Insights into soybean transcriptome reconfiguration under hypoxic stress: Functional, regulatory, structural, and compositional characterization.

Soybean (Glycine max) is one of the major crops worldwide and flooding stress affects the production and expansion of cultivated areas. Oxygen is essential for mitochondrial aerobic respiration to supply the energy demand of plant cells. Because oxygen diffusion in water is 10,000 times lower than in air, partial (hypoxic) or total (anoxic) oxygen deficiency is important component of flooding. Even when oxygen is externally available, oxygen deficiency frequently occurs in bulky, dense or metabolically active tissues such as phloem, meristems, seeds, and fruits. In this study, we analyzed conserved and divergent root transcriptional responses between flood-tolerant Embrapa 45 and flood-sensitive BR 4 soybean cultivars under hypoxic stress conditions with RNA-seq. To understand how soybean genes evolve and respond to hypoxia, stable and differentially expressed genes were characterized structurally and compositionally comparing its mechanistic relationship. Between cultivars, Embrapa 45 showed less up- and more down-regulated genes, and stronger induction of phosphoglucomutase (Glyma05g34790), unknown protein related to N-terminal protein myristoylation (Glyma06g03430), protein suppressor of phyA-105 (Glyma06g37080), and fibrillin (Glyma10g32620). RNA-seq and qRT-PCR analysis of non-symbiotic hemoglobin (Glyma11g12980) indicated divergence in gene structure between cultivars. Transcriptional changes for genes in amino acids and derivative metabolic process suggest involvement of amino acids metabolism in tRNA modifications, translation accuracy/efficiency, and endoplasmic reticulum stress in both cultivars under hypoxia. Gene groups differed in promoter TATA box, ABREs (ABA-responsive elements), and CRT/DREs (C-repeat/dehydration-responsive elements) frequency. Gene groups also differed in structure, composition, and codon usage, indicating biological significances. Additional data suggests that cis-acting ABRE elements can mediate gene expression independent of ABA in soybean roots under hypoxia.

Characterization of Soybean Genetically Modified for Drought Tolerance in Field Conditions.

Drought is one of the most stressful environmental factor causing yield and economic losses in many soybean-producing regions. In the last decades, transcription factors (TFs) are being used to develop genetically modified plants more tolerant to abiotic stresses. Dehydration responsive element binding (DREB) and ABA-responsive element-binding (AREB) TFs were introduced in soybean showing improved drought tolerance, under controlled conditions. However, these results may not be representative of the way in which plants behave over the entire season in the real field situation. Thus, the objectives of this study were to analyze agronomical traits and physiological parameters of AtDREB1A (1Ab58), AtDREB2CA (1Bb2193), and AtAREB1 (1Ea2939) GM lines under irrigated (IRR) and non-irrigated (NIRR) conditions in a field experiment, over two crop seasons and quantify transgene and drought-responsive genes expression. Results from season 2013/2014 revealed that line 1Ea2939 showed higher intrinsic water use and leaf area index. Lines 1Ab58 and 1Bb2193 showed a similar behavior to wild-type plants in relation to chlorophyll content. Oil and protein contents were not affected in transgenic lines in NIRR conditions. Lodging, due to plentiful rain, impaired yield from the 1Ea2939 line in IRR conditions. qPCR results confirmed the expression of the inserted TFs and drought-responsive endogenous genes. No differences were identified in the field experiment performed in crop season 2014/2015, probably due to the optimum rainfall volume during the cycle. These field screenings showed promising results for drought tolerance. However, additional studies are needed in further crop seasons and other sites to better characterize how these plants may outperform the WT under field water deficit.

Seasonal and vertical distribution of Phyllophaga cuyabana (Moser)(Coleoptera: Melolonthidae) in the soil profile.

Phyllophaga cuyabana (Moser) temporal and vertical distribution patterns were evaluated in the soil profile, in order to subsidize methodology for population sampling, aiming at its management. In insect surveys carried out during three years, in Boa Esperança County, State of Parana, Brazil, Phyllophaga cuyabana was univoltine, with little overlap of the larval stages. Population peaked during December-February, but declined during the colder months, when larvae were in diapause. Different developmental stages exploited distinct soil depths. Eggs and early first instars tended to concentrate between 5 cm and 10 cm deep, but they spread more uniformly through the soil profile, reaching depths up to 30 cm, as they developed. Adults and eggs occurred in the spring (October to December) when active larvae also started to be observed; feeding larvae occurred up to late-April between 0 to 15 cm deep. Diapausing larvae and pupae were observed from early fall to early spring, mostly from 15 cm to 30 cm deep. Throughout the year, the number of insects in the soil (up to 40 cm deep) showed a positive functional relationship with air temperature and evapotranspiration. The relationship of percent distribution of larvae in the soil profile and soil temperature, however, was positive only above 10 cm. To estimate the insect population from November to April, samples can be collected until 20 cm deep; from May to October, however, samplings should be deeper, up to 30 cm.

Seasonal and vertical distribution of Phyllophaga cuyabana (Moser)(Coleoptera: Melolonthidae) in the soil profile / Distribuição estacional e vertical de Phyllophaga cuyabana (Moser) (Coleoptera: Melolonthidae) no perfil do solo

Phyllophaga cuyabana (Moser) temporal and vertical distribution patterns were evaluated in the soil profile, in order to subsidize methodology for population sampling, aiming at its management. In insect surveys carried out during three years, in Boa Esperança County, State of Parana, Brazil, Phyllophaga cuyabana was univoltine, with little overlap of the larval stages. Population peaked during December-February, but declined during the colder months, when larvae were in diapause. Different developmental stages exploited distinct soil depths. Eggs and early first instars tended to concentrate between 5 cm and 10 cm deep, but they spread more uniformly through the soil profile, reaching depths up to 30 cm, as they developed. Adults and eggs occurred in the spring (October to December) when active larvae also started to be observed; feeding larvae occurred up to late-April between 0 to15 cm deep. Diapausing larvae and pupae were observed from early fall to early spring, mostly from 15 cm to 30 cm deep. Throughout the year, the number of insects in the soil (up to 40 cm deep) showed a positive functional relationship with air temperature and evapotranspiration. The relationship of percent distribution of larvae in the soil profile and soil temperature, however, was positive only above 10 cm. To estimate the insect population from November to April, samples can be collected until 20 cm deep; from May to October, however, samplings should be deeper, up to 30 cm.

O padrão de distribuição temporal e vertical de Phyllophaga cuyabana (Moser) foi avaliado no perfil do solo para subsidiar amostragens da população do inseto visando ao seu manejo. Em levantamentos populacionais realizados durante três anos, em de Boa Esperança, PR, P. cuyabana foi univoltina, com baixa sobreposição de estádios de desenvolvimento. Houve um pico populacional no verão (dezembro a fevereiro) e um declínio nos meses frios, quando as larvas estavam em diapausa. O inseto, nos distintos estágios de desenvolvimento, explorou diferentes profundidades do solo. Ovos e larvas no início do primeiro instar concentraram-se entre 5 cm e 10 cm de profundidade e, ao se desenvolverem, atingiram 30 cm de profundidade. Adultos e ovos ocorreram na primavera (outubro a dezembro), quando começaram a ser observadas as larvas ativas em amostras realizadas entre zero e 15 cm de profundidade. Larvas em diapausa e pupas foram observadas em maior concentração entre 15 e 30 cm de profundidade, do inicio do outono ao início da primavera. O número de insetos no solo (até 40 cm de profundidade) mostrou relação funcional positiva com a temperatura do ar e com a evapotranspiração. Entretanto, a relação da distribuição percentual de larvas no perfil do solo com a temperatura do solo foi positiva apenas para profundidades de zero a 10 cm. Para estimar a população de corós de novembro a abril, as amostragens podem ser feitas até 20 cm de profundidade, porém de maio a outubro a profundidade das amostragens deve atingir 30 cm.

Differential gene expression and mitotic cell analysis of the drought tolerant soybean (Glycine max L. Merrill Fabales, Fabaceae) cultivar MG/BR46 (Conquista) under two water deficit induction systems

Drought cause serious yield losses in soybean (Glycine max), roots being the first plant organ to detect the water-stress signals triggering defense mechanisms. We used two drought induction systems to identify genes differentially expressed in the roots of the drought-tolerant soybean cultivar MG/BR46 (Conquista) and characterize their expression levels during water deficit. Soybean plants grown in nutrient solution hydroponically and in sand-pots were submitted to water stress and gene expression analysis was conducted using the differential display (DD) and real time polymerase chain reaction (PCR) techniques. Three differentially expressed mRNA transcripts showed homology to the Antirrhinum majus basic helix-loop-helix transcription factor bHLH, the Arabidopsis thaliana phosphatidylinositol transfer protein PITP and the auxin-independent growth regulator 1 (axi 1). The hydroponic experiments showed that after 100 min outside the nutrient solution photosynthesis completely stopped, stomata closed and leaf temperature rose. Both stress induction treatments produced significant decrease in the mitotic indices of root cells. Axi 1, PITP and bHLH were not only differentially expressed during dehydration in the hydroponics experiments but also during induced drought in the pot experiments. Although, there were differences between the two sets of experiments in the time at which up or down regulation occurred, the expression pattern of all three transcripts was related. Similar gene expression and cytological analysis results occurred in both systems, suggesting that hydroponics could be used to simulate drought detection by roots growing in soil and thus facilitate rapid and easy root sampling.

Natural Occurrence of the Entomopathogenic Fungi Metarhizium, Beauveria and Paecilomyces in Soybean Under Till and No-till Cultivation Systems / Ocorrência Natural de Fungos Entomopatogênicos Metarhizium, Beauveria e Paecilomyces na Cultura da Soja sob Plantio Direto e Convencional

A ocorrência de fungos entomopatogênicos Metarhizium, Beauveria e Paecilomyces foi estudada em condições de semeadura direta e convencional da soja. Foi determinada a densidade de unidades formadoras de colônia por g de solo e por cm² de folíolos de soja. Verificou-se que no solo sob semeadura direta ocorreu maior incidência dos entomopatógenos, mas sobre os folíolos essa diferença não ocorreu, proporcionando as mesmas possibilidades de infecção nos insetos da parte aérea suscetíveis que ocorrem nas duas condições de cultivo.

The occurrence of the entomopathogenic fungi Metarhizium, Beauveria and Paecilomyces in soybean under till and no-till cultivation systems was studied. The number of colony forming units per g of soil and per cm² of leaf area was determined. Soil under no-till system showed a higher incidence of entomopathogenic fungi, but no difference in fungal density was observed on leaves. Thus the probability of infection for susceptible leaf inhabiting insects should be similar in both systems.