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The effects of plant growth regulators, BA (N⁶-Benzyladenine) and IAA (indole -3- acetic acid) on tissue culture of Nymphaea ‘jongkolnee’ were studied at Faculty of Agriculture and Natural Resources, Rajamangala University of Technology Tawan-ok between January and March 2014. The cleaned explants were cultured in liquid Murashige and Skoog medium supplemented with 0, 2, 3, 4 and 5 mgl^-1 BA combined with 0 and 2 mgl^-1 IAA. Complete randomized design with 9 treatments and 7 replications was used in this experiment. The concentration of growth regulators in the medium highly significantly influenced to shoot number, while leaf number was significantly different. After 30 days of incubation, the result found that the maximum number of shoots (2.29 shoots per explant) was observed on medium with 5 mgl^-1BA + 2 mgl^-1IAA. The maximum number of leaf (9.43 leaves) was found on medium with free growth regulator. The concentration of growth regulators in the medium had no influence on shoot length.