离子交换色谱法分离青枯菌及其色谱峰的鉴定

 采用离子交换色谱法对世界上分布最广、危害最重且最难防治的植物致病菌青枯菌进行了色谱表征。在以0.02 mol/L哌嗪-盐酸缓冲液(pH 8.0)为流动相、梯度洗脱条件下,青枯菌在SuperQ-650 C色谱柱(200 mm×4.6 mm i.d.)上被分离为3个不同的色谱峰。根据这3个组分的形态和生理生化性质,确定青枯菌原液和分离得到的3个组分都属于青枯菌的生化型Ⅲ型。通过2,3,4-三苯基氯化四氮唑(TTC)平板鉴定两个洗脱峰菌体的致病性及电镜观察二者的运动性,发现第一洗脱峰菌体的致病性弱于第二洗脱峰菌体而运动性强于后者。实验表明,通过离子交换色谱可以把常规微生物方法无法分离的青枯菌的不同状态区分出来。分离结果将为阐明青枯菌的多态性及传代培养过程中致病性逐渐减弱的内在机理,最终研制出特效的青枯菌抗菌剂具有重要意义。

Characterization and Identification of Ralstonia solanacearum by Ion-Exchange Chromatography

Ralstonia solanacearum, a widely distributed and economically important plant pathogen, was characterized by ion-exchange chromatography (IEC). Ralstonia solanacearum was chromatographed on a TOYOPEARL SuperQ-650 C column (200 mm x 4. 6 mm i. d. ) with gradient elution by A (0.02 mol/L piperazidine-chlorhydric acid buffer (pH 8.0) ) and B (A + 1 mol/L NaCl). The pure culture of R. solanacearum was separated into three fractions on a SuperQ-650 C column. They were found all belong to R. solanacearum after the fractions were identified by other biochemical methods. Because of their ability to oxidize 3 disaccharides (lactose, maltose and cellobiose) and 3 hexose alcohols (mannitol, sorbitol and dulcitol), they are classified as biovar III of R. solanacearum. Once the mobility was scaled using microscope and the pathogenetic ability was measured with 2,3 ,4-triphenyltetrazolium chlorid (TTC) medium, the two fractions were in different states. The results are very important to elucidate the multi-states of R. solanacearum and the mechanism of R. solanacearum's pathogenetic mutation.