Exudates Of Seeds Research Articles

A method to determine antifungal activity in seed exudates by nephelometry

BackgroundOne of the levers towards alternative solutions to pesticides is to improve seed defenses against pathogens, but a better understanding is needed on the type and regulation of existing pathways during germination. Dormant seeds are able to defend themselves against microorganisms during cycles of rehydration and dehydration in the soil. During imbibition, seeds leak copious amounts of exudates. Here, we developed a nephelometry method to assay antimicrobial activity (AA) in tomato seed exudates as a proxy to assess level of defenses.ResultsA protocol is described to determine the level of AA against the nonhost filamentous fungus Alternaria brassicicola in the exudates of tomato seeds and seedlings. The fungal and exudate concentrations can be adjusted to modulate the assay sensitivity, thereby providing a large window of AA detection. We established that AA in dormant seeds depends on the genotype. It ranged from very strong AA to complete absence of AA, even after prolonged imbibition. AA depends also on the stages of germination and seedling emergence. Exudates from germinated seeds and seedlings showed very strong AA, while those from dormant seeds exhibited less activity for the same imbibition time. The exudate AA did not impact the growth of a pathogenic fungus host of tomato, Alternaria alternata, illustrating the adaptation of this fungus to its host.ConclusionsWe demonstrate that our nephelometry method is a simple yet powerful bioassay to quantify AA in seed exudates. Different developmental stages from dormant seed to seedlings show different levels of AA in the exudate that vary between genotypes, highlighting a genetic diversity x developmental stage interaction in defense. These findings will be important to identify molecules in the exudates conferring antifungal properties and obtain a better understanding of the regulatory and biosynthetic pathways through the lifecycle of seeds, from dormant seeds until seedling emergence.

Combination of Developmental Behaviors and Transcriptome Reveals Differential Response Mechanisms of Phytophthora sojae to Aspartic Acid and Glucose in Seed Exudates.

Isoflavones in soybean seed and root exudates are host-specific signal molecules for Phytophthora sojae to recognize host soybean. G protein and calcium signaling pathway are involved in the chemotaxis of zoospores in the recognition of isoflavones. To investigate the role of host nonspecific signaling molecules (sugars and amino acids) in seed and root exudates in zoospore chemotaxis and mycelial growth, the transcriptome of P. sojae responding to aspartic acid (Asp) and glucose (Glc) was analyzed by the RNA-seq method. We found that the relative in situ concentrations of amino acids and sugars significantly promoted zoospore chemotaxis, as do isoflavones. Transcriptomics showed that both similarity and difference existed in response mechanisms of P. sojae to Asp and Glc. Asp and Glc activated mitogen-activated protein kinase signaling pathway and phosphatidylinositol signaling system but not G-protein signaling pathway, which have been reported to be responsible for zoospore chemotaxis. In addition, ubiquitin-mediated proteolysis and ATP binding cassette transporters were also activated by Asp and Glc. Meanwhile, glutathione signaling pathway uniquely participated in the response of P. sojae to Asp but not involved in the response process to Glc, which is waiting for further study. Our results provide new insights into the molecular mechanism of zoospore response to Asp and Glc.

Maize and Common Bean Seed Exudates Mediate Part of Nonhost Resistance to Phytophthora sojae Prior to Infection.

Phytophthora sojae does not infect nonhost maize (Zea mays) but infects nonhost common bean (Phaseolus vulgaris) under inoculation. Soybean seed exudates participate in mediating host resistance to P. sojae before infection. This study aims to elucidate the role of seed exudates in mediating the nonhost resistance of maize and common bean to P. sojae before infection. The behaviors of P. sojae zoospores in response to the seed exudates were determined using an assay chamber and a concave slide. The proteomes of P. sojae zoospores in response to the seed exudates were analyzed with the tandem mass tag method. The key proteins were quantitatively verified by parallel reaction monitoring. Maize seed exudates exerted a repellent effect on zoospores of P. sojae. This result explains why zoospores sense repelling signaling molecules in maize seed exudates that weaken and strongly inhibit chemotaxis signals in the phosphatidylinositol signaling pathway and arachidonic acid metabolism pathway. Common bean seed exudates did not exhibit any attraction to the zoospores because the guanine nucleotide-binding protein signaling pathway, which is responsible for transmitting chemotactic signals, had no significant change. The proteins protecting the cell membrane structure were significantly downregulated, and the early apoptosis signal glutathione was enhanced in zoospores responding to common bean seed exudates, which resulted in dissolution of the cysts. Maize and common bean seed exudates mediate part of the nonhost resistance to P. sojae via different mechanisms before infection. The immunity of maize to P. sojae is caused by the repellent effect of maize seed exudates on zoospores. Common bean seed exudates participate in mediating nonhost resistance by dissolving the cysts.

The multiplex PCR assay detection of Staphylococcus sciuri antibiotic resistance, mecA gene, and the inhibitory effect of root exudate of Nigella sativa (black seeds) treated with magnetized water.

121 bacterial samples isolated from wounds from both sexes and all age groups were collected from Salahadin General Hospital, Salahadin provenance, Iraq. Only 8 Staphylococcus sciuri (S. sciuri) isolates were identified. The bacterial isolation showed the highest sensitivity to Amoxicillin/Clavulanic acid, Cefotaxime, Methicillin, Streptomycin, and Vancomycin and resistance to all other antibiotics. The root exudates of black seeds were used for 10 and 20 days for both treatments with and without magnetized water, and the exudates were superior when using magnetized water for 20 days. Antibiotic resistance and the mecA gene were investigated, and a multiplex PCR assay was used to detect the mecA gene in S. sciuri. Optimized conditions were used to amplify mecA fragments that encode methicillin resistance.

Inhibition of Protease and Egg Hatching of Haemonchus contortus by Soybean Seed Exudates.

Gastrointestinal nematode infection of small ruminants causes losses in livestock production. Plant compounds show promises as alternatives to commercial anthelmintics that have been exerting selective pressures that lead to the development of drug-resistant parasites. Soybean (Glycine max) is an economical value crop, with a higher protein content compared to other legumes. The objective of this study was to evaluate whether the protease inhibitors exuded from the G. max mature seeds have anthelmintic activity against Haemonchus contortus. To obtain the soybean exudates (SEX), mature seeds were immersed in 100 mM sodium acetate buffer, pH 5.0, at 10 C, for 24 hr. Then the naturally released substances present in SEX were collected and exhaustively dialyzed (cutoff 12 kDa) against distilled water. The dialyzed seed exudates (SEXD) were heated at 100 C for 10 min and centrifuged (12,000 g, at 4 C for 15 min). The supernatant obtained was recovered and designated as the heat-treated exudate fraction (SEXDH). The protein content, protease inhibitor activity, and the effect of each fraction on H. contortus egg hatch rate were evaluated. The inhibition extent of SEX, SEXD, and SEXDH on H. contortus egg proteases was 31.1, 42.9, and 63.8%, respectively. Moreover, SEX, SEXD, and SEXDH inhibited the egg hatching with EC50 of 0.175, 0.175, and 0.241 mg ml-1, respectively. Among the commercial protease inhibitors tested, only EDTA and E-64 inhibited the H. contortus hatch rate (79.0 and 28.9%, respectively). We present evidence demonstrating that soybean exudate proteins can effectively inhibit H. contortus egg hatching. This bioactivity is displayed by thermostable proteins and provides evidence that protease inhibitors are a potential candidate for anthelmintic use.

An Updated Perspective on Sinorhizobium meliloti Chemotaxis to Alfalfa Flavonoids.

The symbiotic interaction between leguminous plants and their cognate rhizobia allows for the fixation of gaseous dinitrogen into bioavailable ammonia. The perception of host-derived flavonoids is a key initial step for the signaling events that must occur preceding the formation of the nitrogen-fixing organ. Past work investigating chemotaxis – the directed movement of bacteria through chemical gradients – of Bradyrhizobium japonicum, Rhizobium leguminosarum, and Rhizobium meliloti discovered chemotaxis to various organic compounds, but focused on chemotaxis to flavonoids because of their relevance to the symbiosis biochemistry. The current work sought to replicate and further examine Sinorhizobium (Ensifer) meliloti chemotaxis to the flavonoids previously thought to act as the principal attractant molecules prior to the initial signaling stage. Exudate from germinating alfalfa seedlings was analyzed for composition and quantities of different flavonoid compounds using mass spectrometry. The abundance of four prevalent flavonoids in germinating alfalfa seed exudates (SEs) was at a ratio of 200:5:5:1 for hyperoside, luteolin, luteolin-7-glucoside, and chrysoeriol. Using quantitative chemotaxis capillary assays, we did not detect chemotaxis of motile S. meliloti cells to these, and two other flavonoids identified in seed exudates. In support of these findings, the flavonoid fraction of seed exudates was found to be an insignificant attractant relative to the more hydrophilic fraction. Additionally, we observed that cosolvents commonly used to dissolve flavonoids confound the results. We propose that the role flavonoids play in S. meliloti chemotaxis is insignificant relative to other components released by alfalfa seeds.

Myracrodruon urundeuva seed exudates proteome and anthelmintic activity against Haemonchus contortus.

Seed exudates are plant-derived natural bioactive compounds consisting of a complex mixture of organic and inorganic molecules. Plant seed exudates have been poorly studied against parasite nematodes. This study was undertaken to identify proteins in the Myracrodruon urundeuva seed exudates and to assess the anthelmintic activity against Haemonchus contortus, an important parasite of small ruminants. M. urundeuva seed exudates (SEX) was obtained after immersion of seeds in sodium acetate buffer. SEX was fractionated with ammonium sulfate at 0–90% concentration to generate the ressuspended pellet (SEXF1) and the supernatant (SEXF2). SEX, SEXF1, and SEXF2 were exhaustively dialyzed against distilled water (cut-off: 12 kDa) and the protein contents determined. Mass spectrometry analyses of SEX, SEXF1, and SEXF2 were done to identify proteins and secondary metabolites. The seed exudates contained protease, protease inhibitor, peptidase, chitinase, and lipases as well as the low molecular weight secondary compounds ellagic acid and quercetin rhamnoside. SEX inhibited H. contortus larval development (LDA) (IC50 = 0.29 mg mL-1), but did not affect larval exsheathment (LEIA). On the other hand, although SEXF1 and SEXF2 inhibited H. contortus LEIA (IC50 = 1.04 and 0.93 mg mL-1, respectively), they showed even greater inhibition efficiency of H. contortus larval development (IC50 = 0.29 and 0.42 mg mL-1, respectively). To the best of our knowledge, this study is the first to show the anthelmintic activity of plant exudates against a gastrointestinal nematode. Moreover, it suggests the potential of exuded proteins as candidates to negatively interfere with H. contortus life cycle.

Rhizosphere‐Scale Quantification of Hydraulic and Mechanical Properties of Soil Impacted by Root and Seed Exudates

Core Ideas We hypothesized that plant exudates gel soil particles and on drying enhance water repellency. This has been carried out using rhizosphere‐scale mechanical and hydraulic measurements. Plant exudates enhanced soil hardness and modulus of elasticity as chia seed > maize root > barley root. Plant exudates caused measureable decreases in soil wetting rates through water repellency. Using rhizosphere‐scale physical measurements, we tested the hypothesis that plant exudates gel together soil particles and, on drying, enhance soil water repellency. Barley (Hordeum vulgare L. cv. Optic) and maize (Zea mays L. cv. Freya) root exudates were compared with chia (Salvia hispanica L.) seed exudate, a commonly used root exudate analog. Sandy loam and clay loam soils were treated with root exudates at 0.46 and 4.6 mg exudate g−1 dry soil and chia seed exudate at 0.046, 0.46, 0.92, 2.3 and 4.6 mg exudate g−1 dry soil. Soil hardness and modulus of elasticity were measured at −10 kPa matric potential using a 3‐mm‐diameter spherical indenter. The water sorptivity and repellency index of air‐dry soil were measured using a miniaturized infiltrometer device with a 1‐mm tip radius. Soil hardness increased by 28% for barley root exudate, 62% for maize root exudate, and 86% for chia seed exudate at 4.6 mg g−1 concentration in the sandy loam soil. For the clay loam soil, root exudates did not affect soil hardness, whereas chia seed exudate increased soil hardness by 48% at 4.6 mg g−1 concentration. Soil water repellency increased by 48% for chia seed exudate and 23% for maize root exudate but not for barley root exudate at 4.6 mg g−1 concentration in the sandy loam soil. For the clay loam soil, chia seed exudate increased water repellency by 45%, whereas root exudates did not affect water repellency at 4.6 mg g−1 concentration. Water sorptivity and repellency were both correlated with hardness, presumably due to the combined influence of exudates on the hydrological and mechanical properties of the soils.

Proteome of Soybean Seed Exudates Contains Plant Defense-Related Proteins Active against the Root-Knot Nematode Meloidogyne incognita.

Several studies have described the effects of seed exudates against microorganisms, but only few of them have investigated the proteins that have defensive activity particularly against nematode parasites. This study focused on the proteins released in the exudates of soybean seeds and evaluated their nematicidal properties against Meloidogyne incognita. A proteomic approach indicated the existence of 63 exuded proteins, including β-1,3-glucanase, chitinase, lectin, trypsin inhibitor, and lipoxygenase, all of which are related to plant defense. The presence of some of these proteins was confirmed by their in vitro activity. The soybean exudates were able to reduce the hatching of nematode eggs and to cause 100% mortality of second-stage juveniles (J2). The pretreatment of J2 with these exudates resulted in a 90% reduction of the gall number in tobacco plants. These findings suggest that the exuded proteins are directly involved in plant defense against soil pathogens, including nematodes, during seed germination.

Influence of Seeds and Roots Extracts and Exudates of Bean Plant on Growth of Some Pathogenic Fungi

Extracts and exudates of seeds and roots of plant play a role in plant health and attracted the pathogen to the host. This study was conducted the effect of extract and exudates from bean seeds and roots on fungal growth of Botrytis cinerea, Macrophomina phaseolina and Rhizoctonia solani. Chemical analysis of extract and exudates was also studied. Seed and root extracts and exudates obtained from Libyan cv. gave the most stimulating effect on the mycelia growth of all tested fungi in a descending order, than those extracted and exuded from seeds and roots of the Giza-6 cv. In vitro chemical analysis of seed and root extracts and exudates of both cultivars indicated that total amino acids were much higher in Libyan cv. than those in Giza-6 cv., while free and total phenol contents were much greater in the seed and root extracts and exudates of Giza-6 cv. compared with those from Libyan cv.

Inoculação do feijoeiro com Rhizobium tropici associada à exsudato de Mimosa flocculosa com diferentes doses de nitrogênio

O presente estudo teve como objetivo avaliar os efeitos da inoculação de rizóbio, associada à adição de exsudatos de sementes de Mimosa flocculosa, e/ou de N-mineral nas características agronômicas do feijoeiro, em sucessão à soja e ao milho. O delineamento experimental foi em blocos casualizados, com quatro repetições, sendo os tratamentos dispostos em parcelas sub-subdivididas. As culturas soja e milho antecessoras à cultura do feijoeiro formaram as parcelas. As subparcelas foram constituídas pelos tratamentos: testemunha sem inoculação, inoculação de Rhyzobium tropici, estirpes CIAT 899 e PRF 81 e a associação deste com os exsudados das sementes de Mimosa flocculosa. As sub-subparcelas constituiram-se de quatro doses de N aplicadas em cobertura: 0, 40, 80 e 120 kg ha-1. A inoculação de rizóbio associada à adição de exsudato de sementes de Mimosa flocculosa contribuiu para o melhor desenvolvimento da parte aérea do feijoeiro. A adubação nitrogenada reduziu, de forma linear, a nodulação do feijoeiro. O milho como cultura antecessora ao feijão contribuiu para o aumento do peso de 100 grãos e a adição de N combinado não contribuiu para o aumento do número de vagens por planta e produtividade de grãos do feijoeiro, independente da dose utilizada.

Response of allelochemicals under pseudo-microgravity in sunflower plant (Helianthus annuus L. cv. Taiyo)

Interactions between organisms and allelochemicals in space have been not well investigated. We employed a 3D-clinorotation device to generate pseudo-microgravity in order to answer this question. If any allelopathy associated biosynthesis, emission, transport and sensing mechanism is found to be gravity dependent, many organisms and ecological systems might exhibit a different behavior under the microgravity experienced in space. The sunflower (Helianthus annuus L.) is known to show strong allelopathic properties against other plant species. The growth of lettuce seedlings, as an index of the allelopathic action, was compared between the 3D-clinostated group and a control group cultivated in normal gravity. The degree of suppression of the lettuce growth by the sunflower plant was found to be less in the experiment group. Sundiversifolide,4,15-dinor-3-hydroxy-1(5)-xanthene-12,8-olide, has been isolated and identified from the exudate of seeds (achenes) of the Taiyo sunflower as a new plant species-specific allelopathic substance (Ohno et al., 2001). The occurrence of this substance was remarkably different under normal gravity vs. the pseudo-microgravity. It was found that sundiversifolide was not a stable compound and it became a non-active substance, which did not have any inhibitory activity on the shoot lengths of lettuce seedlings after adding the extraction from the achenes. These results indicate that the synthesis and release of the allelochemicals are reduced under pseudo-microgravity.

Efeito do deslintamento químico sobre a ocorrência e desenvolvimento de Colletotrichum gossypii associado às sementes de algodoeiro

A primeira fase do trabalho objetivou avaliar o efeito do tempo de deslintamento químico, 1:30 e 4:30 min, sobre o nível de infecção de Colletotrichum gossypii inoculado artificialmente em sementes de algodoeiro (Gossypium hirsutum). Sementes de algodoeiro com línter foram inoculadas com C. gossypii, mediante contato das mesmas com colônias do fungo em placas de Petri de 9 cm de diâmetro, por 30 h. Os parâmetros avaliados foram a ocorrência de fungos e o poder germinativo das sementes. Na segunda fase, o objetivo foi avaliar a influência do exsudato de sementes de algodoeiro, em função do tempo da duração do deslintamento com ácido sulfúrico, considerando-se as frações de sedimentação de sementes e diferentes condições de envelhecimento artificial sobre o desenvolvimento de C. gossypii, em condições de laboratório. As sementes foram deslintadas quimicamente com ácido sulfúrico comercial concentrado pelos períodos de 1:30 e 4:30 min, separadas em frações de sedimentação em água e submetidas ao envelhecimento artificial por 0, 72 e 96 h. Os substratos foram obtidos a partir do exsudato resultante da embebição contendo os eletrólitos lixiviados das sementes. O desenvolvimento de C. gossypii na presença do exsudato das sementes foi avaliado em meio agarizado, através da medição do crescimento micelial e da esporulação do fungo. O deslintamento químico, pelo período de 1:30 min, propiciou aumento do percentual de ocorrência de C. gossypii em sementes não-desinfestadas superficialmente, e o crescimento micelial e a esporulação do fungo foram favorecidos pelo substrato proveniente de sementes deslintadas por 4:30 e 1:30 min, respectivamente.

Levels of ACC and related compounds in exudate and extracts of canola seeds treated with ACC deaminase-containing plant growth-promoting bacteria.

It was previously proposed that plant growth-promoting bacteria that possess 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase could utilize ACC that is present in the exudate of germinating canola seeds. The uptake and cleavage of ACC by these bacteria would lower the level of ACC, and thus ethylene within the plant, and reduce the extent of its inhibition on root elongation. To test part of the above mentioned model, ACC levels were monitored in canola seed tissues and exudate during germination. Lower amounts of ACC were present in the exudate and tissues of seeds treated with the plant growth-promoting bacterium Enterobacter cloacae CAL3, than in control seeds treated with MgSO4. The ACC-related compounds, alpha- and gamma-aminobutyric acids, both known to stimulate ethylene production, were also measured in the canola seed exudate and tissues. Approximately the same levels of alpha-aminobutyric acid were present in the exudates of the bacterium-treated seeds and the control seeds, but the amount of alpha-aminobutyric acid was lower in the tissues of the bacterium-treated seeds than in the control seeds. Smaller quantities of gamma-aminobutyric acid were seen in both the exudate and tissues of the E. cloacae CAL3-treated seeds than in the control seeds.

Levels of ACC and related compounds in exudate and extracts of canola seeds treated with ACC deaminase-containing plant growth-promoting bacteria

Levels of ACC and related compounds in exudate and extracts of canola seeds treated with ACC deaminase-containing plant growth-promoting bacteria

Chemotaxis of Fluorescent Pseudomonads Towards Seed Exudates and Germinating Seeds in Solarized Soil

We investigated the chemotaxis of fluorescent pseudomonads towards seeds exudates and germinating tomato seeds. Chemotaxis of Pseudomonas putida and P.fluorescens in a capillary tube was more pronounced towards exudates originating from seeds that were germinated in solarized soil than towards comparable exudates from seeds that were germinated in nonsolarized soil. Movement of these bacteria was enhanced also towards a mixture of either amino acids or amino acids and sugars, but not of sugars alone (...)

Phenolic acids in groundnut seed exudates

Seed exudates were collected aseptically and extracted for phenolic acids. Paper chromatographic analysis revealed the presence of eleven phenolic compounds.

Studies on the rhizosphere mycoflora of broad bean and cotton

Seed and root exudates from three varieties of both broad bean and cotton grown aseptically were analyzed for amino acids and sugars. Broad bean varieties were found to excrete considerable amounts of amino compounds with 15 different amino acids, while cotton varieties excreted less consisting of 12 amino acids. Five sugars were identified in the seed and root exudates from broad bean varieties, while three were present in the seed and root exudates from cotton varieties. Seed and root exudates stimulated spore germination and growth of fungi isolated from the rhizosphere.