| Summary: | 碩士 === 國立屏東科技大學 === 植物醫學系所 === 104 === In order to establish the suspension cells of rice that have the good abilities of proliferation and plant differentiation, we evaluated effects of the source and texture of callus, repeated establishment culture of suspension cell, medium and the plant growth regulator on proliferation and plant differentiation of suspension cell.Compared effect of organ explants (the seeds and root fragments) on formation rate and texture of callus. The two explants differ slightly in callus formation rate, both up to 90%. But varied on callus texture, all callus induced from the seed were harden nodular; callus induced from the root fragments had three kinds of callus, friable, harden nodular and sticky. In comparison of the number of suspension cell among the different texture of callus, callus induced from the root fragments better than the seeds in the number of suspension cell, which were 2.2×107 and 6.5×106 per gram callus. However, there’s no difference in cell proliferation, and both of them couldn’t continue to proliferate. Further improve the proliferation rate of suspension cell, callus induced from the root fragments proceed repeated establishment culture 1-6 times and subculture respectively. The result showed that repeated establishment culture could raise the proliferation rate of
suspension cell. Among these treatments, repeated establishment culture three times provided the best result in proliferation rate of suspension cell, which were 262.2% (1.6-folded) and 733.3% (6.3-folded) in primary culture and subculture. In the process of subculture, the suspension cells, suspended from the friable and harden nodular callus, differentiated to minicalli easily. The suspension cells repeated establishment culture twice had the best result in differentiation rate of minicalli. The minicalli number of friable callus and harden nodular callus in the first subculture and the second subculture were 9,418 and 2,758, and 8,215 and 2,729 respectively in 50 mL medium. Both of the minicalli could differentiate to plants, the plant differentiation rate about 8-27.7%. Summarize the above-mentioned, the source and texture of callus and repeated establishment culture of suspension cell had a great influence on the abilities of proliferation and plant differentiation of the suspension cells. The best culture conditions established in this study were：(1) In subculture of suspension cell, callus induced from the root fragments offered as the materials, repeated establishment cultured three times on CS-1 medium containing 2 mg/L NAA to establish the suspension cells and transferred to CSW-1 medium, the times of cell proliferation was the highest, which about 1.6-6.3 times. (2) In minicalli differentiation, the suspension cells suspended from the callus, induced from the root fragments, repeated establishment culture twice to proceed subculture, more than 8,215 minicalli differentiated in 50 mL medium. (3) In plant differentiation of minicalli, 10 mg/L NAA or 3 mg/L BA or 2 mg/L NAA and 4 mg/L Kinetin provided the best result.
|
|---|
