2.**Method D – Sterilisation and Cultivation Process of Stevia, Chicory, Bee Balm (Cambridge Scarlet), and Rhodiola krylovii Cells.**

The 400 ml of M and S media volume with Kinetin was prepared using the above reagents for plant cells. A total of 9 ml of solid media was poured into 56 test tubes. Sterile Petri dishes were used for the sterilisation process of seeds. Seeds were washed adequately with an antibacterial soap in flowing water for 10 mins. Seeds were sterilised with 70% ethanol for 2 mins. Seeds were properly washed three times with sterile water. Then, the seeds were sterilised with 3% NaOCl solution for 5 mins. Seeds were later transferred into sterile test tubes containing solid media. Fourteen labelled, sterile test tubes contained Stevia, Chicory, Bee Balm (Cambridge Scarlet), and *Rhodiola krylovii* cells. Seeds were kept in the climate chamber for incubation with no light intensity, temperature 250C (� 1 ° C), and relative humidity of 50%. After 14 days, the percentage of contamination was determined.

The level of percentage contamination for the four sterilisation methods (A, B, C, and D) for seed cultivation on solid media in tubes for callus formation (medium M&S + kinetin) and the cultivation of seeds on solid media in tubes to obtain sterile micro plants (medium M&S) was determined by using the formula:

% of Contamination ¼ ð Þ Number of Contaminated Test tubes with cells *=* ð Þ Total Number of Test tubes with seeds <sup>∗</sup> <sup>100</sup> (1)

	- A. Non-sterile Plant Seedlings

The seeds of Cambridge scarlet, Rhodiola krylovii, and Chicory were put into six Petri dishes. Two Petri dishes for Bee Balm (Cambridge scarlet), two for Rhodiola krylovii, and two Petri dishes for Chicory were prepared. The seeds were unwashed, and 7 ml of distilled water was added to all Petri dishes. The samples were labelled and sealed with paraffin and incubated for 2 weeks for plant growth in a climate chamber for a plant with no light intensity, a temperature of 250C (�1°C), and relative humidity of 50%.

B. Sterile Plant Seedlings

A total of 7 ml of sterile water was poured into six sterile Petri dishes. Seeds of Chicory, Rhodiola krylovii, and Cambridge scarlet were disinfected with 70% ethanol for 2 mins and 3% NaOCl solution for 2 mins. Seeds were rinsed thoroughly with sterile water twice in separate sterilised Petri dishes for 2 mins. The seeds were poured into the six sterilised Petri dishes and sealed with paraffin. Samples were labelled and incubated in the climate chamber for 2 weeks with no light intensity, temperature 25°C (1°C), and relative humidity of 50%.
