13 polymorphic loci and to estimate linkage relationships. Significant nonrandom segregation was consistently detected for three pairs of loci: ADH-1: AAT-2, ADH-1: PGI-1, and LAP-2: 6PG-1. Preliminary estimates of population parameters reveal a relatively high average heterozygosity (H=0.123). This is partitioned into a high amount of genetic variation within local stands, with only approximately 12% of the total heterozygosity resulting from genic differences between stands.Funds for this study were made available under the McIntire-Stennis program of the Forest, Range, and Conservation Experiment Station of the School of Forestry, University of Montana.     

Genetic divergence between two forms of a tongue sole <Emphasis Type="Italic">Cynoglossus interruptus</Emphasis>

Genetic differences between trilinear and dilinear forms of a tongue sole, Cynoglossus interruptus, were examined by use of isozymes. Unequivocal differences were detected between the two forms, including complete replacement of alleles at the FBALD-2*, MDH-3*, PROT-1*, and SOD-1* loci, almost complete replacement at the AAT-3*, AH-1*, AH-2*, EST-3*, G3PDH-3*, GPI-2*, IDHP-1*, and MDH-1* loci, and extreme differences in allelic frequencies at the GPI-1* and PGDH* loci. The genetic distances (D values) between the two forms were 0.4207–0.4353, figures predicatively significant at the specific level. The considerable genetic differences strongly suggested that the two forms represent distinct species. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.