Engineering <i>Pseudomonas protegens</i> Pf‐5 to improve its antifungal activity and nitrogen fixation

Jing, Xuyang; Cui, Qingwen; Li, Xiaochen; Yin, Jia; Ravichandran, Vinothkannan; Deng, Peng; Fu, Jun; Tu, Qiang; Wang, Hailong; Bian, Xiaoying; Zhang, Youming

doi:10.1111/1751-7915.13335

Cited by 50 publications

(47 citation statements)

References 61 publications

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“…Analysis conducted when the plants were uprooted showed that the P. protegens MP12 strain is able to colonize the rhizosphere. This property appears to be consistent with the origin of P. protegens MP12 [ 18 ] and the previous identification of other root-colonizing strains from the same species [ 42 ]. This strain’s ability to colonize the rhizoplane underlines the importance of the isolation source in the selection of efficient and beneficial plant bacteria.…”

Section: Discussionsupporting

confidence: 89%

“…The ability of the MP12 strain to colonize grapevines grown in vitro and vine cuttings suggests that this strain may be able to completely partially avoid a plant’s defensive system. On the other hand, the presence of potentially pathogenic bacterial strains in the vine nursery soil [ 42 ] may trigger a plant’s defensive system and mechanisms [ 36 ], thereby also affecting the persistence of the MP12 strain in plants.…”

Section: Discussionmentioning

confidence: 99%

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In Vivo Endophytic, Rhizospheric and Epiphytic Colonization of Vitis vinifera by the Plant-Growth Promoting and Antifungal Strain Pseudomonas protegens MP12

et al. 2021

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An evaluation was conducted of the colonization of Pseudomonas protegens MP12, a plant-growth promoting and antagonistic strain, inoculated in vine plants during a standard process of grapevine nursery propagation. Three in vivo inoculation protocols (endophytic, rhizospheric, and epiphytic) were implemented and monitored by means of both culture-dependent and independent techniques. Endophytic treatment resulted in the colonization of the bacterium inside the vine cuttings, which spread to young leaves during the forcing period. Microscopy analysis performed on transformed dsRed-tagged P. protegens MP12 cells confirmed the bacterium’s ability to penetrate the inner part of the roots. However, endophytic MP12 strain was no longer detected once the plant materials had been placed in the vine nursery field. The bacterium also displayed an ability to colonize the rhizosphere and, when the plants were uprooted at the end of the vegetative season, its persistence was confirmed. Epiphytic inoculation, performed by foliar spraying of cell suspension, was effective in controlling artificially-induced Botrytis cinerea infection in detached leaves. The success of rhizospheric and leaf colonization in vine plants suggests potential for the future exploitation of P. protegens MP12 as biofertilizer and biopesticide. Further investigation is required into the stability of the bacterium’s colonization of vine plants under real-world conditions in vineyards.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

In Vivo Endophytic, Rhizospheric and Epiphytic Colonization of Vitis vinifera by the Plant-Growth Promoting and Antifungal Strain Pseudomonas protegens MP12

et al. 2021

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“…Engineering microbial agents for the biocontrol of plant diseases can provide new ways to use natural microorganisms as microbial inoculums, thereby can be used as an eco‐friendly alternative to pesticides (Jing et al ., 2020). Inactivation of the negatively‐acting transcriptional regulator or sensor kinase enhanced the production of antimicrobial secondary metabolites, which contributed to improving the antimicrobial activity to act as a biocontrol agent (Kim et al ., 2003; Jing et al ., 2020). In this study, the biological control effect on rice grain rot using Txn‐degrading bacteria was not evaluated, so future studies on plant disease control incorporating microbial biotechnology are needed.…”

Section: Discussionmentioning

confidence: 99%

A novel toxoflavin‐quenching regulation in bacteria and its application to resistance cultivars

Choi

Lee

Park

et al. 2021

Microbial Biotechnology

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The toxoflavin (Txn), broad host range phytotoxin produced by a variety of bacteria, including Burkholderia glumae, is a key pathogenicity factor of B. glumae in rice and field crops. Two bacteria exhibiting Txn-degrading activity were isolated from healthy rice seeds and identified as Sphingomonas adhaesiva and Agrobacterium sp. respectively. The genes stdR and stdA, encoding proteins responsible for Txn degradation of both bacterial isolates, were identical, indicating that horizontal gene transfer occurred between microbial communities in the same ecosystem. We identified a novel Txnquenching regulation of bacteria, demonstrating that the LysR-type transcriptional regulator (LTTR) StdR induces the expression of the stdA, which encodes a Txn-degrading enzyme, in the presence of Txn as a coinducer. Here we show that the bacterial StdR Txn -quenching regulatory system mimics the ToxR Txn -mediated biosynthetic regulation of B. glumae. Substrate specificity investigations revealed that Txn is the only coinducer of StdR and that StdA has a high degree of specificity for Txn. Rice plants expressing StdA showed Txn resistance. Collectively, bacteria mimic the mechanism of Txn biosynthesis regulation, employ it in the development of a Txn-quenching regulatory system and share it with neighbouring bacteria for survival in rice environments full of Txn.

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“…Many Pseudomonads are PGPR and have been used in commercial fertilizer products (e.g. P. fluorescens 41,42 and P. protegens 43,44 ). Development of high-throughput genetic tools to engineer additional functionality or improve performance of Pseudomonad and other PGPR isolates has substantially lagged behind those of related strains used for industrial fuels and chemicals production (e.g.…”

Section: Development Of a Conditional-replication Plasmid For Transiementioning

confidence: 99%

The SAGE genetic toolkit enables highly efficient, iterative site-specific genome engineering in bacteria

Elmore

Francis

et al. 2020

Preprint

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Sustainable enhancements to crop productivity and increased resilience to adverse conditions are critical for modern agriculture, and application of plant growth promoting rhizobacteria (PGPR) is a promising method to achieve these goals. However, many desirable PGPR traits are highly regulated in their native microbe, limited to certain plant rhizospheres, or insufficiently active for agricultural purposes. Synthetic biology can address these limitations, but its application is limited by availability of appropriate tools for sophisticated, high-throughput genome engineering that function in environments where selection for DNA maintenance is impractical. Here we present an orthogonal, Serine-integrase Assisted Genome Engineering (SAGE) system, which enables iterative, site-specific integration of up to 10 different DNA constructs at efficiency on par or better than replicating plasmids. SAGE does not require use of replicating plasmids to deliver recombination machinery, and employs a secondary serineintegrase to excise and recycle selection markers. Furthermore, unlike the widely utilized pBBR1 origin, DNA transformed using SAGE is stable without selection. We highlight SAGE's utility by constructing a 287-member constitutive promoter library with a ~40,000-fold dynamic range in P. fluorescens SBW25. We show that SAGE functions robustly in diverse ⍺and "-proteobacteria, thus providing evidence that it will be broadly useful for engineering industrial or environmental bacteria.

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Engineering Pseudomonas protegens Pf‐5 to improve its antifungal activity and nitrogen fixation

Cited by 50 publications

References 61 publications

In Vivo Endophytic, Rhizospheric and Epiphytic Colonization of Vitis vinifera by the Plant-Growth Promoting and Antifungal Strain Pseudomonas protegens MP12

In Vivo Endophytic, Rhizospheric and Epiphytic Colonization of Vitis vinifera by the Plant-Growth Promoting and Antifungal Strain Pseudomonas protegens MP12

A novel toxoflavin‐quenching regulation in bacteria and its application to resistance cultivars

The SAGE genetic toolkit enables highly efficient, iterative site-specific genome engineering in bacteria

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