Induction of phytoalexins and proteins related to pathogenesis in plants treated with extracts of cutaneous secretions of southern Amazonian Bufonidae amphibians.
ABSTRACT: Cutaneous secretions produced by amphibians of the family Bufonidae are rich sources of bioactive compounds that can be useful as new chemical templates for agrochemicals. In crop protection, the use of elicitors to induce responses offers the prospect of durable, broad-spectrum disease control using the plant's own resistance. Therefore, we evaluated the potential of methanolic extracts of cutaneous secretions of two species of amphibians of the family Bufonidae found in the Amazon biome-Rhaebo guttatus (species 1) and Rhinella marina (species 2)-in the synthesis of phytoalexins in soybean cotyledons, bean hypocotyls, and sorghum mesocotyls. Additionally, changes in the enzyme activity of ?-1,3-glucanase, peroxidase (POX), and polyphenol oxidase (PPO) and in the total protein content of soybean cotyledons were determined. In the soybean cultivar 'TMG 132 RR', our results indicated that the methanolic extract of R. guttatus cutaneous secretions suppressed glyceollin synthesis and ?-1,3-glucanase activity and increased POX and PPO activities at higher concentrations and total protein content at a concentration of 0.2 mg/mL. On the other hand, the methanolic extract of R. marina cutaneous secretions induced glyceollin synthesis in the soybean cultivars 'TMG 132 RR' and 'Monsoy 8372 IPRO' at 0.1-0.2 mg/mL and 0.2 mg/mL, respectively. The methanolic extract of R. marina cutaneous secretions also increased the specific activity of POX and PPO in 'Monsoy 8372 IPRO' and 'TMG 132 RR', respectively, and decreased the activity of ?-1,3-glucanases in 'Monsoy 8372 IPRO'. At 0.3 mg/mL, it stimulated phaseolin synthesis. The extracts did not express bioactivity in the synthesis of deoxyanthocyanidins in sorghum mesocotyls. The study in soybean suggests that the bioactivity in defense responses is influenced by cultivar genotypes. Therefore, these results provide evidence that extracts of cutaneous secretions of these amphibians species may contribute to the bioactivity of defense metabolites in plants.
Project description:Photosynthetic efficiency has become the target of several breeding programs since the positive correlation between photosynthetic rate and yield in soybean suggests that the improvement of photosynthetic efficiency may be a promising target for new yield gains. However, studies on combining ability of soybean genotypes for physiological traits are still scarce in the literature. The objective of this study was to estimate the combining ability of soybean genotypes based on F2 generation aiming to identify superior parents and segregating populations for physiological traits. Twenty-eight F2 populations resulting from partial diallel crossings between eleven lines were evaluated in two crop seasons for the physiological traits: photosynthesis, stomatal conductance, internal CO2 concentration, and transpiration. General combining ability (GCA) of the parents and specific combining ability (SCA) of the F2 populations were estimated. Our findings reveal the predominance of additive effects in controlling the traits. The genotype TMG 7062 IPRO is the most promising parent for programs aiming at photosynthetic efficiency. We have also identified other promising parents and proposed cross-breeding with higher potential for obtaining superior lines for photosynthetic efficiency.
Project description:Priming can improve plant innate capability to deal with the stresses caused by both biotic and abiotic factors. In this study, the effect of DL-?-amino-n-butyric acid (BABA) against Aphis glycines Matsumura, the soybean aphid (SA) was evaluated. We found that 25 mM BABA as a root drench had minimal adverse impact on plant growth and also efficiently protected soybean from SA infestation. In both choice and non-choice tests, SA number was significantly decreased to a low level in soybean seedlings drenched with 25 mM BABA compared to the control counterparts. BABA treatment resulted in a significant increase in the activities of several defense enzymes, such as phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO), chitinase (CHI), and ?-1, 3-glucanase (GLU) in soybean seedlings attacked by aphid. Meanwhile, the induction of 15 defense-related genes by aphid, such as AOS, CHS, MMP2, NPR1-1, NPR1-2, and PR genes, were significantly augmented in BABA-treated soybean seedlings. Our study suggest that BABA application is a promising way to enhance soybean resistance against SA.
Project description:BACKGROUND:From initial seed germination through reproduction, plants continuously reprogram their transcriptional repertoire to facilitate growth and development. This dynamic is mediated by a diverse but inextricably-linked catalog of regulatory proteins called transcription factors (TFs). Statistically quantifying TF binding site (TFBS) abundance in promoters of differentially expressed genes can be used to identify binding site patterns in promoters that are closely related to stress-response. Output from today's transcriptomic assays necessitates statistically-oriented software to handle large promoter-sequence sets in a computationally tractable fashion. RESULTS:We present Marina, an open-source software for identifying over-represented TFBSs from amongst large sets of promoter sequences, using an ensemble of 7 statistical metrics and binding-site profiles. Through software comparison, we show that Marina can identify considerably more over-represented plant TFBSs compared to a popular software alternative. CONCLUSIONS:Marina was used to identify over-represented TFBSs in a two time-point RNA-Seq study exploring the transcriptomic interplay between soybean (Glycine max) and soybean rust (Phakopsora pachyrhizi). Marina identified numerous abundant TFBSs recognized by transcription factors that are associated with defense-response such as WRKY, HY5 and MYB2. Comparing results from Marina to that of a popular software alternative suggests that regardless of the number of promoter-sequences, Marina is able to identify significantly more over-represented TFBSs.
Project description:Sunflower broomrape is a noxious parasitic weed which has caused severe damage to crop ecosystems. Trap crops can release a mixture of allelochemicals to induce the germination of sunflower broomrape. We studied the allelopathic effects of soybean on sunflower broomrape. Fourteen common soybean cultivars were grown in pots. Samples were collected from soybean plants and rhizosphere soil at five growth stages (V1, V3, V5, R2, and R4). The allelopathic effects of soybean reached highest at the V3 stage. Methanolic extracts of soybean roots induced higher broomrape germination than methanolic extracts of stems or leaves. The germination rates induced by root extracts (10-fold dilution) were positively correlated with germination rates induced by stem (10-fold dilution) and leaf extracts (10-fold dilution). The broomrape germination rates induced by root extracts were also positively correlated with soybean nodule diameter and dry weight. The results indicated that soybeans could induce sunflower broomrape germination. We conclude that soybean has the potential to be used as a trap crop for sunflower broomrape.
Project description:The susceptibility of soybean genotype to Agrobacterium infection is a key factor for the high level of genetic transformation efficiency. The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process. This study selected three genotypes (Williams 82, Shennong 9 and Bert) with high transformation efficiency, which presented better susceptibility to Agrobacterium infection, and three low transformation efficiency genotypes (General, Liaodou 16 and Kottman), which showed a relatively weak susceptibility. Gibberellin (GA) levels and soybean GA20ox2 and CYP707A2 transcripts of high-efficiency genotypes increased and were higher than those of low-efficiency genotypes; however, the opposite performance was shown in abscisic acid (ABA). Higher zeatin riboside (ZR) content and DNA quantity, and relatively higher expression of soybean IPT5, CYCD3 and CYCA3 were obtained in high-efficiency genotypes. High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71. GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively.
Project description:Two new compounds-a trioxilin and a sulfoquinovosyl diacylglycerol (SQDG)-were isolated from the methanolic extract of the heterotrophic dinoflagellate Oxyrrhis marina cultivated by feeding on dried yeasts. The trioxilin was identified as (4Z,8E,13Z,16Z,19Z) -7(S),10(S),11(S)-trihydroxydocosapentaenoic acid (1), and the SQDG was identified as (2S)-1-O-hexadecanosy-2-O-docosahexaenoyl-3-O-(6-sulfo-?-d-quinovopyranosyl)-glycerol (2) by a combination of nuclear magnetic resonance (NMR) spectra, mass analyses, and chemical reactions. The two compounds were associated with docosahexaenoic acid, which is a major component of O. marina. The two isolated compounds showed significant nitric oxide inhibitory activity on lipopolysaccharide-induced RAW264.7 cells. Compound 2 showed no cytotoxicity against hepatocarcinoma (HepG2), neuroblastoma (Neuro-2a), and colon cancer (HCT-116) cells, while weak cytotoxicity was observed for compound 1 against Neuro-2a cells.
Project description:BACKGROUND: Plant polyphenol oxidases (PPOs) are enzymes that typically use molecular oxygen to oxidize ortho-diphenols to ortho-quinones. These commonly cause browning reactions following tissue damage, and may be important in plant defense. Some PPOs function as hydroxylases or in cross-linking reactions, but in most plants their physiological roles are not known. To better understand the importance of PPOs in the plant kingdom, we surveyed PPO gene families in 25 sequenced genomes from chlorophytes, bryophytes, lycophytes, and flowering plants. The PPO genes were then analyzed in silico for gene structure, phylogenetic relationships, and targeting signals. RESULTS: Many previously uncharacterized PPO genes were uncovered. The moss, Physcomitrella patens, contained 13 PPO genes and Selaginella moellendorffii (spike moss) and Glycine max (soybean) each had 11 genes. Populus trichocarpa (poplar) contained a highly diversified gene family with 11 PPO genes, but several flowering plants had only a single PPO gene. By contrast, no PPO-like sequences were identified in several chlorophyte (green algae) genomes or Arabidopsis (A. lyrata and A. thaliana). We found that many PPOs contained one or two introns often near the 3' terminus. Furthermore, N-terminal amino acid sequence analysis using ChloroP and TargetP 1.1 predicted that several putative PPOs are synthesized via the secretory pathway, a unique finding as most PPOs are predicted to be chloroplast proteins. Phylogenetic reconstruction of these sequences revealed that large PPO gene repertoires in some species are mostly a consequence of independent bursts of gene duplication, while the lineage leading to Arabidopsis must have lost all PPO genes. CONCLUSION: Our survey identified PPOs in gene families of varying sizes in all land plants except in the genus Arabidopsis. While we found variation in intron numbers and positions, overall PPO gene structure is congruent with the phylogenetic relationships based on primary sequence data. The dynamic nature of this gene family differentiates PPO from other oxidative enzymes, and is consistent with a protein important for a diversity of functions relating to environmental adaptation.
Project description:In response to attack by bacterial pathogens, soybean (Gylcine max) leaves accumulate isoflavone aglucones, isoflavone glucosides, and glyceollins. In contrast to pathogens, the dynamics of related insect-inducible metabolites in soybean leaves remain poorly understood. In this study, we analyzed the biochemical responses of soybean leaves to Spodoptera litura (Lepidoptera: Noctuidae) herbivory and also S. litura gut contents, which contain oral secretion elicitors. Following S. litura herbivory, soybean leaves displayed an induced accumulation of the flavone and isoflavone aglycones 4',7-dihyroxyflavone, daidzein, and formononetin, and also the isoflavone glucoside daidzin. Interestingly, foliar application of S. litura oral secretions also elicited the accumulation of isoflavone aglycones (daidzein and formononetin), isoflavone 7-O-glucosides (daidzin, ononin), and isoflavone 7-O-(6'-O-malonyl-?-glucosides) (malonyldaidzin, malonylononin). Consistent with the up-regulation of the isoflavonoid biosynthetic pathway, folair phenylalanine levels also increased following oral secretion treatment. To establish that these metabolitic changes were the result of de novo biosynthesis, we demonstrated that labeled (13C9) phenylalanine was incorporated into the isoflavone aglucones. These results are consistent with the presence of soybean defense elicitors in S. litura oral secretions. We demonstrate that isoflavone aglycones and isoflavone conjugates are induced in soybean leaves, not only by pathogens as previously demonstrated, but also by foliar insect herbivory.
Project description:Purpose:The iPro Cube is a small portable point-of-care device designed to analyse salivary markers of stress in a user-friendly manner (e.g., fast, convenient). Our aim was to test the reliability and validity of the iPro Cube to measure salivary cortisol and ?-amylase as compared to the common laboratory standard method (ELISA immunoassay) prior to and after moderate intensity exercise. Methods:The study was a repeated measures, pre-registered design, and statistical framework that incorporated prior knowledge directly into the estimation process. Twenty-nine individuals (age = 27.4 ± 6.6 y; body-mass = 70.8 ± 11.3 kg; height = 1.74 ± 0.92 m; 18 males) completed a single PWC75%HRmax, with repeated measures of salivary cortisol and -amylase pre, immediately post, and 30 min post-exercise. Results:Correlation between the iPro Cube and laboratory-based assessments of salivary cortisol was moderate-to-large (0.53 > r < 0.81) across all three testing points. In contrast, correlation between the iPro Cube and laboratory-based assessments of -amylase was small-to-moderate (0.25 > r < 0.46). We found a large correlation between duplicate samples of iPro Cube cortisol assessment (0.75 > r < 0.82), and a moderate-to-large correlation for -amylase (0.51> r < 0.77). Conclusions:The iPro Cube is capable of taking measures of salivary cortisol that are moderately correlated to values obtained via ELISA immunoassay, however the unit underestimates salivary cortisol and overestimates salivary -amylase at rest and post-moderate intensity exercise. It is recommended that researchers continue using standard laboratory techniques to assess these salivary stress markers.
Project description:Soybean cyst nematode (SCN) is a severe soil borne disease. The control of this disease is still a worldwide problem in agriculture. In this study, we found that application of potassium (K) fertilizer could decrease the occurrence of SCN at two field sites. Furthermore, the application of K could suppress Heterodera glycines with the activation of Phenylalanine Ammonia Lyase (PAL) and Polyphenol Oxidase (PPO) expression via pot experiments in a greenhouse. The release of cinnamic, ferulic and salicylic acids was significantly enhanced by K application of 3 mM, and each of three acids can dramatically constrain Heterodera glycines in vitro. This research indicated that K induce multiple mechanisms to improve the resistance of soybean against SCN and provide a new strategy to control SCN in fields with nutrient application.