Fang Tian1, Yanqin Ding, Hui Zhu, Liangtong Yao, FengLi Jin, Binghai Du. 1. College of Life Sciences, Shandong Agricultural University, Key Laboratory of Agricultural Microbiology, Shandong Agricultural University, Tai'an 271018, China. tianfang2001@126.com
Abstract
OBJECTIVE: To screen antagonistic bacteria from rhizosphere of tobacco against Phytophthora parasitica var. nicotianae (Breda de Hann) Tucker and to analyze its antagonistic activity. METHODS: The antagonistic bacteria were screened by dual culture with P. parasitica var. nicotianae (Breda de Hann) Tucker on low iron (2 micromol/L FeCl3) Sucrose-L-Asparagine (SA) plate. The chrome azurol S (CAS) assay was used to detect the siderophore production and its affinity to ferric ion. The strain was identified by using morphological, biochemical and physiological characteristics, 16S rRNA sequence homology, phylogenetics and specific species molecular analysis. The siderophore was isolated by column chromatography on Amberlite XAD-2 and analyzed by spectrophotometer. Antagonistic activity of the siderophore was confirmed by incubation of P. parasitica var. nicotianae (Breda de Hann) Tucker in liquid SA medium with different concentration of siderophore and FeCl3. RESULTS: We obtained an antagonistic bacterium G-229-21T against P. parasitica var. nicotianae (Breda de Hann) Tucker. The strain produced high-affinity siderophore under low iron conditions and was identified as Pseudomonas mediterranea. This strain produced carboxylate-type siderophore against P. parasitica var. nicotianae (Breda de Hann) Tucker. The suppression ratio was up to 92.3% under low iron conditions (0.16 micromol/L-10 micromol/L FeCl3). However, it was only 2.0% under Fe-replete conditions (100 micromol/L FeCl3). CONCLUSION: P. mediterranea G-229-21T could produce high-affinity carboxylate-type siderophore against P. parasitica var. nicotianae (Breda de Hann) Tucker under low iron conditions.
OBJECTIVE: To screen antagonistic bacteria from rhizosphere of tobacco against Phytophthora parasitica var. nicotianae (Breda de Hann) Tucker and to analyze its antagonistic activity. METHODS: The antagonistic bacteria were screened by dual culture with P. parasitica var. nicotianae (Breda de Hann) Tucker on low iron (2 micromol/L FeCl3) Sucrose-L-Asparagine (SA) plate. The chrome azurol S (CAS) assay was used to detect the siderophore production and its affinity to ferric ion. The strain was identified by using morphological, biochemical and physiological characteristics, 16S rRNA sequence homology, phylogenetics and specific species molecular analysis. The siderophore was isolated by column chromatography on Amberlite XAD-2 and analyzed by spectrophotometer. Antagonistic activity of the siderophore was confirmed by incubation of P. parasitica var. nicotianae (Breda de Hann) Tucker in liquid SA medium with different concentration of siderophore and FeCl3. RESULTS: We obtained an antagonistic bacterium G-229-21T against P. parasitica var. nicotianae (Breda de Hann) Tucker. The strain produced high-affinity siderophore under low iron conditions and was identified as Pseudomonas mediterranea. This strain produced carboxylate-type siderophore against P. parasitica var. nicotianae (Breda de Hann) Tucker. The suppression ratio was up to 92.3% under low iron conditions (0.16 micromol/L-10 micromol/L FeCl3). However, it was only 2.0% under Fe-replete conditions (100 micromol/L FeCl3). CONCLUSION:P. mediterranea G-229-21T could produce high-affinity carboxylate-type siderophore against P. parasitica var. nicotianae (Breda de Hann) Tucker under low iron conditions.