Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (2): 182-191.DOI: 10.11983/CBB19169

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

The Application of Double-barreled Particle Bombardment for Transient Gene Expression in Arabidopsis

Hua Zhao1,Guangda Shao1,2,Wenxin Gao1,2,Biao Gu1,2,*()   

  1. 1 State Key Laboratory of Crop Stress Biology for Arid Areas, Northwest A&F University, Yangling 712100, China
    2 College of Plant Protection, Northwest A&F University, Yangling 712100, China
  • Received:2019-08-28 Accepted:2019-11-28 Online:2020-03-01 Published:2020-02-12
  • Contact: Biao Gu

Abstract:

Transient gene expression is a favorite tool used for functional analysis of target genes in plant. Of three techniques applied for genetic transformation in the model plant Arabidopsis thaliana, biolistic delivery system was less used than protoplast- or Agrobacterium-mediated transformation. This is mainly due to the smaller leaf size of Arabidopsis, the complicated procedure of bombardment and the limited efficiency and consistency of gene expression. Here, we report applications of an optimized double-barreled particle bombardment system for transient transformation in Arabidopsis, which displayed high expression level of GFP and GUS reporter genes in leaf epidermal cells. By introducing the parallel control in the same shoot by co-bombardment, gene expression efficiency and consistency were dramatically improved, which allows quantitative analysis of target genes with several replicates. Furthermore, cell death inducers BAX, Avh238 or ATR13/Rpp13, were co-expressed with GUS in Arabidopsis rosette leaves and led to strong necrosis phenotypes visualized by significant reduction of number of GUS spots. On the contrary, Avrblb1/RB gene pair triggered strong cell death in Nicotiana benthaminana, but not in A. thaliana. Therefore, this time-saving protocol is an alternative to quantitatively evaluate biological functions of the gene of interest and high-throughput screening of immune suppressors in Arabidopsis and its mutants.

Key words: transient expression, double-barreled gene gun, Arabidopsis, effector gene, disease-resistance gene