Analysis of Trace Elements During Different Developmental Stages of Somatic Embryogenesis in Plantago ovata Forssk Using Energy Dispersive X-ray Fluorescence

Abstract

Energy dispersive X-ray fluorescence (ED-XRF) technique has been used for the determination of trace element profile during different developmental stages of somatic embryogenic callus of an economically important medicinal plant, Plantago ovata Forssk. Somatic embryogenesis is a plant tissue culture-based technique, which is used for plant regeneration and crop improvement. In the present investigation, elemental content was analysed using ED-XRF technique during different developmental stages and also determine the effect of additives--casein hydrolysate and coconut water on the trace elemental profile of embryogenic callus tissue of P. ovata. Subsequent experiments showed significant alteration in the concentration of K, Ca, Mn, Fe, Zn, Cu, Br, and Sr in both the embryogenic and non-embryogenic callus. Higher K, Ca, Fe, Cu, and Zn accumulation was in embryogenic tissue stage compared to other stages, suggesting these elements are crucial for successful embryogenesis. The results suggest that this information could be useful for formulating a media for in vitro embryo induction of P. ovata.

Formulation of Nutrient Medium for In Vitro Somatic Embryo Induction in Plantago ovata Forsk

“…As shown in Table 2, MS medium contained 27.80 mg/L of Fe, mainly in FeSO 4 form, whereas the formulated medium contained 151.51 mg/L of iron. The ED-XRF results obtained demonstrate that during the somatic embryogenic stage, about five times more Fe accumulated than during the non-embryogenic stages of P. ovata [25]. Thus, it could be said that the formulated medium, because of its five times higher iron concentration, substantially increased the rate and the frequency of embryo and plantlet regeneration of P. ovata.…”

“…According to the elemental analysis data of ED-XRF [25], a nutrient medium was formulated for the induction of high-frequency somatic embryogenesis and regeneration of P. ovata. To obtain the optimal composition of the culture medium for P. ovata, three nutrient media with different combinations of macro-and microelements were formulated (FM1, FM2, and FM3), and all of them were different from the normally used MS [27] medium with respect to their macro-and micronutrient composition (Tables 1 and 2).…”

“…Based on the results of ED-XRF analyses, the concentrations of macro-and microelements were altered (Tables 1 and 2). The results of ED-XRF showed that more (about five times) iron was accumulated in the stage of somatic embryogenesis as compared to non-embryogenic callus stage of P. ovata [25]; therefore, in the formulated medium, iron was added five times more as compared to MS medium (Table 2). Similarly, other elemental concentrations were also changed in the new nutrient medium.…”

“…Based on the result of ED-XRF analyses [25], we have tried to formulate a nutrient medium for the induction of high-frequency embryogenesis of P. ovata. The results of the present investigation showed that three nutrient media (FM1, FM2, and FM3) with different combination of macro-and microelements were suitable for the induction of somatic embryogenesis, while FM3 was the most suitable as compared to MS medium (Figs.…”

“…Using energy-dispersive X-ray fluorescence (ED-XRF) technique, we already determined the trace elemental constituents during the different developmental stages of somatic embryogenesis in P. ovata and evaluated the role of trace elements during embryogenesis [25]. The results suggest that the information on the accumulation of elements during developmental stages in vitro could be useful for formulating a medium for the induction of high-frequency embryogenesis of P. ovata.…”