Abstract
Dragon’s head plant (Lallemantia iberica), is a flowering species belongs to the mint family (Lamiaceae). The species contains valuable essential oils, mucilage and oil which are used in pharmaceutical and food industries. Tissue culture is a feasible strategy to attain large‐scale production of plantlets with a huge potential to produce plants with superior quality. The objective of this study was to develop a simple and efficient method for regeneration and transformation of L. iberica. To reach this goal, the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyledon segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine). According to the results, cotyledonary nodes showed the best regeneration response. The maximum rate of regeneration (and number of induced shoots was achieved in 1 mg l−1 BAP in combination with 0.05 mg l−1 NAA from the cotyledonary nodes. Additionally, through the optimized regeneration technique Agrobacterium-mediated transformation of L. iberica was successfully accomplished. Gene transfer was assessed on leaf samples from regenerated plantlets under a fluorescent microscope to detect the GFP signals. Moreover, transgene integration and its expression were confirmed by PCR and RT-PCR analysis, respectively. The establishment of these efficient regeneration and genetic transformation methods paved the way for further application such as plant improvement, functional analysis and gene editing.
Highlights
Lallemantia iberica, known generally as dragon’s head, is a herb belonging to the mint family (Lamiaceae), originated from Caucasus and Middle East regions and widely distributed in Europe and some western areas of Asia[1]
Cotyledonary node, cotyledon, hypocotyl and leaf explants were examined on 16 different plant growth regulator (PGR) combinations (Table 1)
No shoots or calli were obtained on cotyledon and leaf explants in the presence of the 16 combinations, hypocotyls produced a few shoots only on the medium supplemented with 2 mg l−1 BAP + 0.1 mg l−1 naphthaleneacetic acid (NAA) (Fig. 2, b and c)
Summary
Lallemantia iberica, known generally as dragon’s head, is a herb belonging to the mint family (Lamiaceae), originated from Caucasus and Middle East regions and widely distributed in Europe and some western areas of Asia[1]. Its seed mucilage is used in the treatment of nervous, hepatica and renal d iseases[4] This herb is mainly harvested for its seeds since they contain valuable oil compounds around 30% drying weight holding high amount of valuable omega 3 fatty acid and alpha-linolenic acid (67–74%) which has many applications in pharma and food industries[4,5]. Genetic transformation enables the introduction of foreign genes into L. iberica or efficient modification of target genes to create new cultivar with desirable traits such as higher valuable secondary metabolites or omega 3 fatty acid and alpha-linolenic acid. In vitro regeneration and genetic transformation are important tools to enhance the production of secondary metabolites and engineer the valuable oil content in plants such as L. iberica. For the first time an appropriate technique for in vitro regeneration and gene delivery using Agrobacterium are presented This allows genetic improvements to be put into practice for Lallemantia iberica. The major factors which affect the regeneration and transformation such as, type of explants, concentrations or combinations of PGRs, acetosyringone, Agrobacterium inoculation duration, have been successfully optimized and a viable regeneration and transformation protocol well established
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