植物有机营养研究中供试种子组合灭菌方法比较

Studies on combinative sterilization method to seed for plant organic nutrient experiments

植物有机营养试验中供试无菌苗通常由种子获得 ,因此 ,种子灭菌非常重要 .采用多种消毒剂( 70 %酒精 ,10 % H2 O2 ,2 %次氯酸钠和 0 .1% Hg Cl2 等 )对几种作物种子进行组合灭菌 .结果表明 ,灭菌效果与作物种类、种子贮存时间及不同消毒剂组合等有很大关系 .采用 70 %酒精浸 1min→无菌水冲3次→ 2 %次氯酸钠浸 5min→无菌水冲 5次→ 0 .1% Hg Cl2 浸 5min→无菌水冲 6次的组合灭菌程序 ,可取得较好的灭菌效果 ;若种子对 2 %次氯酸钠敏感 ,则可用 10 % H2 O2

Obtaining sterile seedlings from seeds is an important step for plant organic nutrition experiments. The plant seeds of Chinese cabbage, Pai tsai, green gram, rice and wheat were sterilized with different combinations of three sterilizing solutions (70% ethanol, 0.1% HgCl 2 and 2% NaClO or 10% H 2O 2) in the experiments. The results showed that the effectiveness of combinative sterilization to plant seed was affected by plant species and seed storage time. The combinative sterilization procedure recommended was the following. ①Seeds were firstly immersed in ethanol (70% V/V) for 1 min and then rinsed three times with sterile distilled water. ②Secondly immersed in sodium hypochlorite solution (2% NaClO active constituents) for 5 min and then rinsed five times with sterile distilled water. ③Finally immersed in mercuric chloride (0.1% HgCl 2 W/V) for 5 min and then rinsed with sterile distilled water six times. If the plant seeds were very sensitive to 2% NaClO, it would be advisable to use 10% H 2O 2 (v/v) instead of 2% NaClO.