Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach

Dang Viet Cao, Reniel S. Pamplona, Jiwon Kim, Young Kyoung Oh, Seok Keun Cho, Jongcheol Ahn, Seong Wook Yang, Key Zung Riu, Kyung Hwan Boo

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The Agrobacterium-mediated transient assay is a relatively rapid technique and a promising approach for assessing the expression of a gene of interest. Despite the successful application of this transient expression system in several plant species, it is not well understood in spinach. In this study, we analyzed various factors, including infiltration method, Agrobacterium strain and density, and co-infiltration of an RNA silencing suppressor (p19), that affect transient expression following agroinfiltration in spinach. To evaluate the effects of these factors on the transient expression system, we used the β-glucuronidase (GUS) reporter gene construct pB7WG2D as a positive control. The vacuum-based infiltration method was much more effective at GUS gene expression than was the syringe-based infiltration method. Among the three Agrobacterium strains examined (EHA105, LBA4404, and GV2260), infiltration with the GV2260 strain suspension at a final optical cell density (OD600) of 1.0 resulted in the highest gene expression. Furthermore, co-expression of suppressor p19 also increased the efficiency and duration of gene expression and protein accumulation. The results indicate that the use of optimized conditions for transient gene expression could be a simple, rapid, and effective tool for functional genomics in spinach.

Original languageEnglish
Pages (from-to)397-405
Number of pages9
JournalPlant Biotechnology Reports
Volume11
Issue number6
DOIs
Publication statusPublished - 2017 Dec 1

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Agrobacterium
Spinacia oleracea
spinach
Gene Expression
gene expression
Glucuronidase
agroinfiltration
syringes
Syringes
Vacuum
Genomics
RNA Interference
RNA interference
methodology
Reporter Genes
reporter genes
Suspensions
Cell Count
heterologous gene expression
genomics

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Plant Science

Cite this

Cao, D. V., Pamplona, R. S., Kim, J., Oh, Y. K., Cho, S. K., Ahn, J., ... Boo, K. H. (2017). Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach. Plant Biotechnology Reports, 11(6), 397-405. https://doi.org/10.1007/s11816-017-0457-4
Cao, Dang Viet ; Pamplona, Reniel S. ; Kim, Jiwon ; Oh, Young Kyoung ; Cho, Seok Keun ; Ahn, Jongcheol ; Yang, Seong Wook ; Riu, Key Zung ; Boo, Kyung Hwan. / Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach. In: Plant Biotechnology Reports. 2017 ; Vol. 11, No. 6. pp. 397-405.
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Cao, DV, Pamplona, RS, Kim, J, Oh, YK, Cho, SK, Ahn, J, Yang, SW, Riu, KZ & Boo, KH 2017, 'Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach', Plant Biotechnology Reports, vol. 11, no. 6, pp. 397-405. https://doi.org/10.1007/s11816-017-0457-4

Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach. / Cao, Dang Viet; Pamplona, Reniel S.; Kim, Jiwon; Oh, Young Kyoung; Cho, Seok Keun; Ahn, Jongcheol; Yang, Seong Wook; Riu, Key Zung; Boo, Kyung Hwan.

In: Plant Biotechnology Reports, Vol. 11, No. 6, 01.12.2017, p. 397-405.

Research output: Contribution to journalArticle

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AU - Cho, Seok Keun

AU - Ahn, Jongcheol

AU - Yang, Seong Wook

AU - Riu, Key Zung

AU - Boo, Kyung Hwan

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