Agrobacterium tumefaciens-mediated transformation of a hevein-like gene into asparagus leads to stem wilt resistance

PLoS One. 2019 Oct 7;14(10):e0223331. doi: 10.1371/journal.pone.0223331. eCollection 2019.

Abstract

Asparagus stem wilt, is a significant and devastating disease, typically leading to extensive economic losses in the asparagus industry. To obtain transgenic plants resistant to stem wilt, the hevein-like gene, providing broad spectrum bacterial resistance was inserted into the asparagus genome through Agrobacterium tumefaciens-mediated transformation. The optimal genetic transformation system for asparagus was as follows: pre-culture of embryos for 2 days, inoculation using a bacterial titre of OD600 = 0.6, infection time 10 min and co-culturing for 4 days using an Acetosyringone concentration of 200 μmol/L. Highest transformation frequencies reached 21% and ten transgenic asparagus seedlings carrying the hevein-like gene were identified by polymerase chain reaction. Moreover, integration of the hevein-like gene in the T1 generation of transgenic plants was confirmed by southern blot hybridization. Analysis showed that resistance to stem wilt was enhanced significantly in the transgenic plants, in comparison to non- transgenic plants. The results provide additional data for genetic improvement and are of importance for the development of new disease-resistant asparagus varieties.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / genetics*
  • Agrobacterium tumefaciens / pathogenicity
  • Antimicrobial Cationic Peptides / genetics*
  • Antimicrobial Cationic Peptides / metabolism
  • Asparagus Plant / genetics*
  • Asparagus Plant / microbiology
  • Disease Resistance*
  • Fungi / pathogenicity
  • Gene Transfer Techniques*
  • Plant Lectins / genetics*
  • Plant Lectins / metabolism
  • Transformation, Genetic
  • Transgenes*

Substances

  • Antimicrobial Cationic Peptides
  • Plant Lectins
  • hevein

Grants and funding

This work was sponsored by the Special Fund for Agro-scientific Research in the Public Interest (201003074) and Technology Project of Hainan (ZDXM2015047). And the funders had no role in study design, design, data collection and analysis, decision to publish, or preparation of the manuscript.