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Transient transformation is extremely useful for rapid in vivo assessment of gene function, especially for fruit-related genes. Grape berry, while an important fruit crop, is recalcitrant to transient transformation, due to the high turgor pressure in its mesocarp cells that limits the ability of Agrobacterium to penetrate into the tissue. It is urgent to establish a simple transient transformation system for rapid analysis of gene function. In this study, different injection methods, grape genotypes, and developmental stages were tested in order to develop a rapid and efficient Agrobacterium-mediated transient transformation methodology for grape berries. Two injection methods, namely punch injection and direct injection, were evaluated using the β-glucuronidase (GUS) gene and by x-gluc tissue staining and 4-methylumbelliferyl-β-D-glucuronide fluorescence analysis. The results indicated that there were no significant differences on transformation effects between the two methods, but the latter was more suitable because of its simplicity and convenience. Six grape cultivars ('Hanxiangmi', 'Moldova', 'Zijixin', 'Jumeigui', 'Shine-Muscat', and 'A17') were tested for transient transformation. 'Hanxiangmi', 'Moldova', and 'Zijixin' grape berries were not suitable for agroinfiltration due to frequently fruit cracking, browning, and formation of scar skin. The fruit integrity rates of 'Jumeigui', 'Shine-Muscat', and 'A17' berries were all above 80%, and GUS activity was detected in the berries of the three cultivars 3-14 days after injection with the Agrobacterium culture, while higher GUS activities were observed in the 'Jumeigui' berries. The levels of GUS activity in injected berries at 7-8 weeks after full blooming (WAFB) were more than twice at 6 WAFB. In subsequent assays, the over-expression of MYB transcription factor VvMYB44 via transient transformation accelerated the anthocyanin accumulation and fruit coloring through raising the expression levels of VvLAR1, VvUFGT, VvLDOX, VvANS, and VvDFR, which verified the effectiveness of this transformation system. These experiments finally identified the reliable grape cultivars and suitable operational approach for transient transformation and further indicated that this Agrobacterium-mediated transient transformation system was efficient and suitable for the elucidation of gene function in grape berries.
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NanoImpact
December 2024
In Vitro Toxicology Group, Faculty of Medicine, Health and Life Sciences, Institute of Life Sciences, Swansea University Medical School, Singleton Park, Swansea SA2 8PP, UK. Electronic address:
Multi-walled carbon nanotubes (MWCNTs) are a desirable class of high aspect ratio nanomaterials (HARNs) owing to their extensive applications. Given their demand, the growing occupational and consumer exposure to these materials has warranted an extensive investigation into potential hazards they may pose towards human health. This study utilised both the in vitro mammalian cell gene mutation and the cytokinesis-blocked micronucleus (CBMN) assays to investigate genotoxicity in human lymphoblastoid (TK6) and 16HBE14o human lung epithelial cells, following exposure to NM-400 and NM-401 MWCNTs for 24 h.
View Article and Find Full Text PDFSci Rep
December 2024
Department of Mechanical Design, Faculty of Engineering, Helwan University, P.O. Box 11718, Mataria, Helmeiat-Elzaton, Cairo, Egypt.
Water hammer (WH) is a phenomenon characterized by the rapid opening or closing of valves or pumps in pipelines, resulting in a disruptive noise, intense vibrations, and potential damage to pipes, fittings, structures, and even human safety. While WH arresters are commonly employed to mitigate this issue in smaller plumbing systems, alternative solutions are required for larger applications like power plants. Researchers have proposed the utilization of pipe materials with a low modulus of elasticity in areas prone to WH events, as these materials possess the capability to absorb a significant portion of the resulting vibrations.
View Article and Find Full Text PDFJ Plant Physiol
December 2024
Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, College of Horticulture & Landscape Architecture, Northeast Agricultural University, Harbin, 150030, China; National-Local Joint Engineering Research Center for Development and Utilization of Small Fruits in Cold Regions, Northeast Agricultural University, Harbin, 150030, China. Electronic address:
MYB transcription factors exert crucial functions in enhancing plant stress tolerance, which is impacted by soil drought and salinity. In our study, the R2R3-type MYB transcription factor gene LcMYB5 from blue honeysuckle (Lonicera caerulea L.) was successfully cloned and identified, and confirmed its nuclear localization.
View Article and Find Full Text PDFBMC Plant Biol
December 2024
College of Tea/Agrobioengineering Sciences, Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang, 550025, China.
Background: Chitinases (CHIs) are glycosidases that degrade chitin, playing critical roles in plant responses to both abiotic and biotic stress. Despite their importance, the CHI family's systematic analysis and evolutionary pattern in F. tataricum (Tartary buckwheat) yet to be explored.
View Article and Find Full Text PDFACS Pharmacol Transl Sci
December 2024
Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, Tennessee 38163, United States.
Acute cerebral ischemia is a leading cause of death and disability, particularly among old adults. The narrow therapeutic window and risk of hemorrhagic transformation largely limit patient eligibility for the current treatment. The neuroinflammatory signaling pathway involving the prostaglandin E2 (PGE) receptor subtype EP2 has now been clarified to contribute to the secondary neurotoxicity following ischemic stroke.
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