A genomic journey across the past, present, and future of South American maize.

Maize (Zea mays ssp. mays) diverged from one of its wild relatives, the teosinte Zea mays ssp. parviglumis, in the lowlands of southwest Mexico approximately 9000 years ago. Following this divergence, maize rapidly expanded throughout the Americas, becoming a staple food. This dispersal was accompanied by significant demographic and selective changes, leading to the development of numerous local varieties with a complex evolutionary history that remains incompletely understood. In recent years, genomic advances have challenged traditional models of maize domestication and spread to South America. At least three distinct genetic lineages associated with different migratory waves have been described: ancestral Andean, ancestral Lowland, and Pan-American. Additionally, the significant role of the teosinte Zea mays ssp. mexicana, in the evolution of modern maize has been recently uncovered. Genomic studies have shed light into highland adaptation processes, revealing largely independent adaptation events in Meso- and South America. As new evidence emerges, the regional complexity underlying maize diversity and the need for comprehensive, multi-scale approaches become evident. In the face of climate change and evolving agricultural landscapes, the conservation of native maize in South America is of growing interest, with genomics serving as an invaluable tool for identifying and preserving the genetic variability of locally adapted germplasm.

Large scale sampling of Mexican maize landraces for the presence of transgenes.

The presence and levels of transgenic maize in Mexico and the effect this could have on local landraces or closely related species such as teosinte has been the subject of several previous reports, some showing contrasting results. Cultural, social and political factors all affect maize cultivation in Mexico and although since 1998 there has been a moratorium on the commercial cultivation of transgenic maize, Mexico imports maize, mainly from the USA where transgenic cultivars are widely grown. Additionally extensive migration between rural areas in Mexico and the USA and customs of seed exchange between farmers may also play an unintentional role in the establishment of transgenic seed. A comprehensive study of all Mexican maize landraces throughout the country is not feasible, however this report presents data based on analysis of 3204 maize accessions obtained from the central region of Mexico (where permits have never been authorized for cultivation of transgenic maize) and the northern region (where for a short period authorization for experimental plots was granted). The results of the study confirm that transgenes are present in all the geographical areas sampled and were more common in germplasm obtained in the northern region. However, there was no evidence that regions where field trials had been authorized showed higher levels of transgene presence or that the morphology of seed lots harboring transgenic material was significantly modified in favor of expected transgenic phenotypes.

In search of better snacks: ohmic-heating nixtamalized flour and amaranth addition increase the nutraceutical and nutritional potential of vegetable-enriched tortilla chips.

BACKGROUND: Nixtamalized flour snacks such as tortilla chips are widely consumed across the world, but they are nutritionally poor and contribute to obesity and other non-communicable diseases. The production of healthy versions of such snacks, by incorporating vegetables and improving the quality of the flours used in their formulation, could help address these nutritional challenges. This study compared the fortification of baked tortilla chips with vegetable leaf powders (kale and wild amaranth at 0%, 4%, 8%, and 16% w/w) and using two types of nixtamalized flour: traditional (TNF) and with ohmic heating (OHF). RESULTS: Overall, the use of OHF increased 1.88 times the fibre in enriched and non-enriched snacks with respect to TNF, but the latter had 1.85 times more protein. Addition of 16% of vegetable powders increased protein (kale = 1.4-fold; amaranth = 1.3-fold) and dietary fibre (kale = 1.52-fold; amaranth = 1.7-fold). Amaranth enrichment improved total phenolic content (TPC) and total flavonoid content (TFC) of chips at least 1.2 and 1.63 times, respectively. OHF chips also had higher bound TPC than TNF ones, regardless of vegetable addition. Combinations of OHF with 16% amaranth produced chips 1.74-fold higher in antioxidant capacity than non-enriched ones, due to increased content of phenolics such as ferulic acid. CONCLUSION: This work showed that tortilla chips made using nixtamalized flour produced with assisted ohmic heating, alone or in combination with wild amaranth leaf powder, could be used in the production of healthy maize snacks to enhance their prospective antioxidant activity and nutritional value. © 2023 Society of Chemical Industry.

Effect of Storage Technologies on Postharvest Insect Pest Control and Seed Germination in Mexican Maize Landraces.

Smallholder farmers who grow maize landraces face important challenges to preserve their seed biodiversity from one season to another. This study was carried out in the central highlands of Mexico to compare the effectiveness of two seed storage practices-specifically, polypropylene woven bags (farmers' conventional practice) vs. hermetic containers-for minimizing seed losses and maintaining germination. Four Mexican landraces were stored for three and six months. Data on moisture content and kernel damage were collected at the beginning and the end of the storage period. Pest-free samples collected were also analyzed for seed germination. Moisture content was below 13% overall and was not significantly affected by storage technology or storage time. Samples from the polypropylene woven bags suffered significant damage from Sitophilus zeamais and Prostephanus truncatus, with the percentages of insect damage and weight loss reaching 61.4% and 23.4%, respectively. Losses were minimal in seed stored in hermetic containers, with a maximum insect damage of 4.1% and weight loss of 2.2%. Overall, the germination rate of samples stored in these airtight containers was greater than 90%. This study provides additional evidence on the effectiveness of hermetic containers at maintaining Mexican landraces' seed quantity and quality during storage in smallholder conditions in central Mexico.

Causes and consequences of DNA content variation in Zea / Causas y consecuencias de la variación del contenido de ADN en Zea

ABSTRACT Cytogenetic evidence indicates that Zea, which comprises maize (Z. mays ssp. mays) and its wild relatives, is an allopolyploid genus. Our research group has carried out numerous cytogenetic studies on Zea species, mainly focused on native Argentinian and Bolivian maize landraces. We found a wide inter- and intraspecific genome size variation in the genus, with mean 2C-values ranging between 4.20 and 11.36 pg. For the maize landraces studied here, it varied between 4.20 and 6.75 pg. The objectives of this work are to analyze the causes of genome size variation and to discuss their adaptive value in Zea. This variation is mainly attributed to differences in the heterochromatin located in the knobs and to the amount of interspersed DNA from retrotransposons. Polymorphisms in presence or absence of B-chromosomes (Bs) and the population frequency of Bs are also a source of genome size variation, with doses ranging between one and eight in the landraces analyzed here. Correlation analysis revealed that the percentage of heterochromatin is positively correlated with genome size. In addition, populations cultivated at higher altitudes, which are known to be precocious, have smaller genome sizes than do those growing at lower altitudes. This information, together with the positive correlation observed between the length of the vegetative cycle and the percentage of heterochromatin, led us to propose that it has an adaptive role. On the other hand, the negative relationship found between Bs and heterochromatic knobs allowed us to propose the existence of an intragenomic conflict between these elements. We hypothesize that an optimal nucleotype may have resulted from such intranuclear conflict, where genome adjustments led to a suitable length of the vegetative cycle for maize landraces growing across altitudinal clines.

RESUMEN La evidencia citogenética indica que el género Zea, el maíz (Z. mays ssp. mays) y sus parientes silvestres, posee un origen alopoliploide. Nuestro grupo de investigación ha realizado numerosos estudios en especies de Zea, principalmente en maíces nativos de Argentina y Bolivia. En este género, hallamos una amplia variación inter e intraespecífica en el tamaño del genoma, con valores 2C medios que oscilan entre 4,20 y 11,36 pg. El valor 2C medio de los maíces nativos estudiados varió entre 4,20 y 6,75 pg. Los objetivos de este trabajo son analizar las causas de la variación del tamaño del genoma en Zea y discutir su valor adaptativo. Esta variación se atribuye principalmente a las diferencias en la heterocromatina de los knobs y en la cantidad de ADN intercalado de los retrotransposones. Otras fuentes de variación son los polimorfismos para presencia/ausencia de cromosomas B (Bs) y para la frecuencia poblacional de Bs en las razas analizadas, con dosis que oscilan entre uno y ocho Bs. El porcentaje de heterocromatina se correlaciona positivamente con el tamaño del genoma. Las poblaciones cultivadas en altitudes altas, que son precoces, tienen tamaños de genoma más pequeños que las que crecen en bajas altitudes. Esta información, junto con la correlación positiva observada entre la duración del ciclo vegetativo y el porcentaje de heterocromatina, nos llevó a proponer el rol adaptativo de la heterocromatina. Por otro lado, la relación negativa encontrada entre Bs y knobs heterocromáticos nos permitió proponer la existencia de un conflicto intragenómico entre estos elementos. Hipotetizamos que de este conflicto intranuclear habría resultado el nucleotipo óptimo, donde ajustes genómicos condujeron a una duración adecuada del ciclo vegetativo en las razas de maíz que crecen a lo largo de clines altitudinales.

Functional characterization of culturable fungi from microbiomes of the "conical cobs" Mexican maize (Zea mays L.) landrace.

Mexican maize landraces, produced for local consumption, are adapted to different environmental conditions, and their yield is affected by abiotic and biotic factors, including the use of agrochemicals. The search for sustainable alternatives to agrochemicals includes the study of the culturable microbial communities. In this study, the fungal communities associated with 2 Mexican maize landraces reddish and bluish "conical cobs" were found to be comprised of Ascomycota fungi, represented by 89 strains within 6 orders (Pleosporales, Hypocreales, Onygenales, Capnodiales, Helotiales, and Eurotiales) and 16 genera. Cellulases and metallophores production were the primary enzymatic products and plant growth-promoting activities were detected among the isolates. Penicillium, Didymella, and Fusarium strains had the most active enzymatic and plant growth promoting activities, however, Aspergillus sp. HES2-2.2, Talaromyces sp. RS1-7, and Penicillium sp. HFS3-3 showed antagonistic activity against the four phytopathogenic Fusarium strains Fusarium oxysporum, Fusarium sambucinum, Fusarium fujikuroi and Fusarium incarnatum-equiseti and also a high and diverse production of enzymatic and plant growth promoting activities; here we identified fungal strains as candidates to promote maize growth.

Plastome genomics in South American maize landraces: chloroplast lineages parallel the geographical structuring of nuclear gene pools.

BACKGROUND AND AIMS: The number of plastome sequences has increased exponentially during the last decade. However, there is still little knowledge of the levels and distribution of intraspecific variation. The aims of this study were to estimate plastome diversity within Zea mays and analyse the distribution of haplotypes in connection with the landrace groups previously delimited for South American maize based on nuclear markers. METHODS: We obtained the complete plastomes of 30 South American maize landraces and three teosintes by means of next-generation sequencing (NGS) and used them in combination with data from public repositories. After quality filtering, the curated data were employed to search for single-nucleotide polymorphisms, indels and chloroplast simple sequence repeats. Exact permutational contingency tests were performed to assess associations between plastome and nuclear variation. Network and Bayesian phylogenetic analyses were used to infer evolutionary relationships among haplotypes. KEY RESULTS: Our analyses identified a total of 124 polymorphic plastome loci, with the intergenic regions psbE-rps18, petN-rpoB, trnL_UAG-ndhF and rpoC2-atpI exhibiting the highest marker densities. Although restricted in number, these markers allowed the discrimination of 27 haplotypes in a total of 51 Zea mays individuals. Andean and lowland South American landraces differed significantly in haplotype distribution. However, overall differentiation patterns were not informative with respect to subspecies diversification, as evidenced by the scattered distribution of maize and teosinte plastomes in both the network and Bayesian phylogenetic reconstructions. CONCLUSIONS: Knowledge of intraspecific plastome variation provides the framework for a more comprehensive understanding of evolutionary processes at low taxonomic levels and may become increasingly important for future plant barcoding efforts. Whole-plastome sequencing provided useful variability to contribute to maize phylogeographic studies. The structuring of haplotype diversity in the maize landraces examined here clearly reflects the distinction between the Andean and South American lowland gene pools previously inferred based on nuclear markers.

Assessment of potential impacts associated with gene flow from transgenic hybrids to Mexican maize landraces.

Genetically modified (GM) maize has been grown and safely consumed on a global scale since its commercialization in 1996. However, questions have been raised about the potential impact that GM maize could have on native maize landraces in Mexico, which is the center of origin and diversity of maize. This research was conducted to evaluate potential changes to maize landraces in an unlikely event of transgene introgression. For this study, two GM traits that confer insect protection and herbicide tolerance in maize (MON 89034 and MON 88017), designated as VT3Pro, were introgressed into two Mexican landraces, Tuxpeño and Tabloncillo. Field trials were conducted across four environments to assess phenotypic characteristics, plant response to stressors, and kernel composition of landraces with and without VT3Pro traits. Furthermore, materials from four backcrossing generations were analyzed for segregation of these GM traits. Generally, no significant differences were observed between landraces with and without VT3Pro traits for the evaluated characteristics and the segregation analysis showed that GM traits, when introgressed into landraces, followed Mendelian principles. These results support the conclusion that, if inadvertently introgressed into landraces, VT3Pro traits are not expected to alter phenotypic or kernel characteristics, plant response to stressors (except for targeted insect protection and herbicide tolerance traits) and would segregate like any endogenous gene. These results should be taken into consideration when discussing benefits and risks associated with commercial production of GM maize hybrids in the centers of origin and diversity of maize.

Are cytological parameters of maize landraces (Zea mays ssp. mays) adapted along an altitudinal cline?

The Northwestern Argentina (NWA) highland region is one of the southernmost areas of native maize cultivation. We studied variations of different cytological parameters, such as DNA contents, presence/absence of B chromosomes (Bs), and number and sequence composition of heterochromatic knobs in ten accessions of four maize landraces growing along a broad altitudinal cline in NWA. The aim of this work was to assess variations in cytological parameters and their relationship with the crop altitude of cultivation, in an adaptive context. The A-DNA content of the A chromosome complements showed 40% of difference between the lowest (4.5 pg) and the highest (6.3 pg) 2C value. This variation could be attributed to differences in number and size of heterochromatic knobs. Fluorescent in situ hybridization studies revealed the sequence composition of each knob, with a higher proportion of knobs composed of 180-bp repeats rather than TR-1 repeats, in all accessions. We also found numerical polymorphisms and the highest frequency of Bs reported in maize to this date. These results lead us to propose that the frequencies and doses of Bs are influenced by the landrace genotypical make-up. The Bs might be maintained in higher frequencies in those accessions having lower heterochromatin content, so as to preserve an optimal nucleotype. Furthermore, selective forces acting along the altitudinal gradient might be modulating the cytological parameters studied, as suggested by the significant correlations found among them.

An SSR-based approach incorporating a novel algorithm for identification of rare maize genotypes facilitates criteria for landrace conservation in Mexico.

As maize was domesticated in Mexico around 9,000 years ago, local farmers have selected and maintained seed stocks with particular traits and adapted to local conditions. In the present day, many of these landraces are still cultivated; however, increased urbanization and migration from rural areas implies a risk that this invaluable maize germplasm may be lost. In order to implement an efficient mechanism of conservation in situ, the diversity of these landrace populations must be estimated. Development of a method to select the minimum number of samples that would include the maximum number of alleles and identify germplasm harboring rare combinations of particular alleles will also safeguard the efficient ex-situ conservation of this germplasm. To reach this goal, a strategy based on SSR analysis and a novel algorithm to define a minimum collection and rare genotypes using landrace populations from Puebla State, Mexico, was developed as a "proof of concept" for methodology that could be extended to all maize landrace populations in Mexico and eventually to other native crops. The SSR-based strategy using bulked DNA samples allows rapid processing of large numbers of samples and can be set up in most laboratories equipped for basic molecular biology. Therefore, continuous monitoring of landrace populations locally could easily be carried out. This methodology can now be applied to support incentives for small farmers for the in situ conservation of these traditional cultivars.

Dissecting maize diversity in lowland South America: genetic structure and geographic distribution models.

BACKGROUND: Maize landraces from South America have traditionally been assigned to two main categories: Andean and Tropical Lowland germplasm. However, the genetic structure and affiliations of the lowland gene pools have been difficult to assess due to limited sampling and the lack of comparative analysis. Here, we examined SSR and Adh2 sequence variation in a diverse sample of maize landraces from lowland middle South America, and performed a comprehensive integrative analysis of population structure and diversity including already published data of archaeological and extant specimens from the Americas. Geographic distribution models were used to explore the relationship between environmental factors and the observed genetic structure. RESULTS: Bayesian and multivariate analyses of population structure showed the existence of two previously overlooked lowland gene pools associated with Guaraní indigenous communities of middle South America. The singularity of this germplasm was also evidenced by the frequency distribution of microsatellite repeat motifs of the Adh2 locus and the distinct spatial pattern inferred from geographic distribution models. CONCLUSION: Our results challenge the prevailing view that lowland middle South America is just a contact zone between Andean and Tropical Lowland germplasm and highlight the occurrence of a unique, locally adapted gene pool. This information is relevant for the conservation and utilization of maize genetic resources, as well as for a better understanding of environment-genotype associations.

Characterization of corn landraces planted grown in the campos gerais region (Paraná, Brazil) for industrial utilization

This work has the objective of characterizing twenty corn landraces grown in the Campos Gerais region (Paraná State) in relation to its chemical composition (moisture, ash, protein, ether extract, dietary fiber and starch) and physical properties (weight of 1000 grains, real density, flotation index, granulometry and color). In addition, also the lab scale processing of the kernels from the varieties was carried out for producing starch; starch purity was evaluated by measuring its protein contamination. Amylose contents and viscoamylograph profile were also evaluated. The results showed that the evaluated landraces have differences in chemical composition as well as in pericarp/endosperm/germ proportions and consequently it should have different industrial applications and interest for plant breeding.

Esse trabalho teve o objetivo de caracterizar vinte variedades de milho crioulo cultivadas na região dos Campos Gerais (Estado do Paraná) em relação a sua composição química (umidade, cinzas, proteína, extrato etéreo, fibra alimentar e amido) e propriedades físicas (peso de 1000 grãos, densidade real, índice de flotação, granulometria e cor) Além disso, foi feito o processamento dos grãos em escala de laboratório para a extração do amido, sendo mensurado o teor de proteína. Foram avaliados os conteúdos de amilose e o perfil viscoamilográfico. Os resultados mostraram que os milhos apresentaram diferentes composições químicas e proporções pericarpo/endosperma/ gérmen e consequentemente podem ter diferentes aplicações industriais e interesse ao melhoramento de plantas.