Evaluation of Methods for Extracting Xylella fastidiosa DNA from the Glassy-Winged Sharpshooter

Bextine, Blake; Tuan, S.-J.; Shaikh, Harris; Blua, Matthew J.; Miller, Thomas A.

doi:10.1093/jee/97.3.757

Cited by 18 publications

(13 citation statements)

References 30 publications

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“…There were 1,601 insects dissected to remove eyes, legs, abdomen, and wings under Stereo Star (Reichert ScientiÞc Instruments) dissecting scope to prevent potential PCR inhibition in the following procedures. Each specimen was placed in a 2 ml screw top sterilized tube preloaded with two 5 mm glass beads and submerged in liquid nitrogen for 20 s immediately before being ground using a Mini-bead Beater (BioSpec Products, Inc., Bartlesville, OK) for 20 s at low speed (2,500 rpm) (Bextine 2004). X. fastidiosa DNA was extracted using a modiÞed Qiagen DNeasy Blood and Tissue Kit.…”

Section: Methodsmentioning

confidence: 99%

Cicadomorpha Insects Associated With Bacterial Leaf Scorch Infected Oak in Central New Jersey

Zhang¹,

Lashomb²,

Gould³

et al. 2011

Environ Entomol

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Potential insect vectors for transmission of oak leaf scorch caused by Xylella fastidiosa Wells et al., in pin and red oaks in New Jersey were surveyed by placing yellow sticky card traps in tree canopies and fogging with pyrethrin insecticide during 2002-2006. Thirty-seven Cicadomorpha species were collected from 20 genera in Membracidae, Cicadellidae, Aphrophoridae, and Clastopteridae. Of the 12,880 potential vectors collected, 91.4% were Membracidae, 6.9% were Cicadellidae, and 1.7% were Aphrophoridae and Clastopteridae. Fogging collected more insect species and individuals than sticky card collections. Sticky card sampling, done more frequently and at a larger number of locations provided similar community structure information as fogging. Sticky card collections of the dominant treehopper species, Ophiderma definita Woodruff were male biased when females were gravid. O. definita populations peaked in early June, comprised 68.2% of the total collection, and were more abundant in pin oaks than red oaks. Graphocephala versuta (Say) peaked in mid-July, comprising 6.2% of the total collection. Higher Cicadomorpha populations were observed in asymptomatic oak canopies than in neighboring X. fastidiosa infected oaks. Individual insect specimens collected from oaks were subjected to a X. fastidiosa DNA assay by polymerase chain reaction amplification. The average X. fastidiosa positive rate was 13.89% for all specimens tested. Eleven treehopper species, six leafhopper species, and four spittlebug species tested DNA positive for X. fastidiosa.

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Section: Methodsmentioning

confidence: 99%

Cicadomorpha Insects Associated With Bacterial Leaf Scorch Infected Oak in Central New Jersey

Zhang¹,

Lashomb²,

Gould³

et al. 2011

Environ Entomol

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“…Each insect was then washed in 95% ethanol and then in deionized water to remove any residual orange oil. Insect heads were removed (Bextine et al 2004), and a silica-based DNA extraction was performed to test for the presence of X. fastidiosa. …”

Section: Methodsmentioning

confidence: 99%

“…Vacuum pressure was applied to the plate four times for two minutes each (Bextine 2004). With X. fastidiosa cells dislodged during vacuum extraction, the heads were discarded and the vacuum solution was retained.…”

Section: Methodsmentioning

confidence: 99%

Detection and Analysis of the Bacterium,Xylella fastidiosa, in Glassy-Winged Sharpshooter,Homalodisca vitripennis, Populations in Texas

Hail

Mitchell²,

Lauzière³

et al. 2010

Journal of Insect Science

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The glassy-winged sharpshooter, Homalodisca vitripeninis Germar (Hemiptera: Cicadellidae), is a xylophagous insect that is an endemic pest of several economically important plants in Texas. H. vitripennis is the main vector of Xylella fastidiosa Wells (Xanthomonadales: Xanthomonadaceae), the bacterium that causes Pierce's disease of grapevine and can travel long distances putting much of Texas grape production at risk. Understanding the movement of H. vitripennis populations capable of transmitting X. fastidiosa into Pierce's-disease-free areas is critical for developing a management program for Pierce's disease. To that end, the USDA-APHIS has developed a program to sample vineyards across Texas to monitor populations of H. vitripennis. From this sampling, H vitripennis collected during 2005 and 2006 over the months of May, June, and July from eight vineyards in different regions of Texas were recovered from yellow sticky traps and tested for the presence of X. fastidiosa. The foregut contents were vacuum extracted and analyzed using RT-PCR to determine the percentage of H. vitripennis within each population that harbor X. fastidiosa and have the potential to transmit this pathogen. H. vitripennis from vineyards known to have Pierce's disease routinely tested positive for the presence of X. fastidiosa. While almost all H. vitripennis collected from vineyards with no history of Pierce's disease tested negative for the presence of the pathogen, three individual insects tested positive. Furthermore, all three insects were determined, by DNA sequencing, to be carrying a strain of X. fastidiosa homologous to known Pierce's disease strains, signifying them as a risk factor for new X. fastidiosa infections.

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“…), and optimisation of DNA extraction protocols is necessary when dealing with the small numbers of bacterial cells found in insect mouthparts (Bextine et al . ). For PCR applications, DNA extraction protocols typically require partial dissection of insects to utilise only the head where bacterial cells are located and column or matrix extraction steps to remove inhibitory compounds (Bextine et al .…”

Section: Assessing Vector Abundance and Diversitymentioning

confidence: 97%

Sharpshooters: a review of what moves Xylella fastidiosa

et al. 2019

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Sharpshooters (Hemiptera: Cicadellidae: Cicadellinae) and spittlebugs (Hemiptera: Aphrophoridae: Aphrophorinae) have a worldwide distribution and are often associated with many crops. Because the geographic range of sharpshooters and spittlebugs often overlaps with the range of the plant pathogenic bacterium Xylella fastidiosa, the importance of these groups of insects in spreading X. fastidiosa rises to the level of a major socioeconomic problem. Managing diseases caused by X. fastidiosa is challenging for several reasons. First, multiple sequence types of the pathogen have been identified, with sequence type affecting host plant specificity. Second, multiple vector species may be present in an agroecosystem, and the host range of vector species is often broad. Although multiple vector control options such as biological, chemical, physical and cultural controls have been described, disease control programs must take into account biological and ecological parameters of locally dominant X. fastidiosa sequence types and vector species to develop suitable control strategies. Our review demonstrates how fundamental principles of vector reproductive biology, behaviour, nutrition and population and community ecology have influenced research on insect vectors and provides insights into priority research areas.

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Evaluation of Methods for Extracting Xylella fastidiosa DNA from the Glassy-Winged Sharpshooter

Cited by 18 publications

References 30 publications

Cicadomorpha Insects Associated With Bacterial Leaf Scorch Infected Oak in Central New Jersey

Cicadomorpha Insects Associated With Bacterial Leaf Scorch Infected Oak in Central New Jersey

Detection and Analysis of the Bacterium,Xylella fastidiosa, in Glassy-Winged Sharpshooter,Homalodisca vitripennis, Populations in Texas

Sharpshooters: a review of what moves Xylella fastidiosa

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