The Preliminary Research On Microtubers Of Sagittaria Trifolia Linn. Var. Sinensis Makino

The shoot tips of Sagittaria trifolia Linn. var. sinensis Makino were used as explant in the study to establish an effective mass propagation technique of tissue culture. We gained the favorable induction medium for shoot tips, and studied the best plant growth regulator proportion for regeneration medium. Meanwhile, we preliminarily studied the major influence factors to microtubers of Sagittaria trifolia Linn. var. sinensis. The main results were as the following:1. The best surface sterilization method for inoculation shoot tips was immersed in 72% streptomycin sulphate for 20 minutes, then dipped in 70% ethanol for 30s, and finally agitated in 0.1% HgCl₂ for 2 minutes. The effect was excellent and the survival rate could up to 53.7% and the pollution rate reduced to 43.9%.2. Effective rapid propagation system in vitro of Chinese Arrowhead was set up. Through regulating the concentration of 6-BA and NAA in MS medium, We found out that the favorable induction medium for shoot tips was MS + 6-BA 1.0 mg/L + NAA 0.3mg/L, and the differentiation rate was 88.1%; The optimum medium for regeneration was MS + 6-BA 3.0 mg/L + NAA 0.1mg/L, and the best multiplicative period was 6 weeks, the propagation coefficient could up to 8.87.3. Through orthogonal experiment, carried on research on sucrose concentration. 6-BA:NAA, temperature and illumination time 4 factors to the influence of height of plantlet, microrhizome and microtuber. The results showed that only sucrose concentration could influence to height of plantlet significantly, and with the rising of sucrose concentration, height of plantlet presented the downward trend; Both sucrose concentration and temperature 2 factors could influence to number of microrhizome per plantlet significantly, when we used 30g/L or 60g/L sucrose concentration and 22℃temperature, we got much more microrhizome; All of 4 factors could influence to ratio of microtuber and number of microtuber per plantlet significantly, and the combination of sucrose concentration=60g/L, 6-BA:NAA=0:0, temperature=22℃, illumination time =0h was the best combination to induce tubers in vitro of Sagittaria trifolia Linn. var. sinensis Makino, then we could gain the highest value of both ratio of microtuber and number of microtuber per plantlet. 4. Transplantation experiment of planlets showed that the fresh weight on per tuber formed by plantlets increased 57.63% compared with by seedlings.