Native plants play a crucial role in landscape ecosystems due to their adaptability, yet propagating certain species through traditional methods can pose challenges. Tissue culture emerges as a promising alternative for efficient propagation, demanding an optimized culture medium for desirable proliferation. Efficient proliferation of newly introduced plant species in tissue culture often necessitates meticulous experimentation with varying salt compositions to optimize mineral concentrations and growth regulator levels within the culture medium. This study aimed to employ Response Surface Methods (RSM) to assess the effects of different mineral combinations, including nitrogen (N), calcium (Ca), potassium (K), and phosphorus (P), along with a cytokinin (zeatin), on the shoot proliferation and growth of little-leaf mockorange in tissue culture. Forty-six treatment combinations were designed, with zeatin concentrations of 0.82, 1.095, or 1.37 µM, and varying levels of N (22.5, 30, or 37.5 mM), Ca (1.125, 1.5, or 1.88 mM), P (0.31, 0.625, or 0.94 mM), and K (5, 10, or 15 mM). These treatments were evaluated for their impact on axillary shoot formation, shoot length, and shoot dry weight. Response surface analyses revealed optimal concentrations of N (32.5 to 35 mM), Ca (1.5 mM), and P (0.625 mM), while the influence of potassium on responses was found to be nonsignificant in the regression models, hence, K concentrations were limited to linear trends in the analysis. Notably, medium supplemented with 1.1 µM zeatin demonstrated a significant positive impact on shoot proliferation. The RSM model demonstrated the feasibility of determining optimal concentrations of zeatin, N, Ca, and P in a single experiment, facilitating the in vitro growth of little-leaf mockorange shoots. This study underscores the efficacy of RSM in tissue culture experiments, offering a time- and cost-effective alternative to conducting multiple separate experiments and providing valuable insights into medium optimization. Such findings contribute to the advancement of tissue culture techniques, particularly in propagating native plant species critical for landscape sustainability.
