A genomic analysis of transcytosis in the pea aphid, <i>Acyrthosiphon pisum</i>, a mechanism involved in virus transmission

Tamborindeguy, Cécilia; Monsion, Baptiste; Brault, Véronique; Hunnicutt, Laura E.; Ju, Ho-Jong; Nakabachi, Atsushi; Fleet, E. Van

doi:10.1111/j.1365-2583.2009.00956.x

Cited by 35 publications

(41 citation statements)

References 88 publications

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“…The propagation of YDVs in their plant hosts has been investigated and elaborated using the prevailing absolute and relative quantification methods [15], [16], but rarely has research focused on fluctuation in the virus titre and accumulation in the vector aphid during the acquisition access period (AAP), latent period or inoculation access period (IAP), all pivotal issues that affect rates of transmission, duration of transmissibility and epidemiology of the plant virus in the field. With the entry of the virus into the vector aphid, many proteins are incorporated via a transcytosis mechanism associated with virus transmission [17], [18], [19], [20], followed by up- or down-regulation of the expression of numerous genes [21]. Using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), we can rapidly assess the influence of virus entry on molecular functions and biological processes and on the real-time concentration of virus [22], [23].…”

Section: Introductionmentioning

confidence: 99%

“…In a recent study, some ACT genes (ACT1 to ACT4) and one GAPDH gene, together with many other genes from A. pisum were considered to be potentially related to the transmission of Pea enation mosaic virus and Soybean dwarf virus [21]. Here, we selected these two doubtful HKG genes, ACT1 and GAPDH, as well as EF-1α and 18S rRNA which were always used as reference genes for normalization of target genes in other aphids [42], [47], as four candidate reference gene for relative RT-qPCR analysis in R. padi .…”

Section: Introductionmentioning

confidence: 99%

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Sequencing and Validation of Reference Genes to Analyze Endogenous Gene Expression and Quantify Yellow Dwarf Viruses Using RT-qPCR in Viruliferous Rhopalosiphum padi

Liu

Mar

et al. 2014

PLoS ONE

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The bird cherry-oat aphid (Rhopalosiphum padi), an important pest of cereal crops, not only directly sucks sap from plants, but also transmits a number of plant viruses, collectively the yellow dwarf viruses (YDVs). For quantifying changes in gene expression in vector aphids, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is a touchstone method, but the selection and validation of housekeeping genes (HKGs) as reference genes to normalize the expression level of endogenous genes of the vector and for exogenous genes of the virus in the aphids is critical to obtaining valid results. Such an assessment has not been done, however, for R. padi and YDVs. Here, we tested three algorithms (GeNorm, NormFinder and BestKeeper) to assess the suitability of candidate reference genes (EF-1α, ACT1, GAPDH, 18S rRNA) in 6 combinations of YDV and vector aphid morph. EF-1α and ACT1 together or in combination with GAPDH or with GAPDH and 18S rRNA could confidently be used to normalize virus titre and expression levels of endogenous genes in winged or wingless R. padi infected with Barley yellow dwarf virus isolates (BYDV)-PAV and BYDV-GAV. The use of only one reference gene, whether the most stably expressed (EF-1α) or the least stably expressed (18S rRNA), was not adequate for obtaining valid relative expression data from the RT-qPCR. Because of discrepancies among values for changes in relative expression obtained using 3 regions of the same gene, different regions of an endogenous aphid gene, including each terminus and the middle, should be analyzed at the same time with RT-qPCR. Our results highlight the necessity of choosing the best reference genes to obtain valid experimental data and provide several HKGs for relative quantification of virus titre in YDV-viruliferous aphids.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Sequencing and Validation of Reference Genes to Analyze Endogenous Gene Expression and Quantify Yellow Dwarf Viruses Using RT-qPCR in Viruliferous Rhopalosiphum padi

Liu

Mar

et al. 2014

PLoS ONE

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“…During the process of transmission, luteovirids cross two epithelial cell layers in the aphid vector: the gut, at the level of the hindgut and posterior midgut, and the accessory salivary glands [15,31]. Because non-transmissible luteovirids may fail to cross either of these cellular epithelia in non-aphid vectors, these tissues have been referred to as ‘transmission barriers’ [31].…”

Section: Resultsmentioning

confidence: 99%

Localization, Concentration, and Transmission Efficiency of Banana bunchy top virus in Four Asexual Lineages of Pentalonia aphids

Watanabe

Greenwell

Bressan

2013

Viruses

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Banana bunchy top virus (BBTV) is the most destructive pathogenic virus of banana plants worldwide. The virus is transmitted in a circulative non-propagative manner by the banana aphid, Pentalonia nigronervosa Coquerel. In this work, we examined the localization, accumulation, and transmission efficiency of BBTV in four laboratory-established lineages of Pentalonia aphids derived from four different host plants: taro (Colocasia esculenta), heliconia (Heliconia spp.), red ginger (Alpinia purpurata), and banana (Musa sp.). Mitochondrial sequencing identified three and one lineages as Pentalonia caladii van der Goot, a recently proposed species, and P. nigronervosa, respectively. Microsatellite analysis separated the aphid lineages into four distinct genotypes. The transmission of BBTV was tested using leaf disk and whole-plant assays, both of which showed that all four lineages are competent vectors of BBTV, although the P. caladii from heliconia transmitted BBTV to the leaf disks at a significantly lower rate than did P. nigronervosa. The concentration of BBTV in dissected guts, haemolymph, and salivary glands was quantified by real-time PCR. The BBTV titer reached similar concentrations in the guts, haemolymph, and salivary glands of aphids from all four lineages tested. Furthermore, immunofluorescence assays showed that BBTV antigens localized to the anterior midguts and the principal salivary glands, demonstrating a similar pattern of translocations across the four lineages. The results reported in this study showed for the first time that P. caladii is a competent vector of BBTV.

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“…Thus, the proteins involved in endocytosis, vesicle transport and exocytosis (dynamins, together with NSF, alpha-SNAP, SNAREs, Munc18, Rab, annexins, etc.) are expected to play important roles in virus transmission (Tamborindeguy et al, 2009). Indeed, Dyn proteins have been demonstrated to be essential for the infection of various viruses in animal hosts (Cherry & Perrimon, 2004;Marsh & Helenius, 2006).…”

Section: Discussionmentioning

confidence: 99%

Expansion of genes encoding a novel type of dynamin in the genome of the pea aphid, Acyrthosiphon pisum

Nakabachi

Miyagishima

2010

Insect Molecular Biology

Self Cite

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Abstracti mb_941 165..174Screening of the entire genome of the pea aphid, Acyrthosiphon pisum, detected 15 genes for putative dynamin superfamily proteins -self-assembling large GTPases that are involved in the fission and fusion of membranes. In addition to a single gene each for Dyn, Drp1, and Opa1, orthologues that are common in Metazoa, 12 genes encoding a novel type of dynamin were found. Phylogenetic analyses showed that these novel-class genes are monophyletic. Quantitative reverse transcription-PCR demonstrated that expressions of four novel-class dynamin genes are highly up-regulated in the midgut, through which aphids take in phloem-sap diets and plant viruses. As this type of dynamin is absent from all other fully sequenced organisms, they may function in processes unique to aphids.

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A genomic analysis of transcytosis in the pea aphid, Acyrthosiphon pisum, a mechanism involved in virus transmission

Cited by 35 publications

References 88 publications

Sequencing and Validation of Reference Genes to Analyze Endogenous Gene Expression and Quantify Yellow Dwarf Viruses Using RT-qPCR in Viruliferous Rhopalosiphum padi

Sequencing and Validation of Reference Genes to Analyze Endogenous Gene Expression and Quantify Yellow Dwarf Viruses Using RT-qPCR in Viruliferous Rhopalosiphum padi

Localization, Concentration, and Transmission Efficiency of Banana bunchy top virus in Four Asexual Lineages of Pentalonia aphids

Expansion of genes encoding a novel type of dynamin in the genome of the pea aphid, Acyrthosiphon pisum

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