Genome Diversity and Intra- and Interspecies Recombination Events in <i>Grapevine fanleaf virus</i>

Mekuria, Tefera A.; Gutha, Linga R.; Martín, Robert R.; Naidu, Rayapati A.

doi:10.1094/phyto-99-12-1394

Cited by 50 publications

(30 citation statements)

References 48 publications

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“…Total RNA extracted from petiole samples collected from individual source grapevines, originally used for NGS, was used as template in one step-single tube RT-PCR for the detection of GFLV, Grapevine rupestris stem pitting-associated virus (GRSPaV), HpSVd, GSYVd-1, CEVd, and Citrus exocortis Yucatan viroid (CEYVd) using species-specific primers (Table 2) and conditions described earlier [25], [26], [27]. For the detection of GRLaV, DNA was isolated [28] from leaf tissue collected from the same grapevines and used as a template in PCR assays.…”

Section: Methodsmentioning

confidence: 99%

A Leafhopper-Transmissible DNA Virus with Novel Evolutionary Lineage in the Family Geminiviridae Implicated in Grapevine Redleaf Disease by Next-Generation Sequencing

et al. 2013

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A graft-transmissible disease displaying red veins, red blotches and total reddening of leaves in red-berried wine grape (Vitis vinifera L.) cultivars was observed in commercial vineyards. Next-generation sequencing technology was used to identify etiological agent(s) associated with this emerging disease, designated as grapevine redleaf disease (GRD). High quality RNA extracted from leaves of grape cultivars Merlot and Cabernet Franc with and without GRD symptoms was used to prepare cDNA libraries. Assembly of highly informative sequence reads generated from Illumina sequencing of cDNA libraries, followed by bioinformatic analyses of sequence contigs resulted in specific identification of taxonomically disparate viruses and viroids in samples with and without GRD symptoms. A single-stranded DNA virus, tentatively named Grapevine redleaf-associated virus (GRLaV), and Grapevine fanleaf virus were detected only in grapevines showing GRD symptoms. In contrast, Grapevine rupestris stem pitting-associated virus, Hop stunt viroid, Grapevine yellow speckle viroid 1, Citrus exocortis viroid and Citrus exocortis Yucatan viroid were present in both symptomatic and non-symptomatic grapevines. GRLaV was transmitted by the Virginia creeper leafhopper (Erythroneura ziczac Walsh) from grapevine-to-grapevine under greenhouse conditions. Molecular and phylogenetic analyses indicated that GRLaV, almost identical to recently reported Grapevine Cabernet Franc-associated virus from New York and Grapevine red blotch-associated virus from California, represents an evolutionarily distinct lineage in the family Geminiviridae with genome characteristics distinct from other leafhopper-transmitted geminiviruses. GRD significantly reduced fruit yield and affected berry quality parameters demonstrating negative impacts of the disease. Higher quantities of carbohydrates were present in symptomatic leaves suggesting their possible role in the expression of redleaf symptoms.

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Section: Methodsmentioning

confidence: 99%

A Leafhopper-Transmissible DNA Virus with Novel Evolutionary Lineage in the Family Geminiviridae Implicated in Grapevine Redleaf Disease by Next-Generation Sequencing

et al. 2013

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“…Plants for sampling were selected in such a way that individual grapevines exhibiting typical GLRD symptoms are adjacent to disease-free grapevines in a given row to minimize error in sampling and experimental results due to variations in growing conditions. Each pair of symptomatic and adjacent non-symptomatic grapevines was tested for different grapevine viruses by RT-PCR [75]. Mature leaves at the 4 th and 5 th node from the basal portion of primary canes showing typical symptoms of GLRD from virus-infected vines and comparable leaves from adjacent virus-free vines (Figure 2) were collected at the same time in mid September (representing post- véraison stage of berry development) to minimize variation due to developmental stage of leaves.…”

Section: Methodsmentioning

confidence: 99%

“…Lane M represents DNA molecular weight markers used to estimate the size of virus-specific DNA fragment amplified by RT-PCR. The 546 nucleotide DNA band amplified in test samples (indicated by arrow on the right) represents a portion of the 70-kDa heat-shock protein homolog of GLRaV-3 [32,75]. …”

Section: Supplementary Materialsmentioning

confidence: 99%

Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves

et al. 2010

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BackgroundSymptoms of grapevine leafroll disease (GLRD) in red-fruited wine grape (Vitis vinifera L.) cultivars consist of green veins and red and reddish-purple discoloration of inter-veinal areas of leaves. The reddish-purple color of symptomatic leaves may be due to the accumulation of anthocyanins and could reflect an up-regulation of genes involved in their biosynthesis.ResultsWe examined six putative constitutively expressed genes, Ubiquitin, Actin, GAPDH, EF1-a, SAND and NAD5, for their potential as references for normalization of gene expression in reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Using the geNorm program, a combination of two genes (Actin and NAD5) was identified as the stable set of reference genes for normalization of gene expression data obtained from grapevine leaves. By using gene-specific RT-qPCR in combination with a reliable normalization factor, we compared relative expression of the flavonoid biosynthetic pathway genes between leaves infected with Grapevine leafroll-associated virus 3 (GLRaV-3) and exhibiting GLRD symptoms and virus-free green leaves obtained from a red-fruited wine grape cultivar (cv. Merlot). The expression levels of these different genes ranged from two- to fifty-fold increase in virus-infected leaves. Among them, CHS3, F3'5'H, F3H1, LDOX, LAR1 and MybA1 showed greater than 10-fold increase suggesting that they were expressed at significantly higher levels in virus-infected symptomatic leaves. HPLC profiling of anthocyanins extracted from leaves indicated the presence of cyanidin-3-glucoside and malvidin-3-glucoside only in virus-infected symptomatic leaves. The results also showed 24% higher levels of flavonols in virus-infected symptomatic leaves than in virus-free green leaves, with quercetin followed by myricetin being the predominant compounds. Proanthocyanidins, estimated as total tannins by protein precipitation method, were 36% higher in virus-infected symptomatic leaves when compared to virus-free green leaves.ConclusionsThe results, the first example to our knowledge, showed that modulation of the flavonoid biosynthetic pathway occurred in GLRaV-3-infected leaves of a red-fruited wine grape cultivar (cv. Merlot) leading to de novo synthesis of two classes of anthocyanins. These anthocyanins have contributed to the expression of reddish-purple color of virus-infected grapevine leaves exhibiting GLRD symptoms.

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“…The satRNAs of GFLV-R2 and R6 were 1,140 nt in length and shared higher nucleotide identities to the ArMV-J86 and ArMV-NW large satRNAs than to the GFLV-F13 satRNA [11]. To date, the RNA1 and RNA2 of only four GFLV isolates (GFLV-F13, GFLV-WAPN173, GFLV-WAPN6132 and GFLV-SAPCS3) have been completely sequenced [14,15,16,17]. However, isolates GFLV-F13 and ArMV-NW are the only isolates that have their full genome, as well as their large satRNAs completely sequenced [10,13,14,15,18,19].…”

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Complete Nucleotide Sequence of a South African Isolate of Grapevine Fanleaf Virus and Its Associated Satellite RNA

Lamprecht

Spaltman

Stephan

et al. 2013

Viruses

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The complete sequences of RNA1, RNA2 and satellite RNA have been determined for a South African isolate of Grapevine fanleaf virus (GFLV-SACH44). The two RNAs of GFLV-SACH44 are 7,341 nucleotides (nt) and 3,816 nt in length, respectively, and its satellite RNA (satRNA) is 1,104 nt in length, all excluding the poly(A) tail. Multiple sequence alignment of these sequences showed that GFLV-SACH44 RNA1 and RNA2 were the closest to the South African isolate, GFLV-SAPCS3 (98.2% and 98.6% nt identity, respectively), followed by the French isolate, GFLV-F13 (87.3% and 90.1% nt identity, respectively). Interestingly, the GFLV-SACH44 satRNA is more similar to three Arabis mosaic virus satRNAs (85%–87.4% nt identity) than to the satRNA of GFLV-F13 (81.8% nt identity) and was most distantly related to the satRNA of GFLV-R2 (71.0% nt identity). Full-length infectious clones of GFLV-SACH44 satRNA were constructed. The infectivity of the clones was tested with three nepovirus isolates, GFLV-NW, Arabis mosaic virus (ArMV)-NW and GFLV-SAPCS3. The clones were mechanically inoculated in Chenopodium quinoa and were infectious when co-inoculated with the two GFLV helper viruses, but not when co-inoculated with ArMV-NW.

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Genome Diversity and Intra- and Interspecies Recombination Events in Grapevine fanleaf virus

Cited by 50 publications

References 48 publications

A Leafhopper-Transmissible DNA Virus with Novel Evolutionary Lineage in the Family Geminiviridae Implicated in Grapevine Redleaf Disease by Next-Generation Sequencing

A Leafhopper-Transmissible DNA Virus with Novel Evolutionary Lineage in the Family Geminiviridae Implicated in Grapevine Redleaf Disease by Next-Generation Sequencing

Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves

Complete Nucleotide Sequence of a South African Isolate of Grapevine Fanleaf Virus and Its Associated Satellite RNA

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