Genetic divergence among maize hybrids by morphological descriptors

Two experiments with 25 maize commercial hybrids were carried out in a direct sowing system in Southern Brazil in the harvests of 2009/2010 and 2010/2011. Quantitative descriptors were used with the objective of determining the genetic divergence and the relative contributions of traits among hybrids for extraction of inbred lines. This study was carried out in Oxisol soil using a randomized block design with four replicates. Data were subjected to combined analysis of variance, and based on the multivariate analyses, Tocher and average linkage (UPGMA) cluster analyses, based on generalized distance of Mahalanobis, to quantify divergence in addition to Singh criterion to validate trait with the most contribution. The multivariate methods were consistent with each other, and the weight of 100 grains was the trait that contributed most to the divergence and had similar behavior in grain yield between hybrids in both years. Furthermore, this descriptor representing significant genetic variability for crossings and lines extraction to hybridization between BM 3061, ATL 200 and P 30B39 Y.

Seed quality, chlorophyll content index and leaf nitrogen levels in maize inoculated with Azospirillum brasilense

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Maize leaf phytotoxicity and grain yield are affected by nitrogen source and application method

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Maize and soybeans production in integrated system under no-tillage with different pasture combinations and animal categories

The adoption of no-till system (NTS) combined with crop-livestock integration (CLI) has been a strategy promoted in Brazil, aiming to maximize areas yield and increase agribusiness profitability. This study aimed to evaluate grains yield and phytotechnical attributes from maize and soybean culture by CLI system under NTS after winter annual pure and diversified pastures with the absence or presence of grazing animals. The experiment was installed in Castro (Parana State, Brazil) on in a dystrophic Humic Rhodic Hapludox with a clay texture, using experimental design of randomized complete blocks in 4 x 2 factorial scheme with three replications. Treatments included four pasture combinations (diversified or pure) and animal categories (light and heavy) subjected or not to grazing animals during the winter. During 2008/09 and 2009/10 summers, the area was cultivated with soybeans and maize, respectively, with yield assessment of grains and phytotechnical attributes. Treatments did not alter the yield and weight of a thousand seeds (WTS) of soybeans. In maize culture, the grazing animal during the winter increased the plant population and grains yield, but gave slight decrease in WTS. Pasture combinations (diversified or pure) and animal categories (light and heavy) did not interfere in soybean culture, but benefited the maize crop.

Diurnal and nocturnal herbivore induction on maize elicit different innate response of the fall armyworm parasitoid, Campoletis flavicincta

Herbivore-attacked plants produce specific volatile substances that represent important cues for host finding by natural enemies. The fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is a voracious herbivore and usually feed on maize in all periods of the day. Given that plant needs light to synthesize de novo herbivore-induced volatiles, volatile blend may be changed depending on time of the day the plant is induced, what could interfere in natural enemy foraging. In this sense, the current study aimed to investigate differential attractiveness of maize elicited by fall armyworm regurgitant under light and dark conditions to its specialist larval parasitoid Campoletis flavicincta (Ashmead) (Hymenoptera: Ichneumonidae). All bioassays were conducted in Y-tube olfactometer to assess parasitoid response to odors from undamaged maize, mechanical damage, and regurgitant-treated plants at 0-1, 5-6, and 24-25 h after induction. The results showed that na < ve wasps were attracted to volatiles emitted by nocturnal regurgitant-treated maize at 5-6 h, but not to odors from diurnal regurgitant-treated plants. The differential attractiveness is likely due to blend composition as nocturnal regurgitant-treated plants emit aromatic compounds and the homoterpene (3E)-4,8-dimethyl-1,3,7-nonatriene in larger amounts than diurnal-treated plants.

Functional characterization of sugarcane mustang domesticated transposases and comparative diversity in sugarcane, rice, maize and sorghum

Transposable elements (TEs) account for a large portion of plant genomes, particularly in grasses, in which they correspond to 50%-80% of the genomic content. TEs have recently been shown to be a source of new genes and new regulatory networks. The most striking contribution of TEs is referred as "molecular domestication", by which the element coding sequence loses its movement capacity and acquires cellular function. Recently, domesticated transposases known as mustang and derived from the Mutator element have been described in sugarcane. In order to improve our understanding of the function of these proteins, we identified mustang genes from Sorghum bicolor and Zea mays and performed a phenetic analysis to assess the diversity and evolutionary history of this gene family. This analysis identified orthologous groups and showed that mustang genes are highly conserved in grass genomes. We also explored the transcriptional activity of sugarcane mustang genes in heterologous and homologous systems. These genes were found to be ubiquitously transcribed, with shoot apical meristem having the highest expression levels, and were downregulated by phytohormones. Together, these findings suggest the possible involvement of mustang proteins in the maintenance of hormonal homeostasis.

Inheritance of resistance to anthracnose stalk rot (Colletotrichum graminicola) in tropical maize inbred lines

Generation means was used to study the mode of inheritance of resistance to anthracnose stalk rot in tropical maize. Each population was comprised of six generations in two trials under a randomized block design. Inoculations were performed using a suspension of 105 conidia mL(-1) applied into the stalk. Internal lesion length was directly measured by opening the stalk thirty days after inoculation. Results indicated contrasting modes of inheritance. In one population, dominant gene effects predominated. Besides, additive x dominant and additive x additive interactions were also found. Intermediate values of heritability indicated a complex resistance inheritance probably conditioned by several genes of small effects. An additive-dominant genetic model sufficed to explain the variation in the second population, where additive gene effects predominated. Few genes of major effects control disease resistance in this cross. Heterosis widely differed between populations, which can be attributed to the genetic background of the parental resistant lines.

Germline-Specific MATH-BTB Substrate Adaptor MAB1 Regulates Spindle Length and Nuclei Identity in Maize

Germline and early embryo development constitute ideal model systems to study the establishment of polarity, cell identity, and asymmetric cell divisions (ACDs) in plants. We describe here the function of the MATH-BTB domain protein MAB1 that is exclusively expressed in the germ lineages and the zygote of maize (Zea mays). mab1 (RNA interference [RNAi]) mutant plants display chromosome segregation defects and short spindles during meiosis that cause insufficient separation and migration of nuclei. After the meiosis-to-mitosis transition, two attached nuclei of similar identity are formed in mab1 (RNAi) mutants leading to an arrest of further germline development. Transient expression studies of MAB1 in tobacco (Nicotiana tabacum) Bright Yellow-2 cells revealed a cell cycle-dependent nuclear localization pattern but no direct colocalization with the spindle apparatus. MAB1 is able to form homodimers and interacts with the E3 ubiquitin ligase component Cullin 3a (CUL3a) in the cytoplasm, likely as a substrate-specific adapter protein. The microtubule-severing subunit p60 of katanin was identified as a candidate substrate for MAB1, suggesting that MAB1 resembles the animal key ACD regulator Maternal Effect Lethal 26 (MEL-26). In summary, our findings provide further evidence for the importance of posttranslational regulation for asymmetric divisions and germline progression in plants and identified an unstable key protein that seems to be involved in regulating the stability of a spindle apparatus regulator(s).

Dry spell mitigation to upgrade semi-arid rainfed agriculture : Water harvesting and soil nutrient management for smallholder maize cultivation in Machakos, Kenya

Improvements in on-farm water and soil fertility management through water harvesting may prove key to up-grade smallholder farming systems in dry sub-humid and semi-arid sub-Sahara Africa (SSA). The currently experienced yield levels are usually less than 1 t ha-1, i.e., 3-5 times lower than potential levels obtained by commercial farmers and researchers for similar agro-hydrological conditions. The low yield levels are ascribed to the poor crop water availability due to variable rainfall, losses in on-farm water balance and inherently low soil nutrient levels. To meet an increased food demand with less use of water and land in the region, requires farming systems that provide more yields per water unit and/or land area in the future. This thesis presents the results of a project on water harvesting system aiming to upgrade currently practised water management for maize (Zea mays, L.) in semi-arid SSA. The objectives were to a) quantify dry spell occurrence and potential impact in currently practised small-holder grain production systems, b) test agro-hydrological viability and compare maize yields in an on-farm experiment using combinations supplemental irrigation (SI) and fertilizers for maize, and c) estimate long-term changes in water balance and grain yields of a system with SI compared to farmers currently practised in-situ water harvesting. Water balance changes and crop growth were simulated in a 20-year perspective with models MAIZE1&2. Dry spell analyses showed that potentially yield-limiting dry spells occur at least 75% of seasons for 2 locations in semi-arid East Africa during a 20-year period. Dry spell occurrence was more frequent for crop cultivated on soil with low water-holding capacity than on high water-holding capacity. The analysis indicated large on-farm water losses as deep percolation and run-off during seasons despite seasonal crop water deficits. An on-farm experiment was set up during 1998-2001 in Machakos district, semi-arid Kenya. Surface run-off was collected and stored in a 300m3 earth dam. Gravity-fed supplemental irrigation was carried out to a maize field downstream of the dam. Combinations of no irrigation (NI), SI and 3 levels of N fertilizers (0, 30, 80 kg N ha-1) were applied. Over 5 seasons with rainfall ranging from 200 to 550 mm, the crop with SI and low nitrogen fertilizer gave 40% higher yields (**) than the farmers’ conventional in-situ water harvesting system. Adding only SI or only low nitrogen did not result in significantly different yields. Accounting for actual ability of a storage system and SI to mitigate dry spells, it was estimated that a farmer would make economic returns (after deduction of household consumption) between year 2-7 after investment in dam construction depending on dam sealant and labour cost used. Simulating maize growth and site water balance in a system of maize with SI increased annual grain yield with 35 % as a result of timely applications of SI. Field water balance changes in actual evapotranspiration (ETa) and deep percolation were insignificant with SI, although the absolute amount of ETa increased with 30 mm y-1 for crop with SI compared to NI. The dam water balance showed 30% productive outtake as SI of harvested water. Large losses due to seepage and spill-flow occurred from the dam. Water productivity (WP, of ETa) for maize with SI was on average 1 796 m3 per ton grain, and for maize without SI 2 254 m3 per ton grain, i.e, a decerase of WP with 25%. The water harvesting system for supplemental irrigation of maize was shown to be both biophysically and economically viable. However, adoption by farmers will depend on other factors, including investment capacity, know-how and legislative possibilities. Viability of increased water harvesting implementation in a catchment scale needs to be assessed so that other down-stream uses of water remains uncompromised.

Oviposition by a moth suppresses constitutive and herbivore-induced plant volatiles in maize

Plant volatiles function as important signals for herbivores, parasitoids, predators, and neighboring plants. Herbivore attack can dramatically increase plant volatile emissions in many species. However, plants do not only react to herbivore-inflicted damage, but also already start adjusting their metabolism upon egg deposition by insects. Several studies have found evidence that egg deposition itself can induce the release of volatiles, but little is known about the effects of oviposition on the volatiles released in response to subsequent herbivory. To study this we measured the effect of oviposition by Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) moths on constitutive and herbivore-induced volatiles in maize (Zea mays L.). Results demonstrate that egg deposition reduces the constitutive emission of volatiles and suppresses the typical burst of inducible volatiles following mechanical damage and application of caterpillar regurgitant, a treatment that mimics herbivory. We discuss the possible mechanisms responsible for reducing the plant's signaling capacity triggered by S. frugiperda oviposition and how suppression of volatile organic compounds can influence the interaction between the plant, the herbivore, and other organisms in its environment. Future studies should consider oviposition as a potential modulator of plant responses to insect herbivores. © 2011 Springer-Verlag.

Metabolism of sulphonated anthraquinones in rhubarb, maize and celery: the role of cytochromes P450 and peroxidases

Sulphonated anthraquinones are precursors of many synthetic dyes and pigments, recalcitrant to biodegradation, and thus contaminating many industrial effluents and rivers. In the development of a phytotreatment to remove sulphonated aromatic compounds, rhubarb (Rheum rhaponticum), a plant producing natural anthraquinones, as well as maize (Zea mays) and celery (Apium graveolens), plants not producing anthraquinones, were tested for their ability to metabolise these xenobiotics. Plants were cultivated under hydroponic conditions, with or without sulphonated anthraquinones, and were harvested at different times. Either microsomal or cytosolic fractions were prepared. The monooxygenase activity of cytochromes P450 towards several sulphonated anthraquinones was tested using a new method based on the fluorimetric detection of oxygen consumed during cytochromes P450-catalysed reactions. The activity of cytosolic peroxidases was measured by spectrophotometry, using guaiacol as a substrate. Results indicated that the activity of cytochromes P450 and peroxidases significantly increased in rhubarb plants cultivated in the presence of sulphonated anthraquinones. A higher activity of cytochromes P450 was also detected in maize and celery exposed to the pollutants. In these two plants, a peroxidase activity was also detected, but without a clear difference between the control plants and the plants exposed to the organic contaminants. This research demonstrated the existence in rhubarb, maize and celery of biochemical mechanisms involved in the metabolism and detoxification of sulphonated anthraquinones. Taken together, results confirmed that rhubarb might be the most appropriate plant for the phytotreatment of these organic pollutants.

Natural Variation in Maize Aphid Resistance Is Associated with 2,4-Dihydroxy-7-Methoxy-1,4-Benzoxazin-3-One Glucoside Methyltransferase Activity

Plants differ greatly in their susceptibility to insect herbivory, suggesting both local adaptation and resistance tradeoffs. We used maize (Zea mays) recombinant inbred lines to map a quantitative trait locus (QTL) for the maize leaf aphid (Rhopalosiphum maidis) susceptibility to maize Chromosome 1. Phytochemical analysis revealed that the same locus was also associated with high levels of 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one glucoside (HDMBOA-Glc) and low levels of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside (DIMBOA-Glc). In vitro enzyme assays with candidate genes from the region of the QTL identified three O-methyltransferases (Bx10a-c) that convert DIMBOA-Glc to HDMBOA-Glc. Variation in HDMBOA-Glc production was attributed to a natural CACTA family transposon insertion that inactivates Bx10c in maize lines with low HDMBOA-Glc accumulation. When tested with a population of 26 diverse maize inbred lines, R. maidis produced more progeny on those with high HDMBOA-Glc and low DIMBOA-Glc. Although HDMBOA-Glc was more toxic to R. maidis than DIMBOA-Glc in vitro, BX10c activity and the resulting decline of DIMBOA-Glc upon methylation to HDMBOA-Glc were associated with reduced callose deposition as an aphid defense response in vivo. Thus, a natural transposon insertion appears to mediate an ecologically relevant trade-off between the direct toxicity and defense-inducing properties of maize benzoxazinoids.

Two enzymes responsible for the formation of herbivore-induced volatiles of maize, the methyltransferase AAMT1 and the terpene synthase TPS23, are regulated by a similar signal transduction pathway

Herbivore-induced volatiles play an important role in the indirect defense of plants. After herbivore damage, volatiles are released from the plant and can attract herbivore enemies that protect the plant from additional damage. The herbivore-induced volatile blend is complex and usually consists of mono- and sesquiterpenes, aromatic compounds, and indole. Although these classes of compounds are generally produced at different times after herbivore damage, the release of the terpene (E)-β-caryophyllene and the aromatic ester methyl anthranilate appear to be tightly coordinated. We have studied the herbivore induction patterns of two terpene synthases from Zea mays L. (Poaceae), TPS23 and TPS10, as well as S-adenosyl-L-methionine:anthranilic acid carboxyl methyltransferases (AAMT1), which are critical for the production of terpenes and anthranilate compounds, respectively. The transcript levels of tps23 and aamt1 displayed the same kinetics after damage by the larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae), and showed the same organ-specific and haplotype-specific expression patterns. Despite its close functional relation to TPS23, the terpene synthase TPS10 is not expressed in roots and does not display the haplotype-specific expression pattern. The results indicate that the same JA-mediated signaling cascade maycontrol the production of both the terpene (E)-β-caryophyllene and aromatic ester methyl anthranilate.

Maize Domestication and Anti-Herbivore Defences: Leaf-Specific Dynamics during Early Ontogeny of Maize and Its Wild Ancestors

As a consequence of artificial selection for specific traits, crop plants underwent considerable genotypic and phenotypic changes during the process of domestication. These changes may have led to reduced resistance in the cultivated plant due to shifts in resource allocation from defensive traits to increased growth rates and yield. Modern maize (Zea mays ssp. mays) was domesticated from its ancestor Balsas teosinte (Z. mays ssp. parviglumis) approximately 9000 years ago. Although maize displays a high genetic overlap with its direct ancestor and other annual teosintes, several studies show that maize and its ancestors differ in their resistance phenotypes with teosintes being less susceptible to herbivore damage. However, the underlying mechanisms are poorly understood. Here we addressed the question to what extent maize domestication has affected two crucial chemical and one physical defence traits and whether differences in their expression may explain the differences in herbivore resistance levels. The ontogenetic trajectories of 1,4-benzoxazin-3-ones, maysin and leaf toughness were monitored for different leaf types across several maize cultivars and teosinte accessions during early vegetative growth stages. We found significant quantitative and qualitative differences in 1,4-benzoxazin-3-one accumulation in an initial pairwise comparison, but we did not find consistent differences between wild and cultivated genotypes during a more thorough examination employing several cultivars/accessions. Yet, 1,4-benzoxazin-3-one levels tended to decline more rapidly with plant age in the modern maize cultivars. Foliar maysin levels and leaf toughness increased with plant age in a leaf-specific manner, but were also unaffected by domestication. Based on our findings we suggest that defence traits other than the ones that were investigated are responsible for the observed differences in herbivore resistance between teosinte and maize. Furthermore, our results indicate that single pairwise comparisons may lead to false conclusions regarding the effects of domestication on defensive and possibly other traits.

Belowground herbivore tolerance involves delayed overcompensatory root regrowth in maize

Plants can tolerate leaf-herbivore attack through metabolic reconfigurations that allow for the rapid regrowth of lost leaves. Several studies indicate that root-attacked plants can re-allocate resources to the aboveground parts. However, the connection between tolerance and root regrowth remains poorly understood. We investigated the timing and extent of root regrowth of tolerant and susceptible lines of maize, Zea mays L. (Poaceae), attacked by the western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), in the laboratory and the field. Infested tolerant maize plants produced more root biomass and even overcompensated for the lost roots, whereas this effect was absent in susceptible lines. Furthermore, the tolerant plants slowed growth of new roots in the greenhouse and in the field 4–8 days after infestation, whereas susceptible plants slowed growth of new roots only in the field and only after 12 days of infestation. The quick response of tolerant lines may have enabled them to escape root attack by starving the herbivores and by saving resources for regrowth after the attack had ceased. We conclude that both timing and the extent of regrowth may determine plant tolerance to root herbivory.