School of BioSciences - Research Publications
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ItemAssessing the Efficacy, Acute Toxicity, and Binding Modes of the Agricultural Nitrification Inhibitors 3,4-Dimethyl-1H-pyrazole (DMP) and Dicyandiamide (DCD) with Nitrosomonas europaea(American Chemical Society, 2023-01-25)Nitrification inhibitors have been coformulated with nitrogen fertilizers since the 1970s to modulate the microbiological conversion of nitrogen in agricultural soils. 3,4-Dimethyl-1H-pyrazole (DMP) and dicyandiamide (DCD) are currently the most used commercial nitrification inhibitors, but their mode of action is not well understood. This work seeks to fill this void by assessing for the first time in detail their mechanism of inhibition, efficacy, and acute toxicity with pure cell cultures of Nitrosomonas europaea. Bacterial assays based on the quantification of the nitrite (NO2–) production showed that both inhibitors reversibly target ammonia monooxygenase (AMO), which catalyzes the first step of the nitrification process. Michaelis–Menten kinetics suggest that both DMP and DCD act as uncompetitive inhibitors. Real-time measurements of the oxygen (O2) consumption confirmed the nonmechanistic mode of inhibition and showed that DMP reduced the O2 uptake rate by AMO much more at considerably lower concentrations than DCD, in line with the lower inhibitory efficiency of the latter. Acute toxicity tests revealed that DCD has a 10% higher toxicity than DMP when comparing treatments at the same inhibition efficacy (i.e., DMP at 10 ppm, DCD at 100 ppm), indicating that the inhibition of the nitrification process cannot simply be achieved by increasing the inhibitor concentration. The methods presented in this study could assist the development of more reliable nitrification inhibitors in the future.
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ItemNo Preview AvailableRapid and Inexpensive Assay for Testing the Efficiency of Potential New Synthetic Nitrification Inhibitors(AMER CHEMICAL SOC, 2023-03-20)
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ItemEffects of chitin and chitosan on root growth, biochemical defense response and exudate proteome of Cannabis sativa.(Wiley, 2023-06)Fungal pathogens pose a major threat to Cannabis sativa production, requiring safe and effective management procedures to control disease. Chitin and chitosan are natural molecules that elicit plant defense responses. Investigation of their effects on C. sativa will advance understanding of plant responses towards elicitors and provide a potential pathway to enhance plant resistance against diseases. Plants were grown in the in vitro Root-TRAPR system and treated with colloidal chitin and chitosan. Plant morphology was monitored, then plant tissues and exudates were collected for enzymatic activity assays, phytohormone quantification, qPCR analysis and proteomics profiling. Chitosan treatments showed increased total chitinase activity and expression of pathogenesis-related (PR) genes by 3-5 times in the root tissues. In the exudates, total peroxidase and chitinase activities and levels of defense proteins such as PR protein 1 and endochitinase 2 were increased. Shoot development was unaffected, but root development was inhibited after chitosan exposure. In contrast, chitin treatments had no significant impact on any defense parameters, including enzymatic activities, hormone quantities, gene expression levels and root secreted proteins. These results indicate that colloidal chitosan, significantly enhancing defense responses in C. sativa root system, could be used as a potential elicitor, particularly in hydroponic scenarios to manage crop diseases.
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ItemRoot-TRAPR: a modular plant growth device to visualize root development and monitor growth parameters, as applied to an elicitor response of Cannabis sativa(BMC, 2022-04-09)BACKGROUND: Plant growth devices, for example, rhizoponics, rhizoboxes, and ecosystem fabrication (EcoFAB), have been developed to facilitate studies of plant root morphology and plant-microbe interactions in controlled laboratory settings. However, several of these designs are suitable only for studying small model plants such as Arabidopsis thaliana and Brachypodium distachyon and therefore require modification to be extended to larger plant species like crop plants. In addition, specific tools and technical skills needed for fabricating these devices may not be available to researchers. Hence, this study aimed to establish an alternative protocol to generate a larger, modular and reusable plant growth device based on different available resources. RESULTS: Root-TRAPR (Root-Transparent, Reusable, Affordable three-dimensional Printed Rhizo-hydroponic) system was successfully developed. It consists of two main parts, an internal root growth chamber and an external structural frame. The internal root growth chamber comprises a polydimethylsiloxane (PDMS) gasket, microscope slide and acrylic sheet, while the external frame is printed from a three-dimensional (3D) printer and secured with nylon screws. To test the efficiency and applicability of the system, industrial hemp (Cannabis sativa) was grown with or without exposure to chitosan, a well-known plant elicitor used for stimulating plant defense. Plant root morphology was detected in the system, and plant tissues were easily collected and processed to examine plant biological responses. Upon chitosan treatment, chitinase and peroxidase activities increased in root tissues (1.7- and 2.3-fold, respectively) and exudates (7.2- and 21.6-fold, respectively). In addition, root to shoot ratio of phytohormone contents were increased in response to chitosan. Within 2 weeks of observation, hemp plants exhibited dwarf growth in the Root-TRAPR system, easing plant handling and allowing increased replication under limited growing space. CONCLUSION: The Root-TRAPR system facilitates the exploration of root morphology and root exudate of C. sativa under controlled conditions and at a smaller scale. The device is easy to fabricate and applicable for investigating plant responses toward elicitor challenge. In addition, this fabrication protocol is adaptable to study other plants and can be applied to investigate plant physiology in different biological contexts, such as plant responses against biotic and abiotic stresses.
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ItemModulators or facilitators? Roles of lipids in plant root-microbe interactions(CELL PRESS, 2022-02)Lipids have diverse functions in regulating the plasma membrane's cellular processes and signaling mediation. Plasma membrane lipids are also involved in the plant's complex interactions with the surrounding microorganisms, with which plants are in various forms of symbiosis. The roles of lipids influence the whole microbial colonization process, thus shaping the rhizomicrobiome. As chemical signals, lipids facilitate the stages of rhizospheric interactions - from plant root to microbe, microbe to microbe, and microbe to plant root - and modulate the plant's defense responses upon perception or contact with either beneficial or phytopathogenic microorganisms. Although studies have come a long way, further investigation is needed to discover more lipid species and elucidate novel lipid functions and profiles under various stages of plant root-microbe interactions.
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ItemMicrobial Phosphorus Mobilization Strategies Across a Natural Nutrient Limitation Gradient and Evidence for Linkage With Iron Solubilization Traits.(Frontiers Media SA, 2021)Microorganisms have evolved several mechanisms to mobilize and mineralize occluded and insoluble phosphorus (P), thereby promoting plant growth in terrestrial ecosystems. However, the linkages between microbial P-solubilization traits and the preponderance of insoluble P in natural ecosystems are not well known. We tested the P solubilization traits of hundreds of culturable bacteria representative of the rhizosphere from a natural gradient where P concentration and bioavailability decline as soil becomes progressively more weathered. Aluminum, iron phosphate and organic P (phytate) were expected to dominate in more weathered soils. A defined cultivation medium with these chemical forms of P was used for isolation. A combination of soil chemical, spectroscopic analyses and 16S rRNA gene sequencing were used to understand the in situ ability for solubilization of these predominant forms of P. Locations with more occluded and organic P harbored the greatest abundance of P-mobilizing microorganisms, especially Burkholderiaceae (Caballeronia and Paraburkholderia spp.). Nearly all bacteria utilized aluminum phosphate, however fewer could subsist on iron phosphate (FePO4) or phytate. Microorganisms isolated from phytic acid were also most effective at solubilizing FePO4, suggesting that phytate solubilization may be linked to the ability to solubilize Fe. Significantly, we observed Fe to be co-located with P in organic patches in soil. Siderophore addition in lab experiments reinstated phytase mediated P-solubilization from Fe-phytate complexes. Taken together, these results indicate that metal-organic-P complex formation may limit enzymatic P solubilization from phytate in soil. Additionally, the linked traits of phytase and siderophore production were mostly restricted to specific clades within the Burkholderiaceae. We propose that Fe complexation of organic P (e.g., phytate) represents a major constraint on P turnover and availability in acidic soils, as only a limited subset of bacteria appear to possess the traits required to access this persistent pool of soil P.
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ItemWheat Can Access Phosphorus From Algal Biomass as Quickly and Continuously as From Mineral Fertilizer(FRONTIERS MEDIA SA, 2021-01-28)Algae can efficiently take up excess nutrients from waterways, making them a valuable resource potentially capable of replacing synthesized and mined fertilizers for agriculture. The capacity of algae to fertilize crops has been quantified, but it is not known how the algae-derived nutrients become available to plants. We aimed to address this question: what are the temporal dynamics of plant growth responses to algal biomass? to better propose mechanisms by which plants acquire nutrients from algal biomass and thereby study and promote those processes in future agricultural applications. Data from various sources were transformed and used to reconstruct the nutrient release from the algae Chlorella vulgaris and subsequent uptake by wheat (Triticum aestivum L.) (as reported in Schreiber et al., 2018). Plants had received 0.1x or 1x dried algae or wet algae, or zero, 0.1x or 1x mineral fertilizer calculated from agricultural practices for P application and grown to 55 days in three soils. Contents of P and other nutrients acquired from algae were as high as from mineral fertilizer, but varied based on moisture content and amount of algae applied to soils (by 55 days after sowing plants with 1x mineral fertilizer and 1x dried algae had 5.6 mg P g DWshoot; 2.2-fold more than those with 0 or 0.1x mineral fertilizer, 0.1x dried algae and wet algae, and 1x wet algae). Absolute and relative leaf area growth and estimated P uptake rates showed similar dynamics, indicating that wheat acquires P from algae quickly. A model proposes that algal fertilizer promotes wheat growth after rapid transformation in soil to inorganic nutrients. We conclude theoretically that phosphorus from algal biomass is available to wheat seedlings upon its application and is released gradually over time with minor differences related to moisture content on application. The growth and P uptake kinetics hint at nutrient forms, including N, and biomass stimulation worthy of research to further exploit algae in sustainable agriculture practices. Temporal resolved phenotype analyses in combination with a mass-balance approach is helpful for understanding resource uptake from recycled and biofertilizer sources by plants.
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ItemTranscriptome Profiling Combined With Activities of Antioxidant and Soil Enzymes Reveals an Ability of Pseudomonas sp. CFA to Mitigate p-Hydroxybenzoic and Ferulic Acid Stresses in Cucumber(Frontiers Media, 2020-10-27)Continuous-cropping leads to obstacles in crop productivity by the accumulation of p-hydroxybenzoic acid (PHBA) and ferulic acid (FA). In this study, a strain CFA of Pseudomonas was shown to have a higher PHBA- and FA-degrading ability in media and soil and the mechanisms underlying this were explored. Optimal conditions for PHBA and FA degradation by CFA were 0.2 g/l of PHBA and FA, 37°C, and pH 6.56. Using transcriptome analysis, complete pathways that converted PHBA and FA to acetyl coenzyme A were proposed in CFA. When CFA was provided with PHBA and FA, we observed upregulation of genes in the pathways and detected intermediate metabolites including vanillin, vanillic acid, and protocatechuic acid. Moreover, 4-hydroxybenzoate 3-monooxygenase and vanillate O-demethylase were rate-limiting enzymes by gene overexpression. Knockouts of small non-coding RNA (sRNA) genes, including sRNA 11, sRNA 14, sRNA 20, and sRNA 60, improved the degradation of PHBA and FA. When applied to cucumber-planted soil supplemented with PHBA and FA, CFA decreased PHBA and FA in soil. Furthermore, a reduction of superoxide radical, hydrogen peroxide, and malondialdehyde in cucumber was observed by activating superoxide dismutase, catalase, glutathione peroxidase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate reductase in seedlings, increasing the reduced glutathione and ascorbate in leaves, and inducing catalase, urease, and phosphatase in the rhizosphere. CFA has potential to mitigate PHBA and FA stresses in cucumber and alleviate continuous-cropping obstacles.
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ItemTime-resolution of the shoot and root growth of the model cereal Brachypodium in response to inoculation with Azospirillum bacteria at low phosphorus and temperature(Springer Verlag, 2021-01)A non-invasive plant phenotyping platform, GrowScreen-PaGe, was used to resolve the dynamics of shoot and root growth of the model cereal Brachypodium (Brachypodium distachyon Bd21-3) in response to the plant growth promoting (PGP) bacteria Azospirillum (Azospirillum brasilense Sp245). Inoculated Brachypodium plants had greater early vigor and higher P use efficiency than non-inoculated Brachypodium at low P and low temperature conditions. Root systems were imaged non-invasively at eight time points and data combined with leaf area, shoot biomass and nutrient content from destructive subsamples at 7, 14 and 21 days after inoculation (DAI). Azospirillum colonisation of roots improved Brachypodium shoot and, to a greater degree, root growth in three independent experiments. Inoculation promoted P use efficiency in shoots but not P concentration or uptake, despite increased total root length. Longer roots in inoculated plants arose from twofold faster branch root growth but slower axile root growth, detected at 11 DAI. Analysis of the spatio-temporal phenotypes indicated that the effects of Azospirillum inoculation increased as shoot P concentration declined, but the magnitude depended on the time after inoculation and growth rate of branch roots compared to axile roots. High throughput plant phenotyping platforms allow the details of plant-microorganism symbioses to be resolved, offering insights into the timing of changes in different tissues to allow molecular mechanisms to be determined.