An Effective Method of Ribes spp. Inoculation with Blackcurrant Reversion Virus under In Vitro Conditions

Juškytė, Ana Dovilė; Mažeikienė, Ingrida; Stanys, V.

doi:10.3390/plants11131635

Cited by 3 publications

(7 citation statements)

References 41 publications

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“…Plants in this in vitro inoculation assay were infected with inoculum with three different BRV isolates [ 5 ]. The expression of viral polyprotein encoded by RNA1 (cluster-12591.29271) was detected in virus-inoculated plants.…”

Section: Resultsmentioning

confidence: 99%

“…genotypes of BRV is determined genetically, and markers are created, but the host plant–pathogen interaction at the genetic level has not been studied. De novo transcriptome assembly using NGS was performed for the first time in BRV-infected plants according to the methodology under in vitro conditions created earlier [ 5 ]; this method opened possibilities to study specific and precise host–virus interactions in Ribes spp.…”

Section: Discussionmentioning

confidence: 99%

“…This cultivar was used as a model because the virus may be observed in plant material up to 6–8 days after inoculation and significant expression changes in pathogenesis-related 1 ( PR1 ) were detected at 2 and 4 dpi as stated in previous studies. This gene was used as a molecular marker to indicate blackcurrant’s defense response to BRV infection [ 5 , 24 ]. Therefore, the transcriptomes of cv.…”

Section: Discussionmentioning

confidence: 99%

“…BRV infection was approved by the presence of virus RNA1 and RNA2 in the transcriptome of inoculated samples ( Figure S1 ). According to the data of a previous study [ 5 ], inoculum with three different BRV isolates was used in our experiment. RNA2 strand is responsible for translation of three proteins: X, movement protein (MP), and coat protein (CP).…”

Section: Discussionmentioning

confidence: 99%

“…BRV transmission that uses mechanical inoculation with sap or purified virions is complicated. The new method of transmission of BRV from BRV-positive Ribes to BRV-negative Ribes in vitro through root soaking in the inoculum was created earlier by [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

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De Novo Transcriptome Analysis of R. nigrum cv. Aldoniai in Response to Blackcurrant Reversion Virus Infection

Mažeikienė

Juškytė

Bendokas

et al. 2022

IJMS

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The most damaging pathogen in blackcurrant plantations is mite-transmitted blackcurrant reversion virus (BRV). Some Ribes species have an encoded genetic resistance to BRV. We performed RNA sequencing analysis of BRV-resistant blackcurrant cv. Aldoniai to evaluate the molecular mechanisms related to the BRV infection response. The RNA of virus-inoculated and mock-inoculated microshoots was sequenced, and the transcriptional changes at 2- and 4-days post inoculation (dpi) were analyzed. The accumulation and expression of BRV RNA1 were detected in infected plants. In total, 159,701 transcripts were obtained and 30.7% were unigenes, annotated in 7 databases. More than 25,000 differentially expressed genes (DEGs) according to FPKM were upregulated or downregulated. We observed 221 and 850 DEGs at 2 and 4 dpi, respectively, in BRV-infected microshoots related to the stress response. The proportion of upregulated DEGs at 4 dpi was about 3.5 times higher than at 2 dpi. Pathways of the virus defense response were activated, and key candidate genes were identified. The phenylpropanoid and the cutin, suberine, and wax biosynthesis pathways were activated in infected plants. Our comparative de novo analysis of the R. nigrum transcriptome provides clues not only for understanding the molecular BRV resistance mechanisms but also for breeding BRV-tolerant genotypes.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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De Novo Transcriptome Analysis of R. nigrum cv. Aldoniai in Response to Blackcurrant Reversion Virus Infection

Mažeikienė

Juškytė

Bendokas

et al. 2022

IJMS

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Analysis of R Genes Related to Blackcurrant Reversion Virus Resistance in the Comparative Transcriptome of Ribes nigrum cv. Aldoniai

Juškytė

Mažeikienė

Stanys

2022

Plants

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Blackcurrant reversion virus (BRV) is the most destructive mite-transmitted pathogen in blackcurrants. The understanding of the resistance to BRV is limited, hindering and delaying the selection process. To identify the resistance (R) gene for BRV resistance, a gene expression analysis based on de novo blackcurrant cv. Aldoniai comparative transcriptome analysis (mock- and BRV-inoculated samples at 2 and 4 days post-inoculation (dpi)) was performed. In this study, 111 annotated clusters associated with pathogenesis according to conservative R gene domains were identified. In virus-infected samples, only Cluster-12591.33361 showed significant expression at 4 dpi. The expression profiles of this cluster were significantly associated with the presence of BRV particles in plant tissues, making it a putative R gene in the dominant resistance strategy in the BRV–Ribes nigrum interaction. The newly identified gene R.nigrum_R belongs to the CC-NBS-LRR class and has 63.9% identity with RPM1 in Populus spp. This study provides new insights on dominant putative R genes related to resistance to BRV in R. nigrum, which could aid targeted research and genetic improvement in breeding programs of blackcurrants.

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New Insights into Hop Latent Viroid Detection, Infectivity, Host Range, and Transmission

Atallah,

Yassin,

Verchot

2023

Viruses

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Hop latent viroid (HLVd), a subviral pathogen from the family Pospiviroidae, is a major threat to the global cannabis industry and is the causative agent for “dudding disease”. Infected plants can often be asymptomatic for a period of growth and then develop symptoms such as malformed and yellowing leaves, as well as stunted growth. During flowering, HLVd-infected plants show reduced levels of valuable metabolites. This study was undertaken to expand our basic knowledge of HLVd infectivity, transmission, and host range. HLVd-specific primers were used for RT-PCR detection in plant samples and were able to detect HLVd in as little as 5 picograms of total RNA. A survey of hemp samples obtained from a diseased production system proved sole infection of HLVd (72%) with no coexistence of hop stunt viroid. HLVd was infectious through successive passage assays using a crude sap or total RNA extract derived from infected hemp. HLVd was also highly transmissible through hemp seeds at rates of 58 to 80%. Host range assays revealed new hosts for HLVd: tomato, cucumber, chrysanthemum, Nicotiana benthamiana, and Arabidopsis thaliana (Col-0). Sequence analysis of 77 isolates revealed only 3 parsimony-informative sites, while 10 sites were detected among all HLVd isolates available in the GenBank. The phylogenetic relationship among HLVd isolates allowed for inferring two major clades based on the genetic distance. Our findings facilitate further studies on host–viroid interaction and viroid management.

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An Effective Method of Ribes spp. Inoculation with Blackcurrant Reversion Virus under In Vitro Conditions

Cited by 3 publications

References 41 publications

De Novo Transcriptome Analysis of R. nigrum cv. Aldoniai in Response to Blackcurrant Reversion Virus Infection

De Novo Transcriptome Analysis of R. nigrum cv. Aldoniai in Response to Blackcurrant Reversion Virus Infection

Analysis of R Genes Related to Blackcurrant Reversion Virus Resistance in the Comparative Transcriptome of Ribes nigrum cv. Aldoniai

New Insights into Hop Latent Viroid Detection, Infectivity, Host Range, and Transmission

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