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荧光假单胞菌抗菌特性的检测及抗生素绿脓菌素的分子特征分析

Detection of antimicrobial traits in fluorescent pseudomonads and molecular characterization of an antibiotic pyoluteorin.

作者信息

Vinay J U, Naik M K, Rangeshwaran R, Chennappa G, Shaikh Sohel S, Sayyed R Z

机构信息

Department of Plant Pathology, University of Agricultural Science, Raichur, India.

Department of Microbiology, NBAIR, Bangalore, India.

出版信息

3 Biotech. 2016 Dec;6(2):227. doi: 10.1007/s13205-016-0538-z. Epub 2016 Oct 20.

Abstract

Thirty isolates of fluorescent pseudomonads were obtained from rhizosphere of different crops in Raichur, India. The fluorescent pseudomonad strains were characterized in vitro for biochemical traits, antimicrobial traits, and pyoluteorin antibiotic production. All the isolates that showed fluorescent pigment production under UV light were rod shaped, Gram negative, positive for oxidase, catalase and citrate utilization tests, and negative for indole test. Out of 30 isolates, 07 isolates were positive for HCN production, 15 isolates were positive for HS production, and all the isolates were positive for siderophore production. Among all the isolates, RFP-22 showed the maximum percent inhibition of mycelium (46.66 %) of Rhizoctonia solani, the pathogen, and the remaining isolates showed the moderate to least inhibition of mycelium growth of R. solani. The 16S rRNA analysis confirmed that the antibiotic positive isolates belonged to genus Pseudomonas. The amplification of 779 bp region in isolates RFP- 4 and RFP-19 corresponded to pyoluteorin antibiotic-coding pltB gene. Further characterization of pyoluteorin antibiotic through TLC and TOF-MS analysis confirmed the presence of pyoluteorin at 274.26 (g/mol) peak and 2.10 min retention time. Biochemical and molecular analyses confirmed the antagonism of Pseudomonas and isolate through pyoluteorin production.

摘要

从印度赖丘尔不同作物的根际土壤中分离出30株荧光假单胞菌。对这些荧光假单胞菌菌株进行了体外生化特性、抗菌特性及绿脓菌素抗生素产生情况的鉴定。所有在紫外光下产生荧光色素的分离株均为杆状、革兰氏阴性,氧化酶、过氧化氢酶和柠檬酸盐利用试验呈阳性,吲哚试验呈阴性。30株分离株中,7株HCN产生呈阳性,15株HS产生呈阳性,所有分离株铁载体产生均呈阳性。在所有分离株中,RFP - 22对病原菌立枯丝核菌的菌丝体抑制率最高(46.66%),其余分离株对立枯丝核菌菌丝体生长的抑制作用为中度至最低。16S rRNA分析证实,抗生素阳性分离株属于假单胞菌属。分离株RFP - 4和RFP - 19中779 bp区域的扩增对应于绿脓菌素抗生素编码的pltB基因。通过薄层色谱法(TLC)和飞行时间质谱(TOF - MS)分析对绿脓菌素抗生素进行进一步鉴定,证实了在274.26(g/mol)峰和2.10分钟保留时间处存在绿脓菌素。生化和分子分析证实了假单胞菌及分离株通过产生绿脓菌素具有拮抗作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6194/5073079/2a39be492581/13205_2016_538_Fig1_HTML.jpg

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