Characterization of novel DeoR-family member from the Streptomyces ahygroscopicus strain CK-15 that acts as a repressor of morphological development

Ge, Beibei; Liu, Yan; Liu, Binghua; Zhao, Wenran; Zhang, Kecheng

doi:10.1007/s00253-016-7661-y

Cited by 15 publications

(9 citation statements)

References 23 publications

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“…The nucleoside antibiotic wuyiencin is one such secondary metabolite; it is produced by Streptomyces ahygroscopicus var. wuyiensis, which was first isolated from the natural soil habitat of Wuyi Mountain in China [10]. After being industrially produced (COFCC-R-0903-0070), wuyiencin has been extensively used to control various fungal diseases in vegetables and crops and to enhance their resistance to diverse pathogens.…”

Section: Introductionmentioning

confidence: 99%

Genome-wide transcriptomic analysis of the response of Botrytis cinerea to wuyiencin

et al. 2020

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Grey mould is caused by the ascomycetes Botrytis cinerea in a range of crop hosts. As a biological control agent, the nucleoside antibiotic wuyiencin has been industrially produced and widely used as an effective fungicide. To elucidate the effects of wuyiencin on the transcriptional regulation in B. cinerea, we, for the first time, report a genome-wide transcriptomic analysis of B. cinerea treated with wuyiencin. 2067 genes were differentially expressed, of them, 886 and 1181 genes were significantly upregulated and downregulated, respectively. Functional categorization indicated that transcript levels of genes involved in amino acid metabolism and those encoding putative secreted proteins were altered in response to wuyiencin treatment. Moreover, the expression of genes involved in protein synthesis and energy metabolism (oxidative phosphorylation) and of those encoding ATP-binding cassette transporters was markedly upregulated, whereas that of genes participating in DNA replication, cell cycle, and stress response was downregulated. Furthermore, wuyiencin resulted in mycelial malformation and negatively influenced cell growth rate and conidial yield in B. cinerea. Our results suggest that this nucleoside antibiotic regulates all aspects of cell growth and differentiation in B. cinerea. To summarize, some new candidate pathways and target genes that may related to the protective and antagonistic mechanisms in B. cinerea were identified underlying the action of biological control agents.

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Section: Introductionmentioning

confidence: 99%

Genome-wide transcriptomic analysis of the response of Botrytis cinerea to wuyiencin

et al. 2020

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“…We identified that the wysR3 gene was also a transcriptional regulator belonging to the DeoR family, which had any significant function in sugar metabolism or antibiotic production except morphological development (Liu et al 2015). In addition, we found that the DwysR3 mutant strain can grow quickly to reach a plateau stage of maximum biomass at 60 h, which is 12 h faster than the wild-type strain (Ge et al 2016). As a result, we conducted this present study to evaluate the effect of overexpression of wysR in a DwysR3 mutant strain, and we explore a new method to breeding the genetic engineering strain combined the characteristics of fast morphological development and high yield of wuyiencin.…”

Section: Introductionmentioning

confidence: 92%

“…Positive colonies containing wysR gene fragments showed a 663 bp insert and were further selected for DNA preparation and sequencing. The resulting pSTE-wysR construct was then transferred from E. coli ET12567 (pUZ8002) to the DwysR3 mutant strain (Ge et al 2016) by intergeneric conjugation. The integrative plasmid pSETC cannot replicate in Streptomyces; so stable exconjugants should be obtained only if pSETC has been integrated into the chromosome of the recipient strain.…”

Section: Construction Of Wysr Overexpression Strainmentioning

confidence: 99%

Overexpression of wysR gene enhances wuyiencin production in ΔwysR3 mutant strain of Streptomyces albulus var. wuyiensis strain CK‐15

et al. 2020

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Aim: The aim of the present work was to investigate the overexpression of the wysR gene in Streptomyces albulus var. wuyiensis strain CK-15 based on the DwysR3 mutant strain including the effect on morphological development, wuyiencin production and antibacterial activity. At the same time, we report a new rapid method for producing genetically engineered strains for industrial production of wuyiencin. Methods and Results: We developed a method to create a wysR overexpression strain based on the DwysR3 mutant strain by direct transformation. In this method, the desired gene fragment to be overexpressed was amplified by polymerase chain reaction (PCR) using Phusion High Fidelity DNA polymerase and fused with the linearized pSETC integrative plasmid by Gibson assembly. The resulting recombinant plasmid was transformed into DwysR3 mutant strain by the intergeneric conjugation method. The plasmid was then integrated into the chromosome and the resulting apramycinresistant overexpression strain was confirmed by PCR using the Apra-F and Apra-R primers. Finally, we successfully screened the genetically engineered strain with overexpression of wysR gene in DwysR3 mutant. Conclusion: We can conclude that overexpression of wysR gene in DwysR3 mutant strain proved to be an effective strategy for significantly increasing wuyiencin production together with faster morphological development. Quantitative real-time RT-PCR analysis showed that wysR regulated wuyiencin biosynthesis by modulating other putative regulatory genes and bld, whi, chp, rdl and ram family genes are crucial for the morphological development. Significance and Impact of the Study: Overexpression of wysR gene in the DwysR3 mutant strain named OoWysR strain may increase the efficiency in the industrial fermentation processes for wuyiencin production. The mechanism by which wysR overexpression promotes rapid sporulation and a high yield of wuyiencin production is likely related to modulation of other putative regulatory genes.

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“…Pathway-specific transcriptional regulation, which is the most fundamental level, is mediated by regulatory genes encoded within the respective biosynthetic gene clusters ( Vicente et al, 2015 ). Some regulatory factors affect antibiotic production, while others affect morphological development and production of antibiotics via processes that are sometimes genetically coupled by shared transcriptional regulators ( Hur et al, 2008 ; Ge et al, 2016 ). A few regulatory factors are involved only in morphological development, an example being MtrA, which is crucial for normal development of aerial hyphae of the model strain S. coelicolor ( Cui et al, 2016 ; Zhang et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

“…WysR3, a novel member of the DeoR family of regulators, had no effect on antibiotic production, but acted as a repressor during morphological development. The Δ wysR3 strain grew faster than the WT, reaching the plateau stage of maximum biomass 12 h earlier in the flasks ( Ge et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Identification of wysPII as an Activator of Morphological Development in Streptomyces albulus CK-15

Liu

et al. 2018

Front. Microbiol.

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Wuyiencin is produced by Streptomyces albulus var. wuyiensis and used widely in agriculture to control a variety of fungal diseases, such as cucumber downy mildew, strawberry powdery mildew, and tomato gray mold. As an industrially-produced biopesticide, reducing production costs is very important for popularization of this approach. To obtain a rapidly growing strain that effectively shortens the fermentation time, we investigated the effects of knockout and overexpression of the wysPII gene, a member of the LuxR regulatory gene family, in S. albulus strain CK-15. The ΔwysPII mutant exhibited a reduced rate of growth and sporulation. The time taken to reach the greatest mycelial biomass was approximately 18 h shorter in the ooPII (wysPII overexpressing) strain compared with that of the wild-type (WT) strain. In addition, the time to reach the greatest wuyiencin production was 56 h in the ooPII strain compared with 62 h in the WT strain. Furthermore, wysPII was shown to act as an activator of morphological development without affecting wuyiencin production. Thus, the ooPII strain can be used to reduce costs and increase efficiency in industrial fermentation processes for wuyiencin production.

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Characterization of novel DeoR-family member from the Streptomyces ahygroscopicus strain CK-15 that acts as a repressor of morphological development

Cited by 15 publications

References 23 publications

Genome-wide transcriptomic analysis of the response of Botrytis cinerea to wuyiencin

Genome-wide transcriptomic analysis of the response of Botrytis cinerea to wuyiencin

Overexpression of wysR gene enhances wuyiencin production in ΔwysR3 mutant strain of Streptomyces albulus var. wuyiensis strain CK‐15

Identification of wysPII as an Activator of Morphological Development in Streptomyces albulus CK-15

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