Mature seed samples from 35 populations of Echinochloa were collected in fields of the Eastern Iberian Peninsula and evaluated for nine morphologic traits. Four kernels per population were separately assayed for four isozyme systems. Genetic variability components were studied by correspondence analysis and chi-square distance using the isozyme pattern as variables. Genetic uniformity was found within populations as would be expected from their autogamous mating system. Intraspecific isozyme variability covered a wide spectrum, from none in E. colonum to very high in E. oryzoides. All the populations of E. colonum clustered together based on morphometric and isozymatic characters, and also those of E. oryzicola, but were distinct from the populations of all other species. Populations of E. crus-galli, E. hispidula, and E. oryzoides clustered within species based on morphology, but did not cluster based on isozymatic characters, being mixed in the isozymatic dendrogram. Our results support the hypothesis that E. hispidula and E. oryzoides are infraspecific taxa of E. crus-galli. The large between-population genetic variability may explain the differential response to herbicides within a given species and indicate that weed specialists should study the differences in response to a wide spectrum of Echinochloa populations to several herbicides instead of testing many plants from a small number of populations.

Genetic variability for loci governing enzyme/morphological variants and for herbicide response was determined in 10 populations of the slender wild oat (Avena barbata Pott. ex Link ♯ AVEBA), six populations of wild oat (Avena fatua L. ♯ AVEFA), and three populations of godetia (Clarkia williamsonii Lewis & Lewis). The enzyme loci were identified by starch gel electrophoresis and included peroxidase, 6-phosphogluconate dehydrogenase, esterase, and leucine aminopeptidase for the slender wild oat; peroxidase, esterase, leucine aminopeptidase, and malate dehydrogenase for the wild oat; and esterase, phosphoglucoisomerase, leucine aminopeptidase, acid phosphatase, and glutamate oxaloacetate transaminase for godetia. Morphological loci included lemma and leaf sheath hairiness for the oats. For both the enzymatic and morphological loci, levels of genetic variation for each population were quantified by a polymorphic index. The herbicide barban (4-chloro-2-butynyl 3-chlorophenylcarbamate) was used on the wild oats; bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) was used on godetia. Genetic variation for herbicide response was based on genetic variances calculated from phytotoxicity scores. Populations were ranked from highest to lowest for the polymorphic indices and the genetic variances. Concordance between the rankings was tested by rank correlation. Statistically significant relationships were found between the enzyme/morphological characters and herbicide response in the slender wild oat and the wild oat. For some species, the level of genetic variation for response to herbicides appears to be associated with genetic variation for enzymatic and morphological loci.

In this study, 123 almond (Prunus dulcis (Mill.) D.A. Webb) trees identified among traditional orchards in the Algarve region and 53 trees of the local field collection managed by the regional office of the Portuguese Ministry of Agriculture (DRAALG) were assessed using isozyme, inter-single sequence repeat and simple sequence repeat or microsatellite techniques for the evaluation of genetic diversity and genetic relatedness and identification of new accessions for the field collection. The isozyme analysis allowed the distribution of the 176 plants into 13 different classes of enzyme similarity, while the use of DNA markers increased the distribution of the analysed trees among 140 discriminating DNA patterns. Multiple cases of homonymy and synonymy were identified in the local germplasm. Some traditional varieties, such as Lourencinha, appeared to be relatively homogeneous, while other local denominations, e.g. Galamba, included diverse genotypes. Of the 13 commercial varieties analysed in this study, 11 assembled in one major cluster clearly differentiated from the majority of the local genotypes. These results reinforced the perception that the Algarve traditional germplasm constitutes an important repository of genetic diversity, eventually carrying alleles of high agricultural interest such as the recently identified Phomopsis resistance in the traditional variety Barrinho Grado.

Grasspea (Lathyrus sativus L.) is an annual, herbaceous, drought-resistant legume and staple crop in Asian and African countries. Little is known about the nature and the amount of genetic diversity present in existing grasspea seed collections, yet this information is pivotal for future breeding programmes, such as those striving to reduce high neurotoxin levels present in seeds. Here we report on the level of genetic diversity within a world-wide collection of L. sativus, determined by isozymatic analysis. Although grasspea is generally considered a predominantly self-pollinating species, we found that the population genetic structure of these accessions showed a considerable outcrossing rate of 36%. The identification of a mixed mating system in L. sativus has significant implications for collecting and multiplying genetic resources for conservation and for future breeding purposes. In addition, we determined the genetic closeness of grasspea accessions from different geographical regions around the world. While we noticed an allelic richness in this species that was conserved across the regions, we did not find any evidence of high genetic identity between accessions, even when originating from the same geographical location. Instead, we found that greater genetic variability existed at the intra-regional level than at the inter-regional level.

Isozyme data were used to assess genetic diversity within and among a subdivided population of the salt-tolerant Batini barley landrace. Population diversity and its components were estimated on the basis of 12 isozymes scored on 450 single plants representing seven subpopulations. Two principal components, based on mean gene diversity, Shannon's diversity index, percentage polymorphic loci, genetic identity and genetic distance among subpopulations accounted for 91.7% of total variation and separated the subpopulations into four distinct groups. Contributions to total diversity by individual subpopulations were partitioned into intra- and inter-population components. The level of population differentiation indicates that a large percentage of total genetic diversity was apportioned within subpopulations. The presence of valuable genetic diversity in this landrace was confirmed.

The genetic variation of isozymes [esterase (EST), superoxide dismutase (SOD), malate dehydrogenase (MDH), malic enzyme (ME) and glucose-6-phosphate dehydrogenase (G6PD)] extracted from two different adult tissues (adductor muscle and gill) of Pteria penguin R. and offspring from two stocks of Pinctada martensii D. were analysed by vertical polyacrylamide gel electrophoresis. The result showed an obvious and stable variation in the isozyme phenotypes in two different pearl oyster species. The SOD and EST isozymes from gill and MDH, ME and G6PDH from adductor muscle were species-specific. The electrophoretograms of these isozymes could be used as markers to differentiate the two pearl oysters. Between inbreeding and hybridized stocks of P. martensii, the electrophoresis patterns of isozymes were alike, but the similarities of electrophoretograms in the inbreeding progenies were higher than in the hybridized ones. The characteristic electrophoretograms of isozymes could be applied as molecular markers in the breeding of pearl oysters.

Zymography is an electrophoretic technique by which enzyme activity can be visualized directly on a polyacrylamide gel as discrete bands. A modified, more rapid technique for amylase zymography is described and compared with previously published methods. Whereas previous methods are based on 0.1 M acetate buffer as substrate buffer, our method utilizes 50mM Tris buffer containing Ca2+, Na+, NaN3 and Triton X-100 which helps rapid hydrolysis of the starch and stabilization of the enzyme. The staining procedure, previously requiring overnight incubation of the gel in iodine solution at 4 °C, has been reduced to 5 min at room temperature. Both methods gave rise to comparable levels of enzyme activity on polyacrylamide gels. Our modified method requires 8 h to complete the whole zymographical procedure instead of 18-20 h as in previous methods.

Human liver cytosolic and mitochondrial isozymes of aldehyde dehydrogenase share 70% sequence identity. However, the first 21 residues are not conserved between the human isozymes (15% identity). The three-dimensional structures of the beef mitochondrial and sheep cytosolic forms have virtually identical three-dimensional structures. Here, we solved the structure of the human mitochondrial enzyme and found it to be identical to the beef enzyme. The first 21 residues are found on the surface of the enzyme and make no contact with other subunits in the tetramer. A pair of chimeric enzymes between the human isozymes was made. Each chimera had the first 21 residues from one isozyme and the remaining 479 from the other. When the first 21 residues were from the mitochondrial isozyme, an enzyme with cytosolic-like properties was produced. The other was expressed but was insoluble. It was possible to restore solubility and activity to the chimera that had the first 21 cytosolic residues fused to the mitochondrial ones by making point mutations to residues at the N-terminal end. When residue 19 was changed from tyrosine to a cysteine, the residue found in the mitochondrial form, an active enzyme could be made though the Km for NAD+ was 35 times higher than the native mitochondrial isozyme and the specific activity was reduced by 75%. This residue interacts with residue 203, a nonconserved, nonactive site residue. A mutation of residue 18, which also interacts with 203, restored solubility, but not activity. Mutation to residue 15, which interacts with 104, also restored solubility but not activity. It appears that to have a soluble or active enzyme a favorable interaction must occur between a residue in a surface loop and a residue elsewhere in the molecule even though neither make contact with the active site region of the enzyme.

A new species of Porphyra is described from the south western Cape, South Africa. The gametophyte of Porphyra aeodis sp. nov. grows epiphytically on Aeodes orbitosa (Suhr) Schmitz, and has a seasonal life history that matches that of its host. Although P. aeodis has been confused with P. capensis Kützing in the past, P. aeodis is more similar to the sympatric but epilithic P. saldanhae Stegenga, Bolton et Anderson. There is considerable morphological overlap between P. aeodis and P. saldanhae, although they may be distinguished using a combination of morphological and ecological characters. The taxonomic separation of P. aeodis and P. saldanhae was confirmed using isozyme electrophoresis.

Use of isozymes (including allozymes) in studies of population genetics and systematics of seaweeds has increased sufficiently in the last decade to allow some generalization. Only a single locus has been observed for about half the enzymes analysed in seaweeds, compared with 29% in vascular plants. Compared with higher plants, macroalgal species generally have low amounts of electrophoretically detectable genetic variation; the lowest levels of genetic variation found in natural populations are those reported for seaweeds. Nonetheless, seaweeds show an association between levels of genetic diversity as revealed by isozymes and species-specific attributes, such as mating system and predominance of asexual versus sexual reproduction. In systematic studies, isozymes have revealed cryptic species and identified pairs of sibling taxa. The quaternary structure of enzymes appears to be conserved at the phylum level. With the current availability of improved techniques for enzyme electrophoresis and for data interpretation, we expect future studies utilizing isozyme electrophoresis to provide further insight into population and evolutionary processes in seaweeds.

Spontaneous hybrids between the two diploids Poa infirma Kunth and P. supina Schrader were obtained at the rate of 0·2%. No reciprocal maternal effect was observed. Almost all hybrids survived, grew normally but were sterile. Hybrids were diploid, and resembled in morphology the tetraploid P. annua L. (annual meadow grass). In their isozymes, they showed additivity of the parental patterns, and were very similar to those found in P. annua populations. The ease with which hybrids were obtained and the similarity of their morphological and isozyme pattern to P. annua suggest that the two diploid species are closely related to the allotetraploid species, and are perhaps its parent species.

Populations of the whitefly, Bemisia tabaci collected from the Americas and the Caribbean Basin were examined for non-specific esterases and for the ability to induce characteristic phytotoxic disorders in key assay species as a means of investigating biogeographic diversity. Esterase markers were used to detect polymorphisms among regional B. tabaci populations and to establish the present distribution of B. tabaci biotypes in the region. The A biotype occurred only in contiguous locales in northern Mexico and the southwestern US, while the B biotype was present throughout much of the Caribbean Basin and the US, and in Brazil. Distinct C and D type esterase markers were observed for Costa Rican and Nicaraguan B. tabaci populations, respectively. The 0 or null type population was collected only from Jatropha gossypifolia (L.) in Puerto Rico. Laboratory colonies of the A and the B biotypes were almost equally sensitive to an organophosphate, profenofos. The B biotype was more resistant to a pyrethroid, permethrin, suggesting the existence of a biotype of fi. tabaci, with a history of exposure to pesticides with a pyrethroid-based chemistry. In mating studies involving reciprocal crosses between the A and the B biotypes, very few F, female progeny were produced, indicating either minimal or non-existent reproductive compatibility between these haplo-diploid B. tabaci populations, presently considered to be the same species. Evidence is presented for the recent and widespread introduction, and subsequent spread of the B biotype throughout the US, the Caribbean Basin, and other proximal locations.

Electrophoretic studies of Guatemalan blackflies, Simulium ochraceum, the main vector of Central American onchocerciasis, were performed using isozyme patterns. A total of six larval and adult populations originating from four localities (a non-endemic area, Finca La Ruda, a low-endemic area, Finca Rincon, and two high-endemic areas, Finca Brote and Finca Recreo) were analyzed. The enzymes examined were adenylate kinase (AK), alkaline phosphatase (ALP), glucose phosphate isomerase (GPI), hexokinase (HK), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), malic enzyme (ME), and phosphoglucomutase (PGM). Of these, six enzymes, AK, ALP, HK, IDH, LAP, and ME showed no variation either within or among populations. GPI and PGM were found to be highly polymorphic in all of the six populations. χ2-test for fit to Hardy-Weinberg equilibrium (HWE) revealed that the distribution of the phenotypes of the two polymorphic enzymes was not significantly out of the HWE. A verages of values of proportion of polymorphic loci (P) and heterozygosity per individual (H) of the six populations were 0.234 and 0.084, respectively, each of which was similar to the values reported for different groups of organisms (Nei, 1975). Genetic diversity among populations was measured by FST and GST and both parameters showed that the non-endemic population from the Finca La Ruda (Dl) was greatly differentiated genetically from the other five populations. Estimates of genetic distance (Nei, 1975) showed that the Dl population was furthest from the other populations.