Skip to main content

ORIGINAL RESEARCH article

Front. Plant Sci.
Sec. Plant Biotechnology
Volume 15 - 2024 | doi: 10.3389/fpls.2024.1476754
This article is part of the Research Topic Genetically Modified (GM) and Genome-Edited (GE) Plants For Achieving Sustainable Agriculture View all 10 articles

Expression evaluation of exogenous and endogenous alcohol dehydrogenase genes in transgenic Arabidopsis

Provisionally accepted
Hong Lv Hong Lv *Haoqiang Yu Haoqiang Yu Fengzhong Lu Fengzhong Lu Qingqing Yang Qingqing Yang *Wanchen Li Wanchen Li *Fengling Fu Fengling Fu *
  • Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan Province, China

The final, formatted version of the article will be published soon.

    A lot of endogenous genes, as well as genes from related species, are transformed back into crops for overexpression to improve their corresponding traits. However, almost all of these transgenic events remain at the testing stage. Most of the singular transgenic events of crops approved for commercial release are developed by the transformation and heterologous expression of exogenous genes from distant species. To detect the differences in expression, protein accumulation, and enzyme activity between transformed exogenous and endogenous genes, the coding sequences (DCSs) of the alcohol dehydrogenase (ADH) genes were cloned from dicotyledonous Arabidopsis, monocotyledonous maize, and prokaryotic Escherichia coli, constructed into expression vector pBI121-cMycNY, and used to transform wild-type Arabidopsis, respectively. Three homozygous T3 lines with a single integration site were screened for each of the three transformed genes by antibiotic screening, PCR identification, and genomic DNA resequencing. Real-time quantitative PCR (RT-qPCR) analysis showed that the relative expression levels of the transformed exogenous ZmADH and EcADH genes were ten or tens of times higher than that of the transformed endogenous AtADH gene. After confirming the encoded proteins of these transformed genes by Western blotting, enzyme-linked immunosorbent assay (ELISA) showed that the accumulation levels of the proteins encoded by the transformed genes ZmADH and EcADH were significantly higher than that encoded by the transformed endogenous gene AtADH. Enzyme reaction assay showed that the ADH activities of the T3 lines transformed by the exogenous genes ZmADH and EcADH were also significantly higher than that transformed by the endogenous gene AtAHD as well as the wild-type control. These results indicated that exogenous genes were more conducive to transgenic improvement of crops.

    Keywords: Endogenous gene, enzyme activity, Exogenous gene, Protein accumulation, transgenics

    Received: 06 Aug 2024; Accepted: 27 Dec 2024.

    Copyright: © 2024 Lv, Yu, Lu, Yang, Li and Fu. 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) or licensor 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.

    * Correspondence:
    Hong Lv, Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan Province, China
    Qingqing Yang, Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan Province, China
    Wanchen Li, Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan Province, China
    Fengling Fu, Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan Province, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.