Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach
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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, 2017, p. 397-405.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Optimization of Agrobacterium-mediated transient expression of heterologous genes in spinach
AU - Cao, Dang Viet
AU - Pamplona, Reniel S.
AU - Kim, Jiwon
AU - Oh, Young Kyoung
AU - Cho, Seok Keun
AU - Ahn, Jongcheol
AU - Yang, Seong Wook
AU - Riu, Key Zung
AU - Boo, Kyung Hwan
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Agrobacterium tumefaciens
KW - Spinach
KW - Syringe
KW - Transient assay
KW - Vacuum
U2 - 10.1007/s11816-017-0457-4
DO - 10.1007/s11816-017-0457-4
M3 - Journal article
AN - SCOPUS:85032181097
VL - 11
SP - 397
EP - 405
JO - Plant Biotechnology Reports
JF - Plant Biotechnology Reports
SN - 1863-5466
IS - 6
ER -
ID: 193503701