Bacillus velezensis strain Ag75 as a new multifunctional agent for biocontrol, phosphate solubilization and growth promotion in maize and soybean crops.

Soybean and maize are some of the main drivers of Brazilian agribusiness. However, biotic and abiotic factors are of great concern, causing huge grain yield and quality losses. Phosphorus (P) deficiency is important among the abiotic factors because most Brazilian soils have a highly P-fixing nature. Thus, large amounts of phosphate fertilizers are regularly applied to overcome the rapid precipitation of P. Searching for alternatives to improve the use of P by crops is essential to reduce the demand for P input. The use of multifunctional rhizobacteria can be considered one of these alternatives. In this sense, the objective of the present work was to select and validate bacterial strains with triple action (plant growth promoter, phosphate solubilizer, and biocontrol agent) in maize and soybean, aiming to develop a multifunctional microbial inoculant for Brazilian agriculture. Bacterial strains with high indole acetic acid (IAA) production, phosphate solubilization, and antifungal activity against soil pathogenic fungi (Rhizoctonia solani, Macrophomina phaseolina, and Fusarium solani) were selected from the maize rhizosphere. Then, they were evaluated as growth promoters in maize under greenhouse conditions. Based on this study, strain 03 (Ag75) was selected due to its high potential for increasing biomass (root and shoot) and shoot P content in maize. This strain was identified through genomic sequencing as Bacillus velezensis. In field experiments, the inoculation of this bacterium increased maize and soybean yields by 17.8 and 26.5%, respectively, compared to the control (25 kg P2O5). In addition, the inoculation results did not differ from the control with 84 kg P2O5, indicating that it is possible to reduce the application of phosphate in these crops. Thus, the Ag75 strain has great potential for developing a multifunctional microbial inoculant that combines the ability to solubilize phosphate, promote plant growth, and be a biocontrol agent for several phytopathogenic fungi.

Efficiency of Combining Strains Ag87 (Bacillus megaterium) and Ag94 (Lysinibacillus sp.) as Phosphate Solubilizers and Growth Promoters in Maize.

Increasing phosphorus (P) use efficiency in agricultural systems is urgent and essential to significantly reduce the global demand for this nutrient. Applying phosphate-solubilizing and plant growth-promoting bacteria in the rhizosphere represents a strategy worthy of attention. In this context, the present work aimed to select and validate bacterial strains capable of solubilizing phosphorous and promoting maize growth, aiming to develop a microbial inoculant to be used in Brazilian agriculture. Bacterial strains from the maize rhizosphere were evaluated based on their ability to solubilize phosphate and produce indole acetic acid. Based on these characteristics, 24 strains were selected to be further evaluated under laboratory, greenhouse, and field conditions. Among the selected strains, four (I04, I12, I13, and I17) showed a high potential to increase maize root growth and shoot P content. Strains I13 (Ag87) and I17 (Ag94) were identified by genomic sequencing as Bacillus megaterium and Lysinibacillus sp., respectively. These strains presented superior yield increments relative to the control treatment with 30% P. In addition, combining Ag87 and Ag94 resulted in even higher yield gains, indicating a synergistic effect that could be harnessed in a commercial inoculant for Brazilian agriculture.

Adaptability and Stability of Safflower Genotypes for Oil Production.

The study aimed to analyze the agronomic performance of 11 safflower genotypes using adaptability and stability methods, while identifying safflower genotypes with stable behavior and a high grain yield in different environments of the Brazilian Cerrado. Ten lines and a cultivar of safflower were evaluated in four environments in the Brazilian conditions. Our results revealed the genotypes P30, P35, P9, P11, and P31 to be superior for grain yield and P43, P7, P11, and P31 to be superior for oil content. The lowest Wricke index, an indication of genotype stability, was observed for P9 (0.41%), which is considered the most stable genotype, followed by P35 (1.29%) and P31 (1.98%). For the predictability of the behavior of genotypes in the environments, P7 (80.85%), P35 (86.10%), P31 (85.90%), and P9 (97.42%) were considered predictable genotypes. The genotypes P11 (1045.6 kg ha-1 and 19.7%) and P21 (952.7 kg ha-1 and 20.6%) are recommended for cultivation in this region, considering both their grain yield and oil content. Safflower is viable to use out of season in the Brazilian Cerrado. The crop can generate profits for farmers and be used for oil production in periods of uncertain corn production.

Genome-Wide Association Study Identifies Genomic Regions for Important Morpho-Agronomic Traits in Mesoamerican Common Bean.

The population growth trend in recent decades has resulted in continuing efforts to guarantee food security in which leguminous plants, such as the common bean (Phaseolus vulgaris L.), play a particularly important role as they are relatively cheap and have high nutritional value. To meet this demand for food, the main target for genetic improvement programs is to increase productivity, which is a complex quantitative trait influenced by many component traits. This research aims to identify Quantitative Trait Nucleotides (QTNs) associated with productivity and its components using multi-locus genome-wide association studies. Ten morpho-agronomic traits [plant height (PH), first pod insertion height (FPIH), number of nodules (NN), pod length (PL), total number of pods per plant (NPP), number of locules per pod (LP), number of seeds per pod (SP), total seed weight per plant (TSW), 100-seed weight (W100), and grain yield (YLD)] were evaluated in four environments for 178 Mesoamerican common bean domesticated accessions belonging to the Brazilian Diversity Panel. In order to identify stable QTNs, only those identified by multiple methods (mrMLM, FASTmrMLM, pLARmEB, and ISIS EM-BLASSO) or in multiple environments were selected. Among the identified QTNs, 64 were detected at least thrice by different methods or in different environments, and 39 showed significant phenotypic differences between their corresponding alleles. The alleles that positively increased the corresponding traits, except PH (for which lower values are desired), were considered favorable alleles. The most influenced trait by the accumulation of favorable alleles was PH, showing a 51.7% reduction, while NN, TSW, YLD, FPIH, and NPP increased between 18 and 34%. Identifying QTNs in several environments (four environments and overall adjusted mean) and by multiple methods reinforces the reliability of the associations obtained and the importance of conducting these studies in multiple environments. Using these QTNs through molecular techniques for genetic improvement, such as marker-assisted selection or genomic selection, can be a strategy to increase common bean production.

Genetic Variability of Mineral Content in Different Grain Structures of Bean Cultivars from Mesoamerican and Andean Gene Pools.

Beans (Phaseolus vulgaris L.) are an important source of proteins, carbohydrates, and micronutrients in the diets of millions of people in Latin America and Africa. Studies related to genetic variability in the accumulation and distribution of nutrients are valuable for biofortification programs, as there is evidence that the seed coat and embryo differ in the bioavailability of essential nutrients. In this study, we sought to evaluate the genetic variability of total mineral content in the grain and its constituent parts (seed coat, cotyledon, and embryonic axis) of bean genotypes from Mesoamerican and Andean centers of origin. Grain samples of 10 bean cultivars were analyzed for the content of proteins and minerals (Mg, Ca, K, P, Mn, S, Cu, B, Fe, and Zn) in the whole grains and seed coat, cotyledons, and embryonic axis tissues. Genetic variability was observed among the cultivars for protein content and all evaluated minerals. Moreover, differential accumulation of minerals was observed in the seed coat, cotyledons, and embryonic axis. Except for Ca, which accumulated predominantly in the seed coat, higher percentages of minerals were detected in the cotyledons. Furthermore, 100-grain mass values showed negative correlations with the contents of Ca, Mg, P, Zn, Fe, and Mn in whole grains or in the different grain tissues. In general, the Mesoamerican cultivars showed a higher concentration of minerals in the grains, whereas Andean cultivars showed higher concentrations of protein.

Genome-wide association study for grain mineral content in a Brazilian common bean diversity panel.

KEY MESSAGE: QTNs significantly associated to nine mineral content in grains of common bean were identified. The accumulation of favorable alleles was associated with a gradually increasing nutrient content in the grain. Biofortification is one of the strategies developed to address malnutrition in developing countries, the aim of which is to improve the nutritional content of crops. The common bean (Phaseolus vulgaris L.), a staple food in several African and Latin American countries, has excellent nutritional attributes and is considered a strong candidate for biofortification. The objective of this study was to identify genomic regions associated with nutritional content in common bean grains using 178 Mesoamerican accessions belonging to a Brazilian Diversity Panel (BDP) and 25,011 good-quality single nucleotide polymorphisms. The BDP was phenotyped in three environments for nine nutrients (phosphorus, potassium, calcium, magnesium, copper, manganese, sulfur, zinc, and iron) using four genome-wide association multi-locus methods. To obtain more accurate results, only quantitative trait nucleotides (QTNs) that showed repeatability (i.e., those detected at least twice using different methods or environments) were considered. Forty-eight QTNs detected for the nine minerals showed repeatability and were considered reliable. Pleiotropic QTNs and overlapping genomic regions surrounding the QTNs were identified, demonstrating the possible association between the deposition mechanisms of different nutrients in grains. The accumulation of favorable alleles in the same accession was associated with a gradually increasing nutrient content in the grain. The BDP proved to be a valuable source for association studies. The investigation of different methods and environments showed the reliability of markers associated with minerals. The loci identified in this study will potentially contribute to the improvement of Mesoamerican common beans, particularly carioca and black beans, the main groups consumed in Brazil.

Morpho-agronomic, biochemical and molecular analysis of genetic diversity in the Mesoamerican common bean panel.

The common bean (Phaseolus vulgaris L.) is of great importance to the food and nutritional security of many populations, and exploitation of the crop's genetic diversity is essential for the success of breeding programs. Thus, the aim of the present study was to evaluate the genetic diversity of 215 common bean accessions, which included cultivars, obsolete cultivars, improved lines, and landraces using morpho-agronomic and biochemical traits, and amplified fragment length polymorphism markers (AFLP). Genetic parameters, box plots, Pearson's correlation analysis, and Ward's hierarchical clustering were used to analyze the data. The Jaccard similarity coefficient and neighbour-joining clustering method were used for molecular analysis. A wide variability among the accessions was observed for morpho-agronomic and biochemical traits. Selective accuracy (Ac) and broad-sense heritability (h2) values were high to intermediate for all traits, except seed yield. Ward's hierarchical clustering analysis generated six groups. AFLP analysis also revealed significant differences among the accessions. There was no correlation between the differences based on genetic markers and those based on morpho-agronomic and biochemical data, which indicates that both datasets are important for elucidating the differences among accessions. The results of the present study indicate great genetic diversity among the evaluated accessions.

Development of a Protocol and a Diagrammatic Scale for Quantification of Bacterial Leaf Streak Disease on Young Plants of Maize.

Bacterial leaf streak (BLS), caused Xanthomonas vasicola pv. vasculorum (Xvv), has become a major concern for maize production, mainly in the United States and South America. Therefore, this study aimed to establish a protocol for Xvv inoculation in young maize plants under controlled conditions and to develop and validate a diagrammatic scale for evaluation of maize hybrids in regard to BLS resistance. The study was carried out in three steps: the establishment of a protocol for inoculation of Xvv in young maize plants under controlled conditions; the development and validation of a diagrammatic scale for BLS severity evaluation; and the screening for BLS resistance of 45 hybrids using the proposed protocol for bacterial inoculation and the diagrammatic scale developed in this study. Besides reproducing a more natural Xvv infection, the bacterial suspension spraying without injury inoculation method induced higher disease incidence and severity, as well as reproducibility of results under the experimental conditions established in this study. The proposed diagrammatic scale allowed evaluating BLS severity with up to 97.49% of the leaf area affected by the disease. Further, the use of the diagrammatic scale resulted in an increase of accuracy from 0.909 up to 0.992. The reaction of 45 maize hybrids to BLS allowed establishing six major groups of susceptibility to the disease. The most resistant maize hybrids to BLS formed a group of 13 hybrids, with disease severity below 5%.

Genetic Progress of Seed Yield and Nitrogen Use Efficiency of Brazilian carioca Common Bean Cultivars Using Bayesian Approaches.

Common bean (Phaseolus vulgaris L.) is one of the most important crops worldwide and is considered an essential source of proteins, fibers, and minerals in the daily diet of several countries. Nitrogen (N) is considered the most important nutrient for common bean crop. On the other hand, the reduction of chemical fertilizers is a global challenge, and the development of cultivars with more N use efficiency (NUsE) is considered one of the main strategies to reduce the amount of N fertilizers. Genetic progress of NUsE has been reported in several crops; however, there was still no quantity in common bean. In this study, our goal was to analyze the genetic progress of seed yield (SY) and NUsE-related traits of 40 carioca common bean cultivars release from 1970 to 2017 in eight environments under low (zero) or high N (40 kg ha-1) in top-dressing. Genetic progress, principal component analysis, correlations among traits, and cultivar stability were analyzed using Bayesian approaches. The lowest values of the deviance information criterion (DIC) for the full model tested indicated the presence of the genotype × N × environment interaction for all evaluated traits. Nitrogen utilization efficiency (NUtE) and nitrogen uptake efficiency (NUpE) were the traits that most contributed to discriminate cultivars. The genetic progress of SY under high N (0.53% year-1, 95% HPD = 0.39; 0.65% year-1) was similar to that obtained in low N conditions (0.48% year-1, 95% HPD = 0.31; 0.64% year-1). These results indicate that modern cultivars do not demand more N fertilizers to be more productive. In addition, we observed a high genetic variability for NUsE-related traits, but there was no genetic progress for these variables. SY showed negative correlation with seed protein content (Prot) in both N conditions, and there was no reduction in Prot in modern cultivars. Both modern and old cultivars showed adaptability and stability under contrasting N conditions. Our study contributed to improve our knowledge about the genetic progress of common bean breeding program in Brazil in the last 47 years, and our data will help researchers to face the challenge of increase NUsE and Prot in the next few years.

Azospirillum brasilense promotes increases in growth and nitrogen use efficiency of maize genotypes.

The development of cultivars with an improved nitrogen use efficiency (NUE) together with the application of plant growth-promoting bacteria is considered one of the main strategies for reduction of fertilizers use. In this sense, this study: i) evaluated the effect of Azospirillum brasilense on the initial development of maize genotypes; ii) investigated the influence of A. brasilense inoculation on NUE under nitrogen deficit; and iii) sought for more NUE genotypes with higher responsiveness to A. brasilense inoculation. Twenty-seven maize genotypes were evaluated in three independent experiments. The first evaluated the initial development of maize genotypes with and without A. brasilense (strain Ab-V5) inoculation of seeds on germination paper in a growth chamber. The second and third experiments were carried out in a greenhouse using Leonard pots and pots with substrate, respectively, and the genotypes were evaluated at high nitrogen, low nitrogen and low nitrogen plus A. brasilense Ab-V5 inoculation. The inoculation of seeds with A. brasilense Ab-V5 intensified plant growth, improved biochemical traits and raised NUE under nitrogen deficit. The inoculation of seeds with A. brasilense can be considered an economically viable and environmentally sustainable strategy for maize cultivation.