Isozyme Markers Research Articles

Mapping of quantitative trait loci for resistance to Mycosphaerella pinodes in Pisum sativum subsp. syriacum

Aschochyta blight, caused by Mycosphaerella pinodes, is one of the most economically serious pea pathogens, particularly in winter sowings. The wild Pisum sativum subsp. syriacum accession P665 shows good levels of resistance to this pathogen. Knowledge of the genetic factors controlling resistance to M. pinodes in this wild accession would facilitate gene transfer to pea cultivars; however, previous studies mapping resistance to M. pinodes in pea have never included this wild species. The objective of this study was to identify quantitative trait loci (QTL) controlling resistance to M. pinodes in P. sativum subsp. syriacum and to compare these with QTLs previously described for the same trait in P. sativum. A population formed by 111 F6:7 recombinant inbred lines derived from a cross between accession P665 and a susceptible pea cultivar (Messire) was analysed using morphological, isozyme, RAPD, STS and EST markers. The map developed covered 1214 cM and contained 246 markers distributed in nine linkage groups, of which seven could be assigned to pea chromosomes. Six QTLs associated with resistance to M. pinodes were detected in linkage groups II, III, IV and V, which collectively explained between 31 and 75% of the phenotypic variation depending of the trait. While QTLs MpIII.1 and MpIII.2 were detected both for seedlings and field resistance, MpV.1 and MpII.1 were specific for growth chamber conditions and MpIII.3 and MpIV.1 for field resistance. Quantitative trait loci MpIII.1, MpII.1, MpIII.2 and MpIII.3 may coincide with other QTLs associated with resistance to M. pinodes previously described in P.sativum. Four QTLs associated with earliness of flowering were also identified. While dfIII.2 and dfVI.1, may correspond with other genes and QTLs controlling earliness in P. sativum, dfIII.1 and dfII.1 may be specific to P. sativum subsp. syriacum. Flowering date and growth habit were strongly associated with resistance to M. pinodes in the field evaluations. The relation observed between earliness, growth habit and resistance to M. pinodes is discussed.

Identification of a candidate gene for the wheat endopeptidase Ep-D1 locus and two other STS markers linked to the eyespot resistance gene Pch1

Wheat is prone to strawbreaker foot rot (eyespot), a fungal disease caused by Oculimacula yallundae and O. acuformis. The most effective source of genetic resistance is Pch1, a gene derived from Aegilops ventricosa. The endopeptidase isozyme marker allele Ep-D1b, linked to Pch1, has been shown to be more effective for tracking resistance than DNA-based markers developed to date. Therefore, we sought to identify a candidate gene for Ep-D1 as a basis for a DNA-based marker. Comparative mapping suggested that the endopeptidase loci Ep-D1 (wheat), enp1 (maize), and Enp (rice) were orthologous. Since the product of the maize endopeptidase locus enp1 has been shown to exhibit biochemical properties similar to oligopeptidase B purified from E. coli, we reasoned that Ep-D1 may also encode an oligopeptidase B. Consistent with this hypothesis, a sequence-tagged-site (STS) marker, Xorw1, derived from an oligopeptidase B-encoding wheat expressed-sequence-tag (EST) showed complete linkage with Ep-D1 and Pch1 in a population of 254 recombinant inbred lines (RILs) derived from a cross between wheat cultivars Coda and Brundage. Two other STS markers, Xorw5 and Xorw6, and three microsatellite markers (Xwmc14, Xbarc97, and Xcfd175) were also completely linked to Pch1. On the other hand, Xwmc14, Xbarc97, and Xcfd175 showed recombination in the W7984 x Opata85 RIL population suggesting that recombination near Pch1 is reduced in the Coda/Brundage population. In a panel of 44 wheat varieties with known eyespot reactions, Xorw1, Xorw5, and Xorw6 were 100% accurate in predicting the presence or absence of Pch1 whereas Xwmc14, Xbarc97, and Xcfd175 were less effective. Thus, linkage mapping and a germplasm survey suggest that the STS markers identified here should be useful for indirect selection of Pch1.

A Genomewide Study of Reproductive Barriers Between Allopatric Populations of a Homosporous Fern,Ceratopteris richardii

Biological factors involved in reproductive barriers between two divergent races of Ceratopteris richardii were investigated. We used a combination of spore germination rates, QTL analysis of spore germination rates, and transmission ratio distortion (TRD) of 729 RFLPs, AFLPs, and isozyme markers distributed across the genome on the basis of hybrid populations of 488 doubled haploid lines (DHLs) and 168 F(2)'s. Substantial reproductive barriers were found between the parental races, predominantly in the form of spore inviability (23.7% F(1) spore viability). Intrinsic genetic factors such as Bateson-Dobzhansky-Muller (BDM) incompatibilities involving both nuclear-nuclear and nuclear-cytoplasmic factors and chromosomal rearrangements appear to contribute to intrinsic postzygotic isolation. The genomewide distribution patterns of TRD loci support the hypothesis that reproductive barriers are a byproduct of divergence in allopatry and that the strong reproductive barriers are attributable to a small number of genetic elements scattered throughout the genome.

Gene flow among wild and cultivated sunflower, Helianthus annuus in Argentina

Naturalized populations of wild sunflower, Helianthus annnus, occur in six provinces of central Argentina, usually near sunflower crops. Plants of intermediate morphology indicate that gene flow might take place in both senses. In order to quantify gene flow between cultivated and wild sunflower, an experimental stand of sunflower was sown surrounded by plots of wild plants at increasing distances. Hybridization rate was estimated using a crop specific isozyme marker, and a mean of 7% progenies were crop-wild hybrids. The nearest wild plants (3 m from the cultivar) showed the highest percentage (18%) of gene flow, which was found to decrease with distance, up to 500 m. Pollen flow from wild plants to crop, evaluated through morphological characters in the progenies of cultivated plants from a stand invaded by wild sunflowers, produced 3.75% intermediate plants.

Genetical assessment of diploid progenitors of S. scabra by isozyme, RAPD and STS markers: a possible strategy for improvement of drought tolerant allo-tetraploid S. scabra species

Assessment of genetic variations of two progenitors of S. scabra was investigated employing isozyme, RAPD and STS markers. Earlier reports have conclusively indicated S. seabrana and S. viscosa as two diploid progenitors of allo-tetraploid S. scabra. A total of 476 RAPD bands generated from 32 primers, 104 STS fragments from 17 primer pairs and 73 isoforms from 14 enzyme systems indicated significant sharing of these bands to S. scabra with that of S. seabrana in comparison to its other progenitor i.e., S. viscosa. High frequencies of isozyme band sharing between S. seabrana and S. scabra (0.125–0.30) as well as high genetic similarities (59%) of it with S. seabrana as reveled from dendrogram analysis indicated impact of close proximity of the occurrence of S. seabrana in the development of S. scabra. The proportion of bands shared by RAPD (40%) and STS (40%) markers were also observed high between S. seabrana and S. scabra. However, the numbers of unique bands were less in all these species. More correspondence and sharing of bands by S. seabrana towards S. scabra suggested using selected lines of S. seabrana the genetic base of allo-tetraploid, drought tolerant S. scabra lines can be broaden.

Isozyme markers associated with O 3 tolerance indicate shift in genetic structure of ponderosa and Jeffrey pine in Sequoia National Park, California

Effects of canopy ozone (O 3) exposure and signatures of genetic structure using isozyme markers associated with O 3 tolerance were analyzed in ∼20-, ∼80-, and >200-yr-old ponderosa ( Pinus ponderosa Dougl. ex Laws.) and Jeffrey pine ( Pinus jeffreyi Grev. & Balf.) in Sequoia National Park, California. For both species, the number of alleles and genotypes per loci was higher in parental trees relative to saplings. In ponderosa pine, the heterozygosity value increased, and the fixation index indicated reduction of homozygosity with increasing tree age class. The opposite tendencies were observed for Jeffrey pine. Utilizing canopy attributes known to be responsive to O 3 exposure, ponderosa pine was more symptomatic than Jeffrey pine, and saplings were more symptomatic than old growth trees. We suggest that these trends are related to differing sensitivity of the two species to O 3 exposure, and to higher O 3 exposures and drought stress that younger trees may have experienced during germination and establishment.

Linkage Map Construction and Quantitative Trait Loci Analysis for Bolting Based on a Double Haploid Population of Brassica rapa

AbstractEarly bolting of Chinese cabbage (Brassica rapa L.) during spring cultivation often has detrimental effects on the yield and quality of the harvested products. Breeding late bolting varieties is a major objective of Chinese cabbage breeding programs. In order to analyze the genetic basis of bolting traits, a genetic map of B. rapa was constructed based on amplified fragment‐length polymorphism (AFLP), sequence‐related amplified polymorphism (SRAP), simple sequence repeat (SSR), random amplification of polymorphic DNA (RAPD), and isozyme markers. Marker analysis was carried out on 81 double haploid (DH) lines obtained by microspore culture from F1 progeny of two homozygous parents: B. rapa L. ssp. pekinensis (BY) (an extra‐early bolting Chinese cabbage line) and B. rapa L. ssp. rapifera (MM) (an extra‐late bolting European turnip line). A total of 326 markers including 130 AFLPs, 123 SRAPs, 16 SSRs, 43 RAPDs and 14 isozymes were used to construct a linkage map with 10 linkage groups covering 882 cM with an average distance of 2.71 cM between loci. The bolting trait of each DH line was evaluated by the bolting index under controlled conditions. Quantitative trait loci (QTL) analysis was conducted using multiple QTL model mapping with MapQTL5.0 software. Eight QTLs controlling bolting resistance were identified. These QTLs, accounting for 14.1% to 25.2% of the phenotypic variation with positive additive effects, were distributed into three linkage groups. These results provide useful information for molecular marker‐assisted selection of late bolting traits in Chinese cabbage breeding programs.(Handling editor: Yong‐Biao Xue)

Genetic characterization of a collection of chayote, Sechium edule (Jacq.) Swartz, in Costa Rica by using isozyme markers

We established protocols for the analysis of genetic diversity in chayote (Sechium edule) by using isozyme markers, thereby determining the level of genetic diversity present in 42 accessions of chayote from Costa Rica. We obtained clear and reproducible zymograms for eight enzyme staining systems: PGM, 6-PGD, PGI, IDH, MDH, SOD, SKD, and EST, and were able to score 14 putative loci. Eight of the 14 loci examined were polymorphic. We found 35 distinct multilocus genotypes among these accessions. Five of these multilocus genotypes were homozygous for all loci. In addition, our data also revealed that most of the multilocus genotypes (24) were heterozygous for only one of the eight loci, and the rest were heterozygous for two or three loci (9 and 4 accessions, respectively). Seven multilocus genotypes were found in two different accessions. Dice similarity coefficient was used to study the relationship between accessions. This analysis, based on the presence and absence of alleles, revealed that accessions collected in the same location seldom shared the same multilocus genotype. The value of isozyme polymorphisms as tools to continue studies on the characterization of chayote is discussed.

Development of PCR-based markers from the tomato glutamate oxaloacetate transaminase isozyme gene family as a means of revitalising old isozyme markers and recruiting new ones

Sequences annotated as aspartate aminotransferases (synonymous with glutamate oxaloacetate transaminases) in the SOL Genomics Network unigene database were used to design 10 pairs of PCR primers for genetic marker development. These primer pairs generated nine CAPS markers, two SCAR markers and one SSR marker, which were bin-mapped using a set of tomato introgression lines (IL) derived from Lycopersicon esculentum cv. M82 and Lycopersicon pennellii LA716. Based on their bin locations, these markers are largely dispersed throughout the tomato genome and appear to have tagged all four of the glutamate oxaloacetate transaminase (Got) isozyme marker genes placed on the classical genetic map of tomato. Orthologous relationships with Arabidopsis aspartate aminotransferase (Asp) genes suggest the existence of at least two additional functional Got genes in tomato that have also been tagged by these markers and likewise an additional functional Asp gene in Arabidopsis. The Got-2 isozyme marker has often been used for the marker-assisted breeding of the I-3 gene for Fusarium wilt resistance introgressed from L. pennellii LA716. The Got-2 CAPS marker that we have developed offers a facile PCR-based alternative to the isozyme marker for the marker-assisted breeding of I-3. However, all of the PCR-based markers we have developed have the potential to assist the breeding of linked traits introgressed from wild relatives of tomato.

Genetic Diversity Analysis in Sugarbeet (Beta vulgaris L.) Using Isozymes, RAPD and ISSR Markers

Four elite diploid populations of sugarbeet viz. IISR Comp-1, LKC-11, LS-6 and Ramonskaya-06 were examined using isozymes and molecular markers. Seventy-one bands consisting of 28 isomorphs of 6 isozyme systems viz. superoxide dismutase, guaiacol peroxidase, malate dehydrogenase, amylase, esterase and aspartate amino transferase were resolved. Molecular analysis was performed using 28 RAPD (random amplification of polymorphic DNA) and 4 inter simple sequence repeat (ISSR) primers. Highly polymorphic band profiles with 327 RAPD markers (11.67/primer) of 162–2862 bp and 39 ISSR markers (9.75/primer) of 160–1884 bp were obtained. Mean value of Polymorphic Information Content (PIC) for RAPD and ISSR primers was 0.41 and 0.44 respectively. Based on the binary matrix of presence/absence of bands, Dice coefficient of genetic similarity (GS) matrix was computed for each marker system, which ranged from 0.62–0.76 (mean value 0.726), 0.10–0.74 (mean value 0.27) and 0.12–0.86 (mean value 0.35) for isozyme, RAPD and ISSR markers. There was significant correlation (r=0.908) between the RAPD-GS and ISSR-GS matrices, however, the correlation of isozyme matrix vs. RAPD and ISSR pattern was low. GS coefficients of isozyme, RAPD and ISSR were used to cluster the genotypes based on the UPGMA method and the dendrograms obtained were compared to develop consensus phylogenetic trees using NTSYSpc. The X-axis contained IISR Comp-1 and R-06, and the Y-axis contained LS-6 and LKC-11. The much lower values of average genetic similarity based on RAPD (0.27) and ISSR (0.35) indicated the ability of DNA based markers to detect high degree of polymorphism among these populations suggesting thereby, the possibility of screening a higher number of anonymous loci in sugarbeet using these molecular markers as compared to isozymes to enable the selection of the best parents in order to obtain new genetic combinations.

Genetic Differentiation of Rice and Non‐rice Populations of Magnaporthe grisea from North‐western Himalayas using Native Protein and Isozyme Polymorphisms

AbstractNative protein and isozyme markers were used to analyse the population structure of rice, finger millet (Eleusine coracana), jungle rice (Echinochloa colonum), goosegrass (Eleusine indica) and crabgrass (Digitaria sanguinalis) infecting isolates of Magnaporthe grisea from the north‐western Himalayan region of India. Both protein and isozyme analyses revealed a high level of genetic diversity among different host‐limited populations of the pathogen including those infecting rice. Cluster analysis of protein and isozyme data permitted the grouping of isolates in accordance with their host specificity. A close association was observed between specific esterase and shikimate dehydrogenase (SDH) isozyme electromorphs and the host specificity of the isolates, indicating that these electromorphs might be linked to the genes conditioning host specialization. Cluster analysis indicated the presence of two lineages in M. grisea species complex: first lineage with moderate bootstrap support was constituted by the isolates from rice, finger millet, goosegrass and jungle rice and a second statistically robust lineage was represented by the isolates from crabgrass. Subpopulations of the pathogen attacking rice and weeds in the same field were genetically distinct and the isolates from different hosts never exhibited a same multilocus haplotype. The overall results indicated that M. grisea populations infecting different hosts were genetically isolated and there was no gene flow among rice and non‐rice isolates of the pathogen.

Genetic map-based analysis of genome structure in the homosporous fern Ceratopteris richardii.

Homosporous ferns have extremely high chromosome numbers relative to flowering plants, but the species with the lowest chromosome numbers show gene expression patterns typical of diploid organisms, suggesting that they may be diploidized ancient polyploids. To investigate the role of polyploidy in fern genome evolution, and to provide permanent genetic resources for this neglected group, we constructed a high-resolution genetic linkage map of the homosporous fern model species, Ceratopteris richardii (n = 39). Linkage map construction employed 488 doubled haploid lines (DHLs) that were genotyped for 368 RFLP, 358 AFLP, and 3 isozyme markers. Forty-one linkage groups were recovered, with average spacing between markers of 3.18 cM. Most loci (approximately 76%) are duplicated and most duplicates occur on different linkage groups, indicating that as in other eukaryotic genomes, gene duplication plays a prominent role in shaping the architecture of fern genomes. Although past polyploidization is a potential mechanism for the observed abundance of gene duplicates, a wide range in the number of gene duplicates as well as the absence of large syntenic regions consisting of duplicated gene copies implies that small-scale duplications may be the primary mode of gene duplication in C. richardii. Alternatively, evidence of past polyploidization(s) may be masked by extensive chromosomal rearrangements as well as smaller-scale duplications and deletions following polyploidization(s).

A new QTL for Ascochyta blight resistance in an RIL population derived from an interspecific cross in chickpea

A linkage map in a population of recombinant inbred lines (RILs) derived from an interspecific cross between Cicer arietinum (ILC72) × Cicer reticulatum (Cr5-10), resistant and susceptible to blight, caused by Ascochyta rabiei, respectively, was obtained using RAPD, ISSR, STMS, isozyme (Pdf6) and flower colour (pink/white) markers. The map comprised ten linkage groups and covered a distance of 601.2 cM. When the population was evaluated for reaction to Ascochyta blight under field conditions by determining the Area Under the Disease Progress Curve (AUDPC), the distribution of frequencies was bimodal: most of the lines had an intermediate reaction, fewer were nearly as susceptible as the susceptible parent and none had values close to the resistant parent. A QTL explaining 28% of the variation in resistance was located in linkage group 2 (LG2). Five RAPD markers on this linkage group showed significant association with resistance (OPX04372, UBC881621, OPAI09746, OPAI09352 and OPAC12700) and the major QTL peak lay midway between OPAI09746 and UBC881621 which are 14.1 cM apart. Contrary to other studies, no association of linkage group 4 with resistance was found. The QTL for resistance to Ascochyta blight in this study is therefore different from QTLs for this character reported in other interspecific crosses and may be the same as that reported in linkage group 2 in intraspecific crosses where genes for resistance to races of Fusarium oxysporum f. sp. ciceri, causing wilt, are also located.

Commercial production of Cordyline terminalis (L) Kunth. from shoot apex meristem and assessment for genetic stability of somaclones by isozyme markers

Rapid multiplication of Cordyline terminalis (L) Kunth. was achieved from shoot apex explant on Murashige and Skoog's (MS) medium supplemented with 3% (w/v) sucrose and different concentration of growth regulators at various combinations. On MS medium supplemented with BA in combination with Adenine sulphate and IAA, shoot initiation and multiplication were obtained. Best elongations of shoots were found on 1/2 MS basal medium and shoots were rooted on 1/2 MS medium supplemented with IBA. Rooted plants passed through a hardening phase prior to ex vitro transfer. Clonal propagated plants established in garden soil were uniform and identical to mother plant in respect to growth characteristics and morphology. Isozymic profiles of different micropropagated clones were assessed for their genetic stability. Ten clones were tested for six isozymes. Only a few showed variation with respect to the banding pattern in esterase and superoxide dismutase. In superoxide dismutase, the two polymorphic isoforms (Rf 0.06 and 0.45) appeared in the clone C8 of the plants transferred to the field after 15 subcultural passages. Mobility and intensity of bands were monitored in other isozymes. Isozyme markers may be used as a tool for rapid screening of genetic stability in tissue cultured clones of C. terminalis.

Mating System in a Low-density Natural Population of the Dioecious Wind-pollinated Patagonian Cypress

The mating system of a species is an essential factor that determines the genotypic frequencies of its populations in each generation. Thus, knowledge in this regard is important in the application of practical measures in conservation and forest tree breeding. Austrocedrus chilensis (D.DON) PIC. SER. ET BIZZARRI: is a member of the Cupressaceae native to the Andean-Patagonian Forest of Argentina and Chile with regional economic importance. It is a dioecious tree with pollen and seeds distributed by wind. Isozyme markers were utilized to study the mating system of this species through the survey of individual effective pollen clouds in a natural low-density population. If random mating occurs, effective pollen clouds that pollinate single seed trees are expected to be genetically homogeneous. The classical HardyWeinberg test of panmixia was also performed. Although it failed to reject random mating, the analysis of the pollen clouds showed that this mating pattern does not occur. Evidence of locally distributed heterogeneous pollen clouds is presented. The two methods are critically compared.

Direct comparison between genomic constitution and flavonoid contents in Allium multiple alien addition lines reveals chromosomal locations of genes related to biosynthesis from dihydrokaempferol to quercetin glucosides in scaly leaf of shallot (Allium cepa L.)

The extrachromosome 5A of shallot (Allium cepa L., genomes AA) has an important role in flavonoid biosynthesis in the scaly leaf of Allium fistulosum-shallot monosomic addition lines (FF+nA). This study deals with the production and biochemical characterisation of A. fistulosum-shallot multiple alien addition lines carrying at least 5A to determine the chromosomal locations of genes for quercetin formation. The multiple alien additions were selected from the crossing between allotriploid FFA (female symbol) and A. fistulosum (male symbol). The 113 plants obtained from this cross were analysed by a chromosome 5A-specific PGI isozyme marker of shallot. Thirty plants were preliminarily selected for an alien addition carrying 5A. The chromosome numbers of the 30 plants varied from 18 to 23. The other extrachromosomes in 19 plants were completely identified by using seven other chromosome markers of shallot. High-performance liquid chromatography analyses of the 19 multiple additions were conducted to identify the flavonoid compounds produced in the scaly leaves. Direct comparisons between the chromosomal constitution and the flavonoid contents of the multiple alien additions revealed that a flavonoid 3'-hydroxylase (F3'H) gene for the synthesis of quercetin from kaempferol was located on 7A and that an anonymous gene involved in the glucosidation of quercetin was on 3A or 4A. As a result of supplemental SCAR analyses by using genomic DNAs from two complete sets of A. fistulosum-shallot monosomic additions, we have assigned F3'H to 7A and flavonol synthase to 4A.

Isozyme variation and the conservation genetics of Garry oak

Garry oak ( Quercus garryana Dougl. ex Hook) has a long north–south distribution along the inland Pacific coast. In British Columbia, it is a keystone species in a unique and endangered “Garry oak meadow” ecosystem. Here, we apply isozyme markers to address issues in the conservation and phylogeography of Garry oak. Among 42 populations and 23 gene loci, gene diversity (expected heterozygosity) averaged 0.17, and number of alleles per locus averaged 1.84. These values are about one-half of those found in other white oak species. Using progeny arrays, we found outcrossing rates in two Vancouver Island populations to average 0.96, with no detectable biparental inbreeding; also inbreeding coefficients of the 42 populations averaged near zero (0.025); thus inbreeding is not a significant concern. Cluster analysis of genetic distances identified two major groups of populations: southern Washington – Oregon and Vancouver Island – Gulf Islands; populations within the island region were particularly homogenous. An isolated mainland British Columbia population near Sumas, British Columbia, perhaps of anthropogenic origin, showed the least genetic variability and greatest genetic distance. Generally, geographically isolated populations were more genetically depauperate, which may place handicaps on their survival, but also more genetically distinct, providing a greater evolutionary legacy to the species.

Evaluation of AFLP markers to reveal genetic diversity in Typha

We investigated to what extent DNA-markers can assist species determination in the genus Typha. A set of AFLP markers was used to discriminate samples of the species Typha latifolia and Typha angustifolia collected in Flanders (North Belgium). The T. latifolia samples formed a compact cluster while the T. angustifolia samples were divided into smaller groups. It was not clear whether interspecific hybrids or higher levels of diversity present in the T. angustifolia dataset could account for this. As in previous surveys, using isozyme and VNTR markers, AFLP markers revealed an almost complete lack of genetic variation in Flemish T. latifolia. Despite the low degree of diversity, a significant level of genetic differentiation was found between the T. latifolia samples originating from different river basins. Whether this differentiation has any ecological relevance remains to be investigated. The methodology applied was not able to detect clonal reproduction in T. latifolia. Probably, the low levels of diversity present in this species can account for this, indicating that the usefulness of the methodology applied depends on the level of diversity present in the species studied.

Isozymic variability in a Brazilian collection of annatto (Bixa orellana L.)

The objectives of this work were to optimize the isozyme electrophoresis technique for Bixa orellana, and use isozyme markers for a preliminary survey on the genetic variability in Brazilian annatto germplasm accessions. Collection consisted of seed samples from sixty open pollinated trees, representing two Northern and four Southern geographic provenances. The extraction, electrophoresis, and interpretation of annatto isozymes are described. Three out of the twenty-one identified isozyme loci were polymorphic in the collection. The percentage of polymorphic loci (P = 21.05) and the expected heterozygosity in annatto (H T = 0.064) were low, compared to other tropical woody species. A UPGMA phenogram, constructed with Nei's genetic distances, clearly separated the germplasm provenant from North and Central Brazil. Variability was significantly higher among the accessions from Maranhão. A sharp genetic differentiation was detected between accessions from Maranhão and Pará States, despite their geographical proximity. The distinctive isozyme polymorphism, observed in the accessions from Maranhão, together with reports on local morphological heterogeneity in annatto fruit shape, color, and pubescence, calls for more detailed genetic and taxonomic investigation.

Genetic variability is correlated with population size and reproduction in American wild‐rice (Zizania palustrisvar.palustris, Poaceae) populations

American wild-rice (Zizania palustris var. palustris) has served as a staple for indigenous North Americans for thousands of years, but has had significant habitat losses in recent centuries. We investigated genetic variability among 17 wild-rice populations in northern Wisconsin using 13 isozyme markers. We then compared these genetic patterns to differences in habitat and population characteristics and phenotypic variation in plant growth and reproduction across sites. Wild-rice's mean genetic diversity (0.15) is moderate compared to wind-pollinated outcrossers but lower than the mean (0.20) reported for the Poaceae. Estimated inbreeding coefficients within populations (f) average 0.12 but vary greatly among the populations (from -0.44-0.52), suggesting heterogeneous population histories. Larger populations in larger lakes express higher levels of genetic variability and smaller inbreeding coefficients than smaller or more isolated populations. The number of panicles per plant is also higher in populations with greater genetic variability. Estimated genetic differentiation among the 17 populations (F(ST)) was high (0.30), suggesting limited gene flow among drainages. Wild-rice population size and degree of isolation have opposing effects on its genetic variability, and plant performance is positively associated with genetic variability.