Seed of Ullucus tuberosus—An apomict or not

Summary The paper deals with variations in enzyme pattern in the seeds of ulluco, Ullucus tuberosus. Six seeds per clone, four clones and three enzyme systems (malate dehydrogenase, glutamate oxaloacetate transaminase and 6-fosfoglucose dehydrogenase) were examined. There was variation in enzyme pattern between the seeds of a single clone. It is concluded that ulluco is not an apomictic species but a truly sexual one. Consequences of these findings for breeding programmes of ulluco are pointed out.     

Phenotypic polymorphism of six enzymes in the grasspea (Lathyrus sativus L.)

Summary The phenotype variation in six enzymes, 6-phosphogluconate dehydrogenase (6-PGD), malate dehydrogenase (MDH), peroxidase (PRX), isocitrate dehydrogenase (IDH), galactose dehydrogenase (GD) and glutamate oxaloacetate transaminase (GOT), was investigated using horizontal starch gel electrophoresis in 52 accessions of the grasspea, Lathyrus sativus. Phenotypic polymorphism was observed for all six enzymes. High phenotypic polymorphism (Pj) was observed for PRX and 6-PGD, while there was little polymorphism for GOT, with only two accessions showing variation. There was no correlation between phenotypic polymorphism and region of origin, or groupings of accessions made on the basis of flower colour. Tentative genetic interpretations of banding patterns are given for five of the enzyme systems. The level of apparent heterozygosity was higher than expected in this predominantly autogamous species. The level of variation in the grasspea is discussed in terms of its potential for exploitation through plant breeding.     

Isozyme polymorphism in three gene pools of cultivated chicory (Cichorium intybus L.)

Summary Polymorphism of ten enzymes, acid phosphatase (APH), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), phosphorylase (PP), superoxide dismutase (SOD), malic enzyme (ME), glutamate oxaloacetate transaminase 1 and 2 (GOT-1, GOT-2) and phosphoglucomutase 1 and 2 (PGM-1 and PGM-2), was investigated in three gene pools of cultivated chicory, including six cultivated wild chccory, eight industrial chicory and eight Brussels chicory varieties. LAP, APH, PP and PGM-2 showed high phenotypic polymorphism whilst GOT-1 and ME had poor polymorphism. For three enzyme coding loci Lap, Pgm-1 and Got-2, allele frequencies were determined. Isozyme composition in the three chicory gene pools was significantly different, showing, respectively, high, intermediate and poor average amount of phenotypic polymorphism in cultivated wild chicory, industrial chicory and Brussels chicory. Isozyme variation within and between varieties of the three gene pools is discussed in relation to breeding practices.     

Morphological variations and isoenzyme polymorphism of date palm clones from in vitro culture acclimatized and established on soil in South Morocco

Five clones of date palm (Phoenix dactylifera L.) corresponding to three reputed Moroccan cultivars (BFG, JHL and BSK) and two selected seedling genotypes (S16 and S35), derived from in vitro culture, were acclimatized and planted in Errachidia soil (South Morocco) in 1989. After 10 years of culture, clones were evaluated on the presence/absence of the two principal morphological characters (offshoots, inflorescences). Then, their leaf extracts were subjected to electrophoresis on polyacrylamide gels in order to determine their isoenzyme polymorphism. Inflorescence formation is relatively more frequent in the clones S16 and S35 than in clones JHL and BSK. Clone BFG showed an intermediate situation. High isoenzyme variations were found for the oxidoreductases (peroxidases (POX) and polyphenoloxidases (PPO)), the transferase (glutamate oxaloacetate transaminase (GOT)) and the hydrolases (esterases (EST) and endopeptidases (ENP)) enzymes. The factorial component analysis exhibited a negative correlation between the two principal morphological characters (presence of inflorescences and offshoots formation). Date palm clones typified respectively by high inflorescences and high offshoots formed two separated groups showing each one some isoenzyme characteristics. All morphological and enzyme variations were discussed in respect to the genotype effect on the micropropagation process.     

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     

Assessment of genetic variation among asparagus (Asparagus officinalis L.) populations and cultivars: agromorphological and isozymic data

Summary The morphological variation and the isozymic polymorphism in 19 asparagus accessions currently used in cultivation and breeding are described. Moreover 2 wild populations from Turkey were added in the isozymic study. Characters of ramification height permit to separate accessions usually cultivated for white asparagus and accessions usually cultivated for green asparagus. Starch and polyacrylamide gel electrophoresis was realized to assay 7 enzyme systems. 29 polymorphic bands were taken into account in the study. Four accessions show specific bands for glutamate oxaloacetate transaminase, shikimate dehydrogenase and alcohol dehydrogenase. In particular, the two populations from Turkey and one population traditionally cultivated in local area of Spain are well differentiated from the remaining accessions. Accessions from the United States representing different selections are relatively well separated from each other.     

Electrophoretic Analysis of Eleven Isozyme Systems and their Possible Use as Biochemical Markers in Breeding of Chicory (Cichorium intybus L.)

Polymorphism and ontogeny of 11 chicory (Cichorium intybus L.) enzymatic systems have been analyzed by native polyacrylamide gel electrophoresis, namely: leucine aminopeptidase, phosphoglucomutase, shikimate dehydrogenase, glutamate oxaloacetate transaminase, superoxide dismutase, isocitrate dehydrogenase, esterase, phosphorylase, glucose-6-phosphate dehydrogenase, 6-phos-phogluconate dehydrogenase and glucose phosphate isomerase. The use of these systems as biochemical markers is discussed.     

Correlation of isoenzyme polymorphism and Bayoud-disease resistance in date palm cultivars and progeny

Summary Seedlings of date palm (Phoenix dactylifera L.), obtained from seven cultivars crossed with two males, were analyzed by polyacrylamide gel electrophoresis for esterase (EST), glutamate oxaloacetate transaminase (GOT), endopeptidase (ENP) and alcohol dehydrogenase (ADH) polymorphisms. Eleven, eight, five and two phenotypes were revealed for the enzymes tested, respectively. Seedlings of F₁ populations derived from Bayoud (Fusarium)-resistant and low fruit quality cultivars were characterized by a high electrophoretic polymorphism, when compared with progenies of Bayoud-susceptible and high fruit quality cultivars. In almost all cases, the most frequent electrophoretic phenotypes scored for each enzyme in different F₁ populations, were similar to those of the corresponding parent cultivars. Heterozygous phenotypes have been found for, at least, 3 loci; Got-2, Est-1 and Enp, indicating that such loci could be used to screen for hybrid seedlings. The expected Mendelian segregation of allozymes has been observed for these 3 loci, in many F₁ populations. It seems that progenies of Bayoud-resistant cultivars are characterized by a high level of electrophoretic polymorphism. The estimation of this index and the search for genetic linkage with segregating allozymes, may be biochemical criteria useful as an aid in distinguishing date palm seedling populations derived from Bayoud-resistant cultivars and suitable for breeding programs.     

Geographical distribution of allozyme patterns in shallot (Allium cepa var. ascalonicum Backer) and wakegi onion (A. × wakegi Araki)

Summary Shallot and wakegi were collected in the main islands of Indonesia, and in Japan, Korea, Taiwan, Malaysia, Thailand and Bangladesh. Five isozyme resolutions, phosphoglucomutase (PGM), glutamate oxaloacetate (GOT), glutamate dehydrogenase (GDH), esterase (EST) and peroxidase (POX) were employed for demonstrating inter-and intraspecific differences. A dendrogram separated 189 collected accessions into 25 types of wakegi onion and 18 types of shallot. All accessions of Japan, Korea and Taiwan were determined to be wakegi onion, whereas those of Bangladesh, Malaysia and Thailand were shallot. Twenty-six out of 165 Indonesian accessions indicated wakegi onion distribution in Sumatra, West Java province and in Sulawesi Island. This confirmed that there is mixed-cultivation of the two Allium species with no distinction made between them. Japan and Indonesia had respectively 12 and eight unique types of wakegi onion, while Korea had only one type. West Java showed the most various type of wakegi onion, whereas East Java had many types of shallot. Shallots collected from Bangladesh were distinetly different from those of South East Asian types.     

Independent segregation of two isozyme markers and inter-plant differences in nuclear DNA content in the interspecific backcross (Allium fistulosum L. × A. cepa L.) × A. cepa L.

Summary From backcrosses of three interspecific hybrids (A. fistulosum x A. cepa) with a. cepa 14 diploid and 2 triploid plants were recovered.In this BC1 population introgression of A. fistulosum genetic material into the A. cepa nuclear genome was studied using two isozyme markers: Got-1 and Got-2. Both loci carried two alleles. A. cepa was monomorphic for both markers. A. fistulosum was polymorphic for Got-2. Based on their Got isozyme pattern seven out of the 14 diploid BC1 plants had a recombinant genotype. The loci appeared to be unlinked. Differences were observed in nuclear DNA contents between the diploid BC1 plants, indicating that gametes produced by the interspecific hybrids contained different combinations of chromosomal material from A. cepa and A. fistulosum.Abbreviations Adh alcohol dehydrogenase - Got glutamate oxaloacetate transaminase - Idh isocitrate dehydrogenase - Pgi phosphoglucoisomerase - Pgm phosphoglucomutase     

Cytogenetic and isozymic characterization of anther-panicle culture derived tall fescue aneuploids

Summary Chromosome associations of 44 tall fescue (Festuca arundinacea Schreb., 2n=6x=42) plants derived from anther-panicle culture of Kentucky 31 were evaluated to determine if new genetic stocks could be identified. Seventeen of the plants were euploids (21 II), two were monosomic (20II+1I), 22 were double monosomic (19II+2I) and three were triple monosomic (18II+3I). Zymograms were obtained for phosphoglucoisomerase (PGI), glutamate oxaloacetate transaminase (GOT), acid phosphatase (ACPH), malate dehydrogenase (MDH) and 6-phosphogluconate dehydrogenase (6-PGD). The zymograms were identical for GOT, ACPH, and 6-PGD but different patterns were found for MDH and PGI in some of the double monosomics and euploids from a different piece of callus than the majority of the plants.Contribution of the USDA-ARS in cooperation with the Kentucky Agricultural Experiment Station. Scientific Journal Series No. 86-3-61.     

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 diversity among elite varieties of the aromatic grasses, Cymbopogon martinii

The elite and popular cultivars of Cymbopogon martinii were examined for genomic and expressed molecular diversity through RAPD, enzyme and SDS-PAGE protein polymorphisms. The allelic score at each locus of the enzymes as well as presence and absence profiling in RAPDs, overall occurrence of band types etc. were subjected to computation of gene diversity, expected heterozygosity, allele number per locus, and similarity matrix. These, in turn, provide inputs to derive primary account of allelic variability, genetic bases of the cultivated germplasm, putative need for gene/trait introgression from the wild or geographically diverse habitat etc. in elite selections. ‘PRC1’ possessed highest number of unique bands based on RAPD polymorphism. In variety ‘IW31245E’, diaphorase and glutamate oxaloacetate transaminase isozymes generated two unique bands as dia-III ² and got-II ⁴. ‘RRL(B)77’ exhibited three unique bands; one produced by esterase as allele est-II ¹ and two by malic enzyme (me-III ^1,3). Only one unique band was generated by malic enzyme in variety ‘Trishna’. But sofia had three unique bands, two contributed by diaphorase (dia-II ³ and dia-II ⁴ and one by glutamate oxaloacetate transaminase (got-II ²). SDS-PAGE analysis revealed presence of unique polypeptide fragments (97.7 kDa to 31.6 kDa) in varieties ‘IW31245E’, ‘RRL(B)77’, ‘Tripta’, ‘Trishna’, ‘PRC1’ and var. sofia, generated as a diagnostic marker. In general, molecular distinctions associated with var. motia and var. sofia have been clearly noticed in C. martinii. This revised version was published online in July 2006 with corrections to the Cover Date.     

Isozyme polymorphism in Festuca rubraL.

Summary Six Festuca rubra populations from Europe and Scandinavia were studied for variation at three isozyme loci; phosphoglucoisomerase (PGI-2), glutamate oxaloacetate transaminase (GOT-3) and superoxide dismutase (SOD-1). Seven alleles were found at the Pgi-2 locus, four at the Got-3 locus and five at the Sod-1 locus. Most plants were heterozygous and up to five alleles were found in the same plant at the Pgi-2 locus. Each population could be distinguished by the presence or absence of certain alleles or by differences in the frequencies of the alleles present. Values for the Shannon diversity index were calculated which showed that there was considerable heterogeneity both within and between loci. In general, 53% of this diversity could be attributed to within population variation.     

Genetic diversity of natural populations of Atriplex halimus L. in Morocco: An isoenzyme-based overview

Based on polyacrylamide gel electrophoresis, nine natural populations of Atriplex halimus L., a perennial shrub, collected in different regions of Morocco, were studied for their genetic variation using isoenzyme polymorphism of the highly active enzyme systems: esterases (EST), acid phosphatases (ACP) and glutamate oxaloacetate transaminase (GOT). Different allozyme frequencies from 7 different loci were obtained for all populations of this halophyte species. High levels of genetic diversity were revealed. The mean number of alleles per locus (A = 1.9–2.0), the percentage of polymorphic loci (p = 71.4–85.7) and the mean expected heterozygosity (H_e = 0.339–0.385) showed an important variability in all populations. Gene diversity was essentially explained by the within population component. The between populations differentiation accounted for 8% of the whole diversity (F_ST, averaged over all loci, is 0.08). The relationships among the 9 populations were inferred from the Nei’s genetic distances. Four major groups were formed. The northern population ‘Tanger’, forming a unique group, was highly divergent from the other groups. It appeared that the genetic distance between all groups was related to the geographic distance that separates them. This revised version was published online in August 2006 with corrections to the Cover Date.     

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     

Identification of some Populus deltoides Marsh. x P. nigra L. clones,developed in North America,with the aid of allozymes

Summary Horizontal starch gel electrophoresis was done to assay 10 enzyme systems in root tips of 14 Populus deltoides x P. nigra clones developed by controlled hybridization and selected in North America, and some clones sharing one or both parents. The genotypes of the clones were determined for 31 loci coding for 10 enzyme systems. The interclonal allozyme variability was controlled by 12 loci. Each of the 14 clones had unique 12-locus genotypes, thus could be distinguished from each other. The clones differed from each other on an average of 4.2 loci. The first two Principal Components from Principal Component Analysis of the clonal allozyme genotype data accounted for 48% of the total variation in the 12 variant loci. 6-Pgd-3, Per-2, 6-Pgd-1, Aco-1 and Per-1 were found to be the most discriminating loci for the clones. The ordination of the clones on Principal Component axes 1 and 2 was in general agreement with the origin of the clones.     

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.     

Levels of host plant resistance to banana weevil, Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae), in African Musa germplasm

Forty-five Musa clones, including endemic and introduced cultivars plus hybrids, were evaluated for resistance against the banana weevil, Cosmopolites sordidus, in a field trial in Uganda. The predominant groups of staple crops, East African highland bananas (Musa spp. AAA) and plantains (Musaspp. AAB), as well as plantain-derived hybrids (AAB × AA), showed the highest levels of susceptibility to this pest. These were followed by dessert bananas (Musa spp. AAA), exotic bananas (Musa spp. ABB) and finally diploids of M. acuminata (AA). Hybrids of banana origin were highly resistant. Some East African highland cultivars, especially brewing types (e.g., Kabula, Bagandeseza, Ediirira), showed intermediate levels of resistance. Among the non-highland bananas, high levels of resistance were observed in Yangambi-Km5 (AAA), Cavendish (AAA), Gros Michel (AAA), Kayinja (ABB, Pisang Awak subgroup), Ndiizi (AB, Ney Poovan subgroup)and Kisubi (Ney Poovan subgroup). The highest resistance was observed in banana hybrids TMB2×7197-2, TMB2×8075-7 and the wild banana Calcutta-4 (AA). These were considered the best sources of resistance for a weevil resistance-breeding programme with the two hybrids commonly used as improved male parents. This revised version was published online in July 2006 with corrections to the Cover Date.