Environmental Sustainability

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Browsing Environmental Sustainability by Publisher "Frontiers"

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    DNA-free CRISPR genome editing in raspberry (Rubus idaeus) protoplast through RNP-mediated transfection
    (Frontiers, 2025-06-30) Creeth, Ryan; Thompson, Andrew J.; Kevei, Zoltan
    Protoplast-based systems have been utilised in a wide variety of plant species to enable genome editing without chromosomal introgression of foreign DNA into plant genomes. DNA-free genome editing followed by protoplast regeneration allows elite cultivars to be edited without further genetic segregation, preserving their unique genetic composition and their regulatory status as non-transgenic. However, protoplast isolation presents a barrier to the development of advanced breeding technologies in raspberry and no protocol has been published for DNA-free genome editing in the species. Pre-assembled ribonucleoprotein complexes (RNPs) do not require cellular processing and the commercial availability of Cas9 proteins and synthetic guide RNAs has streamlined genome editing protocols. This study presents a novel high-yielding protoplast isolation protocol from raspberry stem cultures and RNP-mediated transfection of protoplast with CRISPR-Cas9. Targeted mutagenesis of the phytoene desaturase gene at two intragenic loci resulted in an editing efficiency of 19%, though estimated efficiency varied depending on the indel analysis technique. Only amplicon sequencing was sensitive enough to confirm genome editing in a low efficiency sample. To our knowledge, this study constitutes the first use of DNA-free genome editing in raspberry protoplast. This protocol provides a valuable platform for understanding gene function and facilitates the future development of precision breeding in this important soft fruit crop.
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    Feasibility study on using combined tomography and spectroscopy techniques to evaluate the physical and chemical characteristics of organo-mineral fertilisers
    (Frontiers, 2025-06-04) Sakrabani, Ruben; Mosca, Sara; Liptak, Alexander; Burca, Genoveva
    Fertilisers play a key role in agriculture, providing key nutrients needed by crops to ensure a secure food supply. However, with increasing prices and rising environmental concerns, there is a growing need to rely on alternative and sustainable fertiliser sources, introducing the opportunity to use organic amendments to formulate organo-mineral fertilisers (OMF). Despite their environmental advantages, the inherent variability in composition of organic amendments within OMF poses a challenge for their standardization. This study aims to use OMF derived from anaerobic digestate and coupled with carbon capture technologies to analyze for its physical characteristics and chemical composition using neutron computed tomography (NCT), X-ray computed tomography (XCT) and Raman spectroscopy (RS). This is a feasibility study to assess using non-destructive techniques on OMF as previously this has not been explored. This work represents the first attempt to utilize a combination of imaging techniques to investigate on OMF and demonstrates their feasibility for measuring the variability between individual samples. This is a proof-of-concept study which shows that combining NCT and XCT can provide images on how uniformly packed each OMF pellet are. The use of RS is to characterize OMF is more challenging largely due to the high fluorescence background arising from its matrix. This study needs to be further developed to enable image-based analysis using machine learning algorithms to determine characteristics of large batches of OMF. Further development is needed building on this work to quantify OMF pellet characteristics so that it can be confidently used as novel fertilisers in agriculture.
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    Morphological changes of lenticels and their role in gas exchange and sprouting physiology of potato tubers during postharvest storage
    (Frontiers, 2025-06-11) Magwaza, Lembe Samukelo; Bernal, Antonio J. B.; Chope, Gemma A.; Alamar, M. Carmen; Terry, Leon A.
    The application of exogenous gases has been used to suppress sprouting in stored potato tubers. However, their efficacy in extending ecodormancy largely depends on achieving optimal gas exchange between the storage atmosphere and the tuber itself. This study aimed to investigate morphological variations and spatial distribution of lenticels and apical buds and to identify their potential role in tuber respiration rate and sprouting of five potato cultivars (‘Hermes’, ‘Lady Claire’, ‘Lady Rosetta’, ‘Saturna’, and ‘VR808’) during storage. Results revealed a consistent spatial pattern wherein the apical section of potato tubers exhibited significantly higher bud counts compared to lateral and stolon regions. ‘Lady Claire’ stood out as having the highest number of apical buds among the cultivars studied. Digital image analysis showed a seven times higher number of buds surrounding the apical eye and these were generally smaller than those distributed across the skin. ‘Saturna’ displayed double the lenticel density (12 lenticels cm-2) in smaller tubers, suggesting an inverse relationship between tuber size and lenticel density. ‘Lady Claire’ and ‘Saturna’ had respiration rates of 2.75 and 1.9 mL CO2 kg-1 h-1, respectively, and were selected for additional respiration and ethylene efflux analyses. In both cultivars, distinct spatial differences were observed, with the apical section exhibiting a seven-fold increase in lenticel density compared to the lateral and stolon sections. Respiration rate increased five-fold when apical lenticels were blocked, whereas it decreased 30-fold when the apical was the only unblocked section, suggesting differential physiological activity across lenticel locations. The apical sections, with the highest lenticel density, exhibited elevated respiration rates as a stress-induced physiological response upon blockage, compared to the lateral and stolon sections. Lenticels changed their morphology during storage, erupting before bud movement, suggesting lenticel eruption could be used as a pre-symptomatic visual marker of dormancy break. This study highlights the critical role that lenticel morphology and spatial distribution may have in determining potato tuber gas exchange and refining allied storage regimes.
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    Nanomaterials as a new frontier platform: metal-doped and hybrid carbon dots as enzyme mimics for environmental applications
    (Frontiers, 2025-01-01) Yousaf, Aiman; Imran, Muhammad; Farooq Warsi, Muhammad; Alsafari, Ibrahim A.; Khan, Farhan A.; Parra-Saldívar, Roberto; Gutiérrez-Soto, Guadalupe; Iqbal, Hafiz M. N.
    Environmental pollution has become an inexorable problem for the planet Earth. The precise detection and degradation of heavy metals, pesticides, industrial-, pharmaceutical- and personal care- products is needed. Nanotechnology holds great promise in addressing global issues. Over the past decades, nanozymic nanomaterials have exceptionally overcome the intrinsic limitations of natural enzymes. Carbon dots (CDs) exhibit unique structures, surface properties, high catalytic activities, and low toxicity. Different techniques, such as doping or surface passivation, can enhance these exceptional properties. Doping modifies CDs’ electronic, magnetic, optical, and catalytic properties considerably. Metal doping, a more significant strategy, involves the introduction of metallic impurities, which offer insight into enhancing the physicochemical properties of CDs. Metal-doped CDs exhibit higher optical absorbance and catalytic performance than pristine CDs. The literature shows that researchers have utilized various synthetic approaches to fabricate CDs-Metal nanozymes. Researchers have reported the metal-doped and hybrid CDs’ peroxidase, catalase, laccase, and superoxide dismutase-like activities. These metal-doped nanozymes put forward substantial environmental remediations and applications such as sensing, photocatalytic degradation, adsorption, and removal of environmental contaminants. This review thoroughly discussed the metal-based functionalization of CDs, the enzyme-like properties, and the ecological applications of metal-doped and hybrid enzymes. The review also presents the current novelties, remaining challenges, and future directions with key examples.
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    Prolonged heat stress in Brassica napus during flowering negatively impacts yield and alters glucosinolate and sugars metabolism
    (Frontiers, 2025-01-01) Kourani, Mariam; Anastasiadi, Maria; Hammond, John P.; Mohareb, Fady R.
    Oilseed rape (Brassica napus), one of the most important sources of vegetable oil worldwide, is adversely impacted by heatwave-induced temperature stress especially during its yield-determining reproductive stages. However, the underlying molecular and biochemical mechanisms are still poorly understood. In this study, we investigated the transcriptomic and metabolomic responses to heat stress in B. napus plants exposed to a gradual increase in temperature reaching 30°C in the day and 24°C at night for a period of 6 days. High-performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC-MS) was used to quantify the content of carbohydrates and glucosinolates, respectively. Results showed that heat stress reduced yield and altered oil composition. Heat stress also increased the content of carbohydrate (glucose, fructose, and sucrose) and aliphatic glucosinolates (gluconapin and progoitrin) in the leaves but decreased the content of the indolic glucosinolate (glucobrassicin). RNA-Seq analysis of flower buds showed a total of 1,892, 3,253, and 4,553 differentially expressed genes at 0, 1, and 2 days after treatment (DAT) and 4,165 and 1,713 at 1 and 7 days of recovery (DOR), respectively. Heat treatment resulted in downregulation of genes involved in respiratory metabolism, namely, glycolysis, pentose phosphate pathway, citrate cycle, and oxidative phosphorylation especially after 48 h of heat stress. Other downregulated genes mapped to sugar transporters, nitrogen transport and storage, cell wall modification, and methylation. In contrast, upregulated genes mapped to small heat shock proteins (sHSP20) and other heat shock factors that play important roles in thermotolerance. Furthermore, two genes were chosen from the pathways involved in the heat stress response to further examine their expression using real-time RT-qPCR. The global transcriptome profiling, integrated with the metabolic analysis in the study, shed the light on key genes and metabolic pathways impacted and responded to abiotic stresses exhibited as a result of exposure to heat waves during flowering. DEGs and metabolites identified through this study could serve as important biomarkers for breeding programs to select cultivars with stronger resistance to heat. In particular, these biomarkers can form targets for various crop breeding and improvement techniques such as marker-assisted selection.