Citation: ZHENG Xuefang, LIU Bo, ZHU Yujing, CHEN Deju. Optimization of rapid separation conditions of Ralstonia solanacearum using high performance ion-exchange chromatography[J]. Chinese Journal of Chromatography, ;2016, 34(11): 1091-1096. doi: 10.3724/SP.J.1123.2016.06044 shu

Optimization of rapid separation conditions of Ralstonia solanacearum using high performance ion-exchange chromatography

Agricultural Bio-Resources Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China

Received Date: 27 June 2016

Fund Project: Chinese Special Fund for Agro-Scientific Research in the Public Interest (No. 201303015)

Rapid separation of Ralstonia solanacearum with intact cell using high performance ion-exchange chromatography equipped with a UV detector was described. The bacterial growth abilities and cell morphologies in different suspension buffers were compared. The results showed that the bacterial numbers in the balance buffer (0.02 mol/L piperazine-hydrochloric acid), elution buffer (0.02 mol/L piperazine-hydrochloric acid+1 mol/L NaCl) and double distilled water were 6.467×10⁹, 6.267×10⁹ and 6.233×10⁹ cfu/mL, respectively, exhibiting no significant difference. Moreover, the cell morphologies were observed under a transmission electron microscope which showed that the bacterial cells could remain intact. Effects of buffer pH, flow velocity and bacterial cell concentration on the separation efficiency were analyzed. The results showed that the optimum chromatographic separation parameters were as follows: the buffer pH was 8, the flow velocity was 2 mL/min and the bacterial cell concentration was from 1.0×10⁸ cfu/mL to 1.0×10¹⁰ cfu/mL. Under the chromatographic separation conditions, the separation time was shortened and the separation efficiency was improved. Therefore, the chromatographic method developed here may be a promising tool for the rapid separation of Ralstonia solanacearum and other bacteria.
- high performance ion-exchange chromatography,
- separation,
- Ralstonia solanacearum,
- cell surface properties
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