Short-term impact of soybean management on ammonia oxidizers in a Brazilian savanna under restoration as revealed by coupling different techniques

Acknowledgments The authors are very grateful to Mr. Fabiano Bielefeld Nardotto, owner of the Tabapuã dos Pireneus farm, for allowing our free movement around the farm and collection of soil samples, as well as providing information about soybean cultivation. The authors also thank Dr. Plínio de Camargo, who performed the isotopic analysis in the CENA laboratory at the University of São Paulo (USP). This work was supported by grants from the National Council of Technological and Scientific Development (CNPq), Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES), and Foundation for Research Support of Distrito Federal (FAP-DF).

The expansion of soybean cultivation in Campeche, México: Problematic and perspectives

The soybean is the grain in which greater food dependency has Mexico, reason why as of 2008, the government has promoted his culture, granting excellent subsidies, as much to producers as to buyers of the grain, thus contributing to a recent process of expansion in certain states, as it happens in Campeche. The objetive of this article is the analysis of the characteristics and effects of those supports, as well as of the rest of factors that until today they have taken to the producers of the mentioned state to initiate or to expand the cultivation of the soybean. The findings of the investigation reveal that although the producers have improved their levels of income, the process is vulnerable, as it depends on variables like the governmental supports, the international prices of the soybean and exchange rate. Although the study of the negative effects of genetically modified soybeans (GM) in other areas (environment, biodiversity, deforestation, human and animal health) is not the purpose of this investigation, some information will be provided, as on the conflict between soybean producers and beekeepers in the state of Campeche.

Analytical strategies for the early quality and safety assurance in the global feed chain. Approaches for nitrogen adulterants in soybean meal and mineral and transformer oils in vegetable oils.

In the past decade, several major food safety crises originated from problems with feed. Consequently, there is an urgent need for early detection of fraudulent adulteration and contamination in the feed chain. Strategies are presented for two specific cases, viz. adulterations of (i) soybean meal with melamine and other types of adulterants/contaminants and (ii) vegetable oils with mineral oil, transformer oil or other oils. These strategies comprise screening at the feed mill or port of entry with non-destructive spectroscopic methods (NIRS and Raman), followed by post-screening and confirmation in the laboratory with MS-based methods. The spectroscopic techniques are suitable for on-site and on-line applications. Currently they are suited to detect fraudulent adulteration at relatively high levels but not to detect low level contamination. The potential use of the strategies for non-targeted analysis is demonstrated.

Determination of soybean cultivar resistance to soybean cyst nematode with quantitative polymerase chain reaction

Heterodera glycines, the soybean cyst nematode, is the major pathogen of Glycine max (soybean). Effective management of this pathogen is contingent on the use of resistant cultivars, thus screening for resistant cultivars is essential. The purpose of this research was to develop a method to assess infection of soybean roots by H. glycines with real-time quantitative Polymerase Chain Reaction (qPCR), a prelude to differentiation of resistance levels in soybean cultivars. Two experiments were conducted. In the first one, a consistent inoculation method was developed using to provide active second-stage juveniles (J2). Two-day-old soybean roots were infested with 0 and 1000 J2/mL. Twenty-four hours after infestation, the roots were surface sterilized and DNA was extracted with the DNA FastKit (MP Biomedicals, Santa Ana, CA)). For the qPCR assay, primer pair for single copy gene HgSNO, which codes for a protein involved in the production of vitamin B6, was selected for H. glycines DNA amplification within soybean roots. In the second experiment, compatible Lee 74, incompatible Peking and cultivars with different levels of resistance to H. glycines were inoculated with 0 and 1,000 J2/seedlings. Twenty-four hours post inoculation they were transplanted into pasteurized soil. Subsequently they were harvested at 1, 7, 10, 14 and 21 days post inoculation for DNA extraction. With the qPCR assay, the time needed to differentiate highly resistant cultivars from the rest was reduced. Quantification of H. glycines infection by traditional means (numbers of females produced in 30 days) is a time-consuming practice; the qPCR method can replace the traditional one and improve precision in determining infection levels.

Genomic and biochemical investigation of soybean antioxidant metabolism in response to growth at elevated carbon dioxide and elevated ozone

Metabolism in an environment containing of 21% oxygen has a high risk of oxidative damage due to the formation of reactive oxygen species. Therefore, plants have evolved an antioxidant system consisting of metabolites and enzymes that either directly scavenge ROS or recycle the antioxidant metabolites. Ozone is a temporally dynamic molecule that is both naturally occurring as well as an environmental pollutant that is predicted to increase in concentration in the future as anthropogenic precursor emissions rise. It has been hypothesized that any elevation in ozone concentration will cause increased oxidative stress in plants and therefore enhanced subsequent antioxidant metabolism, but evidence for this response is variable. Along with increasing atmospheric ozone concentrations, atmospheric carbon dioxide concentration is also rising and is predicted to continue rising in the future. The effect of elevated carbon dioxide concentrations on antioxidant metabolism varies among different studies in the literature. Therefore, the question of how antioxidant metabolism will be affected in the most realistic future atmosphere, with increased carbon dioxide concentration and increased ozone concentration, has yet to be answered, and is the subject of my thesis research. First, in order to capture as much of the variability in the antioxidant system as possible, I developed a suite of high-throughput quantitative assays for a variety of antioxidant metabolites and enzymes. I optimized these assays for Glycine max (soybean), one of the most important food crops in the world. These assays provide accurate, rapid and high-throughput measures of both the general and specific antioxidant action of plant tissue extracts. Second, I investigated how growth at either elevated carbon dioxide concentration or chronic elevated ozone concentration altered antioxidant metabolism, and the ability of soybean to respond to an acute oxidative stress in a controlled environment study. I found that growth at chronic elevated ozone concentration increased the antioxidant capacity of leaves, but was unchanged or only slightly increased following an acute oxidative stress, suggesting that growth at chronic elevated ozone concentration primed the antioxidant system. Growth at high carbon dioxide concentration decreased the antioxidant capacity of leaves, increased the response of the existing antioxidant enzymes to an acute oxidative stress, but dampened and delayed the transcriptional response, suggesting an entirely different regulation of the antioxidant system. Third, I tested the findings from the controlled environment study in a field setting by investigating the response of the soybean antioxidant system to growth at elevated carbon dioxide concentration, chronic elevated ozone concentration and the combination of elevated carbon dioxide concentration and elevated ozone concentration. In this study, I confirmed that growth at elevated carbon dioxide concentration decreased specific components of antioxidant metabolism in the field. I also verified that increasing ozone concentration is highly correlated with increases in the metabolic and genomic components of antioxidant metabolism, regardless of carbon dioxide concentration environment, but that the response to increasing ozone concentration was dampened at elevated carbon dioxide concentration. In addition, I found evidence suggesting an up regulation of respiratory metabolism at higher ozone concentration, which would supply energy and carbon for detoxification and repair of cellular damage. These results consistently support the conclusion that growth at elevated carbon dioxide concentration decreases antioxidant metabolism while growth at elevated ozone concentration increases antioxidant metabolism.

Soybean ferritin expression in saccharomyces cerevisiae modulates iron accumulation and resistance to elevated iron concentrations.

Fungi, including the yeast Saccharomyces cerevisiae, lack ferritin and use vacuoles as iron storage organelles. This work explored how plant ferritin expression influenced baker's yeast iron metabolism. Soybean seed ferritin H1 (SFerH1) and SFerH2 genes were cloned and expressed in yeast cells. Both soybean ferritins assembled as multimeric complexes, which bound yeast intracellular iron in vivo and, consequently, induced the activation of the genes expressed during iron scarcity. Soybean ferritin protected yeast cells that lacked the Ccc1 vacuolar iron detoxification transporter from toxic iron levels by reducing cellular oxidation, thus allowing growth at high iron concentrations. Interestingly, when simultaneously expressed in ccc1Δ cells, SFerH1 and SFerH2 assembled as heteropolymers, which further increased iron resistance and reduced the oxidative stress produced by excess iron compared to ferritin homopolymer complexes. Finally, soybean ferritin expression led to increased iron accumulation in both wild-type and ccc1Δ yeast cells at certain environmental iron concentrations.

Soybean Stem Fly, Melanagromyza sojae (Diptera: Agromyzidae), in the New World: detection of high genetic diversity from soybean fields in Brazil

Soybean Stem Fly (SSF), Melanagromyza sojae (Zehntner), belongs to the family Agromyzidae and is highly polyphagous, attacking many plant species of the family Fabaceae, including soybean and other beans. SSF is regarded as one of the most important pests in soybean fields of Asia (e.g., China, India), North East Africa (e.g., Egypt), parts of Russia, and South East Asia. Despite reports of Agromyzidae flies infesting soybean fields in Rio Grande do Sul State (Brazil) in 1983 and 2009 and periodic interceptions of SSF since the 1940s by the USA quarantine authorities, SSF has not been officially reported to have successfully established in the North and South Americas. In South America, M. sojae was recently confirmed using morphology and its complete mitochondrial DNA (mtDNA) was characterized. In the present study, we surveyed the genetic diversity of M. sojae, collected directly from soybean host plants, using partial mtDNA cytochrome oxidase I (COI) gene, and provide evidence of multiple (>10) maternal lineages in SSF populations in South America, potentially representing multiple incursion events. However, a single incursion involving multiple-female founders could not be ruled out. We identified a haplotype that was common in the fields of two Brazilian states and the individuals collected from Australia in 2013. The implications of SSF incursions in southern Brazil are discussed in relation to the current soybean agricultural practices, highlighting an urgent need for better understanding of SSF population movements in the New World, which is necessary for developing effective management options for this significant soybean pest. © FUNPEC-RP.

Soybean lunasin mediates colon carcinogenesis by inducing apoptosis and preventing outgrowth of metastasis

Colorectal cancer (CRC) is the third most common cancer worldwide. Various factors such as age, lifestyle and dietary patterns affect the risk of having CRC. Epidemiological studies showed a chemopreventive effect of soy consumption against CRC. However, which component(s) of soybean is associated with this reduced risk is not yet fully delineated. The objective of this research was to evaluate the anti-colon cancer potential of lunasin isolated from defatted soybean flour using in vitro and in vivo models of CRC. Lunasin was isolated from defatted soybean flour by a combination of different chromatographic and ultrafiltration techniques. The anti-colon cancer potential of lunasin was determined using different human colon cancer cell lines in vitro and a CRC liver metastasis model in vivo. Lunasin caused cytotoxicity to different human colon cancer cells with an IC50 value of 13.0, 21.6, 26.3 and 61.7 µM for KM12L4, RKO, HCT-116 and HT-29 human colon cancer cells, respectively. This cytotoxicity correlated with the expression of the α5 integrin on human colon cancer cells with a correlation coefficient of 0.78. The mechanism involved in the cytotoxic effect of lunasin was through cell cycle arrest and induction of the mitochondrial pathway of apoptosis. In KM12L4 human colon cancer cells, lunasin caused a G2/M phase arrest increasing the percentage of cells at G2/M phase from 12% (PBS-treated) to 24% (treated with 10 µM lunasin). This arrest was attributed to the capability of lunasin to increase the expression of cyclin dependent kinase inhibitors p21 and p27. At 10 µM, lunasin increased the expression of p21 and p27 in KM12L4 colon cancer cells by 2.2- and 2.3-fold, respectively. Flow cytometric analysis showed that lunasin at 10 µM increased the percentage of cells undergoing apoptosis from 13.6% to 24.7%. This is further supported by fluorescence microscopic analysis of KM12L4 cells treated with 10 µM lunasin showing chromatin condensation and DNA fragmentation. The mechanism involved is through modification of proteins involved in the mitochondrial pathway of apoptosis in KM12L4 cells as 10 µM lunasin reduced the expression of the anti-apoptotic Bcl-2 protein by 2-fold and increased the expression of the pro-apoptotic proteins Bax, cytochrome c and nuclear clusterin by 2.2-, 2.1- and 2.3- fold, respectively. This led to increased expression and activity of the executioner of apoptosis, caspase-3 by 1.8- and 2.3-fold, respectively. This pro-apoptotic property of lunasin can be attributed to its capability to internalize into the cytoplasm and nucleus of colon cancer cells 24 h and 72 h after treatment, respectively. In addition, lunasin mediated metastasis of colon cancer cells in vitro by inhibiting the focal adhesion kinase activation thereby reducing expression of extracellular regulated kinase and nuclear factor kappa B and finally inhibiting migration of colon cancer cells. In KM12L4 colon cancer cells, 10 µM lunasin resulted in the reduction of phosphorylation of focal adhesion kinase and extracellular regulated kinase by 2.5-fold, resulting in the reduced nuclear translocation of p50 and p65 NF-κB subunits by 3.8- and 1.4-fold, respectively. In an in vivo model of CRC liver metastasis, daily intraperitoneal administration of lunasin at 4 mg/kg body weight resulted in the inhibition of KM12L4 liver metastasis as shown by the reduction of the number of liver metastases from 28 (PBS-treated) to 14 (lunasin-treated, P = 0.047) and reduction in tumor burden as measured by liver weight/body weight from 0.13 (PBS-treated) to 0.10 (lunasin-treated, P = 0.039). Moreover, lunasin potentiated the anti-metastatic effect of the chemotherapeutic drug oxaliplatin given at 5 mg/kg body weight twice per week. Lunasin and oxaliplatin combination resulted in a more potent inhibition of outgrowth of KM12L4 cell metastases to the liver reducing the number of liver metastases by 6-fold and reducing the tumor burden in the liver by 3-fold when compared to PBS-treated group. This can be attributed by the capability of lunasin and oxaliplatin to reduce expression of proliferating cell nuclear antigen in liver-tumor tissue as measured by immunohistochemical staining. The results of this research for the first time demonstrated the anti-colon cancer potential of lunasin isolated from defatted soybean flour which might contribute to the chemopreventive effect of soybean in CRC as seen in different epidemiological studies. In conclusion, lunasin isolated from defatted soybean flour mediated colon carcinogenesis by inducing apoptosis and preventing outgrowth of metastasis. We suggest that the results of this research serve as a basis for further study on the chemopreventive effect of lunasin against CRC and a possible adjuvant role for lunasin in therapy of patients with metastatic CRC.

Analyses of the community compositions of root rot pathogenic fungi in the soybean rhizosphere soil

Soybean ( Glycine max [L.] Merr.) root rot is an important disease of soybean under continuous cropping, and root rot is widely distributed throughout the world. This disease is extremely harmful, and it is difficult to prevent and control. The study aimed to elucidate the composition of root rot pathogenic fungal communities in the continuous cropping of soybean. In this study, we employed PCRDGGE technology to analyze the communities of root rot pathogenic fungi in soybean rhizosphere soil subjected to continuous cropping during a season with a high incidence of root rot in Heilongjiang province, China, the main soybean producing area in China. The results of 13 DGGE bands were analyzed by phylogenetic revealed that the predominant root rot pathogenic fungi in rhizosphere soil in the test area were Pythium ultimum and Fusarium species. The results of cluster analysis showed that the duration of continuous cropping, the soybean variety and the plant growth stage all had significant effects on the diversity of root rot pathogenic fungi in rhizosphere soil.

Development of Dichelops melacanthus and its egg parasitoid Telenomus podisi reared on Bt-soybean MON 87701 x MON 89788 and its near conventional isoline under different temperatures.

Dichelops melacanthus was studied under controlled conditions (60 ± 10% RH and 14/10 h L/D photoperiod), and three constant temperatures (19, 25, and 31 ± 2 °C). Fresh pods of MON 87701 × MON 89788 soybeans and its near non-Bt isoline (A5547) were supplied to nymphs and adults. The biology of T. podisi was studied in the same controlled RH conditions, but only at the standard temperature of 25 ± 2°C. Overall, the development of D. melacanthus was better at higher temperatures, which accelerated the development of the stink bug without affecting adult biological parameters. No influence of Bt-soybeans on the biology of the pest was observed in any temperature studied, which shows that D. melacanthus is not affected by this transgenic soybean. The egg parasitoid T. podisi also was not harmed when it parasitized eggs of the pest fed with MON 87701 × MON 89788 soybeans, with similar results to those obtained in non-Bt isogenic soybeans. Thus, this study demonstrates that D. melacanthus is favored at high temperatures (31 ± 2 °C), and that neither did MON 87701 × MON 89788 soybean pods affect the development of the pest nor its parasitoid T. podisi.

Asian soybean rust in Brazil: past, present, and future.

ABSTRACT: Asian soybean rust, caused by the fungus Phakopsora pachyrhizi, is the most severe disease of the crop and can cause yield losses of up to 90%. The disease was first reported in Brazil in 2001. Epidemics of the disease are common in the country, where the fungus can survive year?round. Regulatory measures to reduce the inoculum between seasons and avoid late-season soybean have been adopted to manage the disease. Disease control has relied mainly on chemical control, but a lower sensibility of the fungus to fungicides has been reported in Brazil. Major?resistance genes have been mapped and incorporated into the cultivars. With the reduced efficacy of the fungicides, the adoption of integrated measures to control the disease will be important for the sustainability of the crop. This review presents the main changes in the soybean crop system caused by the introduction of the fungus in Brazil, the current management strategies adopted to avoid losses, and the new trends that, together with biotechnological strategies, can improve management in the future. RESUMO: A ferrugem?asiática da soja, causada pelo fungo Phakopsora pachyrhizi, é a doença mais severa da cultura e pode causar perdas de produtividade de até 90%. A doença foi relatada pela primeira vez no Brasil em 2001. Epidemias da doença são comuns no País, onde o fungo pode sobreviver durante todo o ano. Medidas regulatórias para reduzir o inóculo entre safras e evitar a semeadura tardia de soja têm sido adotadas para manejar a doença. O controle da doença tem se baseado principalmente no controle químico, mas uma menor sensibilidade do fungo aos fungicidas tem sido relatada no Brasil. Genes de resistência têm sido mapeados e incorporados às cultivares. Por causa da redução da eficiência dos fungicidas, a adoção de medidas integradas para o controle da doença será importante para a sustentabilidade da cultura. Este artigo de revisão apresenta as principais mudanças no sistema de produção da soja causadas pela introdução do fungo no Brasil, as medidas de controle atualmente usadas para evitar perdas, e as novas tendências que, juntas com estratégias biotecnológicas, podem melhorar o manejo da doença no futuro.

A pigeonpea gene confers resistance to Asian soybean rust in soybean.

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.