Development of tissue culture and virus-induced gene silencing for Calendula officinalis

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Title: Development of tissue culture and virus-induced gene silencing for Calendula officinalis
Author: Liu, Yanbo
Other contributor: Helsingin yliopisto, Maatalous-metsätieteellinen tiedekunta, Maataloustieteiden laitos
University of Helsinki, Faculty of Agriculture and Forestry, Department of Agricultural Sciences
Helsingfors universitet, Agrikultur- och forstvetenskapliga fakulteten, Institutionen för lantsbruksvetenskaper
Publisher: Helsingfors universitet
Date: 2011
Language: eng
Thesis level: master's thesis
Discipline: Växtproduktions biologi (trädgårdsvetenskap)
Plant Production Biology (Horticulture)
Kasvintuotannon biologia (puutarhatiede)
Abstract: Calendula officinalis is grown widely as an ornamental plant across Europe. It belongs to the large. Asteraceae family. In this study, the aim was to explore the possibilities to use Calendula officinalis as a new model organism for flower development and secondary mechanism studies in Asteraceae. Tissue culture of Calendula officinalis was established using nine different cultivars. Murashige & Skoog (MS) medium with four different combinations of plant growth regulators were tested. Of all these combinations, the medium containing 1mg/l BAP, 0.1 mg/l IAA, and 1mg/l Zeatin achieved highest frequency of adventitious shoot regeneration from hypocotyl and cotyledon explants. Virus-induced gene silencing is a recent developed genetic tool for charactering the gene functions in plants, and extends the range of host plants that are not accessible for Agrobacterium transformation. Here, tobacco rattle virus (TRV)-based VIGS technique was tested in calendula (cv. Single Orange). We used TRV carrying Gerbera hybrid phytoene desaturase (PDS) gene fragment to induce PDS silencing in calendula. Vacuum infiltration and syringe infiltration methods both resulted in photo-bleaching phenotypes in leaves, bracts and petals. Loss-of-function phenotypes occurred on calendula 13 days post-infiltration. In conclusion, the data indicates that calendula explants can be regenerated through tissue culture which is a prerequisite for development of stable transformation methods. However, further optimization is still needed to improve the frequency. In addition, VIGS was applied to silence PDS marker gene expression indicating that this method has potential for gene functional studies in future.
Subject: Calendula officinalis
tissue culture
plant growth regulator
PDS gene

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