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[烟草根际产铁载体细菌G-229-21T的筛选、鉴定及拮抗活性]

[Screening, identification and antagonistic activity of a siderophore-producing bacteria G-229-21T from rhizosphere of tobacco].

作者信息

Tian Fang, Ding Yanqin, Zhu Hui, Yao Liangtong, Jin FengLi, Du Binghai

机构信息

College of Life Sciences, Shandong Agricultural University, Key Laboratory of Agricultural Microbiology, Shandong Agricultural University, Tai'an 271018, China.

出版信息

Wei Sheng Wu Xue Bao. 2008 May;48(5):631-7.

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.

摘要

目的

从烟草根际筛选对烟草黑胫病菌(Phytophthora parasitica var. nicotianae (Breda de Hann) Tucker)具有拮抗作用的细菌,并分析其拮抗活性。

方法

在低铁(2 μmol/L FeCl3)蔗糖 - L - 天冬酰胺(SA)平板上,通过与烟草黑胫病菌进行对峙培养筛选拮抗细菌。采用铬天青S(CAS)分析法检测铁载体的产生及其与铁离子的亲和力。利用形态学、生化和生理特性、16S rRNA序列同源性、系统发育学和特定物种分子分析对菌株进行鉴定。通过Amberlite XAD - 2柱色谱法分离铁载体,并用分光光度计进行分析。通过在含有不同浓度铁载体和FeCl3的液体SA培养基中培养烟草黑胫病菌来确认铁载体的拮抗活性。

结果

我们获得了一株对烟草黑胫病菌具有拮抗作用的细菌G - 229 - 21T。该菌株在低铁条件下产生高亲和力铁载体,被鉴定为地中海假单胞菌(Pseudomonas mediterranea)。该菌株产生针对烟草黑胫病菌的羧酸盐型铁载体。在低铁条件下(0.16 μmol/L - 10 μmol/L FeCl3)抑制率高达92.3%。然而,在铁充足条件下(100 μmol/L FeCl3)仅为2.0%。

结论

地中海假单胞菌G - 229 - 21T在低铁条件下可产生针对烟草黑胫病菌的高亲和力羧酸盐型铁载体。

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