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Quorum Quenching Approach for Control of Phytopathogen Pectobacterium carotovorum Subsp. Carotovorum.
紀錄類型:
書目-語言資料,印刷品 : Monograph/item
正題名/作者:
Quorum Quenching Approach for Control of Phytopathogen Pectobacterium carotovorum Subsp. Carotovorum./
作者:
Porusasp, Vesuna Ashtaad.
出版者:
Ann Arbor : ProQuest Dissertations & Theses, : 2022,
面頁冊數:
237 p.
附註:
Source: Dissertations Abstracts International, Volume: 84-08, Section: B.
Contained By:
Dissertations Abstracts International84-08B.
標題:
Microbiology. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30250483
ISBN:
9798374405750
Quorum Quenching Approach for Control of Phytopathogen Pectobacterium carotovorum Subsp. Carotovorum.
Porusasp, Vesuna Ashtaad.
Quorum Quenching Approach for Control of Phytopathogen Pectobacterium carotovorum Subsp. Carotovorum.
- Ann Arbor : ProQuest Dissertations & Theses, 2022 - 237 p.
Source: Dissertations Abstracts International, Volume: 84-08, Section: B.
Thesis (D.Phil.)--Maharaja Sayajirao University of Baroda (India), 2022.
This item must not be sold to any third party vendors.
Quorum sensing is N-acyl homoserine lactones (AHL) mediated cell-to-cell communication mechanism by which many pathogenic Bacteria sense their population density and control pathogenesis by regulating the production of major virulence factors. Phytopathogen such as Pectobacterium carotovorum subsp. carotovorum (Pcc) termed brute force pathogen uses Quorum sensing as a mechanism for regulation of its virulence. In Pectobacterium carotovorum subsp. carotovorum (Pcc) the virulence factors are secretory Plant Cell Wall Degrading Enzymes (PCWDEs) that macerate plant tissues and cause blackleg and soft rot diseases. Any process that inhibits quorum sensing without harming the bacteria is known as Quorum Quenching (QQ). Quorum quenching can be enzymatic or non-enzymatic depending on if AHL stability is affected or AHL production is controlled. A total of 79 bacterial isolates enriched and isolated from various sources were screened AHL degradation which is a requisite for quorum quenching. Two isolates viz. Glutamicibacter nicotianae AI5a and Rhodococcus pyridinivorans AI4 from the 79 were selected on basis of their biocontrol activity against PccBR1. An additional isolate Rhodococcus erythropolis CRD13.3C was used in the studies. The three isolates degraded AHL produced by PccBR1, decreased the enzyme activity of PCWDEs (PNL, PL and PGA) and attenuated and prevented soft rot in an in vitro assay with potato and cucumber as hosts. The QQ enzyme of G. nicotianae AI5a operated over wide temperature and pH range of 20°C to 37°C and 6.5 to 9, respectively. G. nicotianae AI5a did not grow on AHL as sole source of carbon in minimal media, the degraded AHL restored back to its original form on acidification and the C6-AHL showed 2 distinct product peaks when degraded upon HPLC analysis which pointed towards the enzyme being a putative lactonase.A cucumber in planta infection model was developed for phytopathogen Pcc. All 3 isolates G. nicotianae AI5a, R. pyridinivorans AI4 or R. erythropolis CRD13.3C reduced the pathogenicity of PccBR1 in in planta experiments. Further in planta study was undertaken with potato plant as host where three different types of infections were carried out: I) Soil inoculation II) Stem inoculation and III) Leaf and lateral stem inoculation. G. nicotianae AI5a was able to stop the pathogenicity of PccBR1 in all the three cases. Quantitative Real Time PCR was used to quantify G. nicotianae AI5a and PccBR1 pHC60. G. nicotianae AI5a was able to reduce soft rot symptoms emphasised the quorum quenching mechanism of biocontrol under storage conditions for various hosts such as potato, tomato, capsicum and brinjal without killing or affecting the growth of PccBR1, thus confirming its QQ ability. In a Mung bean in planta experiment, population of G. nicotianae AI5a were maintained and it was able to prevent the pathogenicity of PccBR1 without killing the pathogen. Similar trend was observed by qPCR in storage experiments.Phytochemicals are also reported to perform quorum quenching. Usually, they perform non-enzymatic quorum quenching which is also referred to as Quorum sensing inhibition (QSI). This mechanism hampers the production of AHL in a QS mediated pathogen. Twelve phytochemicals were screened for their AHL degradation ability at their respective sub-lethal concentrations against PccBR1. Three selected phytochemicals viz. Eugenol, Carvacrol and Salicylic acid were able to reduce the virulent traits of PccBR1 such as AHL production, motility, PCWDEs and biofilm formation ability at sub-lethal concentrations. The three phytochemicals were also efficient in reducing maceration in potato and cucumber host in in vitro soft rot attenuation assay. Under storage conditions, Eugenol, Carvacrol and Salicylic acid reduced the soft rot in potato host without killing PccBR1 which is a prerequisite for quorum quenching. An in planta experiment using mung bean as host under gnotobiotic conditions also showed that Eugenol, Carvacrol and Salicylic acid decreased the pathogenicity of PccBR1 without reducing the amount of PccBR1 in the plant. Thus, this study demonstrates the various aspects of Quorum quenching strategies using AHL degrading Actinomycetotal isolates G. nicotianae AI5a, R. pyridinivorans AI4 or R. erythropolis CRD13.3C and phytochemicals at sub-lethal concentrations to attenuate the virulence of quorum sensing dependent phytopathogen Pectobacterium carotovorum subsp. carotovorum and highlights their biocontrol potentials for further scale up usage.
ISBN: 9798374405750Subjects--Topical Terms:
591510
Microbiology.
Subjects--Index Terms:
Microbiology and Biotechnology Centre
Quorum Quenching Approach for Control of Phytopathogen Pectobacterium carotovorum Subsp. Carotovorum.
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Quorum sensing is N-acyl homoserine lactones (AHL) mediated cell-to-cell communication mechanism by which many pathogenic Bacteria sense their population density and control pathogenesis by regulating the production of major virulence factors. Phytopathogen such as Pectobacterium carotovorum subsp. carotovorum (Pcc) termed brute force pathogen uses Quorum sensing as a mechanism for regulation of its virulence. In Pectobacterium carotovorum subsp. carotovorum (Pcc) the virulence factors are secretory Plant Cell Wall Degrading Enzymes (PCWDEs) that macerate plant tissues and cause blackleg and soft rot diseases. Any process that inhibits quorum sensing without harming the bacteria is known as Quorum Quenching (QQ). Quorum quenching can be enzymatic or non-enzymatic depending on if AHL stability is affected or AHL production is controlled. A total of 79 bacterial isolates enriched and isolated from various sources were screened AHL degradation which is a requisite for quorum quenching. Two isolates viz. Glutamicibacter nicotianae AI5a and Rhodococcus pyridinivorans AI4 from the 79 were selected on basis of their biocontrol activity against PccBR1. An additional isolate Rhodococcus erythropolis CRD13.3C was used in the studies. The three isolates degraded AHL produced by PccBR1, decreased the enzyme activity of PCWDEs (PNL, PL and PGA) and attenuated and prevented soft rot in an in vitro assay with potato and cucumber as hosts. The QQ enzyme of G. nicotianae AI5a operated over wide temperature and pH range of 20°C to 37°C and 6.5 to 9, respectively. G. nicotianae AI5a did not grow on AHL as sole source of carbon in minimal media, the degraded AHL restored back to its original form on acidification and the C6-AHL showed 2 distinct product peaks when degraded upon HPLC analysis which pointed towards the enzyme being a putative lactonase.A cucumber in planta infection model was developed for phytopathogen Pcc. All 3 isolates G. nicotianae AI5a, R. pyridinivorans AI4 or R. erythropolis CRD13.3C reduced the pathogenicity of PccBR1 in in planta experiments. Further in planta study was undertaken with potato plant as host where three different types of infections were carried out: I) Soil inoculation II) Stem inoculation and III) Leaf and lateral stem inoculation. G. nicotianae AI5a was able to stop the pathogenicity of PccBR1 in all the three cases. Quantitative Real Time PCR was used to quantify G. nicotianae AI5a and PccBR1 pHC60. G. nicotianae AI5a was able to reduce soft rot symptoms emphasised the quorum quenching mechanism of biocontrol under storage conditions for various hosts such as potato, tomato, capsicum and brinjal without killing or affecting the growth of PccBR1, thus confirming its QQ ability. In a Mung bean in planta experiment, population of G. nicotianae AI5a were maintained and it was able to prevent the pathogenicity of PccBR1 without killing the pathogen. Similar trend was observed by qPCR in storage experiments.Phytochemicals are also reported to perform quorum quenching. Usually, they perform non-enzymatic quorum quenching which is also referred to as Quorum sensing inhibition (QSI). This mechanism hampers the production of AHL in a QS mediated pathogen. Twelve phytochemicals were screened for their AHL degradation ability at their respective sub-lethal concentrations against PccBR1. Three selected phytochemicals viz. Eugenol, Carvacrol and Salicylic acid were able to reduce the virulent traits of PccBR1 such as AHL production, motility, PCWDEs and biofilm formation ability at sub-lethal concentrations. The three phytochemicals were also efficient in reducing maceration in potato and cucumber host in in vitro soft rot attenuation assay. Under storage conditions, Eugenol, Carvacrol and Salicylic acid reduced the soft rot in potato host without killing PccBR1 which is a prerequisite for quorum quenching. An in planta experiment using mung bean as host under gnotobiotic conditions also showed that Eugenol, Carvacrol and Salicylic acid decreased the pathogenicity of PccBR1 without reducing the amount of PccBR1 in the plant. Thus, this study demonstrates the various aspects of Quorum quenching strategies using AHL degrading Actinomycetotal isolates G. nicotianae AI5a, R. pyridinivorans AI4 or R. erythropolis CRD13.3C and phytochemicals at sub-lethal concentrations to attenuate the virulence of quorum sensing dependent phytopathogen Pectobacterium carotovorum subsp. carotovorum and highlights their biocontrol potentials for further scale up usage.
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