An efficient Agrobacterium-mediated soybean transformation method using green fluorescent protein as a selectable marker

Plant Signal Behav. 2019;14(7):1612682. doi: 10.1080/15592324.2019.1612682. Epub 2019 May 6.

Abstract

Genetic transformation plays a vital role in gene functional study and molecular breeding of soybean. Conventional soybean transformation methods using chemical selectable markers, such as antibiotic or herbicide resistance genes, rely on the identification of positive transgenic lines at advanced developmental stages, making selection procedure labor intensive and time consuming. Utilization of a visual maker to track the transgene would avoid the uncertainty and blindness in the transformation process. In this research, we used green fluorescent protein (GFP) as the selectable marker to detect transgenics at early stages of soybean development. Positive transformants were detected recurrently during each stage of the process based on visualization of the green fluorescence signal, which help us to discard the non-transgenic ones in each stage to reduce the unnecessary experimental cost and lab space. In addition, the positive transgenic seeds can be identified before planting for early detection of transgene and obtain homozygous lines in advance. The method established in this study is also a useful reference for other plant species.

Keywords: Glycine max; cauliflower mosaic virus 35S promoter; green fluorescent protein; transformation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium / physiology*
  • Fluorescence
  • Genetic Markers
  • Glycine max / genetics*
  • Glycine max / microbiology*
  • Green Fluorescent Proteins / metabolism*
  • Plant Shoots / metabolism
  • Plants, Genetically Modified
  • Seeds / genetics
  • Transformation, Genetic*
  • Transgenes

Substances

  • Genetic Markers
  • Green Fluorescent Proteins

Grants and funding

This work was supported by the Key Transgenic Breeding Program of China [grant number 2014ZX0801005B]; the Fundamental Research Funds for the Central Universities [grant number KYT201801]; the China Scholarship Council; and the Program for Changjiang Scholars and Innovative Research Team in University [grant number IRT_17R55]..