Characterization of isozymes in Salvia columbariae

Summary Salvia columbariae is a herbaceous annual species which has potential for domestication as a new source of industrial oil. Isozyme markers provide a mean by which this process may be facilitated. Isozyme survey of field grown Salvia columbariae plants showed variation for malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphogllucomutase (PGM). Selfed seed was obtained from the field and was grown in the greenhouse for segregation analyses. Electrophoretic results indicated that MDH was variable at zone 1, showing presence or absence of a band. The observed segregation ratio was not significantly different from expected ratio for Pgi-4 and Pgm-3 isozymes, indicating monogenic control of the two loci. Pgi-4 locus was heterozygous for a null allele. Cross dimerization between the allozyme Pgi-3 and Pgi-4 loci resulted in an intergenic band for this isozyme system.Abbreviations ACO aconitase - ADH alcohol dehydrogenase - DTT-DL dithiothreitol - FDH formate dehydrogenase - GOT glutamate-oxalacetate transaminase - IDH isocitric dehydrogenase - MDH malate dehydrogenase - MNR menadione reductase - 6PGD 6-phosphogluconate dehydrogenase - PGI phosphoglucose isomerase - PGM phosphoglucomutase - PVP-40 polyvinylpyrrolidone - SKDH shikimate dehydrogenase - TPI triosephosphate isomerase     

苹果酸脱氢酶在番茄F1杂种纯度检测中的应用

本文应用垂直板不连续系统的聚丙烯酰胺凝胶电泳（ＰＡＧＥ）法,分析了５个杂交一代种（佳粉一号、佳粉二号、佳粉十号、佳粉十五号、双抗二号）发芽肿种苗的五种同工酶（苹果酸脱氢酶（ＭＤＨ）、乙醇脱氢酶（ＡＤＨ）、磷酸葡萄糖变位酶（ＰＧＭ）、酯酶（ＥＳＴ）、异柠檬酸脱氢酶（ＩＤＨ）。发现５个杂交一代种中,３个要交一代种与其双亲的苹果酸脱氢酶（ＭＤＨ）谱带有有明显稳定的区别。两个杂交一代种与其双亲酯（ＥＳＴ）     

Expression of heritable biochemical markers from various pecan tissues

Summary Isozyme expression of malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were examined from 12 tissues of pecan by starch gel electrophoresis. Tissue type, stage of development, and extraction method had little effect on isozyme expression of MDH or PGM but did influence PGI expression. Additional PGI bands were observed from seed tissue and soaked pollen. A direct relationship was observed between PGI expression from seed tissue and eventual expression from the germinated seedling. Additional PGI bands from soaked pollen were thought to be produced from structural changes and not due to more efficient extraction. The genotype of immature pecan seeds could be determined for Mdh-1, Pgi-2, and Pgm-1 provided cellular endosperm had developed. Watery endosperm did not contain detectable levels of the study enzymes. The advantages of uniform isozyme expression among various tissues and ability to determine the genotype of immature pecan seed are discussed.     

Genetic and linkage analysis of isozyme loci in Daucus carota L.

Summary Electrophoretic polymorphisms of eight enzyme systems were studied in leaves of Daucus carota ssp. sativus in order to identify additional isozyme loci and generate first linkage groups of genetic markers. The genetic analysis of aconitase (ACO), leucin aminopeptidase (LAP), menadione reductase (MDR), phosphoglucomutase (PGM), 6-phosphogluconic dehydrogenase (6-PGD), shikimate dehydrogenase (SKD), and triose phosphate isomerase (TPI) zymograms resulted in the identification of 8 isozyme marker loci, designated as Aco-1, Lap-1, Pgm-1, Pgm-3, 6-Pgd-2, Skd-1, Tpi-1, and Tpi-2. All loci segregated with codominant alleles and encoded for monomers (ACO, LAP, PGM, SKD), and dimers (6-PGD, TPI), respectively. MDR enzymes of the variable region MDR-2 appeared to be identical with Dia-2 isozymes. Tests of joint segregation for pairwise comparisons of all 14 isozyme marker loci now available in carrots indicate that 12 loci are linked in 4 linkage groups (marked K1 to K4) in the following order: Aco-1, Pgi-1, and Dia-3 (K1), Tpi-2, Got-2, and Lap-1 (K2), Got-3 and Tpi-1 (K3) and Pgm-1, Pgm-3, 6-Pgd-2 and Skd-1 (K4). Dia-2 and Got-1 remained unlinked. The possible duplication of a PGM locus and a 6-PGD locus is discussed.     

Use of isozyme patterns in characterization of potato mutant lines selected for agronomic quantitative traits

Summary Seven varieties and 57 spontaneous or induced in vitro mutant lines (20 macromutant and 37 micromutant events) of potato were tested by starch gel electrophoresis for ADH, GOT, PGI, PGM, ACO, IDH, MDH and 6PGDH isozymes in tuber extracts. The data showed that in contrast to variety comparisons, the isozyme patterns rarely differentiate mutant lines which have altered morphological traits. But trying to identify isozyme differences in mutants can still be useful for a chimeric structure for GOT-2 alleles in a mutant from Atlantic and a new tuber specific locus for 6PGDH in mutants from Russet Burbank were found.Abbreviations ACO aconitase - ADH alcohol dehydrogenase - GOT glutamate oxaloacetate transaminase - IDH isocitric acid dehydrogenase - MDH malate dehydrogenase - 6PGDH 6-phosphogluconate dehydrogenase - PGI phosphoglucoisomerase - PGM phosphoglucomutase - SGE starch gel electrophoresis - EMS ethyl metanesulfonate     

Genetic Analysis of Isozyme Loci in Chives (Allium schoenoprasum L.)

The inheritance ot nine isozyme systems in selfed progenies of chives was studied elcctrophoretically. The isozyme systems studied were malate-dehydrogenase (MDH), phosphoglucoisomerase (PGI), triosephosphate-isomerase (TP1), diaphorase (D1A), menadione-reductase (MDR), esterase (EST), glutamate-oxalacetate-transaminase (GOT), hexokinase (HK) and superoxide-dismutase (SOD). Segregation results of Goodness-of-Fit tests against the expected 1:2:1 and 3 : 1 ratios for codominant and dominant inheritance, respectively, led to the identification of 16 isozyme loci whose banding patterns are given in this paper. The first results on linkage relationships between isozyme loci and between the Mdr-3-locus and a male sterility gene are given. These results and further studies will be used to mark important characters in chives.     

Genetic control of isozymes in eggplant and its wild relatives

Summary This study was conducted to elucidate the inheritance and linkage relationships of isozymes in aspartate aminotransferase (AAT, EC 2.6.1.1), alcohol dehydrogenase (ADH, EC 1.1.1.1), phosphogluconate dehydrogenase (PGD, EC 1.1.1.43), phosphoglucomutase (PGM, EC 2.7.5.1) and shikimate dehydrogenase (SKDH, EC 1.1.1.25) in eggplant and its wild relatives. Segregating populations were generated by backcrossing of hybrids among the species. Evidence of Mendelian inheritance was obtained for seven loci: Aat-1, Adh-1, Adh-2, Pgd-1, Pgm-1, Pgm-2 and Skdh-1. Twenty-one pairs of loci were tested for independent assortment, suggesting three linked pairs, Aat-2 with Pgd-2 (R=0.35±0.07), Adh-2 with Pgm-1 (R=0.33±0.07) and Pgd-2 with Pgm-2 (R=0.32±0.06).Abbreviations AAT aspartate aminotransferase - ADH alcohol dehydrogenase - PGD phosphogluconate dehydrogenase - PGM phosphoglucomutase - SKDH shikimate dehydrogenase     

Meiotic and Isozymic Analyses of Tall Fescue × Giant Fescue Hybrids and Amphiploids1

The meiotic behavior of three tall fescue (Festuca arundinacea, 2n = 6x = 42) genotypes, giant fescue (F. gigantea, 2n = 6x = 42), and their reciprocal F₁ hybrids and C₁, amphiploids was evaluated to determine the parental genomic relationships. Isozyme banding patterns were used to confirm the parental identity of the hybrids and amphiploids. At meta-phase I, the parents had predominantly bivalent pairing. The hybrids had an average of 9.51 I, 16.02 II, 0.12 III, 0.02 IV, and the amphiploids had 2.17 I, 38.82 II, 0.60 III, 0.58 IV, 0.01 V—VIII. The prevalence of bivalent pairing in both hybrids and amphiploids suggested a homoeologous relationship between the six genomes, with four of the six being more closely related. Bivalent pairing in the amphiploids indicated genetic regulation of chromosome pairing. Zymograms were obtained for acid phosphatase (ACPH), alcohol dehydrogenase (ADH), glutamate oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), 6-phosphogluconate dehydrogenase (6-PGD) and phosphoglucoisomerase (PGI). The three tall fescue and giant fescue parents had different zymograms for ACPH, MDH, 6-PGD and PGI; thus, the tall fescue parents of the hybrids and amphiploids could be determined based on the banding patterns of these four enzymes. Phenotypes were determined for ACPH-1, PGI-2 and 6-PGD-1. ACPH-1 may be used to follow the introgression of giant fescue chromatin into a certain tall fescue genotype.     

Genetic Analysis of DIA, GOT and PGI Isozyme Loci in Daucus carota L. ssp. sativas

Isozymes are useful biochemical markers in plant breeding. This paper presents the first results on the inheritance of three enzyme systems: diaphorase (DIA), glutamate oxalacetate transaminase (GOT) and phosphoglucoisomerase (PGI) from leaves of Daucus carota L. ssp. sativus and the linkage relationships among these isozyme loci, marked Dia, Got, and Pgi. Methods of extraction and electrophoresis are described. Segregation ratios indicated that the DIA system is controlled by three genes: Dia-1, Dia-2 and Dia-3, Dia-1 as invariant. Two codominant alleles and a null allele were found for Dia-2, and four codominant alleles for Dia-3-GOT isozymes are encoded by three genes; Got-1, Got-2 and Got-3. An active allele and a null allele were observed for Got-1, two codominant alleles for Got-2 and Got-3, respectively. PGI isozymes are controlled by a single gene, Pgi-1, with two codominant alleles. Altogether one invariant isozyme locus and six polymorphic loci are postulated. Test of joint segregation suggested a linkage of Dia-3 and Pgi-1. The recombination value is 0.04 ± 0.009.     

Expression of Introduced Genes after Tuber Propagation of Transgenic Potato Plants

The linkage relationship between eighteen isozyme loci and the morphological markers hypocotyl colour (R-r), monogerm character (M-m), pollen fertility (X) and stem fasciation (Verb.) are tested. Three linkage groups could be set up, involving all morphological marker loci and eight of the isozyme loci. Est-2, R-r, Fdp-2, Got2 and Icd-1 belong to linkage group I, linkage group II includes the loci Fas-fas M-m, Est-3 and Aco-1, linkage group III contains the loci X, Mdh-1 and Est-5. When analysing the inheritance of isozymes and RFLPs, deviations are usually found in some lines from the expected frequencies of a 3 : 1 or 1 : 2 : 1 segregation at single marker loci. In many cases these data can still be used for the estimation of recombination values with linked loci under the control of selection. Procedures to estimate linkage in such cases are given and applied to experimental data in Beta vulgaris.     

Inheritance of isozymes in peach x Prunus kansuensis and peach x Prunus davidiana hybrids

Summary Isozyme variation and inheritance were investigated with starch gel electrophoresis in peach (Prunus persica L. Batsch) x P. kansuensis Rehd. and peach x P. davidiana (Carr.) Franch. interspecific hybrids. Of five enzyme systems surveyed for polymorphism, four systems were identified as polymorphic [isocitrate dehydrogenase (IDH, EC 1.1.1.41), phosphoglucomutase (PGM, EC 2.7.5.1), aspartate aminotransferase (AAT, EC 2.6.1.1), and 6 phosphogluconate dehydrogenase (PGD, EC 1.1.1.44)] and may be useful as genetic markers in future cultivar and rootstock development. Analysis of progenies segregating for pairs of loci suggests a possible linkage between the loci coding for Aat-1 and Pgd-2. Independent assortment was observed for isozyme loci Idh/Pgm-2, Idh/Aat-1, Idh/Pgd-2, Pgm-2/Aat-1, Pgm-2/Pgd-2, and Aat-2/Aat-1. The red leaf locus, Gr, assorted independently of the isozyme loci: Idh, Pgm-2, Aat-1, and Pgd-2.     

Linkage maps for nine isozyme and four marker loci in sugarbeet (Beta vulgaris L.)

Summary An analysis of linkage in sugarbeet (Beta vulgaris L.) was conducted for nine isozyme loci, Ak ₁, Gdh ₂, Idh ₁, Lap, Mdh ₁, Mdh ₃, Pgi ₂, Pgm ₁,and Skdh ₂,and four marker loci, annuality (B), red hypocotyl-color (R), pollen fertility restorer (X), and monogermity (m). Four linkage groups were identified; R-B-Idh ₁, Gdh ₂-Mdh ₁, Ak ₁ -Lap, and Mdh ₃-Pgm ₁.In addition, X was linked to Mdh ₁and Skdh ₂with a recombination value of 13.4% and 34.7%, respectively, and m was linked to Pgm ₁with a recombination value of 35.8%. Pgi ₂was inherited independently of the four linkage groups. This locus showed a skewed ratio in F₂ progeny of a cross between self-compatible and self-incompatible lines and the allele derived from self-incompatible parents decreased markedly. On the other hand, the expected segregation ratio was observed in the backcrossed progeny and also in F₂ progeny of a cross between self-compatible lines. The results obtained suggest that Pgi ₂may be linked to a self-compatibility locus (S ^f)and the two loci may be assigned to an additional linkage group.     

New Isozyme Markers in Vicia faba: Inheritance and Linkage

Results on the inheritance of 6 enzyme systems: LDH, PGM, FDH, SKD, SOD, AAT from seeds of Vicia faba and the linkage relationships among these isozyme loci are presented. The allozymes at each one of these loci behaved in a codominant manner and segregated in the expected Mendelian ratio. Linkage tests between these loci showed that they segregate independently.     

Genetic variation for isozyme genes and proteins in spanish primitive cultivars and wild subspecies of Lens

Summary An analysis of the variability for genes encoding seven isozyme systems and storage proteins in a collection of cultivated and wild accessions of Lens is reported. The collection, which is part of the Spanish INIA Cenebank, contains the ssp. culinaris, orientalis, odemensis, nigricans and ervoides, and presents a high degree of genetic diversity both within and between the accessions. A total 25 loci were examined; of these, 18 were polymorphic (the 7 genes encoding storage proteins, and the following isozyme loci: Acp-1, Acp-2, Cpx-1, Cpx-2, Aat-p, Aat-m, Lap-1, Mdh-2, Mdh-3, Mdh-4 and 6pgd-p) and 7 were monomorphic (Aat-mb, Aat-c, Mdh-1, Mdh-5, 6pgd-2, Pgm-c and Pgm-p). The phylogenetic relationships between subspecies were analyzed using the allelic frequencies. The study suggests that orientalis and odemensis share more biochemical characters than the other subspecies, and that those subspecies keep an intermediate position between Lens culinaris and Lens nigricans.     

Genetic analysis and nomenclature for seven isozyme systems in Brassica nigra, B. oleracea and B. campestris

General criteria for the assignment of names to enzyme systems, regions of activity, isozyme loci and allozymes have been lacking in crucifer species. This paper proposes a standard nomenclature for seven isozyme systems in the three diploid species of U's triangle: Brassica nigra, B. oleracea and B. campestris. Gel/electrode buffers, which provided the best resolution for seven isozyme systems, acid phosphatase (APS), aconitase (ACO), leucine aminopeptidase (LAP), 6-phosphogluconate dehydrogenase (6 PGD), phosphoglucoisomerase (PGI), phosphoglucomutase (PGM), and triosephosphate isomerase (TPI), were proposed as standards. Isozyme genetic analysis was determined for B. oleracea and B. campestris from previous studies and by segregation of selfed progenies of heterozygous B. nigra plants. Several populations were studied and 148 allozymes at the 18 loci observed were described for the three species. Their relative mobility was studied using a pure line of oilseed rape as reference. The comparison of the different alleles within and between the species is discussed.     

Comparison of isoenzymes in Prunus avium separated by two different electrophoretic techniques

Isoenzyme variation for seven systems revealed by two different electrophoretic procedures was compared in Prunus avium. Fourteen cultivars and 14 wild selections were analysed for acid phosphatase (ACP), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), malate dehydrogenase (MDH), phosphoglucomutase (PGM), shikimate dehydrogenase (SKD) and superoxide dismutase (SOD). Extracts were separated by isoelectric focusing (IEF) and by polyacrylamide gel electrophoresis (PAGE). For the eight loci that had been described previously in these enzyme systems on the basis of IEF analysis, we compared the variation revealed with IEF and PAGE. Similar variation was revealed for Acp‐1 and Pgm‐1, and the alleles revealed by PAGE could be identified directly with those reported for IEF. For Lap‐1, Mdh‐1 and Skd‐1, variation was seen with IEF but not with PAGE. For Mdh‐2, PAGE revealed additional variation not revealed by IEF. For Idh‐1, different patterns of variation were revealed by PAGE and IEF, and both procedures would be needed to genotype cherry accessions. We were unable to detect variation corresponding to that reported previously for Sod‐1 with either technique. The implications of these findings for allele labelling, for studies of genetic diversity and for linkage analysis are discussed.     

Chloroplast DNA diversity in eggplant (Solanum melongena) and its related species S. incanum and S. marginatum

Summary The potential of isozymes for distinguishing asparagus varieties was carried out by a survey on 21 varieties using 10 enzyme systems: GOT, SkDH, DIA, PGM, MDH, IDH, PGD, ACP, PGI, MR and ADH. Only 3 enzymes, SkDH, GOT and PGM, showed useful polymorphisms. The varieties were found heterogeneous according to their genetic structure: open pollinated varieties were more heterogeneous than clonal hybrids; the F1 hybrid and the vitroclones were homogeneous. As expected from the narrow genetic basis of the varieties, only a few alleles per isozyme locus were present. Moreover, for each enzyme, one allele or type was predominant so that the discriminating power of the method was low. However some of the varieties could be identified and different applications of the results are presented.Abbreviations D.U.S.- Distinction-Uniformity-Stability - ACO- aconitase - ACP- acid phosphatase - ADH- alcohol dehydrogenase - CAT- catalase - DIA- diaphorase - END- endopeptidase - GOT- glutamate oxaloacetate transaminase - IDH- isocitrate dehydrogenase - MDH- malate dehydrogenase - MR- menadione reductase - PGI- phosphoglucoisomerase - PGM- phosphoglucomutase - PGD- phosphoglucose dehydrogenase - POX- peroxidase - SkDH- shikimate dehydrogenase     

Isozyme electrophoretic characterization of 29 related cultivars of lily (Lilium spp.)

Isozyme banding patterns (IBPs) were studied for cultivars of lily (Lilium spp.) by means of horizontal starch‐gel electrophoresis (SGE). An array of continuous histidine‐citrate buffer systems at eight ranges of pH and four extraction buffers were tested. On the basis of this survey, the extraction buffer two (Eb‐2) and the buffer system E at pH 7.7 were found to be suitable for detection of lily isozymes. Using the SGE technique, IBP in catalase (CAT; EC 1.11.1.6), esterase (EST; EC 3.1.1.1), malate dehydrogenase (MDH; EC 1.1.1.37), malic enzyme (MAL; EC 1.1.1.40), peroxidase (POX; EC 1.11.1.7), phosphoglucomutase (PGM; EC 2.7.5.1), phosphoglucose isomerase (PGI; EC 5.3.1.9) and 6‐phosphogluconate dehydrogenase (PGD; EC 1.1.1.44) were assayed. In total 29 cultivars were tested in this study: nine were analysed for all eight enzyme systems, 16 cultivars for seven systems, three for six, and one for five enzyme systems. Some IBP were identified as section‐specific biochemical markers. Eight enzymes systems were analysed by constructing a dendogram using the unweighted pair group method, arithmetic average (UPGMA) cluster analysis. The analysis indicated that the lily cultivars could be separated from other Lilium species, except for two L. x formonlogi cultivars:‘Hakuba’ and ‘Hakuko’ which could not be distinguished from each other by the isozyme patterns assayed here. This study shows that isozymes can provide useful biochemical markers for lily cultivar identification and to estimate the phylogenetic relationships among those cultivars.     

Construction of a linkage map for watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai) using random amplified polymorphic DNA (RAPD)

Summary A linkage map for watermelon (Citrullus lanatus) was constructed on the basis of RADP, ribosomal DNA restriction fragment length polymorphism (RFLP), isozyme, and morphological markers using F₁BC1. A segregating population of 78 individuals was the result of a backcross of a cultivated inbred line (H-7; Citrullus lanatus; 2n=22) and a wild form (SA-1; C. lanatus; 2n=22), in which the latter was the recurrent (male) parent. A total of 69 RAPD, one RFLP, one isozyme, and three morphological markers was found to segregate in the BC1 population. Linkage analysis revealed that 62 loci could be mapped to 11 linkage groups that extended more than 524 centimorgans (cM), while 12 loci segregated independently of all other markers. The locus for exocarp color was linked to two RAPD markers within a region of 5 cM on linkage group 4. The locus for flesh color was linked to a RAPD marker within a region of 30 cM on linkage group 6. The isozyme marker GOT was located on the linkage group 1. Linkage group 2 contained a locus for ribosomal DNA within 5 cM of a RAPD marker. Half of the RAPD markers on the linkage group 7 displayed severely distorted segregation. The construction of linkage map using molecular markers is necessary for the breeding of watermelon to introduce useful gene of wild watermelon efficiently. However the linkage map that was constructed for the most part on the basis of RAPD markers could not cover significant parts of the genome, the linkage map provides breeders of watermelons the possibility of tagging useful agronomic traits, as well as the gene for exocarp color.Abbreviations RAPD random amplified polymorphic DNA - RFLP restriction fragment length polymorphism - GOT glutamate oxaloacetate transaminase - MDH malate dehydrogenase - ACP acid phosphatase - 6PGH 6-phosphogluconate dehydrogenase     

Genetic control of isozymes in Carica papaya L.

The genetic control of isozymes was investigated in papaya. Out of six buffer systems tested, Histidine-citrate pH 6.5 gave the most consistent and greatest number of well-resolved enzymes. Nine enzyme systems (ACO, IDH, MDH, PGM, PGI, SKD, TPI, UGP, 6-PGD) out of 28 were resolved. Based on the initial screening of 131 accessions of papaya from Central and South America for electromorph variants, parents were selected for various combinations of controlled crosses. These controlled crosses were then used to elucidate the inheritance of isozymes in papaya. Evidence of Mendelian inheritance was obtained for eight out of nine polymorphic loci: Aco-1, Aco-2, Pgi-2, Pgm-1, Skd, Tpi-1, Tpi-2, and Ugp. No controlled cross was available to study the genetic control of the polymorphic locus Pgm-2. Because they were not variable in all accessions included in this study, the loci Idh, Mdh-1, Mdh-2, and 6-Pgd, were also not examined by segregation analysis. This revised version was published online in August 2006 with corrections to the Cover Date.