Materials and Methods
Sample Collection
The root adhering soil (RAS) samples were collected from wheat crop grown field at 60 days of growth. The plants were sampled from different locations in the four districts viz., Varanasi, Mau, Ballia, and Ghazipur of Uttar Pradesh (UP) (23520 to 30250 N latitude and 7730 to 84390 E longitude), India. Physicochemical parameters of soil were analyzed and most of the soils have electrical conductivity (ECe) C8.5, pH ranging from 8.0 to 9.5, and organic carbon from 0.27 to 2.29%. The plants were carefully removed from the soil and the roots with adherent soil were put in plastic bags for isolation of rhizospheric bacteria.
Isolation of Rhizobacteria
For the isolation of rhizospheric bacteria, the roots were shaken to remove excess soil and 10 g of closely associated rhizospheric soil from each sample was added to 90 ml of sterile water and shaken for 30 min on a mechanical rotary shaker. Ten fold dilutions were made and plated on to five media, Jensen’s N free medium, King’s B (KB) medium,
nutrient agar (NA), trypticase soy agar (TSA), and soil extract agar (SEA) [28]. Bacterial cultures were maintained on the respective slants and stored at 4C till further use