Associative bacteria influence maize (Zea mays L.) growth, physiology and root anatomy under different nitrogen levels.

Despite the great diversity of plant growth-promoting bacteria (PGPB) with potential to partially replace the use of N fertilisers in agriculture, few PGPB have been explored for the production of commercial inoculants, reinforcing the importance of identifying positive plant-bacteria interactions. Aiming to better understand the influence of PGPB inoculation in plant development, two PGPB species with distant phylogenetic relationship were inoculated in maize. Maize seeds were inoculated with Bacillus sp. or Azospirillum brasilense. After germination, the plants were subjected to two N treatments: full (N+) and limiting (N-) N supply. Then, anatomical, biometric and physiological analyses were performed. Both PGPB species modified the anatomical pattern of roots, as verified by the higher metaxylem vessel element (MVE) number. Bacillus sp. also increased the MVE area in maize roots. Under N+ conditions, both PGPB decreased leaf protein content and led to development of shorter roots; however, Bacillus sp. increased root and shoot dry weight, whereas A. brasilense increased photosynthesis rate and leaf nitrate content. In plants subjected to N limitation (N-), photosynthesis rate and photosystem II efficiency increased in maize inoculated with Bacillus sp., whilst A. brasilense contained higher ammonium, amino acids and total soluble sugars in leaves, compared to the control. Plant developmental and metabolical patterns were switched by the inoculation, regardless of the inoculant bacterium used, producing similar as well as distinct modifications to the parameters studied. These results indicate that even non-diazotrophic inoculant strains can improve the plant N status as result of the morpho-anatomical and physiological modifications produced by the PGPB.

Reference genes for quantitative real-time polymerase chain reaction studies in soybean plants under hypoxic conditions.

Quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful tool used to measure gene expression. However, because of its high sensitivity, the method is strongly influenced by the quality and concentration of the template cDNA and by the amplification efficiency. Relative quantification is an effective strategy for correcting random and systematic errors by using the expression level of reference gene(s) to normalize the expression level of the genes of interest. To identify soybean reference genes for use in studies of flooding stress, we compared 5 candidate reference genes (CRGs) with the NormFinder and GeNorm programs to select the best internal control. The expression stability of the CRGs was evaluated in root tissues from soybean plants subjected to hypoxic conditions. Elongation factor 1-beta and actin-11 were identified as the most appropriate genes for RT-qPCR normalization by both the NormFinder and GeNorm analyses. The expression profiles of the genes for alcohol dehydrogenase 1, sucrose synthase 4, and ascorbate peroxidase 2 were analyzed by comparing different normalizing combinations (including no normalization) of the selected reference genes. Here, we have identified potential genes for use as references for RT-qPCR normalization in experiments with soybean roots growing in O2-depleted environments, such as flooding-stressed plants.

Characterization of Injury Caused by Edessa meditabunda (F.), Chinavia impicticornis (Stål), and Piezodorus guildinii (West.) (Hemiptera: Pentatomidae) to Soybean.

Field cage trials were carried out in Ponta Grossa, Paraná, season 2010/2011, aiming to characterize and evaluate the injury caused by Edessa meditabunda (F.), Chinavia impicticornis (Stål), and Piezodorus guildinii (West.) (Hemiptera: Pentatomidae) to soybean. Non-infested plants were compared with infested plants with 2 and 3 adults/m (number of adult specimens per linear meter) at the R5.1-R9, R5.3-R9, R6-R9, and R7-R9 soybean growing stages. The following variables were analyzed: foliar retention, seed germination, and injury levels estimated by the tetrazolium test. Foliar retention index ranged from 2.3 to 4.1 in plants infested with P. guildinii and from 2.6 to 3.3 in plants infested with E. meditabunda and C. impicticornis, respectively. The lowest seed germination ranged from 66.5 to 76.5% and was observed in seeds from plants infested with 2 adults of C. impicticornis at R5.3-R9 and 3 adults of P. guildinii (R5.1-R9) and E. meditabunda (R7-R9). The highest average percentage of injury level for seeds occurred with seeds from plants infested at R5.1 with 2 or 3 adults/m of P. guildinii, ranging from 22.6 to 25.0% and from 63.6 to 68.0% in the first and second trials, respectively. Edessa meditabunda caused the highest damage to the seeds of plants infested at R7 (34.0 and 41.4%), and 2 adults/m of C. impicticornis caused the highest number of damaged seeds (54.0%) in plants infested at R6. Although both species were less harmful than P. guildinii, they could be a constraint factor for grain and seed commercialization.

Biological characteristics of Anticarsia gemmatalis (Lepidoptera: Noctuidae) for three consecutive generations under different temperatures: understanding the possible impact of global warming on a soybean pest.

Climate changes can affect the distribution and intensity of insect infestations through direct effects on their life cycles. Experiments were carried out during three consecutive generations to evaluate the effect of different temperatures (25°C, 28°C, 31°C, 34°C and 37±1°C) on biological traits of the velvetbean caterpillar Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Noctuidae). The insects were fed on artificial diet and reared in environmental chambers set at 14 h photophase. The developmental cycle slowed with the increase in the temperature, within the 25°C to 34°C range. Male and female longevities were reduced with an increase in temperature from 25°C to 28°C. Egg viability was highest at 25°C, and the sex ratio was not influenced by temperature, in the three generations. There was no interactive effect between development time and temperature on pupal weight. The results suggested that the increase in the temperature negatively impacted A. gemmatalis development inside the studied temperature range, indicating a possible future reduction of its occurrence on soybean crops, as a consequence of global warming, mainly considering its impact on tropical countries where this plant is cropped. A. gemmatalis was not able to adapt to higher temperatures in a three-generation interval for the studied temperature range. However, a gradual increase and a longer adaptation period may favor insect selection and consequently adaptation, and must be considered in future studies in this area. Moreover, it is important to consider that global warming might turn cold areas more suitable to A. gemmatalis outbreaks. Therefore, more than a future reduction of A. gemmatalis occurrence due to global warming, we might expect changes regarding its area of occurrence on a global perspective.

Molecular, anatomical and physiological properties of a genetically modified soybean line transformed with rd29A:AtDREB1A for the improvement of drought tolerance.

We evaluated the molecular, anatomical and physiological properties of a soybean line transformed to improve drought tolerance with an rd29A:AtDREB1A construct. This construct expressed dehydration- responsive element binding protein DREB1A from the stress-inducible rd29A promoter. The greenhouse growth test included four randomized blocks of soybean plants, with each treatment performed in triplicate. Seeds from the non-transformed soybean cultivar BR16 and from the genetically modified soybean P58 line (T(2) generation) were grown at 15% gravimetric humidity for 31 days. To induce water deficit, the humidity was reduced to 5% gravimetric humidity (moderate stress) for 29 days and then to 2.5% gravimetric humidity (severe stress). AtDREB1A gene expression was higher in the genetically modified P58 plants during water deficit, demonstrating transgene stability in T(2) generations and induction of the rd29A promoter. Drought-response genes, including GmPI-PLC, GmSTP, GmGRP, and GmLEA14, were highly expressed in plants submitted to severe stress. Genetically modified plants had higher stomatal conductance and consequently higher photosynthetic and transpiration rates. In addition, they had more chlorophyll. Overexpression of AtDREB1A may contribute to a decrease in leaf thickness; however, a thicker abaxial epidermis was observed. Overexpression of AtDREB1A in soybean appears to enhance drought tolerance.

Cloning and quantitative expression analysis of drought-induced genes in soybean.

We determined the expression levels of DREB transcription factor (Gmdreb1) and of the genes Gmgols, Gmpip1b, Gmereb, and Gmdefensin in drought-tolerant (MG/BR46-Conquista) and drought-sensitive (BR16) genotypes of soybean, during drought. The trial was carried out in a controlled-environment chamber, set up to provide drought conditions. Sequences of Arabidopsis thaliana DREB-family proteins were used to build a phylogenetic tree through the alignment of the conserved regions near the AP2 domain. We found that Gmdreb1 is similar to Atrap2.1, which is located near the AtDREB1 and AtDREB2 families. The amplified fragment was cloned and sequenced; alignment with the sequence available at Genbank showed total similarity. Expression analysis showed that under drought: a) Gmdreb1 expression increased in leaves and roots of both genotypes and expression level changes occurred that were correlated with the length of the water-deficit period; b) there were increased expression levels of Gmdefensin in roots of MG/BR46; c) expression of Gmgols increased in leaves and roots of the two genotypes; d) Gmpip1b expression generally increased, except in roots of BR16, and e) the same was found for Gmereb, except in roots of MG/BR46.