假单胞菌HT66的PhzI-PhzR调控系统的功能研究

doi:10.3969/J.ISSN.1671-9964.2016.01.002

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Abstract

Pseudomonas chlororaphis HT66 is a significant growth-promoting rhizobacteriaum in plants.It is reportedly the highest phenazine-1-carboxamide(PCN)producing wild type microbe studied thus far. Based on the whole-genome sequence analysis, we find that two genes phzⅠ and phzR are located just upstream of the phenazine biosynthesis gene cluster.Plate assays with Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NTL4 were used to analyze the signals of strain HT66.With the addition of the fermentation broth extracts, strain NTL4 produced blue pigment, but the CV026 did not produce purple pigment.Then, using a gene knock-out method, we constructed ΔphzⅠ and ΔphzR HT66 mutants.The results show that both the ΔphzⅠ and ΔphzR mutants are unable to produce PCN.And the plate assay of ΔphzⅠ culture extracts shows no blue pigment is produced by strain NTL4.It suggests that phenazine biosynthesis is regulated by the quorum sensing system and AHL signals in strain HT66.Additionally, the ΔphzⅠ and ΔphzR mutants have some changes in the morphology of colonies and antifungal activity in vitro.Moreover, compared with wild type, the swimming motility of ΔphzⅠ mutant decreases greatly, while the swarming motility has no significant change.

Key words： Pseudomonas chlororaphis PhzI-PhzR system quorum sensing

Received: 25 March 2015 Published: 08 March 2016

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	Articles by authors
	Yi OUYANG
	Hua-song PENG
	Wei WANG
	Hong-bo HU
	Xue-hong ZHANG

Cite this article:

Yi OUYANG,Hua-song PENG,Wei WANG, et al. Function of thePhzI-PhzR System in Pseudomonas chlororaphis HT66[J]. Journal of Shanghai Jiaotong University (Agricultural Sciences), 2016, 34(1): 5-10.

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http://www.qk.sjtu.edu.cn/jsjtua/EN/10.3969/J.ISSN.1671-9964.2016.01.002 OR http://www.qk.sjtu.edu.cn/jsjtua/EN/Y2016/V34/I1/5

List of primers and sequences

Confirmation of ΔphzⅠ and ΔphzR mutants by PCR

Note:(a)M:DNA marker;1:ΔphzI mutant;2:strain HT66;(b)M:DNA marker;1 and 2:ΔphzR mutant;3:strain HT66

PCN production by strain HT66 and ΔphzⅠ, ΔphzR mutants

Detection of AHL signals by plate assaywith Agrobacterium tumefaciens NTL4

Note: 1: HT66;2:ΔphzⅠ mutant;3:acetonitrile

Swimming and swarming motility of strain HT66 and ΔphzI mutant

Colonial morphology change among strain HT66, ΔphzⅠ and ΔphzR

Effects of antifungal activity of HT66 and its mutants on the growth of Pythium ultimum, Pythium ultimum (left);HT66 and its mutants(right)

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