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      Cloning and Functional Characterization of a Pericarp Abundant Expression Promoter ( AhGLP17-1P) From Peanut ( Arachis hypogaea L.)

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          Abstract

          Peanut ( Arachis hypogaea L.) is an important oil and food legume crop grown in tropical and subtropical areas of the world. As a geocarpic crop, it is affected by many soil-borne diseases and pathogens. The pericarp, an inedible part of the seed, acts as the first layer of defense against biotic and abiotic stresses. Pericarp promoters could drive the defense-related genes specific expression in pericarp for the defense application. Here, we identified a pericarp-abundant promoter ( AhGLP17-1P) through microarray and transcriptome analysis. Besides the core promoter elements, several other important cis-elements were identified using online promoter analysis tools. Semiquantitative and qRT-PCR analyses validated that the AhGLP17-1 gene was specifically expressed only in the pericarp, and no expression was detected in leaves, stem, roots, flowers, gynophore/peg, testa, and embryo in peanut. Transgenic Arabidopsis plants showed strong GUS expression in siliques, while GUS staining was almost absent in remaining tissues, including roots, seedlings, leaf, stem, flowers, cotyledons, embryo, and seed coat confirmed its peanut expressions. Quantitative expression of the GUS gene also supported the GUS staining results. The results strongly suggest that this promoter can drive foreign genes’ expression in a pericarp-abundant manner. This is the first study on the functional characterization of the pericarp-abundant promoters in peanut. The results could provide practical significance to improve the resistance of peanut, and other crops for seed protection uses.

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          A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures

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            Floral dip: a simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana

            The Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor-intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens, 5% sucrose and 500 microliters per litre of surfactant Silwet L-77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.
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              Protein Identification and Analysis Tools on the ExPASy Server

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                Author and article information

                Contributors
                Journal
                Front Genet
                Front Genet
                Front. Genet.
                Frontiers in Genetics
                Frontiers Media S.A.
                1664-8021
                20 January 2022
                2021
                : 12
                : 821281
                Affiliations
                [1] 1 College of Plant Protection , Fujian Agriculture and Forestry University (FAFU) , Fuzhou, China
                [2] 2 Fujian Provincial Key Laboratory of Plant Molecular and Cell Biology , Oil Crops Research Institute , Fujian Agriculture and Forestry University (FAFU) , Fuzhou, China
                [3] 3 Center of Legume Crop Genetics and Systems Biology , College of Agriculture , Fujian Agriculture and Forestry University (FAFU) , Fuzhou, China
                Author notes

                Edited by: Aamir Raina, Aligarh Muslim University, India

                Reviewed by: Farhat Abbas, South China Agricultural University, China

                Muhammad Azhar Nadeem, Sivas University of Science and Technology, Turkey

                *Correspondence: Weijian Zhuang, weijianz@ 123456fafu.edu.cn

                This article was submitted to Plant Genomics, a section of the journal Frontiers in Genetics

                Article
                821281
                10.3389/fgene.2021.821281
                8811503
                35126474
                2b55ce4a-60f6-4a2d-94dc-5021ffcbed91
                Copyright © 2022 Sharif, Chen, Deng, Ali, Khan, Zhang, Xie, Chen, Cai, Yang, Zhuang, Raza and Zhuang.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 24 November 2021
                : 31 December 2021
                Funding
                Funded by: National Natural Science Foundation of China , doi 10.13039/501100001809;
                Award ID: U1705233 31601337
                Funded by: Fujian Agriculture and Forestry University , doi 10.13039/501100008766;
                Categories
                Genetics
                Original Research

                Genetics
                cis-elements,gus staining,pathogens,tissue-specific expression,transgenic arabidopsis
                Genetics
                cis-elements, gus staining, pathogens, tissue-specific expression, transgenic arabidopsis

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