scholarly journals Genetically Encoded CRISPR components Yield Efficient Gene Editing in the Invasive Pest,Drosophila suzukii

2021 ◽  
Author(s):  
Nikolay P. Kandul ◽  
Esther J. Belikoff ◽  
Junru Liu ◽  
Anna Buchman ◽  
Fang Li ◽  
...  

AbstractOriginally from Asia,Drosophila suzukii(Matsumura, 1931, Diptera:Drosophilidae) is presently a global pest of economically important soft-skinned fruits. Also commonly known as spotted wingDrosophila(SWD), it is largely controlled through repeated applications of broad-spectrum insecticides. There is a pressing need for a better understanding of SWD biology and for developing alternative environmentally-friendly methods of control. The RNA-guided Cas9 nuclease has revolutionized functional genomics and is an integral component of several recently developed genetic strategies for population control of insects. Here we have developed transgenic strains that encode three different terminators and four different promoters to express Cas9 in both the soma and/or germline of SWD. The Cas9 lines were evaluated through genetic crossing to transgenic lines that encode single guide RNAs targeting the conserved X-linkedyellowbody andwhiteeye genes. We find that several Cas9/gRNA lines display very high editing capacity. Going forward, these tools will be instrumental for evaluating gene function in SWD and may provide tools useful for the development of new genetic strategies for control of this invasive species.

2021 ◽  
Vol 14 (7) ◽  
Author(s):  
Abdulsalam I. Isiaku ◽  
Zuobing Zhang ◽  
Vahid Pazhakh ◽  
Harriet R. Manley ◽  
Ella R. Thompson ◽  
...  

ABSTRACT Zebrafish are an important model for studying phagocyte function, but rigorous experimental systems to distinguish whether phagocyte-dependent effects are neutrophil or macrophage specific have been lacking. We have developed and validated transgenic lines that enable superior demonstration of cell-autonomous neutrophil and macrophage genetic requirements. We coupled well-characterized neutrophil- and macrophage-specific Gal4 driver lines with UAS:Cas9 transgenes for selective expression of Cas9 in either neutrophils or macrophages. Efficient gene editing, confirmed by both Sanger and next-generation sequencing, occurred in both lineages following microinjection of efficacious synthetic guide RNAs into zebrafish embryos. In proof-of-principle experiments, we demonstrated molecular and/or functional evidence of on-target gene editing for several genes (mCherry, lamin B receptor, trim33) in either neutrophils or macrophages as intended. These new UAS:Cas9 tools provide an improved resource for assessing individual contributions of neutrophil- and macrophage-expressed genes to the many physiological processes and diseases modelled in zebrafish. Furthermore, this gene-editing functionality can be exploited in any cell lineage for which a lineage-specific Gal4 driver is available. This article has an associated First Person interview with the first author of the paper.


2021 ◽  
Vol 4 (5) ◽  
pp. 739-751 ◽  
Author(s):  
Nikolay P. Kandul ◽  
Esther J. Belikoff ◽  
Junru Liu ◽  
Anna Buchman ◽  
Fang Li ◽  
...  

2021 ◽  
Vol 485 ◽  
pp. 118942
Author(s):  
Alberto Maceda-Veiga ◽  
Sergio Albacete ◽  
Miguel Carles-Tolrá ◽  
Juli Pujade-Villar ◽  
Jan Máca ◽  
...  

Author(s):  
Eugene V. Gasanov ◽  
Justyna Jędrychowska ◽  
Michal Pastor ◽  
Malgorzata Wiweger ◽  
Axel Methner ◽  
...  

AbstractCurrent methods of CRISPR-Cas9-mediated site-specific mutagenesis create deletions and small insertions at the target site which are repaired by imprecise non-homologous end-joining. Targeting of the Cas9 nuclease relies on a short guide RNA (gRNA) corresponding to the genome sequence approximately at the intended site of intervention. We here propose an improved version of CRISPR-Cas9 genome editing that relies on two complementary guide RNAs instead of one. Two guide RNAs delimit the intervention site and allow the precise deletion of several nucleotides at the target site. As proof of concept, we generated heterozygous deletion mutants of the kcng4b, gdap1, and ghitm genes in the zebrafish Danio rerio using this method. A further analysis by high-resolution DNA melting demonstrated a high efficiency and a low background of unpredicted mutations. The use of two complementary gRNAs improves CRISPR-Cas9 specificity and allows the creation of predictable and precise mutations in the genome of D. rerio.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Michelle T. Fountain ◽  
Amir Badiee ◽  
Sebastian Hemer ◽  
Alvaro Delgado ◽  
Michael Mangan ◽  
...  

Abstract Spotted wing drosophila, Drosophila suzukii, is a serious invasive pest impacting the production of multiple fruit crops, including soft and stone fruits such as strawberries, raspberries and cherries. Effective control is challenging and reliant on integrated pest management which includes the use of an ever decreasing number of approved insecticides. New means to reduce the impact of this pest that can be integrated into control strategies are urgently required. In many production regions, including the UK, soft fruit are typically grown inside tunnels clad with polyethylene based materials. These can be modified to filter specific wavebands of light. We investigated whether targeted spectral modifications to cladding materials that disrupt insect vision could reduce the incidence of D. suzukii. We present a novel approach that starts from a neuroscientific investigation of insect sensory systems and ends with infield testing of new cladding materials inspired by the biological data. We show D. suzukii are predominantly sensitive to wavelengths below 405 nm (ultraviolet) and above 565 nm (orange & red) and that targeted blocking of lower wavebands (up to 430 nm) using light restricting materials reduces pest populations up to 73% in field trials.


Weed Science ◽  
1993 ◽  
Vol 41 (2) ◽  
pp. 232-238 ◽  
Author(s):  
Ian M. Heap ◽  
Bruce G. Murray ◽  
Heather A. Loeppky ◽  
Ian N. Morrison

Resistance to aryloxyphenoxypropionate and cyclohexanedione herbicides was identified in four wild oat populations from western Canada. Populations UM1, UM2, and UM3 originated from northwestern Manitoba and UM33 from south-central Saskatchewan. Field histories indicated that these populations were exposed to repeated applications of diclofop-methyl and sethoxydim over the previous 10 yr. The populations differed in their levels and patterns of cross-resistance to these and five other acetyl-CoA carboxylase inhibitors (ACCase inhibitors). UM1, UM2, and UM3 were resistant to diclofop-methyl, fenoxaprop-p-ethyl, and sethoxydim. In contrast, UM33 was resistant to the aryloxyphenoxy propionate herbicides but not to sethoxydim. The dose of sethoxydim required to reduce growth of UM1 by 50% was 150 times greater than for a susceptible population (UM5) or UM33 based on shoot dry matter reductions 21 d after treatment. This population differed from UM2 and UM3 that had R/S ratios of less than 10. In the field UM1 also exhibited a very high level of resistance to sethoxydim. In contrast to susceptible plants that were killed at the recommended dosage, shoot dry matter of resistant plants treated at eight times the recommended dosage was reduced by only 27%. In growth chamber experiments none of the four populations was cross-resistant to herbicides from five different chemical families.


2020 ◽  
Vol 173 ◽  
pp. 107389 ◽  
Author(s):  
Nils Hiebert ◽  
Tessa Carrau ◽  
Merle Bartling ◽  
Andreas Vilcinskas ◽  
Kwang-Zin Lee

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Fernanda Colombari ◽  
Lorenzo Tonina ◽  
Andrea Battisti ◽  
Nicola Mori

Abstract Survival and parasitism activity of Trichopria drosophilae Perkins adults, a cosmopolitan parasitoid of Drosophila spp., were studied under laboratory conditions using five constant temperatures at the lower range known for this enemy, from 4 to 20°C in 4°C increments. Drosophila suzukii Matsumura, an invasive pest of small fruits, was used as a host. Commercially available adult parasitoids were provided with 1) food and D. suzukii pupae; 2) food and no D. suzukii pupae; 3) no food and no pupae. The results show that adult females of T. drosophilae lived longer than males, and both generally benefitted from food supply. The highest level of survival was observed between 8 and 12°C for fed insects, irrespective of whether they were offered host pupae or not. The absence of food led to the highest mortality, but the parasitoid demonstrated considerably resistance to prolonged starvation. Successful parasitism increased steadily with temperature and reached the highest value at 20°C. Conversely, D. suzukii emergence rate was high after exposure of pupae to parasitoids at 4°C, while pupal mortality increased strongly with temperature until 12°C. The findings indicate that T. drosophilae is well adapted to the relatively cold conditions experienced in early spring and in autumn or at high elevations, when the host pupae could be largely available. The long lifespan of the adults and the ability to parasitize the host at low temperature make T. drosophilae potentially useful for the biocontrol of D. suzukii.


2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Michael A Reeve ◽  
M Lukas Seehausen

Abstract The fruit fly Drosophila suzukii has recently become an invasive pest insect of significant economic impact in Europe and the USA. In contrast to other Drosophila species, D. suzukii is able to infest intact fruit by means of a saw-like ovipositor, which allows females to deposit eggs beneath the skin of the fruit. Classical biological control using the parasitoid wasp Ganaspis cf. brasiliensis is currently being researched as an environmentally sustainable option for the control of D. suzukii. In particular, the host specificity of this parasitoid has been assessed for populations from different regions in China and Japan. In order to study the relationship between the differences in specificity and molecular variations, we have adapted a matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method, originally developed for use with plant material, to discriminate between example populations of G. cf. brasiliensis. We have employed a combination of principal component analysis and blind-tested comparison between reference sample MALDI-TOF MS spectra and test sample spectra to discriminate, on the basis of the acid-soluble insect protein spectra generated, between four populations of G. cf. brasiliensis (originally collected from Tokyo and Hasuike in Japan and Dali and Ximing in China). MALDI-TOF MS analysis is able to discriminate with 100% accuracy between populations G. cf. brasiliensis. The Chinese populations were observed to be similar, but the Tokyo population is slightly different and the Hasuike population is significantly different from the other populations. The Tokyo population appears more closely related to the Chinese populations than the Hasuike population, even though both originate from Japan.


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