Background: Drought stress seriously affects the cotton fiber development. Universal stress protein gene isolated from native species has the promising tolerance role against these stresses.
Objectives: This study aimed to clone, characterize, and genetically transform the GUSP1 gene in local cotton and to observe its expression in transgenic plants under drought stress.
Materials And Methods: Universal Stress Protein (GUSP1) gene from was cloned in pCEMBIA (-) 1301plant expression vector by replacing Hygromycin and GUS exon with GUSP1-GFP fusion fragment. The construct was transformed into and transient expression assay was confirmed by agro-infiltration of leaves and green fluorescence under a confocal microscope. Gene integration and expression in transgenic plants was observed through Southern blot and real-time PCR analyses. Cellular localization was observed through a confocal microscope and the copy number of the transgene was observed in progeny plants.
Results: Transformation efficiency was 1.9%. Developmental and spatial expression of GUSP1 was observed through Real-time PCR in stem, root, leaf, inflorescence, and seeds of transgenic plants at the vegetative and flowering stage. Integration of GUSP1 revealed a fragment of approximately 500 bp in Southern Blot analyses. Localization of GUSP1 was detected in the intact leaf of transgenic plants through GFP fluorescence in midrib, guard cells of stomata, and trichomes. Single gene copy was detected in the chromosome of transgenic seeds.
Conclusion: GUSP1 has cloned from native species of local cotton and its integration and expression in transgenic plants confirmed that the role of GUSP1 will provide direction to breed economically important cotton varieties.
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http://dx.doi.org/10.30498/IJB.2020.138051.2312 | DOI Listing |
Bio Protoc
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Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya.
Agrobacterium-mediated gene transformation method is a vital molecular biology technique employed to develop transgenic plants. Plants are genetically engineered to develop disease-free varieties, knock out unsettling traits for crop improvement, or incorporate an antigenic protein to make the plant a green factory for edible vaccines. The method's robustness was validated through successful transformations, demonstrating its effectiveness as a standard approach for researchers working in plant biotechnology.
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January 2025
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.
An endoplasmic reticulum-localized Cu transporter, PhHMA5II1, interacts with copper chaperones and plays an important role in Cu detoxification in petunia. Copper (Cu) is an essential element for plant growth but toxic when present in excess. In this study we present the functional characterization of a petunia (Petunia hybrida) P-type heavy-metal ATPases (HMAs), PhHMA5II1.
View Article and Find Full Text PDFInt J Biol Macromol
January 2025
College of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, China; Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address:
Sucrose is an important factor affecting plant growth and fruit quality, but the molecular regulatory mechanism of sucrose biosynthesis in longan is not yet understood. Here, we characterized a transcription factor, DlbHLH68, positively regulates sucrose accumulation in longan. Subcellular localization and transcriptional activity analysis indicated that DlbHLH68 is a nuclear transcriptional activator.
View Article and Find Full Text PDFPlant Sci
January 2025
Anhui Province Key Laboratory of Forest Resources and Silviculture, School of Forestry and Landscape Architecture, AnHui Agricultural University, HeFei 230036, PR China. Electronic address:
Trichome development and anthocyanin accumulation are regulated by a complex regulatory network, the MBW complexes consist of MYB, bHLH, and WD40 transcription factors. In this study, two sequences, named PaTTG1.1, and PaTTG1.
View Article and Find Full Text PDFInt J Mol Sci
January 2025
Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, National-Local Joint Engineering Research Center for Development and Utilization of Small Fruits in Cold Regions, College of Horticulture & Landscape Architecture, Northeast Agricultural University, Harbin 150030, China.
Iron stress adversely impacts plants' growth and development. Transcription factors (TFs) receive stress signals and modulate plant tolerance by influencing the expression of related functional genes. In the present study, we investigated the role of an apple bHLH transcription factor in the tolerance to iron stresses.
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