Elimination of TSWV from Impatiens hawkerii Bull. and regeneration of virus-free plant

Snežana Milošević¹ · Angelina Subotić*¹ · Aleksandra Bulajić² · Ivana Djekić² · Sladjana Jevremović¹ · Ana Vučurović² · Branka Krstić²

¹ Department of Plant Physiology, Institute for Biological Research Siniša Stanković, University of Belgrade, Belgrade, Serbia
² Department of Phytopathology, Faculty of Agriculture, University of Belgrade, Belgrade- Zemun, Serbia

*Corresponding author: subotic.angelina@gmail.com

Financial support: This study was supported by the Serbian Ministry of Science and Technological Development (Project NºTR-23010-A).

Keywords: meristem culture, New Guinea Impatiens, Tomato spotted wilt virus, virus- free plants.

The possibility for obtaining virus free plants from Impatiens hawkerii Bull. shoots infectedwith Tomato spotted wilt virus (TSWV) through meristem-tip culture was examined. TSWV presence in I. hawkerii plants was detected by DAS-ELISA and RT-PCR and identification of the virus was confirmed by sequencing one of the chosen isolate (GenBank Accesion CQ132190). Meristem-tip explants (0.3-1.5 mm) from virus-infected shoots are cultured on MS media supplemented with different concentrations of the cytokinins, CPPU or TDZ (0.01-1.0 µM), respectively. Using this system, a large number of in vitro shoots could be produced from a single explant. Also, cytokinins showed a stimulatory effect on the length, fresh and dry weights of the newly formed shoots. Plant pigments content in I. hawkerii shoots increased significantly in the presence of cytokinins. Rooting of shoots was spontaneous on the same media. Rooted plantlets were transferred to soil where 97% successfully acclimatized. By DAS-ELISA and RT-PCR, 80% of the in vitro plantlets were shown to be a virus-free. Considering these, the present protocol seems to be an efficient method for in vitro generation of virus-free I. hawkerii plantlets by meristem tip cultures.