3.1.3. Seed Inoculation with Bacterial Endophyte and Growth of Chenopodium Sprouts

The Chenopodium seeds (C. ambrosoides, C. ficifolium, and C. botrys) were brought from the Agricultural Research Centre, Giza, Egypt. They were surface sterilized (2.5% sodium hypochlorite, 5 min) and rinsed thoroughly in sterile distilled water. Afterwards, sterilized Chenopodium seeds were soaked in a liquid suspension of inoculants at 25% concentrations (2.5 × 10⁻⁷ cfu.mL⁻¹) for 6 h at room temperature, and the control was soaked in distilled water. The control and treated Chenopodium seeds were transferred to the sterilized trays filled with vermiculite and irrigated every day. The size of each tray was (8 cm × 12 cm × 4 cm), where sprouts of each treatment were grown in different trays. The Chenopodium sprouts were cultured in growth chambers at 24 °C, photosynthetically active radiation (PAR) of 200 µmol m⁻² s⁻¹, 16 h light/8 h dark cycle, and 60% relative humidity. After eight days of cultivation, 30 sprouts from each tray of 8 × 12 × 4 cm in size (a biological replicate) were harvested and weighed to determine the fresh sprout weight. Finally, the sprouts were frozen in liquid nitrogen at −196 °C and stored for further analyses. At least four biological replicates were used for evaluation of chemical and biological attributes. The percentage of water content of the sprouts was estimated by drying fresh sprouts in an oven at 70 °C for 3 days. After determination of the dried weight of the sprouts, the water content was measured as a percentage. The fresh sprout weight (FW) was expressed as biomass (g/sprout).