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.     

Teosinte inflorescence phytolith assemblages mirror Zea taxonomy

Molecular DNA analyses of the New World grass (Poaceae) genus Zea, comprising five species, has resolved taxonomic issues including the most likely teosinte progenitor (Zea mays ssp. parviglumis) of maize (Zea mays ssp. mays). However, archaeologically, little is known about the use of teosinte by humans both prior to and after the domestication of maize. One potential line of evidence to explore these relationships is opaline phytoliths produced in teosinte fruit cases. Here we use multidimensional scaling and multiple discriminant analyses to determine if rondel phytolith assemblages from teosinte fruitcases reflect teosinte taxonomy. Our results indicate that rondel phytolith assemblages from the various taxa, including subspecies, can be statistically discriminated. This indicates that it will be possible to investigate the archaeological histories of teosinte use pending the recovery of appropriate samples.     

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.     

Spontaneous hybridization between maize and teosinte

The closest wild relatives of maize, Zea mays ssp. mays are various Zea taxa known as "teosinte." Hybrids between maize and the teosinte taxon, Zea mays ssp. mexicana, often occur when the 2 are sympatric in Mexico. Measuring the spontaneous hybridization rate of the 2 taxa would shed light on the mechanisms contributing to the evolution and persistence of these hybrid swarms. We conducted a series of field experiments in Riverside, CA, to measure the natural hybridization rates between maize and 2 teosinte taxa, Z. m. ssp. mexicana and Zea mays ssp. parviglumis. We planted teosinte within and near maize plantations. Hybrids were identified by progeny testing for a maize-specific herbicide resistance allele and a teosinte-specific allozyme allele. Hybridity was confirmed by growing putative hybrid progeny to maturity to evaluate whether they had the characteristic morphology of maize x teosinte hybrids. We found that maize and Z. m. ssp. mexicana naturally hybridize at a low rate (<1%), whereas Z. m. ssp. parviglumis hybridizes with the crop at a high rate (>50%).     

GISHGenomic in situ hybridization reveals cryptic genetic differences between maize and its putative wild progenitor Zea mays subsp. parviglumis.

The aim of this paper is to test with genomic in situ hybridization the genomic affinities between maize and its putative progenitor Zea mays subsp. parviglumis. Blocking procedures were applied for the purpose of improving discrimination among chromosome regions. Unlabeled genomic DNA from Z. mays subsp. parviglumis as a blocking agent and labeled genomic DNA from maize were hybridized on maize chromosomes. On the other hand, mitotic metaphases from Z. mays subsp. parviglumis were blocked with unlabeled genomic DNA of maize and hybridized with labeled genomic DNA from Z. mays subsp. parviglumis. Both experiments showed that either maize or Z. mays subsp. parviglumis chromosomes have their own unique sequences. This means an unexpected degree of divergence if Z. mays subsp. parviglumis is the only progenitor of maize, a result that is discussed in relation to our previous genomic in situ hybridization observations and to the different scenarios proposed about the origin of maize.     

Inheritance of the Morphological Differences between Maize and Teosinte: Comparison of Results for Two F(2) Populations

Molecular marker loci (MMLs) were employed to map quantitative trait loci (QTLs) in an F(2) population derived from a cross of maize (Zea mays ssp. mays) and its probable progenitor, teosinte (Z. mays ssp. parviglumis). A total of 50 significant associations (putative QTLs) between the MMLs and nine key traits that distinguish maize and teosinte were identified. Results from this analysis are compared with our previous analysis of an F(2) population derived from a cross of a different variety of maize and another subspecies of teosinte (Z. mays ssp. mexicana). For traits that measure the architectural differences between maize and teosinte, the two F(2) populations possessed similar suites of QTLs. For traits that measure components of yield, substantially different suites of QTLs were identified in the two populations. QTLs that control about 20% or more of the phenotypic variance for a trait in one population were detected in the other population 81% of the time, while QTLs that control less than 10% of the variance in one population were detected in the other population only 28% of the time. In our previously published analysis of the maize X ssp. mexicana population, we identified five regions of the genome that control most of the key morphological differences between maize and teosinte. These same five regions also control most of the differences in the maize X ssp. parviglumis population. Results from both populations support the hypothesis that a relatively small number of loci with large effects were involved in the early evolution of the key traits that distinguish maize and teosinte. It is suggested that loci with large effects on morphology may not be a specific feature of crop evolution, but rather a common phenomenon in plant evolution whenever a species invades a new niche with reduced competition.     

A cellular study of teosinte Zea mays subsp. parviglumis (Poaceae) caryopsis development showing several processes conserved in maize

The evolutionary history of maize (Zea mays subsp. mays) is of general interest because of its economic and scientific importance. Here we show that many cellular traits described previously in developing caryopses of maize are also seen in its wild progenitor teosinte (Zea mays subsp. parviglumis). These features, each with a possible role in development, include (1) an early programmed cell death in the maternal placento-chalazal (P-C) layer that may lead to increased hydrolytic conductance to the developing seed; (2) accumulation of phenolics and flavonoids in the P-C layer that may be related to antimicrobial activity; (3) formation of wall ingrowths in the basal endosperm transfer layer (BETL); (4) localization of cell wall invertase in the BETL, which is attributed to the increased transport capacity of photosynthates to the sink; and (5) endoreduplication in endosperm nuclei suggested to contribute to increased gene expression and greater sink capacity of the developing seed. In maize caryopsis, these cellular traits have been previously attributed to domestication and selection for larger seed size and vigor. Given the conservation of the entire cellular program in developing teosinte caryopses described here, we suggest that these traits evolved independently of domestication and predate human selection pressure.     

Evidence for a natural allelic series at the Maize domestication Locus teosinte branched1

Despite numerous quantitative trait loci and association mapping studies, our understanding of the extent to which natural allelic series contribute to the variation for complex traits is limited. In this study, we investigate the occurrence of a natural allelic series for complex traits at the teosinte branched1 (tb1) gene in natural populations of teosinte (Zea mays ssp. parviglumis, Z. mays ssp. mexicana, and Z. diploperennis). Previously, tb1 was shown to confer large effects on both plant architecture and ear morphology between domesticated maize and teosinte; however, the effect of tb1 on trait variation in natural populations of teosinte has not been investigated. We compare the effects of nine teosinte alleles of tb1 that were introgressed into an isogenic maize inbred background. Our results provide evidence for a natural allelic series at tb1 for several complex morphological traits. The teosinte introgressions separate into three distinct phenotypic classes, which correspond to the taxonomic origin of the alleles. The effects of the three allelic classes also correspond to known morphological differences between the teosinte taxa. Our results suggest that tb1 contributed to the morphological diversification of teosinte taxa as well as to the domestication of maize.     

Molecular Evidence and the Origin and Development of the Domesticated Sunflower (<Emphasis Type="Italic">Helianthus annum</Emphasis>, Asteraceae)

The domesticated sunflower,Helianthus annuus, is an important economic crop, yet molecular data regarding its evolution are limited. Here we review morphological, geographical, archaeological, and molecular evidence pertaining to its origin and development. New isozyme and chloroplast DNA (cpDNA) evidence is also presented.Morphological, geographical, and archaeological evidence has led to the hypothesis that the domesticated sunflower was derived from a wild/weedy form ofH. annuus possibly in the Midwest. Molecular evidence was concordant with this hypothesis. A high degree of enzymatic and cpDNA sequence similarity was observed between wild and domesticatedH. annuus, and domesticatedH. annuus contained a subset of the alleles and cpDNAs found in wildH. annuus. The extensive polymorphism in the wild plants and the virtual monomorphism in cultivated lines for both isozyme and cpDNA phenotypes further suggest a single origin of the domesticated sunflower from a very limited gene pool. In addition, Native American varieties of the domesticated sunflower were genetically more variable than other cultivated lines, possibly indicating that they gave rise to the other cultivated stocks. Molecular evidence did not, however, allow conclusions as to the exact geographic origin of the domesticated sunflower.     

Identification and heritability of fumonisin insensitivity in Zea mays

Landraces of maize (Zea mays ssp. mays) and its wild teosinte relatives (Zea mays spp. parviglumis and mexicana) were surveyed for sensitivity to fumonisin B(1), a phytotoxin produced by the maize pathogen Gibberella moniliformis. Only two of 42 Z. mays samples were highly insensitive to FB(1) (ED(50) = ca. 200 microM). The teosintes and 76% of the maize landraces were moderately or highly sensitive to FB(1) (ED(50) < or = 30 microM), which indicates that FB(1) sensitivity is likely to be an ancestral trait in Z. mays. F(1) generations derived from crosses between FB(1)-sensitive maize inbred B73 and insensitive landraces were significantly less sensitive than B73. Thus, our data indicate that FB(1)-insensitivity is a relatively rare but heritable trait in maize. We also report the sensitivity of maize to other Gibberella toxins - beauvericin, diacetoxyscirpenol, and moniliformin.     

A Bird in the Hand Versus Two in the Bush? The Specialist Leafhopper <Emphasis Type="Italic">Dalbulus maidis</Emphasis> (Hemiptera: Cicadellidae) Does Not Discriminate Against Sub-optimal Host Plants (<Emphasis Type="Italic">Zea</Emphasis> spp.)

The corn leafhopper [Dalbulus maidis (DeLong & Wolcott)] is a specialist on Zea (Poaceae) that coevolved with maize (Zea mays mays) and its teosinte (Zea spp.) relatives. This study tested the hypothesis that host acceptance by females varies among Zea hosts, and is correlated with variation in defensive levels across those hosts. Prior studies revealed differences in plant defenses among Zea hosts and corresponding differences in corn leafhopper performance. Thus, host acceptance was expected to be correlated with defensive levels and offspring performance across Zea hosts, following the hypothesis that offspring performance mediates host preference. In parallel, host acceptance was expected to be correlated with transitions in life history strategy (perennial to annual life cycle), domestication status (wild to domesticated), and breeding intensity (landrace to hybrid variety) in Zea because variation in defensive levels and corn leafhopper performance were shown in prior studies to be correlated with those transitions. The study’s hypotheses were tested by comparing, under no-choice conditions, host acceptance by corn leafhopper of a suite of Zea hosts encompassing those transitions: perennial teosinte (Zea diploperennis), Balsas teosinte (Zea mays parviglumis), and landrace and commercial hybrid maize. The results did not show differences in host acceptance for oviposition or feeding among the hosts. Thus, under no-choice conditions, all Zea hosts may be similarly acceptable for feeding and oviposition, despite marked ovipositional preferences under choice conditions and poorer offspring performance on teosintes relative to maize shown previously. The results suggested also that oviposition frequency per plant by females was not correlated with their offspring’s performance.     

The genome organization and diversification of maize and its allied species revisited: evidences from classical and FISH-GISH cytogenetic analysis

The present review summarizes our classical and molecular cytogenetic investigations in the genus Zea. The results obtained from the meiotic behavior analysis of Zea species and hybrids, confirm the amphiploid nature of all species in the genus, with a basic number of x = 5 chromosomes. All species with 2n = 20 are diploidized allotetraploids, whereas Z. perennis (2n = 40) is an allooctoploid with four genomes somewhat divergent from one another. These analyses also revealed the existence of postzygotic reproductive isolation among Zea species. Our studies using genomic in situ hybridization (GISH) provide evidence about the evolutionary relationships among maize and its allied species, and reveal remarkable genomic divergences. Particularly, knob sequences were not completely shared between taxa previously considered to be closely related. Our data strongly suggest that the teosinte Z. mays parviglumis is not the only progenitor of cultivated maize. Introgression of Tripsacum into cultivated maize cannot be discarded.     

Restriction site variation in the zea chloroplast genome

Nineteen accessions selected from the four species and three subspecies of the genus Zea and one accession from the related genus Tripsacum were surveyed for variation with 21 restriction endonucleases. In all, 580 restriction sites were assayed in each chloroplast (cp)DNA, this representing 2.2% of the genome. Twenty-four of the 580 sites were variable in one or more of the cpDNAs. The number of nucleotide substitutions per site (p) between Zea and Tripsacum (0.0056) approximates that between other closely related angiosperm genera. The range in values of p among Zea species (0.0003-0.0024) is on the lower end of the range reported for other angiosperm genera. Analysis of the distribution of restriction site mutations throughout the genome indicated that the inverted repeat evolves more slowly than either the small or large unique sequence regions. Parsimony phylogenetic analysis of the restriction site data produced a tree consistent with isoenzymatic and morphological measures of affinity among the species. Chloroplast DNA analysis was not useful in discriminating the subspecies within Zea mays. The lack of any detectable differences between the cpDNA of maize (Z. mays subsp. mays) and some teosintes (Z. mays subsps. mexicana and parviglumis ) is consistent with the hypothesis that maize is a domesticated form of teosinte. Comparison of the degree of sequence divergence for Z. mays cpDNA and the Adh1 locus suggests the latter may be evolving at 10 times the rate of the former. Comparison of rates of sequence evolution for the mitochondrial and chloroplast genomes was inconclusive and could not clarify whether these two genomes have dissimilar rates of sequence evolution.     

Facilitated by nature and agriculture: performance of a specialist herbivore improves with host-plant life history evolution, domestication, and breeding

Plant defenses against herbivores are predicted to change as plant lineages diversify, and with domestication and subsequent selection and breeding in the case of crop plants. We addressed whether defense against a specialist herbivore declined coincidently with life history evolution, domestication, and breeding within the grass genus Zea (Poaceae). For this, we assessed performance of corn leafhopper (Dalbulus maidis) following colonization of one of four Zea species containing three successive transitions: the evolutionary transition from perennial to annual life cycle, the agricultural transition from wild annual grass to primitive crop cultivar, and the agronomic transition from primitive to modern crop cultivar. Performance of corn leafhopper was measured through seven variables relevant to development speed, survivorship, fecundity, and body size. The plants included in our study were perennial teosinte (Zea diploperennis), Balsas teosinte (Zea mays parviglumis), a landrace maize (Zea mays mays), and a hybrid maize. Perennial teosinte is a perennial, iteroparous species, and is basal in Zea; Balsas teosinte is an annual species, and the progenitor of maize; the landrace maize is a primitive, genetically diverse cultivar, and is ancestral to the hybrid maize; and, the hybrid maize is a highly inbred, modern cultivar. Performance of corn leafhopper was poorest on perennial teosinte, intermediate on Balsas teosinte and landrace maize, and best on hybrid maize, consistent with our expectation of declining defense from perennial teosinte to hybrid maize. Overall, our results indicated that corn leafhopper performance increased most with the agronomic transition, followed by the life history transition, and least with the domestication transition.     

The role of teosinte glume architecture (tga1) in coordinated regulation and evolution of grass glumes and inflorescence axes

? Hardened floral bracts and modifications to the inflorescence axis of grasses have been hypothesized to protect seeds from predation and/or aid seed dispersal, and have evolved multiple times independently within the family. Previous studies have demonstrated that mutations in the maize (Zea mays ssp. mays) gene teosinte glume architecture (tga1) underlie a reduction in hardened structures, yielding free fruits that are easy to harvest. It remains unclear whether the causative mutation(s) occurred in the cis-regulatory or protein-coding regions of tga1, and whether similar mutations in TGA1-like genes can explain variation in the dispersal unit in related grasses. ? To address these questions TGA1-like genes were cloned and sequenced from a number of grasses and analyzed phylogenetically in relation to morphology; protein expression was investigated by immunolocalization. ? TGA1-like proteins were expressed throughout the spikelet in the early development of all grasses, and throughout the flower of the grass relative Joinvillea. Later in development, expression patterns differed between Tripsacum dactyloides, maize and teosinte (Z. mays ssp. parviglumis). ? These results suggest an ancestral role for TGA1-like genes in early spikelet development, but do not support the hypothesis that TGA1-like genes have been repeatedly modified to affect glume and inflorescence axis diversification.     

A large-scale screen for artificial selection in maize identifies candidate agronomic loci for domestication and crop improvement

Maize (Zea mays subsp mays) was domesticated from teosinte (Z. mays subsp parviglumis) through a single domestication event in southern Mexico between 6000 and 9000 years ago. This domestication event resulted in the original maize landrace varieties, which were spread throughout the Americas by Native Americans and adapted to a wide range of environmental conditions. Starting with landraces, 20th century plant breeders selected inbred lines of maize for use in hybrid maize production. Both domestication and crop improvement involved selection of specific alleles at genes controlling key morphological and agronomic traits, resulting in reduced genetic diversity relative to unselected genes. Here, we sequenced 1095 maize genes from a sample of 14 inbred lines and chose 35 genes with zero sequence diversity as potential targets of selection. These 35 genes were then sequenced in a sample of diverse maize landraces and teosintes and tested for selection. Using two statistical tests, we identified eight candidate genes. Extended gene sequencing of these eight candidate loci confirmed that six were selected throughout the gene, and the remaining two exhibited evidence of selection in the 3' portion of each gene. The selected genes have functions consistent with agronomic selection for nutritional quality, maturity, and productivity. Our large-scale screen for artificial selection allows identification of genes of potential agronomic importance even when gene function and the phenotype of interest are unknown.     

Plant defense against fall armyworm in micro‐sympatric maize (Zea mays ssp. mays) and Balsas teosinte (Zea mays ssp. parviglumis)

Maize [Zea mays L. ssp. mays (Poaceae)] was domesticated from Balsas teosinte (Zea mays ssp. parviglumis Iltis & Doebley) in present‐day Mexico. Fall armyworm, Spodoptera frugiperda JE Smith (Lepidoptera: Noctuidae), is among the most important pests of maize in Mexico and Central America. We compared the strength of plant defenses against fall armyworm between micro‐sympatric landrace maize and Balsas teosinte in the field and laboratory. The field comparison, conducted in Mexico, consisted of comparing the frequency of fall armyworm infestation between young maize and Balsas teosinte plants in dryland agricultural fields in which Balsas teosinte grew as a weed. The laboratory comparison contrasted the performance of fall armyworm larvae provided a diet of leaf tissue excised from maize or Balsas teosinte plants that were intact or had been primed by larval feeding. In the field, maize plants were more frequently infested with fall armyworm than Balsas teosinte plants: over 3 years and three fields, maize was infested at a ca. 1.8‐fold greater rate than Balsas teosinte. In the laboratory, larval growth, but not survivorship, was differently affected by feeding on maize vs. Balsas teosinte, and on primed vs. intact plants. Specifically, survivorship was ca. 98%, and did not differ between maize and Balsas teosinte, nor between primed and intact plants. Larvae grew less on intact vs. primed maize, and similarly on intact vs. primed Balsas teosinte; overall, growth was 1.2‐fold greater on maize compared to Balsas teosinte, and on primed compared to intact plants. Parallel observations showed that the differences in growth could not be attributed to the amount of leaf tissue consumed by larvae. We discuss our results in relation to differences in the strength of plant defenses between crops and their ancestors, the relevance of unmanaged Balsas teosinte introgression in the context of fall armyworm defenses in maize, and whether greater growth of larvae on primed vs. intact plants signifies herbivore offense.     

Molecular evolution of FLORICAULA/LEAFY orthologs in the Andropogoneae (Poaceae)

Members of the grass family (Poaceae) exhibit a broad range of inflorescence structures and other morphologies, making the grasses an interesting model system for studying the evolution of development. Here we present an analysis of the molecular evolution of FLORICAULA/LEAFY-like genes, which are important developmental regulatory loci known to affect inflorescence development in a wide range of flowering plant species. We have focused on sequences from the Andropogoneae, a tribe within the grass family that includes maize (Zea mays ssp. mays) and Sorghum (Sorghum bicolor). The FLORICAULA/LEAFY gene phylogeny we generated largely agrees with previously published phylogenies for the Andropogoneae using other nuclear genes but is unique in that it includes both members of one of the many duplicate gene sets present in maize. The placement of these sequences in the phylogeny suggests that the duplication of the maize FLORICAULA/LEAFY orthologs, zfl1 and zfl2, is a consequence of a proposed tetraploidy event that occurred in the common ancestor of Zea and a closely related genus, Tripsacum. Our data are consistent with the hypothesis that the transcribed regions of the FLORICAULA/LEAFY-like genes in the Andropogoneae are functionally constrained at both nonsynonymous and synonymous sites and show no evidence of directional selection. We also examined conservation of short noncoding sequences in the first intron, which may play a role in gene regulation. Finally, we investigated the genetic diversity of one of the two maize FLORICAULA/LEAFY orthologs, zfl2, in maize and its wild ancestor, teosinte (Z. mays ssp. parviglumis), and found no evidence for selection pressure resulting from maize domestication within the zfl2-coding region.     

Some physical properties of teosinte (Zea mays subsp. parviglumis) pollen

In parts of the world where teosinte and maize are grown in close proximity, there is concern about gene flow between them. Pollen is the primary vehicle for gene flow. Quantifying the biophysical properties of pollen, such as its settling speed and dehydration rate, is important for evaluating outcrossing potential. These properties were measured for teosinte (Zea mays subsp. parviglumis) pollen. Pollen was found to have an average settling speed of 0.165 m s(-1), which agrees well with theoretical values based on the size of the pollen grains. The conductance of the pollen wall for water was derived from the time rate of change of pollen grain size and gave an average conductance of 3.42x10(-4) m s(-1). Water potential, psi, of teosinte pollen was determined at various values of relative water content (dry-weight basis), theta, by using a thermocouple psychrometer and by allowing samples of pollen to come to vapour equilibrium with various saturated salt solutions. Non-linear regression analysis of the data yielded psi (MPa) = -4.13 theta(-1.23) (r2=0.77). Results for conductance and psi were incorporated into a model equation for the rate of water loss from pollen grains, which yielded results that agreed well (r2=0.96) with observations of water loss from pollen grains in air. The data reported here are important building blocks in a model of teosinte pollen movement and should be helpful in establishing the main factors influencing the degree and the direction of pollination between teosinte populations and between maize and teosinte.