This review article presents a consolidated explanation and provides a comprehensive description of various studies, carried out on in vitro culture and hairy root cultures of S. marianum which can be consider an alternative source of flavonolignans. To overcome the constrains of conventional propagation of silybum plant, tissue culture and advanced biotechnology proved to be an influential tool that can complement conventional breeding and accelerate silybum development. The present review is focused on biotechnological tools like in vitro culture, hairy root cultures and genetic fidelity of S. marianum which can be a potent tool for production of secondary metabolites from these cultures.

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Attempt was made to produce transgenic cell lines of flax cv. Blanka tolerant to drought stress. Genetic transformation systems were used to incorporate the DREB2A gene, as the specific gene for drought stress tolerance. In biolistic transformation, hypocotyl segments were bombarded with DREB2A and GFP genes at particle flight distance of 9 cm and rupture disc pressure of 1300 psi. The expression of the gene was observed under a light microscope after 24 and 48 hrs. In Agrobacterium-mediated transformation, the hypocotyl segments were incubated overnight with Agrobacterium culture at five optical density OD600 i.e. 0.2, 0.4, 0.6, 0.8 and 1 for 30 min with occasional stirring. Later, the explants were transferred to a selection regeneration medium supplemented with 50 mg/dm 3 hygromycin and 300 mg/dm 3 cefotaxime and subcultured every two weeks on a new selection medium. Molecular analysis confirmed the expression of the target DREB2A gene in flax genome.

This study was carried out to investigate the effect of mannitol, sorbitol and sucrose as osmotic agents on in vitro conservation of embryogenic cultures of date palm (Phoenix dactylifera, L.) Bartamoda and Sakkoty cultivars. Embryogenic cultures was obtained using MS medium supplemented with 10 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 3 mg/l isopentenyl adenine (2iP). Among the three types of osmotic substances used for slow growth conservation, sucrose at all concentrations gave the highest percentage of survival with Sakkoty cultivar. However, addition of 40 g/l or 60 g/l mannitol and 20 g/l sorbitol showed the highest percentage of survival percentage with Bartamoda cultivar. The different sucrose concentrations caused higher numbers of germinated embryos of the two cultivars compared with mannitol or sorbitol. Also, the number of germinated embryos was increased with increasing the storage periods till the ninth month. Genetic stability was determined using random amplified polymorphic DNA (RAPD) analysis. There were no clear genetic differences between the two osmotic agents used for preservation. The preserved cultures of Sakkoty cultivar gave the high percent of similarity while Bartamoda cultivar gave low percent of similarity. From the obtained results we can recommend using 40 g/l mannitol or 20 g/l sorbitol for in vitro preservation of Bartamoda cultivar of date palm and 20 g/l of sucrose for Sakkoty cultivar.

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Leaf explants of Echinacea purpurea L. taken from aseptically germinated seedlings were inoculated with A. tumefaciens strains EHA105, carrying a binary vector conferring herbicide resistant bar gene and fungal resistant chitinase gene. Glufosinate ammonium-resistant shoots were regenerated on a medium containing BAP and NAA at a concentration of 4.88 and 0.053 μM, respectively. A subsequent transfer of shoots to medium containing BAP was necessary for stem elongation and leaf development. Transgenic Echinacea plants carrying bar and chitinase genes were selected for their resistance to glufosinate ammonium herbicide. Molecular analysis using PCR confirmed the integration of the transgenes into plant genome. This is the first report on genetic transformation of Echinacea plant using bar gene as a selectable marker.

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