Cloning and Conditions Optimization of the Siderophore Producing Gene for Burkholderia pyrrocinia JK-SH007

doi:10.16409/j.cnki.2095-039x.2019.04.001

Abstract

Abstract: Siderophore plays an important role in the plant growth promotion and plant disease control. The effective biocontrol strain Burkholderia pyrrocinia JK-SH007 was isolated from the poplar stem. In order to determine whether JK-SH007 strain can produce siderophore and reveal its biocontrol mechanism, the ability of the strain to produce siderophore was detected by CAS detection, the cepR gene of JK-SH007 was cloned and the sequence was analyzed. On the basis of single factor experiment, response surface method (RSM) was used to optimize the siderophore production conditions of JK-SH007 strain. The results showed that JK-SH007 strain has the capacity to produce siderophore, and the cepR gene size of JK-SH007 was 720 bp. The strain was most closely related to B. pyrrocinia (EU034001), with 99% homology, 97% the sequence similarity was and SNP at 13 sites. Compared with the cepR gene of B. pyrrocinia (EU034001), three amino acids encoded by cepR gene of JK-SH007 were found to be replaced and the consistency was 97%. The experiment of siderophore production showed that the optimal culture time for siderophore producing was 15 h, the pH value of the medium was 8, and the rotating speed was 200 r/min. The optimal carbon and nitrogen sources were glycerin and ammonium chloride. According to PB (Plackett-Burman) experiment, the key factors influencing siderophore producing were pH, carbon and nitrogen. CCD (short for Central Composite Design) experiment showed that the interaction between pH and nitrogen addition was obvious. The optimal solution of siderophore producing for JK-SH007 by Design-Expert was as follows:carbon source at 15.00 g/L, nitrogen source at 10.50 g/L, the culture temperature at 30℃ and the medium starting pH value of 7.36. After optimization, the siderophore production capacity of JK-SH007 increased from 18.59 to 37.86. The culture conditions obtained by RSM were effective and reliable, and the yield of siderophore improved significantly.

Key words: Burkholderia pyrrocinia, siderophore, cepR gene

CLC Number:

S476

CHEN Yanbin, HAN Yanyan, WANG Fei, YE Jianren, REN Jiahong. Cloning and Conditions Optimization of the Siderophore Producing Gene for Burkholderia pyrrocinia JK-SH007[J]. Chinese Journal Of Biological Control, 2019, 35(4): 630-641.

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