Abstract
Agrobacterium rhizogenes is known as a natural tool of genetic engineering in many plant species. For the first time, hairy root induction in Agastache foeniculum using A. rhizogenes, rosmarinic acid content and the effect of different culture media and inoculation methods on hairy root growth rate were investigated. Hairy root culture of A. foeniculum was established by inoculation of the 1-month-old leaf explant with A4 strain of A. rhizogenes and the effectiveness of light – dark conditions and two inoculation methods (immersion and injection) were tested. Furthermore, in immersion method, the effects of inoculation time (3, 5 and 7 min) on root induction were investigated. In the second part of the study, the hairy root culture of A. foeniculum was studied using different basal culture media (MS, 1/2 MS and B5). Rosmarinic acid content in hairy roots and non- transformed roots was analyzed using high-performance liquid chromatography (HPLC). There was no significant difference between various inoculation methods in the ability of hairy roots induction. Observations showed that percentage of hairy root induction was higher when the explants were immersed for 5 min in bacterial suspension. Light conditions displayed the highest hairy root induction rates compared with dark condition. Various culture media are different in terms of types and amounts of nutrients and have influence on growth rate. The maximum growth rate (1.61 g fr wt/50 ml) of hairy roots were obtained in 1/2 MS medium. Rosmarinic acid content in transformed roots (213.42 µg/g dry wt) was significantly higher than non-transformed roots (52.28 µg/ g dry wt).
Highlights
Anise hyssop (Agastache foeniculum (Pursh) Kuntze) is a perennial herbaceous plant belonging to Labiatae family (Omidbaigi and Mahmoodi, 2010)
The effects of two different inoculation methods on induction of hairy roots were evaluated in Trigonella foenum-graecum L.; the results showed that transformation performance was 26 % in injection method and 6 % in co- cultivation method (Akbarian et al, 2011)
Hairy root induction using 1– month- old leaves as explants can be related to the high sensitivity of these explants to bacteria that depends on the physiological status of explants tissue (Pawar and Mathesh wari, 2003)
Summary
Anise hyssop (Agastache foeniculum (Pursh) Kuntze) is a perennial herbaceous plant belonging to Labiatae family (Omidbaigi and Mahmoodi, 2010). This plant is native to the United States and Canada (Mallavarapu et al, 2004) and is grown in all regions of the Mediterranean, in Northen and Central Europe (Omidbaigi et al, 2008). Clinical trials have shown that anise hyssop extract is effective in treatment of heart, lung, cough diseases and induction sweating to reduce fevers (Mallavarapu et al, 2004). Essential oils of this plant have anti-bacterial and anti- fungal properties. The antioxidant activity of A. foeniculum (98.6 mg.g dry plant) depends on the total content of polyphenols (Matei, 2012)
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