Plant MethodsPub Date : 2024-08-09DOI: 10.1186/s13007-024-01246-8
Xinyue Fan, Hongmei Sun
{"title":"Exploring Agrobacterium-mediated genetic transformation methods and its applications in Lilium.","authors":"Xinyue Fan, Hongmei Sun","doi":"10.1186/s13007-024-01246-8","DOIUrl":"10.1186/s13007-024-01246-8","url":null,"abstract":"<p><p>As a typical bulb flower, lily is widely cultivated worldwide because of its high ornamental, medicinal and edible value. Although breeding efforts evolved over the last 10000 years, there are still many problems in the face of increasing consumer demand. The approach of biotechnological methods would help to solve this problem and incorporate traits impossible by conventional breeding. Target traits are dormancy, development, color, floral fragrance and resistances against various biotic and abiotic stresses, so as to improve the quality of bulbs and cut flowers in planting, cultivation, postharvest, plant protection and marketing. Genetic transformation technology is an important method for varietal improvement and has become the foundation and core of plant functional genomics research, greatly assisting various plant improvement programs. However, achieving stable and efficient genetic transformation of lily has been difficult worldwide. Many gene function verification studies depend on the use of model plants, which greatly limits the pace of directed breeding and germplasm improvement in lily. Although significant progress has been made in the development and optimization of genetic transformation systems, shortcomings remain. Agrobacterium-mediated genetic transformation has been widely used in lily. However, severe genotypic dependence is the main bottleneck limiting the genetic transformation of lily. This review will summarizes the research progress in the genetic transformation of lily over the past 30 years to generate the material including a section how genome engineering using stable genetic transformation system, and give an overview about recent and future applications of lily transformation. The information provided in this paper includes ideas for optimizing and improving the efficiency of existing genetic transformation methods and for innovation, provides technical support for mining and identifying regulatory genes for key traits, and lays a foundation for genetic improvement and innovative germplasm development in lily.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"120"},"PeriodicalIF":4.7,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11313100/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141913683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-08-06DOI: 10.1186/s13007-024-01254-8
Vinni Thekkudan Novi, Hamada A Aboubakr, Melanie J Moore, Akli Zarouri, Jennifer Juzwik, Abdennour Abbas
{"title":"A rapid LAMP assay for the diagnosis of oak wilt with the naked eye.","authors":"Vinni Thekkudan Novi, Hamada A Aboubakr, Melanie J Moore, Akli Zarouri, Jennifer Juzwik, Abdennour Abbas","doi":"10.1186/s13007-024-01254-8","DOIUrl":"10.1186/s13007-024-01254-8","url":null,"abstract":"<p><strong>Background: </strong>Oak wilt disease, caused by Bretziella fagacearum is a significant threat to oak (Quercus spp.) tree health in the United States and Eastern Canada. The disease may cause dramatic damage to natural and urban ecosystems without management. Early and accurate diagnosis followed by timely treatment increases the level of disease control success.</p><p><strong>Results: </strong>A rapid assay based on loop mediated isothermal amplification (LAMP) was first developed with fluorescence detection of B. fagacearum after 30-minute reaction time. Six different primers were designed to specifically bind and amplify the pathogen's DNA. To simplify the use of this assay in the field, gold nanoparticles (AuNPs) were designed to bind to the DNA amplicon obtained from the LAMP reaction. Upon inducing precipitation, the AuNP-amplicons settle as a red pellet visible to the naked eye, indicative of pathogen presence. Both infected and healthy red oak samples were tested using this visualization method. The assay was found to have high diagnostic sensitivity and specificity for the B. fagacearum isolate studied. Moreover, the developed assay was able to detect the pathogen in crude DNA extracts of diseased oak wood samples, which further reduced the time required to process samples.</p><p><strong>Conclusions: </strong>In summary, the LAMP assay coupled with oligonucleotide-conjugated gold nanoparticle visualization is a promising method for accurate and rapid molecular-based diagnosis of B. fagacearum in field settings. The new method can be adapted to other forest and plant diseases by simply designing new primers.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"119"},"PeriodicalIF":4.7,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11302832/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141894095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-08-02DOI: 10.1186/s13007-024-01247-7
Valentin Michels, Chunwei Chou, Maximilian Weigand, Yuxin Wu, Andreas Kemna
{"title":"Quantitative phenotyping of crop roots with spectral electrical impedance tomography: a rhizotron study with optimized measurement design.","authors":"Valentin Michels, Chunwei Chou, Maximilian Weigand, Yuxin Wu, Andreas Kemna","doi":"10.1186/s13007-024-01247-7","DOIUrl":"10.1186/s13007-024-01247-7","url":null,"abstract":"<p><strong>Background: </strong>Root systems are key contributors to plant health, resilience, and, ultimately, yield of agricultural crops. To optimize plant performance, phenotyping trials are conducted to breed plants with diverse root traits. However, traditional analysis methods are often labour-intensive and invasive to the root system, therefore limiting high-throughput phenotyping. Spectral electrical impedance tomography (sEIT) could help as a non-invasive and cost-efficient alternative to optical root analysis, potentially providing 2D or 3D spatio-temporal information on root development and activity. Although impedance measurements have been shown to be sensitive to root biomass, nutrient status, and diurnal activity, only few attempts have been made to employ tomographic algorithms to recover spatially resolved information on root systems. In this study, we aim to establish relationships between tomographic electrical polarization signatures and root traits of different fine root systems (maize, pinto bean, black bean, and soy bean) under hydroponic conditions.</p><p><strong>Results: </strong>Our results show that, with the use of an optimized data acquisition scheme, sEIT is capable of providing spatially resolved information on root biomass and root surface area for all investigated root systems. We found strong correlations between the total polarization strength and the root biomass ( <math> <mrow><msup><mi>R</mi> <mn>2</mn></msup> <mo>=</mo> <mn>0.82</mn></mrow> </math> ) and root surface area ( <math> <mrow><msup><mi>R</mi> <mn>2</mn></msup> <mo>=</mo> <mn>0.8</mn></mrow> </math> ). Our findings suggest that the captured polarization signature is dominated by cell-scale polarization processes. Additionally, we demonstrate that the resolution characteristics of the measurement scheme can have a significant impact on the tomographic reconstruction of root traits.</p><p><strong>Conclusion: </strong>Our findings showcase that sEIT is a promising tool for the tomographic reconstruction of root traits in high-throughput root phenotyping trials and should be evaluated as a substitute for traditional, often time-consuming, root characterization methods.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"118"},"PeriodicalIF":4.7,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11297745/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141879236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-08-02DOI: 10.1186/s13007-024-01249-5
Dušan Veličković, Tanya Winkler, Vimal Balasubramanian, Thomas Wietsma, Christopher R Anderton, Amir H Ahkami, Kevin Zemaitis
{"title":"RhizoMAP: a comprehensive, nondestructive, and sensitive platform for metabolic imaging of the rhizosphere.","authors":"Dušan Veličković, Tanya Winkler, Vimal Balasubramanian, Thomas Wietsma, Christopher R Anderton, Amir H Ahkami, Kevin Zemaitis","doi":"10.1186/s13007-024-01249-5","DOIUrl":"10.1186/s13007-024-01249-5","url":null,"abstract":"<p><strong>Background: </strong>Elucidating the intricate structural organization and spatial gradients of biomolecular composition within the rhizosphere is critical to understanding important biogeochemical processes, which include the mechanisms of root-microbe interactions for maintaining sustainable plant ecosystem services. While various analytical methods have been developed to assess the spatial heterogeneity within the rhizosphere, a comprehensive view of the fine distribution of metabolites within the root-soil interface has remained a significant challenge. This is primarily due to the difficulty of maintaining the original spatial organization during sample preparation without compromising its molecular content.</p><p><strong>Results: </strong>In this study, we present a novel approach, RhizoMAP, in which the rhizosphere molecules are imprinted on selected polymer membranes and then spatially profiled using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI). We enhanced the performance of RhizoMAP by combining the use of two thin (< 20 μm) membranes (polyester and polycarbonate) with distinct MALDI sample preparations. This optimization allowed us to gain insight into the distribution of over 500 different molecules within the rhizosphere of poplar (Populus trichocarpa) grown in rhizoboxes filled with mycorrhizae soil. These two membranes, coupled with three different sample preparation conditions, enabled us to capture the distribution of a wide variety of molecules that included phytohormones, amino acids, sugars, sugar glycosides, polycarboxylic acids components of the Krebs cycle, fatty acids, short aldehydes and ketones, terpenes, volatile organic compounds, fertilizers from the soil, and others. Their spatial distribution varies greatly, with some following root traces, others showing diffusion from roots, some associated with soil particles, and many having distinct hot spots along the plant root or surrounding soil. Moreover, we showed how RhizoMAP can be used to localize the origin of the molecules and molecular transformation during root growth. Finally, we demonstrated the power of RhizoMAP to capture molecular distributions of key metabolites throughout a 20 cm deep rhizosphere.</p><p><strong>Conclusions: </strong>RhizoMAP is a method that provides nondestructive, untargeted, broad, and sensitive metabolite imaging of root-associated molecules, exudates, and soil organic matter throughout the rhizosphere, as demonstrated in a lab-controlled native soil environment.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"117"},"PeriodicalIF":4.7,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11297713/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141879237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-07-31DOI: 10.1186/s13007-024-01245-9
Antonio Montagnoli, Andrew T Hudak, Pasi Raumonen, Bruno Lasserre, Mattia Terzaghi, Carlos A Silva, Benjamin C Bright, Lee A Vierling, Bruna N de Vasconcellos, Donato Chiatante, R Kasten Dumroese
{"title":"Correction: Terrestrial laser scanning and low magnetic field digitization yield similar architectural coarse root traits for 32-year-old Pinus ponderosa trees.","authors":"Antonio Montagnoli, Andrew T Hudak, Pasi Raumonen, Bruno Lasserre, Mattia Terzaghi, Carlos A Silva, Benjamin C Bright, Lee A Vierling, Bruna N de Vasconcellos, Donato Chiatante, R Kasten Dumroese","doi":"10.1186/s13007-024-01245-9","DOIUrl":"10.1186/s13007-024-01245-9","url":null,"abstract":"","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"116"},"PeriodicalIF":4.7,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11290004/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141860597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-07-29DOI: 10.1186/s13007-024-01230-2
Myrthe Praat, Zhang Jiang, Joe Earle, Sjef Smeekens, Martijn van Zanten
{"title":"Using a thermal gradient table to study plant temperature signalling and response across a temperature spectrum.","authors":"Myrthe Praat, Zhang Jiang, Joe Earle, Sjef Smeekens, Martijn van Zanten","doi":"10.1186/s13007-024-01230-2","DOIUrl":"10.1186/s13007-024-01230-2","url":null,"abstract":"<p><p>Plants must cope with ever-changing temperature conditions in their environment. In many plant species, suboptimal high and low temperatures can induce adaptive mechanisms that allow optimal performance. Thermomorphogenesis is the acclimation to high ambient temperature, whereas cold acclimation refers to the acquisition of cold tolerance following a period of low temperatures. The molecular mechanisms underlying thermomorphogenesis and cold acclimation are increasingly well understood but neither signalling components that have an apparent role in acclimation to both cold and warmth, nor factors determining dose-responsiveness, are currently well defined. This can be explained in part by practical limitations, as applying temperature gradients requires the use of multiple growth conditions simultaneously, usually unavailable in research laboratories. Here we demonstrate that commercially available thermal gradient tables can be used to grow and assess plants over a defined and adjustable steep temperature gradient within one experiment. We describe technical and thermodynamic aspects and provide considerations for plant growth and treatment. We show that plants display the expected morphological, physiological, developmental and molecular responses that are typically associated with high temperature and cold acclimation. This includes temperature dose-response effects on seed germination, hypocotyl elongation, leaf development, hyponasty, rosette growth, temperature marker gene expression, stomatal conductance, chlorophyll content, ion leakage and hydrogen peroxide levels. In conclusion, thermal gradient table systems enable standardized and predictable environments to study plant responses to varying temperature regimes and can be swiftly implemented in research on temperature signalling and response.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"114"},"PeriodicalIF":4.7,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11285400/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141793066","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A CNN model for early detection of pepper Phytophthora blight using multispectral imaging, integrating spectral and textural information.","authors":"Zhijuan Duan, Haoqian Li, Chenguang Li, Jun Zhang, Dongfang Zhang, Xiaofei Fan, Xueping Chen","doi":"10.1186/s13007-024-01239-7","DOIUrl":"10.1186/s13007-024-01239-7","url":null,"abstract":"<p><strong>Background: </strong>Pepper Phytophthora blight is a devastating disease during the growth process of peppers, significantly affecting their yield and quality. Accurate, rapid, and non-destructive early detection of pepper Phytophthora blight is of great importance for pepper production management. This study investigated the possibility of using multispectral imaging combined with machine learning to detect Phytophthora blight in peppers. Peppers were divided into two groups: one group was inoculated with Phytophthora blight, and the other was left untreated as a control. Multispectral images were collected at 0-h samples before inoculation and at 48, 60, 72, and 84 h after inoculation. The supporting software of the multispectral imaging system was used to extract spectral features from 19 wavelengths, and textural features were extracted using a gray-level co-occurrence matrix (GLCM) and a local binary pattern (LBP). The principal component analysis (PCA), successive projection algorithm (SPA), and genetic algorithm (GA) were used for feature selection from the extracted spectral and textural features. Two classification models were established based on effective single spectral features and significant spectral textural fusion features: a partial least squares discriminant analysis (PLS_DA) and one-dimensional convolutional neural network (1D-CNN). A two-dimensional convolutional neural network (2D-CNN) was constructed based on five principal component (PC) coefficients extracted from the spectral data using PCA, weighted, and summed with 19-channel multispectral images to create new PC images.</p><p><strong>Results: </strong>The results indicated that the models using PCA for feature selection exhibit relatively stable classification performance. The accuracy of PLS-DA and 1D-CNN based on single spectral features is 82.6% and 83.3%, respectively, at the 48h mark. In contrast, the accuracy of PLS-DA and 1D-CNN based on spectral texture fusion reached 85.9% and 91.3%, respectively, at the same 48h mark. The accuracy of the 2D-CNN based on 5 PC images is 82%.</p><p><strong>Conclusions: </strong>The research indicates that Phytophthora blight infection can be detected 48 h after inoculation (36 h before visible symptoms). This study provides an effective method for the early detection of Phytophthora blight in peppers.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"115"},"PeriodicalIF":4.7,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11288097/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141793065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-07-27DOI: 10.1186/s13007-024-01242-y
Xuehan Mei, Kaijie Zhu, Danni Yan, Huihui Jia, Wangyao Luo, Junli Ye, Xiuxin Deng
{"title":"Developing a simple and rapid method for cell-specific transcriptome analysis through laser microdissection: insights from citrus rind with broader implications","authors":"Xuehan Mei, Kaijie Zhu, Danni Yan, Huihui Jia, Wangyao Luo, Junli Ye, Xiuxin Deng","doi":"10.1186/s13007-024-01242-y","DOIUrl":"https://doi.org/10.1186/s13007-024-01242-y","url":null,"abstract":"With the rapid development of single-cell sequencing technology, histological studies are no longer limited to conventional homogenized tissues. Laser microdissection enables the accurate isolation of specific tissues or cells, and when combined with next-generation sequencing, it can reveal important biological processes at the cellular level. However, traditional laser microdissection techniques have often been complicated and time-consuming, and the quality of the RNA extracted from the collected samples has been inconsistent, limiting follow-up studies. Therefore, an improved, simple, and efficient laser microdissection method is urgently needed. We omitted the sample fixation and cryoprotectant addition steps. Instead, fresh samples were embedded in Optimal Cutting Temperature medium within 1.5 ml centrifuge tube caps, rapidly frozen with liquid nitrogen, and immediately subjected to cryosectioning. A series of section thicknesses of citrus rind were tested for RNA extraction, which showed that 18 μm thickness yielded the highest quality RNA. By shortening the dehydration time to one minute per ethanol gradient and omitting the tissue clearing step, the resulting efficient dehydration and preserved morphology ensured high-quality RNA extraction. We also propose a set of laser microdissection parameters by adjusting the laser power to optimal values, reducing the aperture size, and lowering the pulse frequency. Both the epidermal and subepidermal cells from the citrus rind were collected, and RNA extraction was completed within nine hours. Using this efficient method, the transcriptome sequencing of the isolated tissues generated high-quality data with average Q30 values and mapping rates exceeding 91%. Moreover, the transcriptome analysis revealed significant differences between the cell layers, further confirming the effectiveness of our isolation approach. We developed a simple and rapid laser microdissection method and demonstrated its effectiveness through a study based on citrus rind, from which we generated high-quality transcriptomic data. This fast and efficient method of cell isolation, combined with transcriptome sequencing not only contributes to precise histological studies at the cellular level in citrus but also provides a promising approach for cell-specific transcriptome analysis in a broader range of other plant tissues.","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"17 1","pages":""},"PeriodicalIF":5.1,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141778595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-07-27DOI: 10.1186/s13007-024-01237-9
Julia Compart, Ardha Apriyanto, Joerg Fettke
{"title":"Starch phosphorylation—A needle in a haystack","authors":"Julia Compart, Ardha Apriyanto, Joerg Fettke","doi":"10.1186/s13007-024-01237-9","DOIUrl":"https://doi.org/10.1186/s13007-024-01237-9","url":null,"abstract":"Phosphoesterification is the only naturally occurring covalent starch modification identified to date, and it has a major impact on overall starch metabolism. The incorporation of phosphate groups mediated by dikinases [α-glucan, water dikinase (GWD), EC 2.7.9.4; phosphoglucan, water dikinase (PWD), EC 2.7.9.5] massively alters the starch granule properties; however, previous studies did not determine whether the starch-related dikinases bind the phosphate to the glucosyl units within the amylopectin molecules in a specific pattern or randomly. In order to answer this challenging question, a number of approaches were initially pursued until a protocol could be established that enabled a massive step forward in the in vitro analysis of phosphorylated glucan chains obtained from starch. For this purpose, phosphorylation by GWD was investigated, including the final state of phosphorylation i.e., the state of substrate saturation when GWD lacks further free hydroxyl groups at OH-C6 for the catalysis of monophosphate esters. Since the separated phosphorylated glucan chains were required for the analysis, isoamylase digestion was performed to cleave the α-1,6-glycosidic bonds and to allow for the removal of the huge number of existing neutral chains by means of anion exchange chromatography. Via Matrix-Assisted Laser Desorption/Ionization–Time of Flight (MALDI-TOF) MS and MALDI-MS/MS, the phosphorylated α-glucan chains were analysed, and the position of the phosphate group within the chain in relation to the reducing end was determined. Here, we demonstrate a protocol that enables the analysis of phosphorylated oligosaccharides, even in small quantities.","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"62 1","pages":""},"PeriodicalIF":5.1,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141778596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plant MethodsPub Date : 2024-07-25DOI: 10.1186/s13007-024-01221-3
Zubair Ahmed, Muhammad Ikram, Ishaq Khan, Kashif Bashir, Abdul Jabbar Shah, Zahid Hussain, Taous Khan
{"title":"LC-ESI-MS/MS-based molecular networking, antioxidant, anti-glioma activity and molecular docking studies of Clematis graveolens.","authors":"Zubair Ahmed, Muhammad Ikram, Ishaq Khan, Kashif Bashir, Abdul Jabbar Shah, Zahid Hussain, Taous Khan","doi":"10.1186/s13007-024-01221-3","DOIUrl":"10.1186/s13007-024-01221-3","url":null,"abstract":"<p><p>Clematis graveolens Lindl., an indigenous climbing plant found in the Himalayan areas, is used by local communities for the treatment of neck tumors. The objective of this work is to examine the comprehensive metabolomic profile, antioxidant capability, in vitro and in silico anti-glioma effects on U-87 human glioma cell lines of the crude extract and fractions from C. graveolens. Liquid chromatography coupled with mass spectroscopy (LC-MS/MS) was used to establish detailed metabolite profiling of C. graveolens. The assessment of cell cytotoxicity was conducted using MTT cell viability assay on U-87 and BHK-21. Through molecular docking studies, the mode of inhibition and binding interaction between identified compounds and target proteins were also determined to evaluate the in vitro results. The use of LC-MS/MS-based global natural products social (GNPS) molecular networking analysis resulted in the identification of 27 compounds. The crude extract, ethyl acetate fraction, and chloroform fraction exhibited significant inhibitory activity against the U-87 cell lines, with IC<sub>50</sub> values of 112.0, 138.1, and 142.7 µg/mL, respectively. The ethyl acetate fraction exhibited significant inhibitory concentration for 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) activity and the metal chelation activity with IC<sub>50</sub> value of 39.50 µg/mL, 32.27 µg/mL, and 53.46 µg/mL, respectively. The crude extract showed maximum total phenolic, and total flavonoid concentration measuring 338.7 µg GAE/mg, and 177.04 µg QE/mg, respectively. The findings of this study indicate that C. graveolens consists of a diverse range of active phytoconstituents that possess antioxidant and anti-glioma properties.</p>","PeriodicalId":20100,"journal":{"name":"Plant Methods","volume":"20 1","pages":"111"},"PeriodicalIF":4.7,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11271027/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141760319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}