TY - JOUR

T1 - Phenotypic effects and the quantification of transcript abundance in Petunia hybrida 'Fantasy Blue' with virus-induced GA2ox gene silencing

AU - Gargul, Joanna Maria

AU - Mibus, Heiko

AU - Serek, Margrethe

N1 - Funding Information: The authors would like to thank Dr. Merete Albrechtsen (University of Aarhus, Denmark) for kindly providing the TRV vectors, and Prof. em. Bjarne M. Stummann (University of Copenhagen, Denmark) for his valuable advice and critical review of the manuscript. This project was supported by a Ph.D. research grant from the German Academic Exchange Service (DAAD) (Ref.: 323, PKZ: A/06/07504), which is gratefully acknowledged.

PY - 2015/12/14

Y1 - 2015/12/14

N2 - The aim of this study was to establish an efficient virus-induced gene silencing (VIGS) protocol for the gibberellin 2-oxidase gene (GA2ox) in Petunia hybrida 'Fantasy Blue' to measure the reduction in GA2ox transcript levels by quantitative RT-PCR (qRT-PCR) and to investigate the phenotypes of the infected plants. In vitro multiplied Petunia plants were subjected to VIGS using tobacco rattle virus vectors. An 838-bp fragment from the N. tabacum gene (NtGA2ox) was cloned in the TRV2 vector. Control plants were also infected with a TRV2 vector containing a fragment of the E. coli β-glucuronidase (GUS) gene as a nonsense sequence. The abundance of the GA2ox and cyclophylin (for normalization) transcripts was determined by qRT-PCR four weeks after inoculation. Shoot tips with the five youngest leaves were used. Stem lengths were measured weekly from the day of inoculation. After four weeks, significant increases in stem elongation were observed in GA2ox-TRV2-infected plants compared with GUS-TRV2-infected plants. In accordance with this observation, the normalized abundance of the GA2ox transcript in GA2ox-TRV2-infected plants was significantly reduced compared with GUS-TRV2-infected plants.

AB - The aim of this study was to establish an efficient virus-induced gene silencing (VIGS) protocol for the gibberellin 2-oxidase gene (GA2ox) in Petunia hybrida 'Fantasy Blue' to measure the reduction in GA2ox transcript levels by quantitative RT-PCR (qRT-PCR) and to investigate the phenotypes of the infected plants. In vitro multiplied Petunia plants were subjected to VIGS using tobacco rattle virus vectors. An 838-bp fragment from the N. tabacum gene (NtGA2ox) was cloned in the TRV2 vector. Control plants were also infected with a TRV2 vector containing a fragment of the E. coli β-glucuronidase (GUS) gene as a nonsense sequence. The abundance of the GA2ox and cyclophylin (for normalization) transcripts was determined by qRT-PCR four weeks after inoculation. Shoot tips with the five youngest leaves were used. Stem lengths were measured weekly from the day of inoculation. After four weeks, significant increases in stem elongation were observed in GA2ox-TRV2-infected plants compared with GUS-TRV2-infected plants. In accordance with this observation, the normalized abundance of the GA2ox transcript in GA2ox-TRV2-infected plants was significantly reduced compared with GUS-TRV2-infected plants.

KW - Gibberellin 2-oxidase

KW - qRT-PCR

KW - TRV

KW - VIGS

KW - Virus-induced gene silencing

UR - http://www.scopus.com/inward/record.url?scp=84949749745&partnerID=8YFLogxK

U2 - 10.1016/j.scienta.2015.09.039

DO - 10.1016/j.scienta.2015.09.039

M3 - Article

AN - SCOPUS:84949749745

VL - 197

SP - 226

EP - 229

JO - Scientia horticulturae

JF - Scientia horticulturae

SN - 0304-4238

ER -