Development of a Method to Monitor Gene Expression in Single Bacterial Cells During the Interaction With Plants and Use to Study the Expression of the Type III Secretion System in Single Cells of Dickeya dadantii in Potato

Cui, Zhipeng; Yuan, Xiaochen; Yang, Ching‐Hong; Huntley, Regan B.; Sun, Weimin; Wang, Jie; Sundin, George W.; Zeng, Quan

doi:10.3389/fmicb.2018.01429

Cited by 14 publications

(18 citation statements)

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“…The abundance of Ea in each collected stigma sample was quantified by determining the cycle threshold (CT) value of the Ea specific gene amsC (12). Quantitative PCR (qPCR) was performed using a SsoAdvanced universal SYBR Green supermix (Bio-Rad, CA, USA), as described previously (13). The CT values for a 1/10 dilution series of known amsC gene copies of E. amylovora chromosomal DNA was determined to make a standard curve for calculation of copy numbers in stigma samples.…”

Section: Methodsmentioning

confidence: 99%

Temporal and spatial dynamics in the apple flower microbiome in the presence of the phytopathogenErwinia amylovora

Cui

Huntley

Zeng

et al. 2020

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17Plant microbiomes have important roles in plant health and productivity. However, 18 despite flowers being directly linked to reproductive outcomes, little is known about the 19 microbiomes of flowers and their potential interaction with pathogen infection. Here, we 20 investigated the temporal dynamics and spatial traits of the apple stigma microbiome 21 when challenged with a phytopathogen Erwinia amylovora, the causal agent of fire blight 22 disease. We profiled the microbiome from the stigmas of a single flower, greatly 23 increasing the resolution at which we can characterize shifts in the composition of the 24 microbiome. Individual flowers harbored unique microbiomes at the OTU level. 25However, taxonomic analysis of community succession showed a population gradually 26 dominated by bacteria within the families Enterobacteriaceae and Pseudomonadaceae. 27 Flowers inoculated E. amylovora established large populations of the phytopathogen, 28 with pathogen specific gene counts of >3.0 x 10 7 in 90% of the flowers. Yet, only 42% of 29 inoculated flowers later developed fire blight symptoms. This reveals pathogen amount 30 on the stigma is not sufficient to predict disease outcome. Our data demonstrate that 31 apple flowers represent an excellent model in which to characterize how plant 32 microbiomes establish, develop, and interact with biological processes such as disease 33 progression in an experimentally tractable plant organ.34 35 36 37 38 39 40 Flowers, the reproductive organs of angiosperms, play a critical role in the plant's 41 lifecycle. The most important function of flowers is to provide a mechanism for 42 pollination, the union of sperm contained within pollen, to the ovules contained in the 43 ovary. The fertilized ovules produce seeds that will later germinate to become the next 44 generation of plants. Yet, unlike other vegetative organs such as the roots, stems, and 45 leaves that are present through a large part of the plant's lifecycle, flowers develop on 46 mature plants and are typically present for the limited period during bloom. As such, 47 research characterizing the microbiome of the flower is generally less developed than for 48 other plant organs. 49 Flowers of apple (Malus domestica) have been subject to considerable research 50 attention as they are the direct precursors of apple fruits, one of the most consumed fruits 51 worldwide (1). The ephemeral nature of apple flowers, with mature flowers from petal 52 open to petal fall only lasting for 5-10 days in spring, offers a unique environment in 53 which to study community succession (1, 2). During bloom, petals open up in a relatively 54 short period of time, typically within one day, which exposes the internal flower parts to 55 the environment and microorganisms. Several of these internal flower parts exude various 56 types of nutrient-rich secretions including nectar, stigmatic exudate, and pollen exudate, 57 for the purpose of attracting pollinators, and inducing the germination of pollen grains (1, 58 3). These secretio...

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Section: Methodsmentioning

confidence: 99%

Temporal and spatial dynamics in the apple flower microbiome in the presence of the phytopathogenErwinia amylovora

Cui

Huntley

Zeng

et al. 2020

Preprint

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“…Heterogeneity may provide fitness advantages for a bacterial population as a whole [29,30]. Heterogeneity in virulence expression has been observed in other bacterial pathogens [10,11,16], where it was observed that expression of virulence genes imposes a penalty on bacterial growth rate [11]. However, the best-studied example of physiological heterogeneity is the phenomenon of antibiotic persistence.…”

Section: Discussionmentioning

confidence: 99%

“…Phenotypic heterogeneity increases the flexibility and versatility of bacteria as a population. Phenotypic heterogeneity has been observed in many bacteria in various functions such as antibiotic persistence [13], virulence [10,[14][15][16], motility [17], and sporulation [18,19].…”

Section: Many Bacterial Pathogens Have Been Observed To Exhibit Variamentioning

confidence: 99%

“…It is not subject to copyright under 17 USC 105 and is also Vegetative growth and virulence expression are two distinct physiological states for bacterial pathogens [7,8]. Virulence is induced under nutrient limited or stress conditions, whereas vegetative growth occurs under nutrient rich, growth favorable conditions [9,10]. For example, genes encoding the T3SS are induced when bacteria are cultured under a nutrient limited hrp-inducing minimal medium condition but were repressed in nutrient rich media such as Lysogeny Broth [9].…”

Section: Introductionmentioning

confidence: 99%

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Cell-length heterogeneity: a population-level solution to growth/virulence trade-offs in the plant pathogen Dickeya dadantii

Cui

Yang

Kharadi

et al. 2019

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Necrotrophic plant pathogens acquire nutrients from dead plant cells, which requires the disintegration of the plant cell wall and tissue structures by the pathogen. Infected plants lose tissue integrity and functional immunity as a result, exposing the nutrient rich, decayed tissues to the environment. One challenge for the necrotrophs to successfully cause secondary infection (infection spread from an initially infected plant to the nearby uninfected plants) is to effectively utilize nutrients released from hosts towards building up a large population before other saprophytes come. In this study, we observed that the necrotrophic pathogen Dickeya dadantii exhibited heterogeneity in bacterial cell length in an isogenic population during infection of potato tuber. While some cells were regular rod-shape (<10μm), the rest elongated into filamentous cells (>10μm). Short cells tended to occur at the interface of healthy and diseased tissues, during the early stage of infection when active attacking and killing is occurring, while filamentous cells tended to form when large amount of nutrients were released at a later stage of infection. Short cells expressed all necessary virulence factors and motility, whereas filamentous cells did not engage in virulence, were non-mobile and more sensitive to environmental stress. However, compared to the short cells, the filamentous cells displayed elevated metabolism and faster growth, which may benefit the pathogens to build up a large population necessary for the secondary infection. The segregation of the two subpopulations was dependent on differential expression of the alarmone guanosine tetraphosphate (ppGpp). When exposed to fresh tuber tissues or freestanding water, filamentous cells quickly transformed to short virulent cells. The pathogen adaptation of cell length heterogeneity identified in this study presents a model for how some necrotrophs balance virulence and vegetative growth to maximize fitness during infection.

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“…Vegetative growth and virulence expression are two distinct physiological states for bacterial pathogens [7, 8]. Virulence is induced under nutrient limited or stress conditions, whereas vegetative growth occurs under nutrient rich, growth favorable conditions [9, 10]. For example, genes encoding the T3SS are induced when bacteria are cultured under a nutrient limited hrp -inducing minimal medium condition but were repressed in nutrient rich media such as Lysogeny Broth [9].…”

Section: Introductionmentioning

confidence: 99%

Cell-length heterogeneity: a population-level solution to growth/virulence trade-offs in the plant pathogen Dickeya dadantii

et al. 2019

Self Cite

View full text Add to dashboard Cite

Necrotrophic plant pathogens acquire nutrients from dead plant cells, which requires the disintegration of the plant cell wall and tissue structures by the pathogen. Infected plants lose tissue integrity and functional immunity as a result, exposing the nutrient rich, decayed tissues to the environment. One challenge for the necrotrophs to successfully cause secondary infection (infection spread from an initially infected plant to the nearby uninfected plants) is to effectively utilize nutrients released from hosts towards building up a large population before other saprophytes come. In this study, we observed that the necrotrophic pathogen Dickeya dadantii exhibited heterogeneity in bacterial cell length in an isogenic population during infection of potato tuber. While some cells were regular rod-shape (<10μm), the rest elongated into filamentous cells (>10μm). Short cells tended to occur at the interface of healthy and diseased tissues, during the early stage of infection when active attacking and killing is occurring, while filamentous cells tended to form at a later stage of infection. Short cells expressed all necessary virulence factors and motility, whereas filamentous cells did not engage in virulence, were non-mobile and more sensitive to environmental stress. However, compared to the short cells, the filamentous cells displayed upregulated metabolic genes and increased growth, which may benefit the pathogens to build up a large population necessary for the secondary infection. The segregation of the two subpopulations was dependent on differential production of the alarmone guanosine tetraphosphate (ppGpp). When exposed to fresh tuber tissues or freestanding water, filamentous cells quickly transformed to short virulent cells. The pathogen adaptation of cell length heterogeneity identified in this study presents a model for how some necrotrophs balance virulence and vegetative growth to maximize fitness during infection.

show abstract

Development of a Method to Monitor Gene Expression in Single Bacterial Cells During the Interaction With Plants and Use to Study the Expression of the Type III Secretion System in Single Cells of Dickeya dadantii in Potato

Cited by 14 publications

References 35 publications

Temporal and spatial dynamics in the apple flower microbiome in the presence of the phytopathogenErwinia amylovora

Temporal and spatial dynamics in the apple flower microbiome in the presence of the phytopathogenErwinia amylovora

Cell-length heterogeneity: a population-level solution to growth/virulence trade-offs in the plant pathogen Dickeya dadantii

Cell-length heterogeneity: a population-level solution to growth/virulence trade-offs in the plant pathogen Dickeya dadantii

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