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Development of an efficient virus-induced gene silencing method in petunia using the pepper phytoene desaturase (PDS) gene

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Abstract

Virus-induced gene silencing (VIGS) is widely used for functional analysis of genes in plants. Because of the variation in its effectiveness among plant species, the VIGS system is normally optimized using the phytoene desaturase (PDS) gene as a visible indicator. In this study, we optimized an efficient VIGS system for petunias using the PDS gene isolated from pepper (Capsicum annuum) plants as a visible indicator. Application of the CaPDS gene effectively induced gene silencing in the petunia cultivars Mirage Rose, Mirage Pink, and Picobella Blue. However, silencing effectiveness was observed to be limited by genotypes and inoculation methods; apical meristem application was the appropriate method, whereas the highest silencing effectiveness among the cultivars was found in Picobella Blue. In addition, it was found that higher silencing effectiveness was associated with higher degradation of the endogenous PhPDS mRNA. Moreover, in terms of plant age and temperatures, 3-week-old plants grown at 20 °C day/18 °C night showed high silencing effectiveness for all cultivars. Taken together, infection of apical meristem of 3-week-old plants grown at 20 °C day/18 °C night with pTRV2-CaPDS exhibited the highest silencing effectiveness of all cultivars. Therefore, this study indicated the effectiveness of the CaPDS in the VIGS experiment with petunias, as well as the involvement of different factors in the mechanism influencing silencing effectiveness in petunias. These results will enable high-throughput functional analysis of genes associated with commercially important traits in petunias.

Key Message

CaPDS gene effectively induced gene silencing in the petunia cultivars, however, higher silencing effectiveness was observed to be limited by genotypes, inoculation methods, plant age, inoculation temperature, and presence of endogenous PhPDS mRNA.

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Abbreviations

CaMV:

Cauliflower mosaic virus

GOI:

Gene of interest

PTGS:

Post transcriptional gene silencing

PDS:

Phytoene desaturase

siRNA:

Short interference RNA

TRV:

Tobacco rattle virus

VIGS:

Virus-induced gene silencing

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Funding

This work was supported by a grant from the Next-Generation BioGreen 21 Program (Project no. PJ01368505), Rural Development Administration, Republic of Korea.

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Author notes

  1. Aung Htay Naing and Hyun Young Song have equally contributed to this work.

Authors and Affiliations

  1. Department of Horticultural Science, Kyungpook National University, Daegu, 4165122, South Korea

    Aung Htay Naing, Hyun Young Song, Je Min Lee, Ki Byung Lim & Chang Kil Kim

Authors
  1. Aung Htay Naing
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  2. Hyun Young Song
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  3. Je Min Lee
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  4. Ki Byung Lim
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  5. Chang Kil Kim
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Contributions

AHN and CKK designed the experiment. JML generated the construct. HYS performed all experimental steps such as planting of the petunia plants, preparation of inoculum, Agro-infiltration, analysis of qRT-PCR, and detection of total chlorophyll contents. KBL assisted in data analysis and molecular analysis AHN wrote and revised the manuscript. CKK supervised the project. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Chang Kil Kim.

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Communicated by Sergio Rosales-Mendoza.

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Naing, A.H., Song, H.Y., Lee, J.M. et al. Development of an efficient virus-induced gene silencing method in petunia using the pepper phytoene desaturase (PDS) gene. Plant Cell Tiss Organ Cult 138, 507–515 (2019). https://doi.org/10.1007/s11240-019-01646-y

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  • DOI: https://doi.org/10.1007/s11240-019-01646-y

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