Decoding the effects of drought stress on popcorn (Zea mays var. everta) flowering combining proteomics and physiological analysis.

Under conditions of soil water limitation and adequate irrigation, we conducted an investigation into the growth dynamics, gas exchange performance, and proteomic profiles of two inbred popcorn lines-L71, characterized as drought-tolerant, and L61, identified as drought-sensitive. Our goal was to uncover the mechanisms associated with tolerance to soil water limitation during the flowering. The plants were cultivated until grain filling in a substrate composed of perlite and peat within 150cm long lysimeter, subjected to two water conditions (WC): i) irrigated (WW) at lysimeter capacity (LC - 100%), and ii) water-stressed (WS). Under WS conditions, the plants gradually reached 45% of LC and were maintained at this level for 10 days. Irrespective of the WC, L71 exhibited the highest values of dry biomass in both shoot and root systems, signifying its status as the most robust genotype. The imposed water limitation led to early senescence, chlorophyll degradation, and increased anthocyanin levels, with a more pronounced impact observed in L61. Traits related to gas exchange manifested differences between the lines only under WS conditions. A total of 1838 proteins were identified, with 169 differentially accumulated proteins (DAPs) in the tolerant line and 386 DAPs in the sensitive line. Notably, differences in energy metabolism, photosynthesis, oxidative stress response, and protein synthesis pathways were identified as the key distinctions between L71 and L61. Consequently, our findings offer valuable insights into the alterations in proteomic profiles associated with the adaptation to soil water limitation in popcorn.

Early selection for drought tolerance in popcorn based on gene effects estimated in seedlings.

Low rainfall rates are becoming increasingly frequent because of climate change, causing droughts and threatening world food security. For popcorn, drought is the most limiting abiotic factor for plant's growth and development. Thus, the water deficit directly impacts for crop productivity. Based on knowledge of the genetic basis of traits involved in stages of popcorn germination and seedling development under water stress, genotypes with potential for adaptation to adverse growing conditions can be selected early. Therefore, data on genetic effects and combining ability of 10 popcorn parents were compiled to propose breeding strategies for the development of cultivars with greater adaptation to water stress in the early stages. Forty-five diallel hybrids were evaluated under two different water regimes, that is, water stress and full irrigation. This corresponded to a water retention capacity of 25% and 70% of the germination paper. The plants were watered daily as needed for seven days. A range of factors were evaluated, that is, germination traits including the germination speed index and germination on the seventh day; shoot traits including length and dry weight; and root system including length, dry weight, root-to-shoot ratio, maximum root number, root network area, specific and root network length, and root volume. Breeding for drought adaption in the early stages of popcorn development can be successful when hybrids are used, because of the genetic effects of dominance (ϕs). These control the traits evaluated at the seedling stage. The combinations L61 x P2 and L71 x P3 were recommended, in view of the more successful performance estimated for traits related to the shoot and root system.

Exploring the Potential of Heterosis to Improve Nitrogen Use Efficiency in Popcorn Plants.

Nitrogen is crucial for plant growth and development, and improving nitrogen use efficiency (NUE) is a viable strategy for reducing dependence on nitrogen inputs and promoting sustainability. While the benefits of heterosis in corn are well known, the physiological mechanisms underlying this phenomenon in popcorn are less understood. We aimed to investigate the effects of heterosis on growth and physiological traits in four popcorn lines and their hybrids under two contrasting nitrogen conditions. We evaluated morpho-agronomic and physiological traits such as leaf pigments, the maximum photochemical efficiency of PSII, and leaf gas exchange. Components associated with NUE were also evaluated. N deprivation caused reductions of up to 65% in terms of plant architecture, 37% in terms of leaf pigments, and 42% in terms of photosynthesis-related traits. Heterosis had significant effects on growth traits, NUE, and foliar pigments, particularly under low soil nitrogen conditions. N-utilization efficiency was found to be the mechanism favoring superior hybrid performance for NUE. Non-additive genetic effects were predominant in controlling the studied traits, indicating that exploring heterosis is the most effective strategy for obtaining superior hybrids to promote NUE. The findings are relevant and beneficial for agro farmers seeking sustainable agricultural practices and improved crop productivity through the optimization of nitrogen utilization.

Recurrent Interpopulation Selection in Popcorn: From Heterosis to Genetic Gains.

In view of the need to develop new popcorn cultivars and considering the uncertainties in choosing the most appropriate breeding methods to ensure consistent genetic progress, simultaneously for both popping expansion and grain yield, this study addressed the efficiency of interpopulation recurrent selection regarding genetic gains, the study of the response in genetic parameters as well as heterotic effects on the control of the main agronomic traits of popcorn. Two populations were established, Pop1 and Pop2. A total of 324 treatments were evaluated, which consisted of 200 half-sib families (100 from Pop1 and 100 from Pop2), 100 full-sib families from the two populations and 24 controls. The field experiment was arranged in a lattice design with three replications in two environments, in the north and northwest regions of the State of Rio de Janeiro, Brazil. The genotype × environment interaction was partitioned and the genetic parameters, heterosis and predicted gains were estimated by the Mulamba and Mock index, based on selection results in both environments. The genetic parameters detected variability that can be explored in successive interpopulation recurrent selection cycles. Exploring heterosis for GY, PE and yield components is a promising option to increase grain yield and quality. The Mulamba and Mock index was efficient in predicting the genetic gains in GY and PE. Interpopulation recurrent selection proved effective to provide genetic gains for traits with predominantly additive and dominance inheritance.

Additive and Non-Additive Effects on the Control of Key Agronomic Traits in Popcorn Lines under Contrasting Phosphorus Conditions.

Phosphorus is a non-renewable natural resource that will run out of reserves in the upcoming decades, making it essential to understanding the inheritance of nutrient use efficiency for selecting superior genotypes. This study investigated the additive and non-additive effects of commercially relevant traits for the popcorn crop (grain yield-GY, popping expansion-PE, and expanded popcorn volume per hectare-PV) in different conditions of phosphorus (P) availability in two locations in Rio de Janeiro State, Brazil. Six S7 lines previously selected for P use-L59, L70, and P7, efficient and responsive; and L54, L75, and L80, inefficient and non-responsive-were used as testers in crosses with 15 progenies from the fifth cycle of intrapopulation recurrent selection of UENF-14, with adaptation to the North and Northwest regions of Rio de Janeiro State. Using the Griffing diallel analysis, P use efficiency was predominantly additive in the expression of PE, and non-additive effects were prominent for GY and PV. For obtaining genotypes that are efficient for phosphorus use, it is recommended that heterosis with parents that provide additive gene accumulation for PE be explored.

All Are in a Drought, but Some Stand Out: Multivariate Analysis in the Selection of Agronomic Efficient Popcorn Genotypes.

The search for productive germplasm adapted to adverse conditions is an important action to mitigate the harmful effects of climate change. The aim was to identify the yield potential of 50 popcorn inbred lines grown in field conditions, in two crop seasons (CS), and under contrasting water conditions (WC). Morphoagronomic, physiological, and root system traits were evaluated. Joint and individual analyses of variance were performed, in addition to the multivariate GT bip-lot analysis. Expressive reductions between WC were observed in 100-grain weight (100 GW), popping expansion (PE), grain yield (GY), expanded popcorn volume per ha (EPV), row number per ear (RNE), plant height (PH), relative chlorophyll content (SPAD), and nitrogen balance index (NBI). It was found that the SPAD, 100 GW, GY, PE, and grain number per ear (GNE) traits had the most significant impact on the selection of genotypes. Regardless of WC and CS, the ideal lines were L294 and L688 for PE; L691 and L480 for GY; and L291 and L292 for both traits. SPAD, 100 GW, and GNE can contribute to the indirect selection. Our work contributes to understanding the damage caused by drought and the integration of traits for the indirect selection of drought-tolerant popcorn genotypes.

Revealing the differential protein profiles behind the nitrogen use efficiency in popcorn (Zea mays var. everta).

We investigated the proteomic profiles of two popcorn inbred lines, P2 (N-efficient and N-responsive) and L80 (N-inefficient and nonresponsive to N), under low (10% of N supply) and high (100% of N supply) nitrogen environments, associated with agronomic- and physiological-related traits to NUE. The comparative proteomic analysis allowed the identification of 79 differentially accumulated proteins (DAPs) in the comparison of high/low N for P2 and 96 DAPs in the comparison of high/low N for L80. The NUE and N uptake efficiency (NUpE) presented high means in P2 in comparison to L80 at both N levels, but the NUE, NUpE, and N utilization efficiency (NUtE) rates decreased in P2 under a high N supply. DAPs involved in energy and carbohydrate metabolism suggested that N regulates enzymes of alternative pathways to adapt to energy shortages and that fructose-bisphosphate aldolase may act as one of the key primary nitrate responsive proteins in P2. Proteins related to ascorbate biosynthesis and nitrogen metabolism increased their regulation in P2, and the interaction of L-ascorbate peroxidase and Fd-NiR may play an important role in the NUE trait. Taken together, our results provide new insights into the proteomic changes taking place in contrasting inbred lines, providing useful information on the genetic improvement of NUE in popcorn.

Driving Sustainable Popcorn Breeding for Drought Tolerance in Brazil.

Drought currently affects several regions worldwide and tends to be more frequent due to climate change. It might compromise food security and the economic structure related to agribusiness. Popcorn has a crucial role in the Brazilian economy, but the cultivars that adapt to water stress, the most prejudicial abiotic stress for crop productivity, are unknown to date. This deficit of popcorn varieties adapted to heat and drought stresses will become more limiting with climate change. Given this scenario, knowing the genetic basis of agronomic traits under stress conditions is essential in promoting crop productivity and plant adaptation to abiotic stresses. Under two contrasting water conditions (WC) and different crop seasons (CS), we aimed to generate information about the combining ability of 10 popcorn progenitors and 15 hybrids through a partial diallel-mating design. The water stress was initiated at the male pre-anthesis stage. Significant genotype*crop seasons (G*CS), genotype*water condition (G*WC), and genotype*crop seasons*water condition (G*CS*WC) interactions were present. Regardless of CS and WC, non-additive effects controlled grain yield (GY), grain number per row (GN), ear length and diameter (ED), and 100-grain weight, while additive effects were present for popping expansion (PE). For each CS, regardless of WC, the cause-effect of GN (2018) and ED (2020) on GY seems to be an opportunity for indirect selection. Utilizing genetically broad-based hybrids is also a good opportunity for obtaining superior genotypes for GY and PE as it is possible to select inbred lines for both of these traits. We recommend the L76 × L61 hybrid for the Brazilian agribusiness context due to its greater productivity and dominance deviations.

Regional Heritability Mapping of Quantitative Trait Loci Controlling Traits Related to Growth and Productivity in Popcorn (Zea mays L.).

The method of regional heritability mapping (RHM) has become an important tool in the identification of quantitative trait loci (QTLs) controlling traits of interest in plants. Here, RHM was first applied in a breeding population of popcorn, to identify the QTLs and candidate genes involved in grain yield, plant height, kernel popping expansion, and first ear height, as well as determining the heritability of each significant genomic region. The study population consisted of 98 S1 families derived from the 9th recurrent selection cycle (C-9) of the open-pollinated variety UENF-14, which were genetically evaluated in two environments (ENV1 and ENV2). Seventeen and five genomic regions were mapped by the RHM method in ENV1 and ENV2, respectively. Subsequent genome-wide analysis based on the reference genome B73 revealed associations with forty-six candidate genes within these genomic regions, some of them are considered to be biologically important due to the proteins that they encode. The results obtained by the RHM method have the potential to contribute to knowledge on the genetic architecture of the growth and yield traits of popcorn, which might be used for marker-assisted selection in breeding programs.

Water Use Efficiency in Popcorn (Zea mays L. var. everta): Which Physiological Traits Would Be Useful for Breeding?

To ensure genetic gains in popcorn breeding programs carried out under drought conditions knowledge about the response of morphophysiological traits of plants to water stress for the selection of key traits is required. Therefore, the objective was to evaluate popcorn inbred lines with agronomically efficient (P2 and P3) and inefficient (L61 and L63) water use and two hybrids (P2xL61 and P3xL63) derived from these contrasting parents, cultivated under two water regimes (WW watered-WW; and water-stressed-WS) in a greenhouse, replicated five times, where each experimental unit consisted of one plant in a PVC tube. Irrigation was applied until stage V6 and suspended thereafter. Individual and combined analyses of variance were performed and the genotypic correlations and relative heteroses estimated. The water use efficient inbred lines were superior in root length (RL), root dry weight (RDW), and net CO2 assimilation rate (A), which were the characteristics that differentiated the studied genotypes most clearly. High heterosis estimates were observed for RL, SDW, leaf width (LW), leaf midrib length (LL), and agronomic water use efficiency (AWUE). The existence of a synergistic association between root angle and length for the characteristics A, stomatal conductance (gs), and chlorophyll concentration (SPAD index) proved most important for the identification and phenotyping of superior genotypes. Based on the study of these characteristics, the higher AWUE of the previously selected inbred lines could be explained. The results reinforced the importance of root physiological and morphological traits to explain AWUE and the possibility of advances by exploiting heterosis, given the morphophysiological superiority of hybrids in relation to parents.

Supporting Physiological Trait for Indirect Selection for Grain Yield in Drought-Stressed Popcorn.

The identification of traits associated with drought tolerance in popcorn is a contribution to support selection of superior plants under soil water deficit. The objective of this study was to choose morphological traits and the leaf greenness index, measured on different dates, to estimate grain yield (GY) and popping expansion (PE), evaluated in a set of 20 popcorn lines with different genealogies, estimated by multiple regression models. The variables were divided into three groups: morpho-agronomic traits-100-grain weight (GW), prolificacy (PR), tassel length (TL), number of tassel branches, anthesis-silking interval, leaf angle (FA) and leaf rolling (FB); variables related to the intensity of leaf greenness during the grain-filling period, at the leaf level, measured by a portable chlorophyll meter (SPAD) and at the canopy level, calculated as the normalized difference vegetation index (NDVI). The inbred lines were cultivated under two water conditions: well-watered (WW), maintained at field capacity, and water stress (WS), for which irrigation was stopped before male flowering. The traits GY (55%) and PE (28%) were most affected by water restriction. Among the morpho-agronomic traits, GW and PR were markedly reduced (>10%). Under dry conditions, the FA in relation to the plant stalk tended to be wider, the FB curvature greater and leaf senescence accelerated (>15% at 22 days after male flowering). The use of multiple regression for the selection of predictive traits proved to be a useful tool for the identification of groups of adequate traits to efficiently predict the economically most important features of popcorn (GY and PE). The SPAD index measured 17 days after male flowering proved useful to select indirectly for GY, while, among the morphological traits, TL stood out for the same purpose. Of all traits, PR was most strongly related with PE under WS, indicating its use in breeding programs. The exploitation of these traits by indirect selection is expected to induce increments in GY and PE.

Phenotyping Latin American Open-Pollinated Varieties of Popcorn for Environments with Low Water Availability.

Drought is a common abiotic stress in tropical and subtropical regions that limits the growth and development of agricultural crops, mainly impacting grain yield. Acting through plant breeding is the most viable alternative for obtaining genotypes more tolerant of environments with stress. This work aims to select popcorn genotypes for environments with drought and to identify discriminating traits for the evaluation of drought tolerance in popcorn germplasm. Fifteen Latin American populations of popcorn were evaluated in water stress (WS) and well-watered (WW) conditions. The evaluated traits were based in morpho-agronomic, physiological and radicular descriptors. Data were submitted to individual and joint ANOVA and GT Biplot analysis. Variability was detected between populations for all traits in both conditions. The drought caused a reduction of 30.61% and 3.5% in grain yield and popping expansion, respectively. Based in GT biplot analysis, 880POP was the most stable in WS and WW, being indicated as a promising population for cultivation in environments with water limitation. This study is going to allow the establishment of a collection of great importance to maize germplasm and to provide information to facilitate the process of selection in breeding programs focused on drought tolerance.

Comparative Performance of High-Yielding European Wheat Cultivars Under Contrasting Mediterranean Conditions.

Understanding the interaction between genotype performance and the target environment is the key to improving genetic gain, particularly in the context of climate change. Wheat production is seriously compromised in agricultural regions affected by water and heat stress, such as the Mediterranean basin. Moreover, wheat production may be also limited by the nitrogen availability in the soil. We have sought to dissect the agronomic and physiological traits related to the performance of 12 high-yield European bread wheat varieties under Mediterranean rainfed conditions and different levels of N fertilization during two contrasting crop seasons. Grain yield was more than two times higher in the first season than the second season and was associated with much greater rainfall and lower temperatures. However, the nitrogen effect was rather minor. Genotypic effects existed for the two seasons. While several of the varieties from central/northern Europe yielded more than those from southern Europe during the optimal season, the opposite trend occurred in the dry season. The varieties from central/northern Europe were associated with delayed phenology and a longer crop cycle, while the varieties from southern Europe were characterized by a shorter crop cycle but comparatively higher duration of the reproductive period, associated with an earlier beginning of stem elongation and a greater number of ears per area. However, some of the cultivars from northern Europe maintained a relatively high yield capacity in both seasons. Thus, KWS Siskin from the UK exhibited intermediate phenology, resulting in a relatively long reproductive period, together with a high green area throughout the crop cycle.

Comparison of Selection Traits for Effective Popcorn (Zea mays L. var. Everta) Breeding Under Water Limiting Conditions.

Climate change is expected to intensify water restriction to crops, impacting on the yield potential of crops such as popcorn. This work aimed to evaluate the performance of 10 field cultivated popcorn inbred lines during two growing seasons, under well-watered (WW) and water stressed (WS) (ψsoil≥ -1.5 MPa) conditions. Water stress was applied by withholding irrigation in the phenological phase of male pre-anthesis. Additionally, two contrasting inbred lines, P7 (superior line) and L75 (low performer) were compared for grain yield (GY) and expanded popcorn volume (EPV), selected from previous studies, were tested under greenhouse conditions. In the field, no genotype x water condition x crop season (G×WC×CS) interaction was observed, whereas GY (-51%), EPV (-55%) and leaf greenness (SPAD index) measured 17 days after anthesis (DAA) (> -10%) were highly affected by water limitation. In general, root traits (angles, number, and density) presented G×WC×CS interaction, which did not support their use as selection parameters. In relation to leaf senescence, for both WS and WW conditions, the superior inbred lines maintained a stay-green condition (higher SPAD index) until physiological maturity, but maximum SPAD index values were observed later in WW (48.7 by 14 DAA) than in WS (43.9 by 7 DAA). Under both water conditions, negative associations were observed between SPAD index values 15 and 8 days before anthesis DBA), and GY and EPV (r ≥ -0.69), as well as between SPAD index 7, 17, and 22 DAA, and angles of brace root (AB), number of crown roots (NC) and crown root density (CD), in WS (r ≥ -0.69), and AB and CD, in WW (r ≥ -0.70). Lower NC and CD values may allow further root deepening in WS conditions. Under WS P7 maintained higher net photosynthesis values, stomatal conductance, and transpiration, than L75. Additionally, L75 exhibited a lower (i.e., more negative) carbon isotope composition value than P7 under WS, confirming a lower stomatal aperture in L75. In summary, besides leaf greenness, traits related to leaf photosynthetic status, and stomatal conductance were shown to be good indicators of the agronomic performance of popcorn under water constraint.

Limited Nitrogen and Plant Growth Stages Discriminate Well Nitrogen Use, Uptake and Utilization Efficiency in Popcorn.

The extensive use of nitrogen (N) in agriculture has caused negative impacts on the environment and costs. In this context, two pot experiments were performed under different N levels and harvested at different vegetative stages to assess two popcorn inbred lines (P2 and L80) and their hybrid (F1 = P2 × L80) for the N use, uptake and utilization efficiency (with the inclusion and exclusion of root N content); to find the contrasting N levels and vegetative stages that effect nitrogen use efficiency (NUE) and to understand the relationship between the traits related to NUE. The hybrid and P2 were confirmed better than L80 for all the studied traits. NUE is mainly affected by the shoot dry weight, uptake and utilization efficiency. Extremely low and high N levels were found to be more discriminating for N use and dry weight, respectively. At the V6 (six fully expanded leaf) stage, root N content (RNC) should be considered; in contrast, at the VT (tasseling stage) stage, RNC should not be considered for the uptake and utilization efficiency. The genetic parameter performance for N use, uptake, shoot dry weight and N content could favor the achievement of the genetic gain in advanced segregating generations.

Relative importance of gene effects for nitrogen-use efficiency in popcorn.

The objective of this study was to evaluate the effects of additive and non-additive genes on the efficiency of nitrogen (N) use and N responsiveness in inbred popcorn lines. The parents, hybrids and reciprocal crosses were evaluated in a 10x10 triple lattice design at two sites and two levels of N availability. To establish different N levels in the two experiments, fertilization was carried out at sowing, according to soil analysis reports. However, for the experiments with ideal nitrogen availability, N was sidedressed according to the crop requirement, whereas for the N-poor experiments sidedressing consisted of 30% of that applied in the N-rich environment. Two indices were evaluated, the Harmonic Mean of the Relative Performance (HMRP) and Agronomic Efficiency under Low Nitrogen Availability (AELN), both based on grain yield at both N levels. Both additive and non-additive gene effects were important for selection for N-use efficiency. Moreover, there was allelic complementarity between the lines and a reciprocal effect for N-use efficiency, indicating the importance of the choice of the parents used as male or female. The best hybrids were obtained from inbred popcorn lines with contrasting N-use efficiency and N responsiveness.

SNP-based mixed model association of growth- and yield-related traits in popcorn.

The identification of the genes responsible for complex traits is highly promising to accelerate crop breeding, but such information is still limited for popcorn. Thus, in the present study, a mixed linear model-based association analysis (MLMA) was applied for six important popcorn traits: plant and ear height, 100-grain weight, popping expansion, grain yield and expanded popcorn volume per hectare. To this end, 196 plants of the open-pollinated popcorn population UENF-14 were sampled, selfed (S1), and then genotyped with a panel of 10,507 single nucleotide polymorphisms (SNPs) markers distributed throughout the genome. The six traits were studied under two environments [Campos dos Goytacazes-RJ (ENV1) and Itaocara-RJ (ENV2)] in an incomplete block design. Based on the phenotypic data of the S1 progenies and on the genetic characteristics of the parents, the MLMA was performed. Thereafter, genes annotated in the MaizeGDB platform were screened for potential linkage disequilibrium with the SNPs associated to the six evaluated traits. Overall, seven and eight genes were identified as associated with the traits in ENV1 and ENV2, respectively, and proteins encoded by these genes were evaluated for their function. The results obtained here contribute to increase knowledge on the genetic architecture of the six evaluated traits and might be used for marker-assisted selection in breeding programs.

Genetic effects on the efficiency and responsiveness to phosphorus use in popcorn as estimated by diallel analysis.

Agricultural expansion and the need for sustainable cultivation are challenges faced by researchers involved in the generation of new cultivars that can adapt to abiotic stress. Knowledge of the genetic effects of characteristics related to efficiency and responsiveness to phosphorus use must be considered when implementing methods to obtain better genotypes. The aim of this study was to characterize and select popcorn hybrids based on their efficiency and responsiveness to phosphorus use, and estimate their combining abilities and genetic effects via diallel analysis to implement improvement programs for sustainable agriculture. Eight contrasting inbred lines were used to obtain simple hybrids for diallel analysis. Twenty-eight diallelic hybrids plus the popcorn parental lines were evaluated at two different sites under two contrasting environments for soil phosphorus availability (6 × 6 lattice design). Grain yield, popping expansion, and volume of expanded popcorn per hectare were measured. A combined analysis of variance and a test of means were performed. The classification and utilization of the phosphorus use efficiency index, according to the grain yield performance of the hybrids under contrasting environments, was considered. Through model 2 of the Griffing's diallel analysis method, the general and specific combining abilities were estimated, along with their environmental interactions. The best strategy to obtain genotypes that are efficient and responsive to phosphorus involves exploring popcorn hybrids using genitors that result in the accumulation of additive genes that promote popping expansion. Hybrids P7 × L80, P7 × L59, P7 × L76, and P6 × L80 presented promising results and may be evaluated as cultivation options in phosphorus-deficient soils.