采用正交设计快速获得梨无菌外植体的研究

Studies on Getting Bacteria-free Explants of Pear by Applying Orthogonal Design

以“雪青”梨为材料 ,采用L9(34)正交设计 ,设外植体 (A)、消毒剂 (B)和消毒时间 (C) 3因素 ,每因素设 3个水平 :A分别为叶片、茎尖和节段 ;B分别为 2 %次氯酸钠、0 1%升汞和 2 %次氯酸钙 ;C分别为 10min ,5min和 5min .外植体经表面消毒后 ,接种在MS琼脂培养基中 ,于 (2 5± 1)℃ ,2 0 0 0lx光照培养 ,15d后统计成活率 .试验结果表明 :最优水平组合为A1B2 C1,A ,B对试验结果的影响达显著水平 (p≤ 0 0 5 ) ,A不同水平间差异显著 (p<0 0 5 ) .

The pear ‘Xueqing’ was used as test material in this paper, test method was orthogonal design L_9 (3~4). Three factors were explants (A), disinfectants (B) and sterilizing times (C). Explants were leaves, shoot tips and nodes with axillary buds, disinfectants were 2% sodium hypochlorite, 0.1% HgCl_2 and 2% calcium hypochlorite, sterilizing times were 10 min, 15 min and 5 min. Explants were inoculated on MS base medium (supplied with 8 g·L^(-1) agar and 30 g·L^(-1) sucrose, pH=5.8) after them were sterilized, and cultured under (25±1) ℃ and 2 000 lx conditions, then the explants were compiled surviving rate after cultured 15 days. The results showed that the best level combination was A_1B_2C_1, factor A and factor B influenced explants survive significantly (p≤0.05), there was significant difference among different levels of factor A at<0.05 level.