Article Inoculation of rice with nitrogen-fixing bacteria—Problems a...

Dear @Anna Koprivova Please have a look at the links given below, and the attached PDFs.

https://bmcgenomics.biomedcentral.com/articles/10.1186/1471-2164-15-755

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6200043/

First of all streak the bacterial strains kept at -20 °C or -80 °C freezer on freshly prepared nutrient agar plates and incubate them at 28 °C for 1-3 days.

Then, scrape off the cells from the plates and suspend them in MilliQ sterile ultrapure water.

Adjust the concentration of this suspension at 108 cells/ml (OD = at 600 nm) using a spectrophotometer.

Then plants are grown as required age use a needleless syringe to pump inoculum and infiltrate 1 ml of the bacterial suspension inoculum into youngest fully expanded leaves using the needleless syringe.