Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama

Coy, Monique R.; Hoffmann, M.; Gibbard, H. N. Kingdom; Kuhns, Emily H.; Pelz‐Stelinski, Kirsten S.; Stelinski, Lukasz L.

doi:10.1016/j.mimet.2014.04.007

Cited by 53 publications

(43 citation statements)

References 19 publications

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“…Methods reported in the literature for assessing incidence of “ Ca . L. asiaticus” in D. citri and plants vary but most over the past 10–15 years have involved standard, real‐time or nested PCR assays with various procedures including different PCR primer targets (e.g., Coy et al., ; Li et al., ; Morgan et al., ; Stover, Hall, Shatters, & Moore, ; Ukuda‐Hosokawa et al., ). Some PCR procedures may be less sensitive for “ Ca .…”

Section: Resultsmentioning

confidence: 99%

Incidence of “Candidatus Liberibacter asiaticus” in a Florida population of Asian citrus psyllid

Hall

2017

J Applied Entomology

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Section: Resultsmentioning

confidence: 99%

Incidence of “Candidatus Liberibacter asiaticus” in a Florida population of Asian citrus psyllid

Hall

2017

J Applied Entomology

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“…The concentration and purity of the extracted DNA was measured spectrophotometrically using a Nano Drop 2000 (Thermo Fisher Scientific, Waltham, MA). The presence of C Las was assessed by the detection of the 16S rDNA gene by quantitative polymerase chain reaction (qPCR) [52]. In brief, a multiplex Taq Man (Applied Biosystems, Foster City, CA) assay targeting the 16S rDNA of C Las and the D .…”

Section: Methodsmentioning

confidence: 99%

“…The amount of C Las was determined according to the assessed gene copy numbers by calculating the 16S/Wg ratio. The concentrations of reagents, plasmid construction, and reaction protocol are described in Coy et al [52]. …”

Section: Methodsmentioning

confidence: 99%

Infection of an Insect Vector with a Bacterial Plant Pathogen Increases Its Propensity for Dispersal

et al. 2015

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The spread of vector-transmitted pathogens relies on complex interactions between host, vector and pathogen. In sessile plant pathosystems, the spread of a pathogen highly depends on the movement and mobility of the vector. However, questions remain as to whether and how pathogen-induced vector manipulations may affect the spread of a plant pathogen. Here we report for the first time that infection with a bacterial plant pathogen increases the probability of vector dispersal, and that such movement of vectors is likely manipulated by a bacterial plant pathogen. We investigated how Candidatus Liberibacter asiaticus (CLas) affects dispersal behavior, flight capacity, and the sexual attraction of its vector, the Asian citrus psyllid (Diaphorina citri Kuwayama). CLas is the putative causal agent of huanglongbing (HLB), which is a disease that threatens the viability of commercial citrus production worldwide. When D. citri developed on CLas-infected plants, short distance dispersal of male D. citri was greater compared to counterparts reared on uninfected plants. Flight by CLas-infected D. citri was initiated earlier and long flight events were more common than by uninfected psyllids, as measured by a flight mill apparatus. Additionally, CLas titers were higher among psyllids that performed long flights than psyllid that performed short flights. Finally, attractiveness of female D. citri that developed on infected plants to male conspecifics increased proportionally with increasing CLas bacterial titers measured within female psyllids. Our study indicates that the phytopathogen, CLas, may manipulate movement and mate selection behavior of their vectors, which is a possible evolved mechanism to promote their own spread. These results have global implications for both current HLB models of disease spread and control strategies.

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“…The presence of CLas was assessed via the detection of the 16S rRNA gene by qPCR, according to Pelz-Stelinski et al (2010). CLas-specific 16S rDNA (modified from Coy et al, 2014) from plant extracts was amplified using probe-primer sets targeting internal control sequences specific to plant cytochrome oxidase gene regions. Quantitative PCR settings were as follows: (i) 2 min at 50 ∘ C; (ii) 10 min at 95 ∘ C; and (iii) 40 cycles with 15 s at 95 ∘ C and 60 s at 60 ∘ C (data collection).…”

Section: Detection Of Clas In Plant Samplesmentioning

confidence: 99%

Drought stress affects response of phytopathogen vectors and their parasitoids to infection‐ and damage‐induced plant volatile cues

Martini

Stelinski

2017

Ecological Entomology

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1. The response of a phytopathogen vector to pathogen‐induced plant volatiles was investigated, as well as the response of the phytopathogen vector's parasitoid to herbivore‐induced plant volatiles released from plants with and without drought stress. 2. These experiments were performed with Asian citrus psyllid (Diaphorina citri), vector of the plant pathogen Candidatus Liberibacter asiaticus (CLas) and its parasitoid Tamarixia radiata as models. Candidatus Liberibacter asiaticus is the presumed causal pathogen of huanglongbing (HLB), also called citrus greening disease. 3. Diaphorina citri vectors were attracted to headspace volatiles of CLas‐infected citrus plants at 95% of their water‐holding capacity (WHC); such attraction to infected plants was much lower under drought stress. Attraction of the vector to infected and non‐stressed plants was correlated with greater release of methyl salicylate (MeSA) as compared with uninfected and non‐stressed control citrus plants. Drought stress decreased MeSA release from CLas‐infected plants as compared with non‐stressed and infected plants. 4. Similarly, T. radiata was attracted to headspace volatiles released from D. citri‐infested citrus plants at 95% of their WHC. However, wasps did not show preference between headspace volatiles of psyllid‐infested and uninfested plants when they were at 35% WHC, suggesting that herbivore‐induced defences did not activate to recruit this natural enemy under drought stress. 5. Our results demonstrate that herbivore‐ and pathogen‐induced responses are environmentally dependent and do not occur systematically following damage. Drought stress affected both pathogen‐ and herbivore‐induced plant volatile release, resulting in concomitant decreases in behavioural response of both the pathogen's vector and the vector's primary parasitoid.

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Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama

Cited by 53 publications

References 19 publications

Incidence of “Candidatus Liberibacter asiaticus” in a Florida population of Asian citrus psyllid

Incidence of “Candidatus Liberibacter asiaticus” in a Florida population of Asian citrus psyllid

Infection of an Insect Vector with a Bacterial Plant Pathogen Increases Its Propensity for Dispersal

Drought stress affects response of phytopathogen vectors and their parasitoids to infection‐ and damage‐induced plant volatile cues

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