1,335 results on '"Alisdair R. Fernie"'
Search Results
2. Effect of exogenous treatment with zaxinone and its mimics on rice root microbiota across different growth stages
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Teresa Mazzarella, Matteo Chialva, Leonardo Perez de Souza, Jian You Wang, Cristina Votta, Rhowell Tiozon, Patrizia Vaccino, Alessandra Salvioli di Fossalunga, Nese Sreenivasulu, Tadao Asami, Alisdair R. Fernie, Salim Al-Babili, Luisa Lanfranco, and Valentina Fiorilli
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Oryza sativa ,Zaxinone ,MiZax ,Microbial communities ,Rhizosphere ,Shoot metabolism ,Medicine ,Science - Abstract
Abstract Enhancing crops productivity to ensure food security is one of the major challenges encountering agriculture today. A promising solution is the use of biostimulants, which encompass molecules that enhance plant fitness, growth, and productivity. The regulatory metabolite zaxinone and its mimics (MiZax3 and MiZax5) showed promising results in improving the growth and yield of several crops. Here, the impact of their exogenous application on soil and rice root microbiota was investigated. Plants grown in native paddy soil were treated with zaxinone, MiZax3, and MiZax5 and the composition of bacterial and fungal communities in soil, rhizosphere, and endosphere at the tillering and the milky stage was assessed. Furthermore, shoot metabolome profile and nutrient content of the seeds were evaluated. Results show that treatment with zaxinone and its mimics predominantly influenced the root endosphere prokaryotic community, causing a partial depletion of plant-beneficial microbes at the tillering stage, followed by a recovery of the prokaryotic community structure during the milky stage. Our study provides new insights into the role of zaxinone and MiZax in the interplay between rice and its root-associated microbiota and paves the way for their practical application in the field as ecologically friendly biostimulants to enhance crop productivity.
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- 2024
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3. Deciphering the roles of bacterial and fungal communities in the formation and quality of agarwood
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Chen-Chen Fu, Bao-Xing Huang, Shan-Shan Wang, Yu-Chen Song, Dolkar Metok, Yu-Xiang Tan, Tai-Ping Fan, Alisdair R. Fernie, Meisam Zargar, Yan Wang, Mo-Xian Chen, Liang-Wen Yu, and Fu-Yuan Zhu
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Aquilaria sinensis ,Endophytic ,Exophytic ,Fungi ,Bacteria ,Plant tissue ,Biology (General) ,QH301-705.5 - Abstract
Abstract Aquilaria sinensis is a significant resin-producing plant worldwide that is crucial for agarwood production. Agarwood has different qualities depending on the method with which it is formed, and the microbial community structures that are present during these methods are also diverse. Furthermore, the microbial communities of plants play crucial roles in determining their health and productivity. While previous studies have investigated the impact of microorganisms on agarwood formation, they lack comprehensiveness, particularly regarding the properties of the microbial community throughout the entire process from seedling to adult to incense formation. We collected roots, stems, leaves, flowers, fruits and other tissues from seedlings, healthy plants and agarwood-producing plants to address this gap and assess the dominant bacterial species in the microbial community structures of A. sinensis at different growth stages and their impacts on growth and agarwood formation. The bacteria and fungi in these tissues were classified and counted from different perspectives. The samples were sequenced using the Illumina sequencing platform, and sequence analyses and species annotations were performed using a range of bioinformatics tools to assess the plant community compositions. An additional comparison of the samples was conducted using diversity analyses to assess their differences. This research revealed that Listeria, Kurtzmanomyces, Ascotaiwania, Acinetobacter, Sphingobium, Fonsecaea, Acrocalymma, Allorhizobium, Bacillus, Pseudomonas, Peethambara, and Debaryomyces are potentially associated with the formation of agarwood. Overall, the data provided in this article help us understand the important roles played by bacteria and fungi in the growth and agarwood formation process of A. sinensis, will support the theoretical basis for the large-scale cultivation of A. sinensis, and provide a basis for further research on microbial community applications in agarwood production and beyond.
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- 2024
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4. Lipidomics-based association study reveals genomic signatures of anti-cancer qualities of pigmented rice sprouts
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Rhowell Navarro Tiozon, Erstelle Pasion-Uy, Saleh Alseekh, Kristel June D. Sartagoda, Shem Gempesaw, Joel H. G. Tolentino, Alisdair R. Fernie, and Nese Sreenivasulu
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antioxidants ,anticancer ,lipidomics ,lipase ,pigmented rice ,Plant culture ,SB1-1110 - Abstract
IntroductionThe genetic wealth present in pigmented rice varieties offer abundant variation in different sources of antioxidants to meet nutritional security targets among rice-consuming communities. There is limited knowledge of the dynamic changes in the lipidome of rice during germination and the corresponding genes associated with the antioxidant and anti-cancerous properties of lipophilic fractions of pigmented rice sprouts (PRS).MethodsIn this study, we profiled the lipidome of diverse pigmented rice collections of germinated sprouts. Further, we employed Genome-wide association studies (GWAS), gene-set analysis, and targeted association analysis to identify the candidate genes linked to these lipids.ResultsThe genetic analyses revealed 72 candidate genes involved in the regulation of these accumulating lipids in PRS. Marker trait associations (MTA) analysis shown that the combination GGTAAC/ACAAGCTGGGCCC was associated with increased levels of unsaturated lipids and carotenoids, which likely underlie these beneficial effects. This superior MTA combination exhibited potent inhibitory activity against HCT116 and A549 cell lines, with average 1/IC50 values of 0.03 and 0.02 (mL/μg), respectively, compared to the inferior MTAs.DiscussionCollectively, our findings demonstrate that MTAs linked to selected GDSL esterase/lipase (GELP) genes, OsACP1, and lecithin-cholesterol acyltransferase significantly enhance antioxidant and anti-cancer properties, potentially through the mobilization of unsaturated lipids and carotenoids during germination. This study offers valuable insights into the health-promoting potential of germinated rice sprouts as a rich dietary source of antioxidants beneficial to human health.
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- 2025
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5. Genomic evidence of a transposon-conferred adaptive response
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Koki Hayashi and Alisdair R. Fernie
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Biology (General) ,QH301-705.5 - Abstract
Raingeval et al.1 characterize the insertion of the ONSEN transposon in the intron of a flowering repressor, which allows the plant to accelerate its life cycle in response to stress. They show the insertion was positively selected in a herbicide-intense environment demonstrating its role in adaptation to the local environment.
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- 2024
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6. Physiological and molecular insights into the effect of a seaweed biostimulant on enhancing fruit yield and drought tolerance in tomato
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Aakansha Kanojia, Rafe Lyall, Neerakkal Sujeeth, Saleh Alseekh, Félix J. Martínez-Rivas, Alisdair R. Fernie, Tsanko S. Gechev, and Veselin Petrov
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Biostimulant ,Drought stress ,Priming ,Photosynthesis ,Senescence ,Seaweed extract ,Plant ecology ,QK900-989 - Abstract
Tomato is one of the most widely grown vegetable crops in Europe. This study describes an approach for treating tomato plants with an extract of Ascophyllum nodosum (ANF) prior to a stress event, which prepares the plants at the molecular level to respond more effectively to stress conditions, through a process known as molecular priming. Solanum lycopersicum L. cv. Micro-Tom, a dwarf tomato variety, was pre-treated with ANF via foliar spray during the flowering phase and subsequently subjected to drought conditions. ANF-treated plants exhibited enhanced growth, fruit yield, and stress tolerance under both moderate and severe drought conditions compared to untreated plants. Transcriptomic studies in leaves revealed that the priming treatment preserved the photosynthetic machinery, inducing stress-protective genes involved in ascorbate, peroxidase, GABA, glutathione, and flavanol biosynthesis. Simultaneously, the treatment repressed key senescence-related genes associated with ethylene biosynthesis, as well as several WRKY and NAC transcription factors. Metabolome analysis demonstrated that ANF induces drought tolerance by promoting the accumulation of stress-protective primary and secondary metabolites, such as GABA, proline, maltose, ascorbic acid, quercetin, and biotin, which can act as osmoprotectants and free radical scavengers during drought. Combined transcriptome and metabolome analyses suggest that ANF treatment represses the senescence process, maintains photosynthetic activity, and induces the accumulation of protective metabolites and amino acids, promoting plant survival and growth under drought. Overall, this research provides new insights into the molecular mechanisms underlying biostimulant-based molecular priming and offers a knowledge-based approach for the accurate application of this ANF molecular priming agent to increase crop productivity and mitigate yield loss during drought, contributing to food security in the era of climate change.
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- 2024
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7. Autophagy in plants
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Morten Petersen, Tamar Avin-Wittenberg, Diane C. Bassham, Yasin Dagdas, Chudi Fan, Alisdair R. Fernie, Liwen Jiang, Divya Mishra, Marisa S. Otegui, Eleazar Rodriguez, and Daniel Hofius
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Plant autophagy ,regulation and signalling ,endomembrane trafficking ,quality control ,cargo receptors ,development ,Cytology ,QH573-671 - Abstract
Autophagy is a process of cellular self-eating, which allows organisms to eliminate and recycle unwanted components and damaged organelles to maintain cellular homeostasis. It is an important process in the development of eukaryotic organisms. Autophagy plays a critical role in many physiological processes in plants such as nutrient remobilization, cell death, immunity, and abiotic stress responses. Autophagy thus represents an obvious target for generating resilient crops. During plant development, autophagy is also implicated in the differentiation and maturation of various cell types and plant organs, including root cap cells, tracheary elements, gametes, fruits and seeds. Here, we review our current understanding and recent advances of plant autophagy including insight into autophagy regulation and signaling as well as autophagosome membrane biogenesis. In addition, we describe how autophagy contributes to development, metabolism, biotic and abiotic stress tolerance and where the autophagic field is heading in terms of applied research for crop improvement.
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- 2024
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8. The natural variance of Arabidopsis secondary metabolism on extended darkness
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Feng Zhu, Micha Wijesingha Ahchige, Weiwei Wen, Yunjiang Cheng, Saleh Alseekh, and Alisdair R. Fernie
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Science - Abstract
Abstract In plants due to their sessile nature, secondary metabolites are important components against different abiotic and biotic stress, such as extended darkness. For this reason, the variation of secondary metabolite content of the Arabidopsis thaliana HapMap natural population following 0-and 6-d darkness treatment were detected and the raw data of different accessions at two timepoints were deposited in the Zenodo database. Moreover, the annotated secondary metabolites of these samples are presented in this data descriptor, which we believe will be a usefully re-usable resource for future integrative analysis with dark-treated transcripts, proteins or other phenotypic data in order to comprehensively illustrate the multiomic landscape of Arabidopsis in response to the stresses exerted by extended darkness.
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- 2024
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9. Variation in a Poaceae-conserved fatty acid metabolic gene cluster controls rice yield by regulating male fertility
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Chenkun Yang, Shuangqian Shen, Chuansong Zhan, Yufei Li, Ran Zhang, Yuanyuan Lv, Zhuang Yang, Junjie Zhou, Yuheng Shi, Xianqing Liu, Jianxin Shi, Dabing Zhang, Alisdair R. Fernie, and Jie Luo
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Science - Abstract
Abstract A wide variety of metabolic gene clusters exist in eukaryotic genomes, but fatty acid metabolic gene clusters have not been discovered. Here, combining with metabolic and phenotypic genome-wide association studies, we identify a major locus containing a six-gene fatty acid metabolic gene cluster on chromosome 3 (FGC3) that controls the cutin monomer hydroxymonoacylglycerols (HMGs) contents and rice yield, possibly through variation in the transcription of FGC3 members. We show that HMGs are sequentially synthesized in the endoplasmic reticulum by OsFAR2, OsKCS11, OsGPAT6, OsCYP704B2 and subsequently transported to the apoplast by OsABCG22 and OsLTPL82. Mutation of FGC3 members reduces HMGs, leading to defective male reproductive development and a significant decrease in yield. OsMADS6 and OsMADS17 directly regulate FGC3 and thus influence male reproduction and yield. FGC3 is conserved in Poaceae and likely formed prior to the divergence of Pharus latifolius. The eukaryotic fatty acid and plant primary metabolic gene cluster we identified show a significant impact on the origin and evolution of Poaceae and has potential for application in hybrid crop breeding.
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- 2024
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10. Metabolic responses to multi-stress: An update
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Mustafa Bulut, Esra Karakas, and Alisdair R. Fernie
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Multifactorial stress ,Abiotic stress ,Metabolomics ,Branched chain amino acids ,Crops ,Plant ecology ,QK900-989 - Abstract
In recent years, several studies investigating multifactorial stresses have emerged. This shift has been driven by the recognition that one of the primary reasons for the inconsistency between laboratory-based results and field observations of plant responses is that, in natural environments, plants are routinely exposed to a combination of biotic and/or abiotic stresses, which they encounter either simultaneously or sequentially. Within this review, we address current advances in multifactorial studies focusing on metabol(om)ic changes in model as well as cereal crop species. The common consensus is that currently, studies on phenotypic and transcriptomics analysis are prevailing, while metabolic studies are scarce. Despite the need for further studies to validate the findings in this review, two clear biological messages emerge. First, and perhaps unsurprisingly, proline stands out as a universal stress metabolite, closely followed by branched-chain amino acids. Interestingly, while multifactorial stress responses are often considered non-additive and unpredictable, our findings reveal that many metabolic changes are both. Expanding the scope of studies to include more species and a wider range of stresses at the metabolic level will be essential for uncovering additional metabolic reprogramming in response to multifactorial stress. This will provide invaluable insights for developing breeding strategies aimed at future-proofing crops.
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- 2025
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11. Evolution and Domestication of a Novel Biosynthetic Gene Cluster Contributing to the Flavonoid Metabolism and High‐Altitude Adaptability of Plants in the Fagopyrum Genus
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Xu Huang, Yuqi He, Kaixuan Zhang, Yaliang Shi, Hui Zhao, Dili Lai, Hao Lin, Xiangru Wang, Zhimin Yang, Yawen Xiao, Wei Li, Yinan Ouyang, Sun Hee Woo, Muriel Quinet, Milen I. Georgiev, Alisdair R. Fernie, Xu Liu, and Meiliang Zhou
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buckwheat ,evolution ,flavonoid metabolism ,novel gene cluster ,UV resistance ,Science - Abstract
Abstract The diversity of secondary metabolites is an important means for plants to cope with the complex and ever‐changing terrestrial environment. Plant biosynthetic gene clusters (BGCs) are crucial for the biosynthesis of secondary metabolites. The domestication and evolution of BGCs and how they affect plant secondary metabolites biosynthesis and environmental adaptation are still not fully understood. Buckwheat exhibits strong resistance and abundant secondary metabolites, especially flavonoids, allowing it to thrive in harsh environments. A non‐canonical BGC named UFGT3 cluster is identified, which comprises a phosphorylase kinase (PAK), two transcription factors (MADS1/2), and a glycosyltransferase (UFGT3), forming a complete molecular regulatory module involved in flavonoid biosynthesis. This cluster is selected during Tartary buckwheat domestication and is widely present in species of the Fagopyrum genus. In wild relatives of cultivated buckwheat, a gene encoding anthocyanin glycosyltransferase (AGT), which glycosylates pelargonidin into pelargonidin‐3‐O‐glucoside, is found inserted into this cluster. The pelargonidin‐3‐O‐glucoside can help plants resist UV stress, endowing wild relatives with stronger high‐altitude adaptability. This study provides a new research paradigm for the evolutionary dynamics of plant BGCs, and offers new perspectives for exploring the mechanism of plant ecological adaptability driven by environmental stress through the synthesis of secondary metabolites.
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- 2024
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12. Emerging role of jasmonic acid in woody plant development
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Yun-Jing Bao, Jia-Xu Chen, Youjun Zhang, Alisdair R. Fernie, Jianhua Zhang, Bao-Xing Huang, Fu-Yuan Zhu, and Fu-Liang Cao
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Alternative splicing ,Jasmonic acid ,Proteogenomic ,Plant development ,Woody plants ,Agriculture (General) ,S1-972 ,Biochemistry ,QD415-436 ,Chemistry ,QD1-999 - Abstract
Jasmonic acid is a crucial phytohormone that plays a pivotal role, serving as a regulator to balancing plant development and resistance. However, there are analogous and distinctive characteristics exhibited in JA biosynthesis, perception, and signal transduction pathways in both herbaceous and woody plants. Moreover, the majority of research subjects have predominantly focused on the function of JA in model or herbaceous plants. Consequently, there is a significant paucity of studies investigating JA regulation networks in woody plants, particularly concerning post-transcriptional regulatory events such as alternative splicing (AS). This review article aims to conduct a comprehensive summary of advancements that JA signals regulate plant development across various woody species, comparing the analogous features and regulatory differences to herbaceous counterparts. In addition, we summarized the involvement of AS events including splicing factor (SF) and transcripts in the JA regulatory network, highlighting the effectiveness of high-throughput proteogenomic methods. A better understanding of the JA signaling pathway in woody plants has pivotal implications for forestry production, including optimizing plant management and enhancing secondary metabolite production.
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- 2024
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13. M urraya koenigii (L.) Sprengel seeds and pericarps in relation to their chemical profiles: new approach for multidrug resistant Acinetobacter baumannii ventilator-associated pneumonia
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Riham A. El-Shiekh, Rana Elshimy, Asmaa A. Mandour, Hanaa A. H. Kassem, Amal E. Khaleel, Saleh Alseekh, Alisdair R. Fernie, and Mohamed A. Salem
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Murraya koenigii seeds and pericarps ,MDR A. baumannii ,LC/MS/MS ,Chemometrics ,Metabolomics ,Molecular networking ,Agriculture (General) ,S1-972 ,Chemistry ,QD1-999 - Abstract
Abstract Acinetobacter baumannii is without a doubt one of the most problematic bacteria causing hospital-acquired nosocomial infections in today's healthcare system. To solve the high prevalence of multi-drug resistant (MDR) in A. baumannii, we investigated one of the medicinal plants traditionally used as antibacterial agent; namely Murraya koenigii (L.) Sprengel. The total methanolic extracts of seeds and pericarps were prepared and their anti-bacterial activity was assessed using the agar diffusion method and minimum inhibitory concentration (MIC) was then calculated as compared to tigecycline. Then, an in-vivo murine model was established which confirmed the promising activity of M. koenigii seeds in demonstrating anti-bacterial and anti-inflammatory actions. The histopathological study of lungs, scoring of pulmonary lesions, counting of bacterial loads after infection by multi-drug resistant A. baumannii all provided evidence to support these findings. LC–MS/MS profiling coupled to molecular networking and chemometrics detected the presence of carbazole alkaloids, and coumarins as dominate metabolites of the active seed extracts. Positively correlated metabolites to antibacterial potential were 6-(2ʹ,3ʹ-dihydroxy-3-methylbutyl)-8-prenylumbelliferone, scopoline, and 5-methoxymurrayatin. An in-silico study was also performed on the crystal structure of MurF from A. baumannii (PDB ID: 4QF5), the studied structures of the mentioned extracts revealed good docking interaction at the active site suggestive of competition with the ATP ligand. These collective findings suggest that extracts of Murraya koenigii (L.) Sprengel seed is a novel prospective for the discovery of drug candidates against infections caused by MDR A. baumannii.
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- 2024
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14. Genomic insight into the origin, domestication, dispersal, diversification and human selection of Tartary buckwheat
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Yuqi He, Kaixuan Zhang, Yaliang Shi, Hao Lin, Xu Huang, Xiang Lu, Zhirong Wang, Wei Li, Xibo Feng, Taoxiong Shi, Qingfu Chen, Junzhen Wang, Yu Tang, Mark A. Chapman, Mateja Germ, Zlata Luthar, Ivan Kreft, Dagmar Janovská, Vladimir Meglič, Sun-Hee Woo, Muriel Quinet, Alisdair R. Fernie, Xu Liu, and Meiliang Zhou
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Domestication ,Migration ,Artificial selection ,Buckwheat ,Genomics ,Biology (General) ,QH301-705.5 ,Genetics ,QH426-470 - Abstract
Abstract Background Tartary buckwheat, Fagopyrum tataricum, is a pseudocereal crop with worldwide distribution and high nutritional value. However, the origin and domestication history of this crop remain to be elucidated. Results Here, by analyzing the population genomics of 567 accessions collected worldwide and reviewing historical documents, we find that Tartary buckwheat originated in the Himalayan region and then spread southwest possibly along with the migration of the Yi people, a minority in Southwestern China that has a long history of planting Tartary buckwheat. Along with the expansion of the Mongol Empire, Tartary buckwheat dispersed to Europe and ultimately to the rest of the world. The different natural growth environments resulted in adaptation, especially significant differences in salt tolerance between northern and southern Chinese Tartary buckwheat populations. By scanning for selective sweeps and using a genome-wide association study, we identify genes responsible for Tartary buckwheat domestication and differentiation, which we then experimentally validate. Comparative genomics and QTL analysis further shed light on the genetic foundation of the easily dehulled trait in a particular variety that was artificially selected by the Wa people, a minority group in Southwestern China known for cultivating Tartary buckwheat specifically for steaming as a staple food to prevent lysine deficiency. Conclusions This study provides both comprehensive insights into the origin and domestication of, and a foundation for molecular breeding for, Tartary buckwheat.
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- 2024
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15. Non-targeted discovery of high-value bio-products in Nicotiana glauca L: a potential renewable plant feedstock
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Natalia Carreno-Quintero, Takayuki Tohge, Rebecca Van Acker, Lauren S. McKee, Qi Zhou, Antje Bolze, Xiaohui Xing, Merve Özparpucu, Markus Rüggeberg, Thomas Piofczyk, Yaw Koram, Vincent Bulone, Wout Boerjan, Alisdair R. Fernie, and Paul D. Fraser
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Nicotiana glauca ,Metabolite profiling ,Biorefinary ,Bioproducts ,Technology ,Chemical technology ,TP1-1185 ,Biotechnology ,TP248.13-248.65 - Abstract
Abstract The evaluation of plant-based feedstocks is an important aspect of biorefining. Nicotiana glauca is a solanaceous, non-food crop that produces large amounts of biomass and is well adapted to grow in suboptimal conditions. In the present article, compatible sequential solvent extractions were applied to N. glauca leaves to enable the generation of enriched extracts containing higher metabolite content comparing to direct leaf extracts. Typically, between 60 to 100 metabolite components were identified within the fractions. The occurrence of plant fatty acids, fatty acid alcohols, alkanes, sterols and terpenoids was detected by gas liquid chromatography–mass spectrometry (GC–MS) and metabolite identification was confirmed by comparison of physico-chemical properties displayed by available authentic standards. Collectively, co-products such waxes, oils, fermentable sugars, and terpenoids were all identified and quantified. The enriched fractions of N. glauca revealed a high level of readily extractable hydrocarbons, oils and high value co-products. In addition, the saccharification yield and cell wall composition analyses in the stems revealed the potential of the residue material as a promising lignocellulosic substrate for the production of fermentable sugars. In conclusion a multifractional cascade for valuable compounds/commodities has been development, that uses N. glauca biomass. These data have enabled the evaluation of N. glauca material as a potential feedstock for biorefining. Graphical Abstract
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- 2024
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16. Physiological and Microstructure Analysis Reveals the Mechanism by Which Formic Acid Delays Postharvest Physiological Deterioration of Cassava
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Yannian Che, Zhongping Ding, Chen Shen, Alisdair R. Fernie, Xiangning Tang, Yuan Yao, Jiao Liu, Yajie Wang, Ruimei Li, and Jianchun Guo
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formic acid ,shelf life ,cassava ,postharvest ,cell wall ,pectin ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Formic acid is reported to act as a food preservative and feed additive, but its effects on controlling postharvest physiological deterioration (PPD) development in cassava are unclear. In this study, we assessed the effectiveness of different concentrations of formic acid in attenuating PPD occurrence in fresh-cut cassava. The results showed that the concentration of 0.1% (v/v) formic acid could significantly delay the occurrence of PPD, and that the higher the concentration of formic acid supplied, the later the occurrence of PPD symptoms. The physiological and biochemical analysis of 0.5%-formic-acid-treated cassava slices revealed that formic acid decreased the degradation of starch, inhibited the accumulation of hydrogen peroxide (H2O2), malondialdehyde (MDA), and water-soluble pectin in cassava slices with PPD development, and increased the activities of the antioxidant enzymes ascorbate peroxidase (APX) and glutathione reductase (GR). A microscopic observation showed that the formic acid treatment inhibited the enlargement of the intercellular space during the cassava PPD process, which suggests that the formation of an intercellular layer of the cell wall was inhibited by formic acid. This study thus revealed the mechanism used by formic acid to extend the cassava shelf life; however, a detailed evaluation of the possible side effects on, for example, the cyanide content will be needed to categorically ensure the safety of this method.
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- 2024
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17. Growth in fluctuating light buffers plants against photorespiratory perturbations
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Thekla von Bismarck, Philipp Wendering, Leonardo Perez de Souza, Jeremy Ruß, Linnéa Strandberg, Elmien Heyneke, Berkley J. Walker, Mark A. Schöttler, Alisdair R. Fernie, Zoran Nikoloski, and Ute Armbruster
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Science - Abstract
Abstract Photorespiration (PR) is the pathway that detoxifies the product of the oxygenation reaction of Rubisco. It has been hypothesized that in dynamic light environments, PR provides a photoprotective function. To test this hypothesis, we characterized plants with varying PR enzyme activities under fluctuating and non-fluctuating light conditions. Contrasting our expectations, growth of mutants with decreased PR enzyme levels was least affected in fluctuating light compared with wild type. Results for growth, photosynthesis and metabolites combined with thermodynamics-based flux analysis revealed two main causal factors for this unanticipated finding: reduced rates of photosynthesis in fluctuating light and complex re-routing of metabolic fluxes. Only in non-fluctuating light, mutants lacking the glutamate:glyoxylate aminotransferase 1 re-routed glycolate processing to the chloroplast, resulting in photooxidative damage through H2O2 production. Our results reveal that dynamic light environments buffer plant growth and metabolism against photorespiratory perturbations.
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- 2023
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18. Metabolomics and machine learning technique revealed that germination enhances the multi-nutritional properties of pigmented rice
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Rhowell Jr. N. Tiozon, Nese Sreenivasulu, Saleh Alseekh, Kristel June D. Sartagoda, Björn Usadel, and Alisdair R. Fernie
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Biology (General) ,QH301-705.5 - Abstract
Abstract Enhancing the dietary properties of rice is crucial to contribute to alleviating hidden hunger and non-communicable diseases in rice-consuming countries. Germination is a bioprocessing approach to increase the bioavailability of nutrients in rice. However, there is a scarce information on how germination impacts the overall nutritional profile of pigmented rice sprouts (PRS). Herein, we demonstrated that germination resulted to increase levels of certain dietary compounds, such as free phenolics and micronutrients (Ca, Na, Fe, Zn, riboflavin, and biotin). Metabolomic analysis revealed the preferential accumulation of dipeptides, GABA, and flavonoids in the germination process. Genome-wide association studies of the PRS suggested the activation of specific genes such as CHS1 and UGT genes responsible for increasing certain flavonoid compounds. Haplotype analyses showed a significant difference (P
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- 2023
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19. Dynamic metabolite QTL analyses provide novel biochemical insights into kernel development and nutritional quality improvement in common wheat
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Bo Yin, Jingqi Jia, Xu Sun, Xin Hu, Min Ao, Wei Liu, Zhitao Tian, Hongbo Liu, Dongqin Li, Wenfei Tian, Yuanfeng Hao, Xianchun Xia, Nir Sade, Yariv Brotman, Alisdair R. Fernie, Jie Chen, Zhonghu He, and Wei Chen
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mQTL ,wheat kernel development ,gene validation ,nutritional quality ,wheat ,Botany ,QK1-989 - Abstract
Despite recent advances in crop metabolomics, the genetic control and molecular basis of the wheat kernel metabolome at different developmental stages remain largely unknown. Here, we performed widely targeted metabolite profiling of kernels from three developmental stages (grain-filling kernels [FKs], mature kernels [MKs], and germinating kernels [GKs]) using a population of 159 recombinant inbred lines. We detected 625 annotated metabolites and mapped 3173, 3143, and 2644 metabolite quantitative trait loci (mQTLs) in FKs, MKs, and GKs, respectively. Only 52 mQTLs were mapped at all three stages, indicating the high stage specificity of the wheat kernel metabolome. Four candidate genes were functionally validated by in vitro enzymatic reactions and/or transgenic approaches in wheat, three of which mediated the tricin metabolic pathway. Metabolite flux efficiencies within the tricin pathway were evaluated, and superior candidate haplotypes were identified, comprehensively delineating the tricin metabolism pathway in wheat. Finally, additional wheat metabolic pathways were re-constructed by updating them to incorporate the 177 candidate genes identified in this study. Our work provides new information on variations in the wheat kernel metabolome and important molecular resources for improvement of wheat nutritional quality.
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- 2024
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20. Roles and regulation of the RBOHD enzyme in initiating ROS-mediated systemic signaling during biotic and abiotic stress
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Felix Juan Martínez Rivas, Alisdair R. Fernie, and Fayezeh Aarabi
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Reactive oxygen species ,Apoplast ,ROS-producing enzyme ,RBOH ,Stress response ,Signaling ,Plant ecology ,QK900-989 - Abstract
Reactive oxygen species are, at high levels, harmful to plant cells, whereas, at a balanced and controlled level, they are involved in transduction of oxidative signaling. The apoplast, also known as the extracellular matrix plays a vital role in transferring the extracellular signals to internal organelles and the signal from cell to cell. Moreover, having a lower level of antioxidants renders it a favorable compartment for ROS production and maintenance. Multiple routes for H2O2 production exist in the apoplast; however, a prominent source is the NADPH oxidases, also named respiratory burst oxidase homologs (RBOHs). Many stimuli, such as pathogens, extracellular ATP, hormones, and abiotic stresses can trigger the RBOH-dependent ROS production in the apoplast. RBOHD and RBOHF are extensively studied and proposed typical roles in stomatal conductance and pathogen responses. In this review, we provide an update on RBOH enzymes and the regulatory mechanisms controlling the key enzyme of the family, RBOHD. Furthermore, we performed a phylogenetic analysis by including the RBOH protein family of Arabidopsis and some important crop species which have not been studied before, such as Brasssica rapa, Brassica oleracea, Oriza sativa, Glycine max, and Zea mays to characterize the evolutionary relationship of these isoforms in order to better classify them. We further describe how ROS are perceived and transduced inside the cell and how this interlinks with other systemic signals, including hormones, and abiotic stress responses such as those toward light and salt stress.
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- 2024
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21. Identification of gene function based on models capturing natural variability of Arabidopsis thaliana lipid metabolism
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Sandra Correa Córdoba, Hao Tong, Asdrúbal Burgos, Feng Zhu, Saleh Alseekh, Alisdair R. Fernie, and Zoran Nikoloski
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Science - Abstract
Abstract Lipids play fundamental roles in regulating agronomically important traits. Advances in plant lipid metabolism have until recently largely been based on reductionist approaches, although modulation of its components can have system-wide effects. However, existing models of plant lipid metabolism provide lumped representations, hindering detailed study of component modulation. Here, we present the Plant Lipid Module (PLM) which provides a mechanistic description of lipid metabolism in the Arabidopsis thaliana rosette. We demonstrate that the PLM can be readily integrated in models of A. thaliana Col-0 metabolism, yielding accurate predictions (83%) of single lethal knock-outs and 75% concordance between measured transcript and predicted flux changes under extended darkness. Genome-wide associations with fluxes obtained by integrating the PLM in diel condition- and accession-specific models identify up to 65 candidate genes modulating A. thaliana lipid metabolism. Using mutant lines, we validate up to 40% of the candidates, paving the way for identification of metabolic gene function based on models capturing natural variability in metabolism.
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- 2023
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22. Comparison of metabolomic reconfiguration between Columbia and Landsberg ecotypes subjected to the combination of high salinity and increased irradiance
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Clara Segarra-Medina, Lidia S. Pascual, Saleh Alseekh, Alisdair R. Fernie, José L. Rambla, Aurelio Gómez-Cadenas, and Sara I. Zandalinas
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Arabidopsis ecotypes ,Abiotic stress combination ,Metabolomics ,Salinity ,Increased irradiance ,Botany ,QK1-989 - Abstract
Abstract Background Plants growing in the field are subjected to combinations of abiotic stresses. These conditions pose a devastating threat to crops, decreasing their yield and causing a negative economic impact on agricultural production. Metabolic responses play a key role in plant acclimation to stress and natural variation for these metabolic changes could be key for plant adaptation to fluctuating environmental conditions. Results Here we studied the metabolomic response of two Arabidopsis ecotypes (Columbia-0 [Col] and Landsberg erecta-0 [Ler]), widely used as genetic background for Arabidopsis mutant collections, subjected to the combination of high salinity and increased irradiance. Our findings demonstrate that this stress combination results in a specific metabolic response, different than that of the individual stresses. Although both ecotypes displayed reduced growth and quantum yield of photosystem II, as well as increased foliar damage and malondialdehyde accumulation, different mechanisms to tolerate the stress combination were observed. These included a relocation of amino acids and sugars to act as potential osmoprotectants, and the accumulation of different stress-protective compounds such as polyamines or secondary metabolites. Conclusions Our findings reflect an initial identification of metabolic pathways that differentially change under stress combination that could be considered in studies of stress combination of Arabidopsis mutants that include Col or Ler as genetic backgrounds.
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- 2023
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23. Selection and adaptive introgression guided the complex evolutionary history of the European common bean
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Elisa Bellucci, Andrea Benazzo, Chunming Xu, Elena Bitocchi, Monica Rodriguez, Saleh Alseekh, Valerio Di Vittori, Tania Gioia, Kerstin Neumann, Gaia Cortinovis, Giulia Frascarelli, Ester Murube, Emiliano Trucchi, Laura Nanni, Andrea Ariani, Giuseppina Logozzo, Jin Hee Shin, Chaochih Liu, Liang Jiang, Juan José Ferreira, Ana Campa, Giovanna Attene, Peter L. Morrell, Giorgio Bertorelle, Andreas Graner, Paul Gepts, Alisdair R. Fernie, Scott A. Jackson, and Roberto Papa
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Science - Abstract
Common bean has two distinct domestication centers in Mesoamerica and in the Andes. The authors show that the Andean is the first gene pool successfully introduced in Europe and identify signature of pervasive introgression among gene pools and of selection for flowering underlying adaptation.
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- 2023
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24. Coordinated reprogramming of renal cancer transcriptome, metabolome and secretome associates with immune tumor infiltration
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Piotr Poplawski, Saleh Alseekh, Urszula Jankowska, Bozena Skupien-Rabian, Roksana Iwanicka-Nowicka, Helena Kossowska, Anna Fogtman, Beata Rybicka, Joanna Bogusławska, Anna Adamiok-Ostrowska, Karolina Hanusek, Jan Hanusek, Marta Koblowska, Alisdair R. Fernie, and Agnieszka Piekiełko-Witkowska
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Renal cancer ,SPARC ,Secretome ,Metabolome ,CAFs ,Immune infiltration ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 ,Cytology ,QH573-671 - Abstract
Abstract Background Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cancer. The molecules (proteins, metabolites) secreted by tumors affect their extracellular milieu to support cancer progression. If secreted in amounts detectable in plasma, these molecules can also serve as useful, minimal invasive biomarkers. The knowledge of ccRCC tumor microenvironment is fragmentary. In particular, the links between ccRCC transcriptome and the composition of extracellular milieu are weakly understood. In this study, we hypothesized that ccRCC transcriptome is reprogrammed to support alterations in tumor microenvironment. Therefore, we comprehensively analyzed ccRCC extracellular proteomes and metabolomes as well as transcriptomes of ccRCC cells to find molecules contributing to renal tumor microenvironment. Methods Proteomic and metabolomics analysis of conditioned media isolated from normal kidney cells as well as five ccRCC cell lines was performed using mass spectrometry, with the following ELISA validation. Transcriptomic analysis was done using microarray analysis and validated using real-time PCR. Independent transcriptomic and proteomic datasets of ccRCC tumors were used for the analysis of gene and protein expression as well as the level of the immune infiltration. Results Renal cancer secretome contained 85 proteins detectable in human plasma, consistently altered in all five tested ccRCC cell lines. The top upregulated extracellular proteins included SPARC, STC2, SERPINE1, TGFBI, while downregulated included transferrin and DPP7. The most affected extracellular metabolites were increased 4-hydroxy-proline, succinic acid, cysteine, lactic acid and downregulated glutamine. These changes were associated with altered expression of genes encoding the secreted proteins (SPARC, SERPINE1, STC2, DPP7), membrane transporters (SLC16A4, SLC6A20, ABCA12), and genes involved in protein trafficking and secretion (KIF20A, ANXA3, MIA2, PCSK5, SLC9A3R1, SYTL3, and WNTA7). Analogous expression changes were found in ccRCC tumors. The expression of SPARC predicted the infiltration of ccRCC tumors with endothelial cells. Analysis of the expression of the 85 secretome genes in > 12,000 tumors revealed that SPARC is a PanCancer indicator of cancer-associated fibroblasts’ infiltration. Conclusions Transcriptomic reprogramming of ccRCC supports the changes in an extracellular milieu which are associated with immune infiltration. The proteins identified in our study represent valuable cancer biomarkers detectable in plasma.
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- 2023
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25. Non-targeted metabolomics and chemometrics for saffron (Crocus sativus L.) authentication and adulteration detection in relation to its anticholinesterase activity
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Inas Y. Younis, Engy Mohsen, Rana M. Ibrahim, Alisdair R. Fernie, Saleh Alseekh, and Mohamed A. Salem
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Saffron ,Crocins ,LC-MS ,Metabolomics ,Authenticity ,Adulteration ,Food processing and manufacture ,TP368-456 - Abstract
Saffron (Crocus sativus L.), is a spice of vulnerable medical importance. It is often adulterated to lower its quality and efficacy. The present study sought to detect the level of adulteration in five traded saffron collected from different localities using a high-performance liquid chromatography coupled with high resolution tandem mass spectrometry (UPLC-HR-MS/MS) technique. In total, 62 metabolites were identified in the five extracts namely from monoterpene aldehydes, flavonoid derivatives, apocarotenoids and carotenoids classes as well as chemical markers of adulteration. One common adulterant; Gardenia jasminoides Ellis was detected through the simultaneous determination of its specific constituents including geniposide, shanzhiside and major jasminosides. Also, one synthetic dye; auramine-O was detected in one marketed saffron product. The anticholinesterase activity of the five marketed saffron was also evaluated. Furthermore, saffron samples that contain high crocin levels possessed a promising anticholinesterase activity. It was detected for the first time that auramine-O dye is sometimes used as a potential adulterant of saffron. In spite of the grand researches on saffron, it still lacks an accurate evaluation of the commercial saffron products. The results of our study showed that high adulteration rate of trade saffron not only reduces saffron efficacy as a herbal remedy but may also simultaneously represent a health risk.
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- 2023
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26. Metabolite profiling of Arabidopsis mutants of lower glycolysis
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Youjun Zhang and Alisdair R. Fernie
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Science - Abstract
Measurement(s) Metabolite profiling of Arabidopsis mutants of enzymes of lower glycolysis Technology Type(s) Gas chromatography–mass spectrometry Factor Type(s) Metabolite profiling of lower glycolysis Sample Characteristic - Organism Arabidopsis mutants of enzymes of lower glycolysis Sample Characteristic - Environment plant grow under 8 h light (22 °C)/16 h dark (18 °C) in growth chamber at a light intensity of 120–150 μmol m-2 s-1 Sample Characteristic - Location 2Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany
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- 2022
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27. Phenology and foraging bias contribute to sex‐specific foraging patterns in the rare declining butterfly Argynnis idalia idalia
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Matthew W. Chmielewski, Skyler Naya, Monica Borghi, Jen Cortese, Alisdair R. Fernie, Mark T. Swartz, Konstantina Zografou, Brent J. Sewall, and Rachel B. Spigler
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flower visitation ,intraspecific variation ,long‐term ecological data ,nectar chemistry ,plant–pollinator interactions ,sex‐specific foraging ,Ecology ,QH540-549.5 - Abstract
Abstract Variation in pollinator foraging behavior can influence pollination effectiveness, community diversity, and plant–pollinator network structure. Although effects of interspecific variation have been widely documented, studies of intraspecific variation in pollinator foraging are relatively rare. Sex‐specific differences in resource use are a strong potential source of intraspecific variation, especially in species where the phenology of males and females differ. Differences may arise from encountering different flowering communities, sex‐specific traits, nutritional requirements, or a combination of these factors. We evaluated sex‐specific foraging patterns in the eastern regal fritillary butterfly (Argynnis idalia idalia), leveraging a 21‐year floral visitation dataset. Because A. i. idalia is protandrous, we determined whether foraging differences were due to divergent phenology by comparing visitation patterns between the entire season with restricted periods of male–female overlap. We quantified nectar carbohydrate and amino acid contents of the most visited plant species and compared those visited more frequently by males versus females. We demonstrate significant differences in visitation patterns between male and female A. i. idalia over two decades. Females visit a greater diversity of species, while dissimilarity in foraging patterns between sexes is persistent and comparable to differences between species. While differences are diminished or absent in some years during periods of male–female overlap, remaining signatures of foraging dissimilarity during implicate mechanisms other than phenology. Nectar of plants visited more by females had greater concentrations of total carbohydrates, glucose, and fructose and individual amino acids than male‐associated plants. Further work can test whether nutritional differences are a cause of visitation patterns or consequence, reflecting seasonal shifts in the nutritional landscape encountered by male and female A. i. idalia. We highlight the importance of considering sex‐specific foraging patterns when studying interaction networks, and in making conservation management decisions for this at‐risk butterfly and other species exhibiting strong intraspecific variation.
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- 2023
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28. Heterosis for capsacinoids accumulation in chili pepper hybrids is dependent on parent-of-origin effect
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Emmanuel Rezende Naves, Federico Scossa, Wagner L. Araújo, Adriano Nunes-Nesi, Alisdair R. Fernie, and Agustin Zsögön
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Medicine ,Science - Abstract
Abstract Heterosis for agronomic traits is a widespread phenomenon that underpins hybrid crop breeding. However, heterosis at the level of cellular metabolites has not yet been fully explored. Some metabolites are highly sought after, like capsaicinoids found in peppers of the Capsicum genus, which confer the characteristic pungent (‘hot’) flavour of the fruits. We analysed the metabolic profile of the fruit placenta and pericarp of inter- and intra-specific hybrids of two species of Capsicum peppers, C. chinense (cv. Habanero and cv. Biquinho) and C. annuum var. annuum (cv. Jalapeño and cv. Cascadura Ikeda) in complete diallel crosses with reciprocals. The parents and hybrids were grown in a glasshouse and the profile of primary metabolites (sugars, amino acids and organic acids) and capsaicinoids was generated via gas chromatography–time of flight-mass spectrometry (GC–TOF-MS) and ultra-performance liquid chromatography coupled to a mass spectrometer (UPLC-MS), respectively. We found considerable heterotic effects specifically for capsaicinoids accumulation in the fruit placenta of the hybrids, including those derived from non-pungent parents. Furthermore, a large fraction of fruit primary metabolism was influenced by the specific cross combination, with marked parent-of-origin effects, i.e. whether a specific genotype was used as the pistillate or pollen parent. The differences in metabolite levels between the hybrids and their parents provide a snapshot of heterosis for primary and secondary metabolites and may contribute to explain the manifestation of whole-plant heterotic phenotypes.
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- 2022
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29. Three types of genes underlying the Gametophyte factor1 locus cause unilateral cross incompatibility in maize
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Yuebin Wang, Wenqiang Li, Luxi Wang, Jiali Yan, Gang Lu, Ning Yang, Jieting Xu, Yuqing Wang, Songtao Gui, Gengshen Chen, Shuyan Li, Chengxiu Wu, Tingting Guo, Yingjie Xiao, Marilyn L. Warburton, Alisdair R. Fernie, Thomas Dresselhaus, and Jianbing Yan
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Science - Abstract
Unilateral cross incompatibility (UCI) is a type of prezygotic reproductive isolation, which is associated with multiple loci in maize. Here, the authors use genetic analysis to separate the Ga1 locus into two functional components and identify seven linked genes encoding three types of proteins.
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- 2022
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30. A pan-Zea genome map for enhancing maize improvement
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Songtao Gui, Wenjie Wei, Chenglin Jiang, Jingyun Luo, Lu Chen, Shenshen Wu, Wenqiang Li, Yuebin Wang, Shuyan Li, Ning Yang, Qing Li, Alisdair R. Fernie, and Jianbing Yan
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Pan-Zea genome ,Gene presence/absence variation (gPAV) ,Structural variation (SV) ,Narrow-sense heritability ,GWAS ,Biology (General) ,QH301-705.5 ,Genetics ,QH426-470 - Abstract
Abstract Background Maize (Zea mays L.) is at the vanguard facing the upcoming breeding challenges. However, both a super pan-genome for the Zea genus and a comprehensive genetic variation map for maize breeding are still lacking. Results Here, we construct an approximately 6.71-Gb pan-Zea genome that contains around 4.57-Gb non-B73 reference sequences from fragmented de novo assemblies of 721 pan-Zea individuals. We annotate a total of 58,944 pan-Zea genes and find around 44.34% of them are dispensable in the pan-Zea population. Moreover, 255,821 common structural variations are identified and genotyped in a maize association mapping panel. Further analyses reveal gene presence/absence variants and their potential roles during domestication of maize. Combining genetic analyses with multi-omics data, we demonstrate how structural variants are associated with complex agronomic traits. Conclusions Our results highlight the underexplored role of the pan-Zea genome and structural variations to further understand domestication of maize and explore their potential utilization in crop improvement.
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- 2022
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31. The diversity of quinoa morphological traits and seed metabolic composition
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Iman Tabatabaei, Saleh Alseekh, Mohammad Shahid, Ewa Leniak, Mateusz Wagner, Henda Mahmoudi, Sumitha Thushar, Alisdair R. Fernie, Kevin M. Murphy, Sandra M. Schmöckel, Mark Tester, Bernd Mueller-Roeber, Aleksandra Skirycz, and Salma Balazadeh
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Science - Abstract
Measurement(s) metabolites Technology Type(s) LC-MS mass spectrometry
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- 2022
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32. Azacytidine arrests ripening in cultivated strawberry (Fragaria × ananassa) by repressing key genes and altering hormone contents
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Félix Juan Martínez-Rivas, Rosario Blanco-Portales, Francisco Javier Molina-Hidalgo, José Luis Caballero, Leonardo Perez de Souza, Saleh Alseekh, Alisdair R. Fernie, Juan Muñoz-Blanco, and Antonio Rodríguez-Franco
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Azacytidine ,Demethylation ,Ripening ,m5-cytosine ,Abscisic acid ,Botany ,QK1-989 - Abstract
Abstract Background Strawberry ripening involves a number of irreversible biochemical reactions that cause sensory changes through accumulation of sugars, acids and other compounds responsible for fruit color and flavor. The process, which is strongly dependent on methylation marks in other fruits such as tomatoes and oranges, is highly controlled and coordinated in strawberry. Results Repeated injections of the hypomethylating compound 5-azacytidine (AZA) into green and unripe Fragaria × ananassa receptacles fully arrested the ripening of the fruit. The process, however, was reversible since treated fruit parts reached full maturity within a few days after AZA treatment was stopped. Transcriptomic analyses showed that key genes responsible for the biosynthesis of anthocyanins, phenylpropanoids, and hormones such as abscisic acid (ABA) were affected by the AZA treatment. In fact, AZA downregulated genes associated with ABA biosynthetic genes but upregulated genes associated with its degradation. AZA treatment additionally downregulated a number of essential transcription factors associated with the regulation and control of ripening. Metabolic analyses revealed a marked imbalance in hormone levels, with treated parts accumulating auxins, gibberellins and ABA degradation products, as well as metabolites associated with unripe fruits. Conclusions AZA completely halted strawberry ripening by altering the hormone balance, and the expression of genes involves in hormone biosynthesis and degradation processes. These results contradict those previously obtained in other climacteric and fleshly fruits, where AZA led to premature ripening. In any case, our results suggests that the strawberry ripening process is governed by methylation marks.
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- 2022
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33. Metabolomics-based profiling for quality assessment and revealing the impact of drying of Turmeric (Curcuma longa L.)
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Mohamed A. Salem, Riham A. El-Shiekh, Alisdair R. Fernie, Saleh Alseekh, and Ahmed Zayed
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Medicine ,Science - Abstract
Abstract Turmeric, the rhizomes of Curcuma longa L., is one of the top selling spices, food preservatives, and food colorants. In addition, it exhibits health promoting benefits owing to its unique phytochemical composition. Nevertheless, it is commonly subjected to heat drying, hence, the dried powder is the most used form and can easily be adulterated with allied species. Therefore, our research aimed to profile the phytochemical composition and investigate the impact of drying of turmeric. Extraction and fractionation followed by LC- and GC–MS analysis resulted in the identification of a total of 161 metabolites belonged to various phytochemical classes. Moreover, multivariate data analysis identified curcuminoids, terpecurcumins, and organic acids as potential markers for drying. Based on the applied analytical techniques in combination with chemometrics, these investigations have succeeded to provide good coverage of the metabolome of turmeric in both fresh and dried forms.
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- 2022
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34. The metabolic changes that effect fruit quality during tomato fruit ripening
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Feng Zhu, Weiwei Wen, Yunjiang Cheng, and Alisdair R. Fernie
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Tomato ,Ripening ,Metabolites regulation ,Fruit quality ,Plant culture ,SB1-1110 ,Botany ,QK1-989 - Abstract
Abstract As the most valuable organ of tomato plants, fruit has attracted considerable attention which most focus on its quality formation during the ripening process. A considerable amount of research has reported that fruit quality is affected by metabolic shifts which are under the coordinated regulation of both structural genes and transcriptional regulators. In recent years, with the development of the next generation sequencing, molecular and genetic analysis methods, lots of genes which are involved in the chlorophyll, carotenoid, cell wall, central and secondary metabolism have been identified and confirmed to regulate pigment contents, fruit softening and other aspects of fruit flavor quality. Here, both research concerning the dissection of fruit quality related metabolic changes, the transcriptional and post-translational regulation of these metabolic pathways are reviewed. Furthermore, a weighted gene correlation network analysis of representative genes of fruit quality has been carried out and the potential of the combined application of the gene correlation network analysis, fine-mapping strategies and next generation sequencing to identify novel candidate genes determinants of fruit quality is discussed.
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- 2022
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35. The AtMYB60 transcription factor regulates stomatal opening by modulating oxylipin synthesis in guard cells
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Fabio Simeoni, Aleksandra Skirycz, Laura Simoni, Giulia Castorina, Leonardo Perez de Souza, Alisdair R. Fernie, Saleh Alseekh, Patrick Giavalisco, Lucio Conti, Chiara Tonelli, and Massimo Galbiati
- Subjects
Medicine ,Science - Abstract
Abstract Stomata are epidermal pores formed by pairs of specialized guard cells, which regulate gas exchanges between the plant and the atmosphere. Modulation of transcription has emerged as an important level of regulation of stomatal activity. The AtMYB60 transcription factor was previously identified as a positive regulator of stomatal opening, although the details of its function remain unknown. Here, we propose a role for AtMYB60 as a negative modulator of oxylipins synthesis in stomata. The atmyb60-1 mutant shows reduced stomatal opening and accumulates increased levels of 12-oxo-phytodienoic acid (12-OPDA), jasmonic acid (JA) and jasmonoyl-l-isoleucine (JA-Ile) in guard cells. We provide evidence that 12-OPDA triggers stomatal closure independently of JA and cooperatively with abscisic acid (ABA) in atmyb60-1. Our study highlights the relevance of oxylipins metabolism in stomatal regulation and indicates AtMYB60 as transcriptional integrator of ABA and oxylipins responses in guard cells.
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- 2022
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36. Strategies for structure elucidation of small molecules based on LC–MS/MS data from complex biological samples
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Zhitao Tian, Fangzhou Liu, Dongqin Li, Alisdair R. Fernie, and Wei Chen
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LC–MS/MS ,Structure elucidation ,Complex biological samples ,Biotechnology ,TP248.13-248.65 - Abstract
LC–MS/MS is a major analytical platform for metabolomics, which has become a recent hotspot in the research fields of life and environmental sciences. By contrast, structure elucidation of small molecules based on LC–MS/MS data remains a major challenge in the chemical and biological interpretation of untargeted metabolomics datasets. In recent years, several strategies for structure elucidation using LC–MS/MS data from complex biological samples have been proposed, these strategies can be simply categorized into two types, one based on structure annotation of mass spectra and for the other on retention time prediction. These strategies have helped many scientists conduct research in metabolite-related fields and are indispensable for the development of future tools. Here, we summarized the characteristics of the current tools and strategies for structure elucidation of small molecules based on LC–MS/MS data, and further discussed the directions and perspectives to improve the power of the tools or strategies for structure elucidation.
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- 2022
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37. MeGLYI-13, a Glyoxalase I Gene in Cassava, Enhances the Tolerance of Yeast and Arabidopsis to Zinc and Copper Stresses
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Ruimei Li, Fenlian Tang, Yannian Che, Alisdair R. Fernie, Qin Zhou, Zhongping Ding, Yuan Yao, Jiao Liu, Yajie Wang, Xinwen Hu, and Jianchun Guo
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glyoxalase ,cassava ,heavy metal ,zinc ,copper ,overexpression ,Botany ,QK1-989 - Abstract
Although zinc and copper are the two essential nutrients necessary for plant growth, their excessive accumulation in soil not only causes environmental pollution but also seriously threatens human health and inhibits plant growth. The breeding of plants with novel zinc or copper toxicity tolerance capacities represents one strategy to address this problem. Glyoxalase I (GLYI) family genes have previously been suggested to be involved in the resistance to a wide range of abiotic stresses, including those invoked by heavy metals. Here, a MeGLYI-13 gene cloned from a cassava SC8 cultivar was characterized with regard to its potential ability in resistance to zinc or copper stresses. Sequence alignment indicated that MeGLYI-13 exhibits sequence differences between genotypes. Transient expression analysis revealed the nuclear localization of MeGLYI-13. A nuclear localization signal (NLS) was found in its C-terminal region. There are 12 Zn2+ binding sites and 14 Cu2+ binding sites predicted by the MIB tool, of which six binding sites were shared by Zn2+ and Cu2+. The overexpression of MeGLYI-13 enhanced both the zinc and copper toxicity tolerances of transformed yeast cells and Arabidopsis seedlings. Taken together, our study shows the ability of the MeGLYI-13 gene to resist zinc and copper toxicity, which provides genetic resources for the future breeding of plants resistant to zinc and copper and potentially other heavy metals.
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- 2023
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38. High-quality reference genome sequences of two coconut cultivars provide insights into evolution of monocot chromosomes and differentiation of fiber content and plant height
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Shouchuang Wang, Yong Xiao, Zhi-Wei Zhou, Jiaqing Yuan, Hao Guo, Zhuang Yang, Jun Yang, Pengchuan Sun, Lisong Sun, Yuan Deng, Wen-Zhao Xie, Jia-Ming Song, Muhammad Tahir ul Qamar, Wei Xia, Rui Liu, Shufang Gong, Yong Wang, Fuyou Wang, Xianqing Liu, Alisdair R. Fernie, Xiyin Wang, Haikuo Fan, Ling-Ling Chen, and Jie Luo
- Subjects
Biology (General) ,QH301-705.5 ,Genetics ,QH426-470 - Abstract
Abstract Background Coconut is an important tropical oil and fruit crop whose evolutionary position renders it a fantastic species for the investigation of the evolution of monocot chromosomes and the subsequent differentiation of ancient plants. Results Here, we report the assembly and annotation of reference-grade genomes of Cn. tall and Cn. dwarf, whose genome sizes are 2.40 Gb and 2.39 Gb, respectively. The comparative analysis reveals that the two coconut subspecies diverge about 2–8 Mya while the conserved Arecaceae-specific whole-genome duplication (ω WGD) occurs approximately 47–53 Mya. It additionally allows us to reconstruct the ancestral karyotypes of the ten ancient monocot chromosomes and the evolutionary trajectories of the 16 modern coconut chromosomes. Fiber synthesis genes in Cn. tall, related to lignin and cellulose synthesis, are found at a higher copy number and expression level than dwarf coconuts. Integrated multi-omics analysis reveals that the difference in coconut plant height is the result of altered gibberellin metabolism, with both the GA20ox copy number and a single-nucleotide change in the promoter together leading to the difference in plant height between Cn. tall and Cn. dwarf. Conclusion We provide high-quality coconut genomes and reveal the genetic basis of trait differences between two coconuts through multi-omics analysis. We also reveal that the selection of plant height has been targeted for the same gene for millions of years, not only in natural selection of ancient plant as illustrated in coconut, but also for artificial selection in cultivated crops such as rice and maize.
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- 2021
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39. Multi-omics approaches explain the growth-promoting effect of the apocarotenoid growth regulator zaxinone in rice
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Jian You Wang, Saleh Alseekh, Tingting Xiao, Abdugaffor Ablazov, Leonardo Perez de Souza, Valentina Fiorilli, Marita Anggarani, Pei-Yu Lin, Cristina Votta, Mara Novero, Muhammad Jamil, Luisa Lanfranco, Yue-Ie C. Hsing, Ikram Blilou, Alisdair R. Fernie, and Salim Al-Babili
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Biology (General) ,QH301-705.5 - Abstract
Wang et al. report zaxinone as a global regulator of the transcriptome and metabolome, as well as of hormonal and cellular composition of rice roots. This study shows that zaxinone promotes rice growth by enhancing root sugar uptake and metabolism and modulation of cytokinin content, indicating the potential application of this compound in increasing rice performance.
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- 2021
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40. Genomic basis underlying the metabolome-mediated drought adaptation of maize
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Fei Zhang, Jinfeng Wu, Nir Sade, Si Wu, Aiman Egbaria, Alisdair R. Fernie, Jianbing Yan, Feng Qin, Wei Chen, Yariv Brotman, and Mingqiu Dai
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Maize ,Drought tolerance ,Metabolome ,Natural variation ,Stress responses ,Biology (General) ,QH301-705.5 ,Genetics ,QH426-470 - Abstract
Abstract Background Drought is a major environmental disaster that causes crop yield loss worldwide. Metabolites are involved in various environmental stress responses of plants. However, the genetic control of metabolomes underlying crop environmental stress adaptation remains elusive. Results Here, we perform non-targeted metabolic profiling of leaves for 385 maize natural inbred lines grown under well-watered as well as drought-stressed conditions. A total of 3890 metabolites are identified and 1035 of these are differentially produced between well-watered and drought-stressed conditions, representing effective indicators of maize drought response and tolerance. Genetic dissections reveal the associations between these metabolites and thousands of single-nucleotide polymorphisms (SNPs), which represented 3415 metabolite quantitative trait loci (mQTLs) and 2589 candidate genes. 78.6% of mQTLs (2684/3415) are novel drought-responsive QTLs. The regulatory variants that control the expression of the candidate genes are revealed by expression QTL (eQTL) analysis of the transcriptomes of leaves from 197 maize natural inbred lines. Integrated metabolic and transcriptomic assays identify dozens of environment-specific hub genes and their gene-metabolite regulatory networks. Comprehensive genetic and molecular studies reveal the roles and mechanisms of two hub genes, Bx12 and ZmGLK44, in regulating maize metabolite biosynthesis and drought tolerance. Conclusion Our studies reveal the first population-level metabolomes in crop drought response and uncover the natural variations and genetic control of these metabolomes underlying crop drought adaptation, demonstrating that multi-omics is a powerful strategy to dissect the genetic mechanisms of crop complex traits.
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- 2021
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41. Temperature-mediated flower size plasticity in Arabidopsis
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Andrew Wiszniewski, Estefanía Uberegui, Michaela Messer, Gulmairam Sultanova, Monica Borghi, Gustavo Turqueto Duarte, Rubén Vicente, Katelyn Sageman-Furnas, Alisdair R. Fernie, Zoran Nikoloski, and Roosa A.E. Laitinen
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Plant biology ,Plant morphology ,Plant genetics ,Plant physiology ,Science - Abstract
Summary: Organisms can rapidly mitigate the effects of environmental changes by changing their phenotypes, known as phenotypic plasticity. Yet, little is known about the temperature-mediated plasticity of traits that are directly linked to plant fitness such as flower size. We discovered substantial genetic variation in flower size plasticity to temperature both among selfing Arabidopsis thaliana and outcrossing A. arenosa individuals collected from a natural growth habitat. Genetic analysis using a panel of 290 A. thaliana accession and mutant lines revealed that MADS AFFECTING FLOWERING (MAF) 2–5 gene cluster, previously shown to regulate temperature-mediated flowering time, was associated to the flower size plasticity to temperature. Furthermore, our findings pointed that the control of plasticity differs from control of the trait itself. Altogether, our study advances the understanding of genetic and molecular factors underlying plasticity on fundamental fitness traits, such as flower size, in response to future climate scenarios.
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- 2022
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42. Spatially resolved metabolomics reveals variety-specific metabolic changes in banana pulp during postharvest senescence
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Zhibin Yin, Tao Dong, Wenjie Huang, Mingyi Du, Dong Chen, Alisdair R. Fernie, Ganjun Yi, and Shijuan Yan
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Banana ,Mass spectrometry imaging ,Metabolomics ,Monoamine ,Amino acid ,Soluble sugars ,Nutrition. Foods and food supply ,TX341-641 ,Food processing and manufacture ,TP368-456 - Abstract
Banana is one of most popular fruits globally due to health-promoting and disease-preventing effects, yet little is known about in situ metabolic changes across banana varieties. Here, we integrated gold nanoparticle (AuNP)-assisted laser desorption/ionization mass spectrometry imaging (LDI-MSI) and metabolomics to investigate the spatiotemporal distribution and levels of metabolites within Brazil and Dongguan banana pulps during postharvest senescence. Metabolomics results indicated that both postripening stages and banana varieties contribute to metabolite levels. Benefiting from improved ionization efficiency of small-molecule metabolites and less peak interference, we visualized the spatiotemporal distribution of sugars, amino acids (AAs) and monoamines within pulps using AuNP-assisted LDI-MSI for the first time, revealing that AAs and monoamines exclusively accumulated in the middle region near the seed zone. Monosaccharides and di/trisaccharides were generally distributed across entire pulps but exhibited different accumulation patterns. These findings provide a guide for breeding new varieties and improving extraction efficiency of bioactive compounds.
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- 2022
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43. OsGF14b modulates defense signaling pathways in rice panicle blast response
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Shijuan Yan, Qing Liu, Thomas Naake, Wenjie Huang, Mengyu Chen, Qian Kong, Sheng Zhang, Wenyan Li, Xuan Li, Qinjian Liu, Jianyuan Yang, Alisdair R. Fernie, and Bin Liu
- Subjects
Rice ,Panicle blast resistance ,OsGF14b ,Gibberellin ,Phytohormone ,Agriculture ,Agriculture (General) ,S1-972 - Abstract
Rice with panicle-blast resistance is needed for stable rice production. Although we have previously demonstrated that OsGF14b underlies a quantitative trait locus that positively regulates rice panicle blast resistance, the mechanism is still unknown. In this study, a multi-omics approach was used to investigate the possible downstream signaling pathway regulated by OsGF14b. OsGF14b both strongly activated the gibberellin biosynthetic pathway during pathogen infection and reprogrammed the lignin biosynthetic pathway. Reduced lignin accumulation was observed in glumes of OsGF14b-overexpressing plants in comparison with the wild type after pathogen inoculation. OsGF14b activated the auxin and jasmonic acid signaling pathways, but inactivated the salicylic acid signaling pathway. Auxin and jasmonic acid appeared to act independently on OsGF14b-mediated panicle blast resistance. The roles of gibberellin, lignin, and auxin were different from their roles in leaf blast, suggesting that different mechanisms underlie leaf and panicle blast resistance in rice. This study provides a comprehensive catalog of molecular changes that could be targets for future studies of rice panicle blast resistance.
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- 2021
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44. Integrated Characterization of Cassava (Manihot esculenta) Pectin Methylesterase (MePME) Genes to Filter Candidate Gene Responses to Multiple Abiotic Stresses
- Author
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Shijia Wang, Ruimei Li, Yangjiao Zhou, Alisdair R. Fernie, Zhongping Ding, Qin Zhou, Yannian Che, Yuan Yao, Jiao Liu, Yajie Wang, Xinwen Hu, and Jianchun Guo
- Subjects
pectin methylesterase ,cassava ,postharvest physiological deterioration ,gene family ,abiotic stress ,Botany ,QK1-989 - Abstract
Plant pectin methylesterases (PMEs) play crucial roles in regulating cell wall modification and response to various stresses. Members of the PME family have been found in several crops, but there is a lack of research into their presence in cassava (Manihot esculent), which is an important crop for world food security. In this research, 89 MePME genes were identified in cassava that were separated into two types (type-Ⅰ and type-Ⅱ) according to the existence or absence of a pro-region (PMEI domain). The MePME gene members were unevenly located on 17 chromosomes, with 19 gene pairs being identified that most likely arose via duplication events. The MePMEs could be divided into ten sub-groups in type-Ⅰ and five sub-groups in type-Ⅱ. The motif analysis revealed 11 conserved motifs in type-Ⅰ and 8 in type-Ⅱ MePMEs. The number of introns in the CDS region of type-Ⅰ MePMEs ranged between one and two, and the number of introns in type-Ⅱ MePMEs ranged between one and nine. There were 21 type-Ⅰ and 31 type-Ⅱ MePMEs that contained signal peptides. Most of the type-Ⅰ MePMEs had two conserved “RK/RLL” and one “FPSWVS” domain between the pro-region and the PME domain. Multiple stress-, hormone- and tissue-specific-related cis-acting regulatory elements were identified in the promoter regions of MePME genes. A total of five co-expressed genes (MePME1, MePME2, MePME27, MePME65 and MePME82) were filtered from different abiotic stresses via the use of UpSet Venn diagrams. The gene expression pattern analysis revealed that the expression of MePME1 was positively correlated with the degree of cassava postharvest physiological deterioration (PPD). The expression of this gene was also significantly upregulated by 7% PEG and 14 °C low-temperature stress, but slightly downregulated by ABA treatment. The tissue-specific expression analysis revealed that MePME1 and MePME65 generally displayed higher expression levels in most tissues than the other co-expressed genes. In this study, we obtain an in-depth understanding of the cassava PME gene family, suggesting that MePME1 could be a candidate gene associated with multiple abiotic tolerance.
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- 2023
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45. Anti-Obesity Effect of a Tea Mixture Nano-Formulation on Rats Occurs via the Upregulation of AMP-Activated Protein Kinase/Sirtuin-1/Glucose Transporter Type 4 and Peroxisome Proliferator-Activated Receptor Gamma Pathways
- Author
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Mohamed A. Salem, Nora M. Aborehab, Mai M. Abdelhafez, Sameh H. Ismail, Nadine W. Maurice, May A. Azzam, Saleh Alseekh, Alisdair R. Fernie, Maha M. Salama, and Shahira M. Ezzat
- Subjects
obesity ,metabolomics ,green tea ,white tea ,oolong tea ,catechins ,Microbiology ,QR1-502 - Abstract
White, green, and oolong teas are produced from the tea plant (Camellia sinensis (L.) Kuntze) and are reported to have anti-obesity and hypolipidemic effects. The current study aims to investigate the anti-obesity effects of a tea mixture nano-formulation by targeting the AMPK/Sirt-1/GLUT-4 axis in rats. In vitro lipase and α-amylase inhibition assays were used to determine the active sample, which was then incorporated into a nanoparticle formulation subjected to in vivo anti-obesity testing in rats by measuring the expression level of different genes implicated in adipogenesis and inflammation using qRT-PCR. Moreover, metabolomic analysis was performed for each tea extract using LC/ESI MS/MS coupled to chemometrics in an attempt to find a correlation between the constituents of the extracts and their biological activity. The in vitro pancreatic lipase and α-amylase inhibition assays demonstrated more effective activity in the tea mixture than the standards, orlistat and acarbose, respectively, and each tea alone. Thus, the herbal tea mixture and its nanoparticle formulation were evaluated for their in vivo anti-obesity activity. Intriguingly, the tea mixture significantly decreased the serum levels of glucose and triglycerides and increased the mRNA expression of GLUT-4, P-AMPK, Sirt-1, and PPAR-γ, which induce lipolysis while also decreasing the mRNA expression of TNF-α and ADD1/SREBP-1c, thereby inhibiting the inflammation associated with obesity. Our study suggests that the tea mixture nano-formulation is a promising therapeutic agent in the treatment of obesity and may also be beneficial in other metabolic disorders by targeting the AMPK/Sirt-1/Glut-4 pathway.
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- 2023
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46. Global mapping of protein–metabolite interactions in Saccharomyces cerevisiae reveals that Ser-Leu dipeptide regulates phosphoglycerate kinase activity
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Marcin Luzarowski, Rubén Vicente, Andrei Kiselev, Mateusz Wagner, Dennis Schlossarek, Alexander Erban, Leonardo Perez de Souza, Dorothee Childs, Izabela Wojciechowska, Urszula Luzarowska, Michał Górka, Ewelina M. Sokołowska, Monika Kosmacz, Juan C. Moreno, Aleksandra Brzezińska, Bhavana Vegesna, Joachim Kopka, Alisdair R. Fernie, Lothar Willmitzer, Jennifer C. Ewald, and Aleksandra Skirycz
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Biology (General) ,QH301-705.5 - Abstract
Marcin Luzarowski et al. use a new method PROMIS for systematic analysis of protein small molecule interactions in yeast. Their work provides an experimentally-derived catalogue of protein–metabolite complexes in S. cerevisiae and reveals that the Ser-Leu dipeptide activates Pgk1, and alters cellular metabolism.
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- 2021
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47. Ancestral sequence reconstruction - An underused approach to understand the evolution of gene function in plants?
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Federico Scossa and Alisdair R. Fernie
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Ancestral sequence reconstruction ,Evolution ,Genomics ,Metabolism ,Phylogenetics ,Plants ,Biotechnology ,TP248.13-248.65 - Abstract
Whilst substantial research effort has been placed on understanding the interactions of plant proteins with their molecular partners, relatively few studies in plants - by contrast to work in other organisms - address how these interactions evolve. It is thought that ancestral proteins were more promiscuous than modern proteins and that specificity often evolved following gene duplication and subsequent functional refining. However, ancestral protein resurrection studies have found that some modern proteins have evolved de novo from ancestors lacking those functions. Intriguingly, the new interactions evolved as a consequence of just a few mutations and, as such, acquisition of new functions appears to be neither difficult nor rare, however, only a few of them are incorporated into biological processes before they are lost to subsequent mutations. Here, we detail the approach of ancestral sequence reconstruction (ASR), providing a primer to reconstruct the sequence of an ancestral gene. We will present case studies from a range of different eukaryotes before discussing the few instances where ancestral reconstructions have been used in plants. As ASR is used to dig into the remote evolutionary past, we will also present some alternative genetic approaches to investigate molecular evolution on shorter timescales. We argue that the study of plant secondary metabolism is particularly well suited for ancestral reconstruction studies. Indeed, its ancient evolutionary roots and highly diverse landscape provide an ideal context in which to address the focal issue around the emergence of evolutionary novelties and how this affects the chemical diversification of plant metabolism.
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- 2021
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48. Systems biology reveals key tissue-specific metabolic and transcriptional signatures involved in the response of Medicago truncatula plant genotypes to salt stress
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Panagiota Filippou, Xavier Zarza, Chrystalla Antoniou, Toshihiro Obata, Carlos A. Villarroel, Ioannis Ganopoulos, Vaggelis Harokopos, Gholamreza Gohari, Vassilis Aidinis, Panagiotis Madesis, Anastasis Christou, Alisdair R. Fernie, Antonio F. Tiburcio, and Vasileios Fotopoulos
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Salinity ,Metabolomics ,Transcriptomics ,Raffinose pathway ,Biotechnology ,TP248.13-248.65 - Abstract
Salt stress is an important factor limiting plant productivity by affecting plant physiology and metabolism. To explore salt tolerance adaptive mechanisms in the model legume Medicago truncatula, we used three genotypes with differential salt-sensitivity: TN6.18 (highly sensitive), Jemalong A17 (moderately sensitive), and TN1.11 (tolerant). Cellular damage was monitored in roots and leaves 48 h after 200 mM NaCl treatment by measuring lipid peroxidation, nitric oxide, and hydrogen peroxide contents, further supported by leaf stomatal conductance and chlorophyll readings. The salt-tolerant genotype TN1.11 displayed the lowest level of oxidative damage, in contrast to the salt sensitive TN6.18, which showed the highest responses. Metabolite profiling was employed to explore the differential genotype-related responses to stress at the molecular level. The metabolic data in the salt tolerant TN1.11 roots revealed an accumulation of metabolites related to the raffinose pathway. To further investigate the sensitivity to salinity, global transcriptomic profiling using microarray analysis was carried out on the salt-stressed sensitive genotypes. In TN6.18, the transcriptomic analysis identified a lower expression of many genes related to stress signalling, not previously linked to salinity, and corresponding to the TIR-NBS-LRR gene class. Overall, this global approach contributes to gaining significant new insights into the complexity of stress adaptive mechanisms and to the identification of potential targets for crop improvement.
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- 2021
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49. Role of Raf-like kinases in SnRK2 activation and osmotic stress response in plants
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Norma Fàbregas, Takuya Yoshida, and Alisdair R. Fernie
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Science - Abstract
A better understanding of how plants respond to osmotic stress could potentially help improve crop yields. Here Fàbregas et al. review the recent characterization of Raf-like kinases that act in both in ABA-dependent and -independent responses to osmotic stress.
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- 2020
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50. A moonlighting role for enzymes of glycolysis in the co-localization of mitochondria and chloroplasts
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Youjun Zhang, Arun Sampathkumar, Sandra Mae-Lin Kerber, Corné Swart, Carsten Hille, Kumar Seerangan, Alexander Graf, Lee Sweetlove, and Alisdair R. Fernie
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Science - Abstract
Protein-protein interactions are thought to channel substrates between consecutive enzymes during glycolysis. Here the authors show that Arabidopsis phosphoglycerate mutase and enolase can form a substrate-channelling metabolon and also play a moonlighting role in promoting colocalization of chloroplasts and mitochondria.
- Published
- 2020
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