Effects of Cell Source and Culture Method on the Proliferation of Suspension Cells in Oryza sativa var. japonica ‘Taiken 9’

• Main Authors: Wen, Min-Chi, 溫敏琪
• Other Authors: Cheng, Chiu-Chsiung
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/87259955329350277269

碩士 === 國立屏東科技大學 === 植物醫學系所 === 102 === The purpose of this study was to establish the stable culture technology of suspension cells in rice. In order to get the suspension cells that have good abilities in proliferation and differentiation, the mature seeds and the root of in vitro seedling in Oryza sativa var. japonica cv. Taiken 9 were used as the materials. In comparison of the number and proliferation rate of suspension cells of callus induced from the seed and the root. Callus induced from the root had the best result in number of suspension cells per gram callus, about 2.4#westeur024#107cells/ml, which was 2.2-fold higher than the callus induced from the seed. Callus were grouped based on the texture, harden nodule callus (induced from seed), friable callus (induced from root) and sticky callus (induced from root). Compared proliferation rate of the suspension cells, induced from the seed and the root, after primary culture 8 days, which were 34% and -6% respectively, the result indicated that the culture cells induced from the root better than from the seed. Repeated establishment culture could raise proliferation rate of the suspension cell induced from the seed and the root in primary culture, in addition, it could slow down the decrease in the number of suspension cells in the period of subculture. In comparison of proliferation rate of the suspension cells among harden nodule callus (induced from seed), friable callus and sticky callus (induced from root). Sticky callus was the best, the primary culture and the first subculture were 90.6% and 15.2% respectively. When these suspension cells continued to culture the second to the fourth subculture, which could differentiated a vast amount of minicalli, there were 1700, 2600 and 1065 minicalli per 50 ml respectively. It’s found that the suspension cells induced from the sticky callus had the good ability in differentiation. Summarize the above-mentioned, these results indicated that source of the culture cell (organ explant and callus texture), cell density and times of establishment culture had a great influences on the abilities of proliferation and differentiation of the suspension cells. In this study, it’s found that (1) the root section (involve the root tip) of in vitro seedling, 10-day-old after germination, offered as the explants (1 cm), and (2) the induced sticky callus that cultured on CS-1 medium containing 2 mg/l 2,4-D at dark for 14 days as the source of culture cells were the best culture conditions, could obtain the suspension cells that have the good abilities in proliferation and differentiation in Oryza sativa var. japonica cv. Taiken 9.