Patterns of isozyme variation between maize and Mexican annual teosinte

Isozyme variation in 94 accessions of Mexican maize (Zea mays ssp. mays) and 37 collections of Mexican annual teosinte (Z. mays ssp. mexicana and var. parviglumis) are compared. Variety parviglumis (a predominantly wild plant) shows a closer genetic relationship to maize than does ssp. mexicana (a weedy teosinte often found in maize fields). The isozyme data suggest that maize and Z. mays var. parviglumis share a more recent common ancestor than either of these taxa share with other members of the genus Zea. In this sense, the isozyme data support the theory that maize is a domesticated form of teosinte. Isozyme data provide no evidence for independent origin of Mexican maize races from different taxa of teosinte. Isozyme analysis suggests that gene flow between maize and ssp. mexicana exists, but that it is highly restricted and more probably goes from weed into crop. Maize and var. parviglumis are isozymically too similar and too variable to allow patterns of gene flow between them (if any) to be discerned. The maize- teosinte complex does not fit a model applied to some other crops in that (I) weedy teosinte (ssp. mexicana) does not appear to be a hybrid of the wild form (var. parviglumis,) and maize and (2) the weedy form does not act as a genetic bridge between wild form and crop.     

Biochemical systematics and evolution ofzea,tripsacum and related genera

Antibody systems were produced in rabbits by immunization with antigen systems from seed ofZea mays, Z. mexicana andTripsacum dactyloides. Antigen systems were generally ranked in the following order using Zeaantibody systems: Zea > Tripsacum > Elyonurus ≈ Bothriochloa > Coix ≈ Manisuris > Andro-pogon ? Triticum. Tripsacum antibody systems also ranked these antigen systems in a similar order except that Tripsacum > Zea, and Manisuris ≈ Bothriochloa ≈ Elyonurus. Some Tripsacum species produced anomalous results. Serology suggests that Zea and Tripsacum should be placed together in the subtribe Tripsacinae of the tribe Andropogoneae: the tribe Maydeae is probably an unnatural assemblage. Tripsacum and Zea have probably evolved from ancestors with affinities to the subtribes Rottboellinae and Bothriochloeae of the tribe Andropogoneae. The high degree of serological correspondence shown by Elyonurus to Zea suggests a close common ancestry, but Manisuris appears no more similar to Tripsacum than do other genera of the Rottboellinae and Bothriochloeae. By polyacrylamide gel electrophoresis, no differences were found between maize and teosinte from Mexico and north Guatemala. Teosinte from south Guatemala consistently lacked bands present in both maize and Mexican teosinte but shared no greater similarity to Tripsacum and cannot therefore be considered as tripsacoid. The high degree of band homology between maize and Mexican teosinte supports a parental relationship and it is suggested that Mexican teosinte represents the germ plasm from which maize was domesticated. Neither electro-phoretic nor serological results supported the hypothesized hybrid (viz. Zea x Manisuris) origin of Tripsacum.     

Variation within teosinte. III. numerical analysis of allozyme data

Principal components analysis of isozyme allele frequencies at 19 loci revealed 133 electrophoretic variants for 77 accessions of annual teosinte and 1 accession each of diploid and tetraploid perennial teosintes. The majority of alleles were found in low frequency, and many were distributed only in specific locations. Zea luxuriansand the annual Mexican teosintes appeared to be the most distantly related of all teosintes. Z. perennisand Z. diploperennisappeared distinct from each other and from all other teosintes. Teosintes of west Guatemala (Huehuetenango) did not appear especially closely related to Balsas teosintes. Several differences were apparent between Chalco and Central Plateau teosintes;however, these differences were not so extreme as those suggested by chromosome knob data. Nobogame teosintes appeared closely related to Chalco and Central Plateau teosintes. Isozyme data reveal teosinte to be a diverse source of germplasm.     

Variation within teosinte. II. numerical analysis of chromosome knob data

Principal components analysis of 46 chromosome knob positions and B chromosomes for 61 accessions of annual teosinte (in 12 groups) and 207 accessions of maize (in 87 groups) produced results which did not entirely agree with previous groupings. Teosintes of northwest and southeast Guatemala (races Huehuetenango and Guatemala) were widely separated from maize and from all other teosintes; however, teosintes of east Mexico-Distrito Federal (race Chalco) were also widely separated from maize and other teosintes. Teosintes of south Chihuahua (race Nobogame) grouped separately from other teosintes, suggesting that race Nobogame is not simply a northern extreme of race Central Plateau. Teosintes of east Michoacan and west Mexico (recently grouped into the Balsas race) grouped together but separately from other teosintes. The latter teosintes may merit special designation in a racial taxonomic system. Only the Central Plateau teosintes consistently overlapped with maize OTU's for the first few principal components. Even in that case there was no consistent association between the Central Plateau teosintes and any specific race of maize.     

Relationships between maize and teosinte of Mexico and Guatemala: Numerical analysis of allozyme data

Electrophoresis revealed 153 variants at 19 enzyme loci among 77 accessions of annual teosinte, 1 accession of diploid perennial teosinte, 1 accession of tetraploid perennial teosinte, and 43 accessions of maize from Mexico and Guatemala. One hundred and thirty-three (87%) variants were found within teosinte, and 95 (61%) variants were found within maize. Seventy-four (48%) variants were distributed among both teosinte and maize, whereas 58 (38%) variants were found only in teosinte and 20 (13%) variants were found only in maize. The majority of alleles were found in low frequency. A single allele predominated across maize and teosinte at 11 loci, 3 loci showed similar trends and 5 loci showed greater variability for the predominant alleles among maize and teosinte. Principal components analysis revealed little overlap between maize and teosinte, involving a minority of maize collections and Balsas teosintes. No clear evidence could be seen suggesting recent introgression between maize and teosinte. The collections of maize and teosinte were seen to represent a rich and contrasted array of germplasm.     

Molecular Evidence and the Evolution of Maize

In this review, the contributions of isozyme and chloroplast DNA studies to questions surrounding the evolution of maize are summarized. These methods of analysis provide generally strong support for the hierarchical system of classification of Zea proposed by Iltis and Doebley (1980). Molecular evidence is fully congruent with the theory that teosinte is ancestral to maize and suggests thatZ. mays subsp.parviglumis was the ancestral teosinte taxon. Further, these data show that only those populations from the central region of the range of subsp. parviglumis resemble maize in both isozymic and chloroplast DNA constitution. Presuming no major changes in the distribution of subsp. parviglumis since the domestication of maize, these data would place the origin of maize in the Balsas River drainage southwest of Mexico City. Molecular systematic evidence provides no support for theories that maize was domesticated independently several times; however, this type of data can not disprove such theories. Analyses of isozyme and chloroplast DNA diversity in Zea provide evidence of limited gene flow between maize and teosinte, but are not consistent with models that postulate extensive genetic interchange between these taxa. Isozyme studies have added substantially to the understanding of evolutionary relationships among extant races of maize and suggest that there are a small number of major racial complexes in Meso- and North America which have often evolved in response to environmental constraints associated with altitude. Ultimately, molecular genetic studies may allow a resolution of the controversy surrounding the morphological evolution of the maize ear.     

<Emphasis Type="Italic">Mutator</Emphasis>-Based Transposon Display: A Genetic Tool for Evolutionary and Crop-Improvement Studies in Maize

Transposable elements account for up to 85% of the maize genome and have significant implications in crop-improvement and evolutionary analyses. The Mutator (Mu) transposon superfamily, a class of DNA transposons, comprises the most complex and active elements in the maize genome, suggesting a special role in plant evolution. Here, we designed a set of Mu-specific primers based on terminal invert repeats and used a transposon display (TD) method for genotyping. We analyzed the distribution pattern of Mu insertions in teosinte (wild relative), sorghum (distant relative), and domesticated maize accessions (dent, sweet, and waxy). The MU-TD analysis suggested the presence of high polymorphic insertions among the species and subspecies, indicating the utility of the method in studying genetic variation and species relationships. Furthermore, we analyzed 80 maize recombinant inbred line populations. Mu-TD generated an average of 60% Mu-anchored polymorphic fragments in which insertions appeared to be segregating in significantly high numbers. The amplification profile was highly reproducible, confirming the utility of Mu elements as a new set of TD markers for developing high-density genetic maps.     

Relationship between constitutive root aerenchyma formation and flooding tolerance in Zea nicaraguensis

Background and aims

The teosinte Zea nicaraguensis, which is adapted to frequently flooded lowlands, is considered a valuable germplasm resource for the development of flooding-tolerant maize. This species can form constitutive root aerenchyma under well-drained conditions. The objectives of this study were to screen Z. nicaraguensis accessions for the capacity to form constitutive aerenchyma, to obtain progeny with differing degrees of aerenchyma formation, and to compare the flooding tolerance of these progeny.

Methods

We evaluated constitutive aerenchyma formation in the root cortex of seedlings of eight accessions and several segregating populations of Z. nicaraguensis. We also evaluated flooding tolerance in lines selected for high or low degrees of constitutive aerenchyma formation.

Results

Seedlings of the eight accessions showed an extremely wide and continuous range of variation in aerenchyma formation. By phenotypic selection within two accessions, we obtained lines with either high or low degrees of constitutive aerenchyma formation. The lines selected for a higher degree of formation showed relatively high flooding tolerance evaluated by shoot dry weight ratio (flooded:control) than those with a lower degree of formation.

Conclusions

A greater capacity to form constitutive aerenchyma can enhance flooding tolerance.     

Collection and evaluation of pearl millet (Pennisetum) germplasm from Malawi

A germplasm collecting trip to Malawi was launched during March/April 1979, resulting in the collection of 260 traditional cultivars of pearl millet (Pennisetum americanum), 11 intermediate forms, and 6 accessions of wild Pennisetum. Most of the cultivated pearl millet samples were obtained from the hot lowlands of the lower Shire Valley, with a few samples from the cool highlands of Mulanje and Mangochi. A mixture of different types that varied in plant height, maturity, and spike characters were observed in farmers’ fields. In the south, early types with loose, thin, cylindrical spikes and in the north, late-maturing types producing many tillers with stout spikes having long bristles were found. The grain is used to prepare a thick porridge, nsima, or to brew local beer, chimera. When the collection was evaluated at ICRISAT Center, Patancheru, considerable variation was observed for days to 50% flowering and plant height, but not for spike and grain characters. During the rainy season, the majority of the accessions flowered early (70 days), grew very tall (250 cm), and produced thin (22 mm), short (22 cm) spikes with small, obovate to elliptical, corneous grey grain. In the postrainy season, most of the accessions flowered a week earlier accompanied by reduction in plant height. Millet germplasm from Malawi belongs to the race typhoides and serves as a good source of genes for earliness, tillering, and corneous endosperm.     

Molecular phylogeny of the neotropical genus Christensonella (Orchidaceae, Maxillariinae): species delimitation and insights into chromosome evolution

Background and Aims

Species'' boundaries applied within Christensonella have varied due to the continuous pattern of variation and mosaic distribution of diagnostic characters. The main goals of this study were to revise the species'' delimitation and propose a more stable classification for this genus. In order to achieve these aims phylogenetic relationships were inferred using DNA sequence data and cytological diversity within Christensonella was examined based on chromosome counts and heterochromatin patterns. The results presented describe sets of diagnostic morphological characters that can be used for species'' identification.

Methods

Phylogenetic studies were based on sequence data of nuclear and plastid regions, analysed using maximum parsimony and maximum likelihood criteria. Cytogenetic observations of mitotic cells were conducted using CMA and DAPI fluorochromes.

Key Results

Six of 21 currently accepted species were recovered. The results also support recognition of the ‘C. pumila’ clade as a single species. Molecular phylogenetic relationships within the ‘C. acicularis–C. madida’ and ‘C. ferdinandiana–C. neowiedii’ species'' complexes were not resolved and require further study. Deeper relationships were incongruent between plastid and nuclear trees, but with no strong bootstrap support for either, except for the position of C. vernicosa. Cytogenetic data indicated chromosome numbers of 2n = 36, 38 and 76, and with substantial variation in the presence and location of CMA/DAPI heterochromatin bands.

Conclusions

The recognition of ten species of Christensonella is proposed according to the molecular and cytogenetic patterns observed. In addition, diagnostic morphological characters are presented for each recognized species. Banding patterns and chromosome counts suggest the occurrence of centric fusion/fission events, especially for C. ferdinandiana. The results suggest that 2n = 36 karyotypes evolved from 2n = 38 through descendent dysploidy. Patterns of heterochromatin distribution and other karyotypic data proved to be a valuable source of information to understand evolutionary patterns within Maxillariinae orchids.Key words: Chromosome number, Christensonella, Cymbidieae, cytotaxonomy, fluorochrome staining, Maxillaria, Maxillariinae, molecular phylogenetics, species delimitation     

The genetic architecture of complex traits in teosinte (Zea mays ssp. parviglumis): new evidence from association mapping

Previous association analyses showed that variation at major regulatory genes contributes to standing variation for complex traits in Balsas teosinte, the progenitor of maize. This study expands our previous association mapping effort in teosinte by testing 123 markers in 52 candidate genes for association with 31 traits in a population of 817 individuals. Thirty-three significant associations for markers from 15 candidate genes and 10 traits survive correction for multiple testing. Our analyses suggest several new putative causative relationships between specific genes and trait variation in teosinte. For example, two ramosa genes (ra1 and ra2) associate with ear structure, and the MADS-box gene, zagl1, associates with ear shattering. Since zagl1 was previously shown to be a target of selection during maize domestication, we suggest that this gene was under selection for its effect on the loss of ear shattering, a key domestication trait. All observed effects were relatively small in terms of the percentage of phenotypic variation explained (<10%). We also detected several epistatic interactions between markers in the same gene that associate with the same trait. Candidate-gene-based association mapping appears to be a promising method for investigating the inheritance of complex traits in teosinte.     

The origin of cornbelt maize: The isozyme evidence

Historical records show t hat the Midwestern dent corns of the United States originated from hybridization of two landraces, Northern Flint and Southern Dent. We examined the origin of Southern and Midwestern Dents by means of isozyme electrophoresis. Isozyme genotypes were determined for 23 loci in 12 plants each of 32 accessions of Southern Dent. Previously published isozyme data for maize landraces of Mexico and North America and for U.S. Midwestern Dents were included for comparative purposes. The data show that Northern Flint and Southern Dent are among the isozymically most divergent maize landraces. Nei’s genetic identities between populations of these two landraces are very low for conspecific populations (ca. 0.80). Southern Dent of the southeastern U.S. appears closely related to similar dent corns of southern Mexico, supporting a previously published hypothesis that U.S. Southern Dent is largely derived from the dent corns of southern Mexico. The Midwestern Dents, which resulted from crosses of Southern Dent and Northern Flint, are much more like Southern Dent than Northern Flint in their isozyme profile. Similarly, public inbreds show greater affinity to Southern Dent with the exception of sweet corn lines, which resemble Northern Flint in their isozyme allele frequencies. North American public inbreds do not contain appreciable isozymic variation beyond that found in Northern Flint and Southern Dent.     

Karyological studies of Iranian <Emphasis Type="Italic">Allium</Emphasis> L. (Amaryllidaceae) species with focus on sect. <Emphasis Type="Italic">Acanthoprason</Emphasis>. 1. Mitotic chromosomes

Eighteen species and subspecies (34 accessions) of Allium sect. Acanthoprason and 11 species (17 accessions) belonging to other subgenera and sections of Allium were karyologically investigated and include first reports for 12 species. The examined plants of 47 accessions were diploid, three accessions of two species were tetraploid, and in the A. bisotunense accession, we found a mix of di- and triploid individuals. B chromosomes were found in 10 accessions. A basic chromosome number of x = 8 was confirmed for all investigated members of subg. Melanocrommyum and subg. Allium, and x = 9 for Allium tripedale of subg. Nectaroscordum. Idiograms were drawn for each accession, and metaphase images are presented illustrating observed chromosomal variations. Also, karyotype features and asymmetry parameters were calculated for all accessions. Chromosomal aberrations, e.g. aneuploid cells or loss of whole or parts of chromosome arms, were rarely observed. In general, the karyotypes showed low variation in inter- and intrachromosomal asymmetry especially inside of the taxonomic groups, though satellited chromosomes were good markers for subgenera and even specific for two studied sections of subg. Allium. Six different types of satellites were recognized, two of them were newly described: Type P was prevalent in subg. Melanocrommyum, and type O in sect. Codonoprasum. Statistical analyses were performed on five karyological parameters to test correct relationships and also to test previous grouping hypotheses. Although our data confirm distinct karyological characters for the subgenera investigated, the remarkable morphological diversity inside of subg. Melanocrommyum is not mirrored by striking karyological differences.     

Fine mapping of a quantitative resistance gene for gray leaf spot of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) derived from teosinte (<Emphasis Type="Italic">Z. mays</Emphasis> ssp. <Emphasis Type="Italic">parviglumis</Emphasis>)

Key message

In this study we mapped the QTL Qgls8 for gray leaf spot (GLS) resistance in maize to a ~130 kb region on chromosome 8 including five predicted genes.

Abstract

In previous work, using near isogenic line (NIL) populations in which segments of the teosinte (Zea mays ssp. parviglumis) genome had been introgressed into the background of the maize line B73, we had identified a QTL on chromosome 8, here called Qgls8, for gray leaf spot (GLS) resistance. We identified alternate teosinte alleles at this QTL, one conferring increased GLS resistance and one increased susceptibility relative to the B73 allele. Using segregating populations derived from NIL parents carrying these contrasting alleles, we were able to delimit the QTL region to a ~130 kb (based on the B73 genome) which encompassed five predicted genes.

     

Physiological and molecular analysis of aluminum tolerance in selected Kenyan maize lines

Aims

Aluminum (Al) toxicity is an important limitation to maize production in many tropical and sub-tropical acid soil areas. The aim of this study was to survey the variation in Al tolerance in a panel of maize lines adapted for Kenya and look for novel sources of Al tolerance.

Methods

112 Kenyan maize accessions were phenotyped for Al tolerance in solution culture. Several Al tolerance-related parameters including relative net root growth (RNRG), root apex Al accumulation, Al-activated root organic acid exudation, and expression of the maize Al tolerance gene, ZmMATE1, were used to classify Kenyan maize accessions.

Results

Based on RNRG, 42 %, 28 %, and 30 % of the lines were classified as highly tolerant, moderately tolerant and sensitive, respectively. Tolerant accessions accumulated less Al in their root apices compared to sensitive lines. The Kenyan maize line, CON 5, and the Brazilian standard for tolerance, Cateto, exhibited the greatest Al tolerance based on RNRG, but CON 5 had only about 50 % of ZmMATE1 gene expression relative to Cateto. CON 5 also had low root apex Al content and high citrate exudation, suggesting that it may employ a citrate transporter other than ZmMATE1.

Conclusions

We identified a very Al tolerant Kenyan maize line whose Al tolerance may be based in part on a novel tolerance gene. The maize lines identified in this study are useful germplasm for the development of varieties suitable for agriculture on acid soils in Kenya.

     

5.8S rDNA variability in Allium species belonging to the third evolutionary group

Sequence analysis of 5.8S rDNA in 67 accessions of the subgenus Allium and six other subgenera belonging to the third evolutionary group of Allium genus (Friesen et al., 2006) was performed. Nucleotide substitutions in 5.8S rDNA sequences of Allium accessions were identified and studied for the first time. The probable secondary structure of 5.8S rRNA was constructed. It was shown that mutations in 5.8S rDNA do not involve conserved motifs, and they did not significantly affect the secondary structure of the RNA molecule in Allium accessions.     

Using Association Mapping in Teosinte to Investigate the Function of Maize Selection-Candidate Genes

Background

Large-scale screens of the maize genome identified 48 genes that show the putative signature of artificial selection during maize domestication or improvement. These selection-candidate genes may act as quantitative trait loci (QTL) that control the phenotypic differences between maize and its progenitor, teosinte. The selection-candidate genes appear to be located closer in the genome to domestication QTL than expected by chance.

Methods and Findings

As a step toward defining the traits controlled by these genes, we performed phenotype-genotype association mapping in teosinte for 32 of the 48 plus three other selection-candidate genes. Our analyses assayed 32 phenotypic traits, many of which were altered during maize domestication or improvement. We observed several significant associations between SNPs in the selection-candidate genes and trait variation in teosinte. These included two associations that surpassed the Bonferroni correction and five instances where a gene significantly associated with the same trait in both of our association mapping panels. Despite these significant associations, when compared as a group the selection-candidate genes performed no better than randomly chosen genes.

Conclusions

Our results suggest association analyses can be helpful for identifying traits under the control of selection-candidate genes. Indeed, we present evidence for new functions for several selection-candidate genes. However, with the current set of selection-candidate genes and our association mapping strategy, we found very few significant associations overall and no more than we would have found with randomly chosen genes. We discuss possible reasons that a large number of significant genotype-phenotype associations were not discovered.     

Campanula xylocarpa — A new species of the seriesSaxicolae Witasek

In this paper, a newCampanula species is described from Eastern Slovakia. Its morphological, distributional and cytological relationships and ecological preferences are briefly discussed and affinity to the otherSaxicolae examined. The taxon is characterized by the following features: specific combination of constant morphological characters; diploid chromosome number (2n = 34); ecological specialization to the fissures of limestone and dolomitic rocks; relict occurrence in a small, sharply limited area; very late flowering. Geographically, it appears to be the northernmost representative of theSaxicolae.