Rescue of an infection clone of barley yellow dwarf virus -GAV

2021 ◽  
Author(s):  
Xingan Hao ◽  
Shuang Song ◽  
Qinrong Zhong ◽  
Jamal-U-Ddin Hajano ◽  
Jie Guo ◽  
...  
Keyword(s):  
Cdna Clone ◽  
Molecular Mechanisms ◽  
Barley Yellow Dwarf Virus ◽  
Full Length ◽  
Yellow Dwarf ◽  
Biologically Active ◽  
Dwarf Virus ◽  
Full Length Cdna ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus

Barley yellow dwarf virus-GAV (BYDV-GAV) is one of the most prevalent viruses causing yellow dwarf disease in wheat in China. The biology and pathology of BYDV-GAV are well studied; however, gene functions and molecular mechanisms of BYDV-GAV disease development are unclear due to the lack of a reverse genetics system. In this study, a full-length cDNA clone of BYDV-GAV was constructed and expressed using Agrobacterium-mediated inoculation of Nicotiana benthamiana. Virions produced by BYDV-GAV in N. benthamiana were transmitted to wheat by an aphid vector after acquisition via a sandwich feeding method. Infectivity of the cDNA clone in wheat was verified through RT-PCR and Western blot assays, and the recombinant virus elicited typical reddening symptoms in oats and was transmitted between wheat plants. These results confirm the production of biologically active transmissible virions. Using the BYDV-GAV infectious clone, we demonstrate that the viral protein P4 was involved in cell-to-cell movement and stunting symptoms in wheat. This is the first report describing the development of an infectious full-length cDNA clone of BYDV-GAV and provides a useful tool for virus-host-vector interaction studies.

scholarly journals Barley yellow dwarf virus-GAV-derived vsiRNAs are involved in the production of wheat leaf yellowing symptoms by targeting chlorophyll synthase

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Chuan Shen ◽  
Caiyan Wei ◽  
Jingyuan Li ◽  
Xudong Zhang ◽  
Qinrong Zhong ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Barley Yellow Dwarf Virus ◽  
Plant Viruses ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Small Rna Sequencing ◽  
Symptom Development ◽  
Barley Yellow Dwarf ◽  
Leaf Yellowing ◽  
Yellow Dwarf Virus

Abstract Background Wheat yellow dwarf virus disease is infected by barley yellow dwarf virus (BYDV), which causes leaf yellowing and dwarfing symptoms in wheat, thereby posing a serious threat to China's food production. The infection of plant viruses can produce large numbers of vsiRNAs, which can target host transcripts and cause symptom development. However, few studies have been conducted to explore the role played by vsiRNAs in the interaction between BYDV-GAV and host wheat plants. Methods In this study, small RNA sequencing was conducted to profile vsiRNAs in BYDV-GAV-infected wheat plants. The putative targets of vsiRNAs were predicted by the bioinformatics software psRNATarget. RT-qPCR and VIGS were employed to identify the function of selected target transcripts. To confirm the interaction between vsiRNA and the target, 5′ RACE was performed to analyze the specific cleavage sites. Results From the sequencing data, we obtained a total of 11,384 detected vsiRNAs. The length distribution of these vsiRNAs was mostly 21 and 22 nt, and an A/U bias was observed at the 5′ terminus. We also observed that the production region of vsiRNAs had no strand polarity. The vsiRNAs were predicted to target 23,719 wheat transcripts. GO and KEGG enrichment analysis demonstrated that these targets were mostly involved in cell components, catalytic activity and plant-pathogen interactions. The results of RT-qPCR analysis showed that most chloroplast-related genes were downregulated in BYDV-GAV-infected wheat plants. Silencing of a chlorophyll synthase gene caused leaf yellowing that was similar to the symptoms exhibited by BYDV-GAV-inoculated wheat plants. A vsiRNA from an overlapping region of BYDV-GAV MP and CP was observed to target chlorophyll synthase for gene silencing. Next, 5′ RACE validated that vsiRNA8856 could cleave the chlorophyll synthase transcript in a sequence-specific manner. Conclusions This report is the first to demonstrate that BYDV-GAV-derived vsiRNAs can target wheat transcripts for symptom development, and the results of this study help to elucidate the molecular mechanisms underlying leaf yellowing after viral infection.

scholarly journals Barley Yellow Dwarf Virus-GAV-derived vsiRNAs Involved in the Production of Wheat Leaf Yellowing Symptoms by Targeting Chlorophyll Synthase

2020 ◽  
Author(s):  
Chuan Shen ◽  
Caiyan Wei ◽  
Jingyuan Li ◽  
Xudong Zhang ◽  
Qinrong Zhong ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Barley Yellow Dwarf Virus ◽  
Length Distribution ◽  
Plant Viruses ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Small Rna Sequencing ◽  
Barley Yellow Dwarf ◽  
Leaf Yellowing ◽  
Yellow Dwarf Virus

Abstract Background: Wheat yellow dwarf virus disease was infected by barley yellow dwarf virus, caused leaf yellowing and dwarfing symptoms in wheat, posing a serious threat to China's food production. Infection of plant viruses can produce a large number of vsiRNAs, which can target host transcripts for symptoms development. However, few studies were conducted to explore the role of vsiRNAs in the interaction between BYDV-GAV and host wheat plants.Methods: In this study, small RNA sequencing was conducted to profile the vsiRNAs in BYDV-GAV infected wheat plants. The putative targets of vsiRNAs were predicted by bioinformatics software. RT-qPCR and VIGS were used to identify the function of selected target transcripts. To confirm the interaction between vsiRNA and the target, 5’RACE was performed to analyze the specific cleavage sites. Results: From the sequencing data, we obtained a total of 11,384 detected vsiRNAs. The length distribution of these vsiRNAs mostly was 21- and 22-nt along with A/U biased at the 5 ' -terminal. We also found the production region of vsiRNAs had no strand polarity. The vsiRNAs were predicted to target 23,719 wheat transcripts. GO and KEGG enrichment analysis revealed that these targets mostly involved in cell parts and catalytic activity and plant-pathogen interaction. The results of RT-qPCR indicted that most chloroplast-related genes showed down-regulation in BYDV-GAV infected wheat plants. Silencing of a chlorophyll synthase caused leaf yellowing that was similar to the symptom of BYDV-GAV inoculated wheat plants. A vsiRNA from an overlap region of BYDV-GAV MP and CP was found that could target the chlorophyll synthase for gene silencing. Then, 5’RACE validated that vsiRNA8856 could cleavage the chlorophyll synthase transcript in a sequence-specific manner.Conclusion: This is the first report demonstrating that BYDV-GAV-derived vsiRNAs can target wheat transcripts for symptom development, and this study provides new insights into the molecular mechanisms underlying leaf yellowing upon viral infection.

scholarly journals Barley yellow dwarf virus-GAV-derived vsiRNAs are involved in the production of wheat leaf yellowing symptoms by targeting chlorophyll synthase

2020 ◽  
Author(s):  
Chuan Shen ◽  
Caiyan Wei ◽  
Jingyuan Li ◽  
Xudong Zhang ◽  
Qinrong Zhong ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Barley Yellow Dwarf Virus ◽  
Plant Viruses ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Small Rna Sequencing ◽  
Symptom Development ◽  
Barley Yellow Dwarf ◽  
Leaf Yellowing ◽  
Yellow Dwarf Virus

Abstract Background: Wheat yellow dwarf virus disease is infected by barley yellow dwarf virus (BYDV), which causes leaf yellowing and dwarfing symptoms in wheat, thereby posing a serious threat to China's food production. The infection of plant viruses can produce large numbers of vsiRNAs, which can target host transcripts and cause symptom development. However, few studies have been conducted to explore the role played by vsiRNAs in the interaction between BYDV-GAV and host wheat plants.Methods: In this study, small RNA sequencing was conducted to profile vsiRNAs in BYDV-GAV-infected wheat plants. The putative targets of vsiRNAs were predicted by the bioinformatics software psRNATarget. RT-qPCR and VIGS were employed to identify the function of selected target transcripts. To confirm the interaction between vsiRNA and the target, 5’ RACE was performed to analyze the specific cleavage sites.Results: From the sequencing data, we obtained a total of 11,384 detected vsiRNAs. The length distribution of these vsiRNAs was mostly 21 and 22 nt, and an A/U bias was observed at the 5' terminus. We also observed that the production region of vsiRNAs had no strand polarity. The vsiRNAs were predicted to target 23,719 wheat transcripts. GO and KEGG enrichment analysis demonstrated that these targets were mostly involved in cell components, catalytic activity and plant-pathogen interactions. The results of RT-qPCR analysis showed that most chloroplast-related genes were downregulated in BYDV-GAV-infected wheat plants. Silencing of a chlorophyll synthase gene caused leaf yellowing that was similar to the symptoms exhibited by BYDV-GAV-inoculated wheat plants. A vsiRNA from an overlapping region of BYDV-GAV MP and CP was observed to target chlorophyll synthase for gene silencing. Next, 5’ RACE validated that vsiRNA8856 could cleave the chlorophyll synthase transcript in a sequence-specific manner.Conclusion: This report is the first to demonstrate that BYDV-GAV-derived vsiRNAs can target wheat transcripts for symptom development, and the results of this study help to elucidate the molecular mechanisms underlying leaf yellowing after viral infection.

scholarly journals Barley yellow dwarf virus-GAV-derived vsiRNAs are involved in the production of wheat leaf yellowing symptoms by targeting chlorophyll synthase

2020 ◽  
Author(s):  
Chuan Shen ◽  
Caiyan Wei ◽  
Jingyuan Li ◽  
Xudong Zhang ◽  
Qinrong Zhong ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Barley Yellow Dwarf Virus ◽  
Plant Viruses ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Small Rna Sequencing ◽  
Symptom Development ◽  
Barley Yellow Dwarf ◽  
Leaf Yellowing ◽  
Yellow Dwarf Virus

Abstract Background: Wheat yellow dwarf virus disease is infected by barley yellow dwarf virus (BYDV), which causes leaf yellowing and dwarfing symptoms in wheat, thereby posing a serious threat to China's food production. The infection of plant viruses can produce large numbers of vsiRNAs, which can target host transcripts and cause symptom development. However, few studies have been conducted to explore the role played by vsiRNAs in the interaction between BYDV-GAV and host wheat plants.Methods: In this study, small RNA sequencing was conducted to profile vsiRNAs in BYDV-GAV-infected wheat plants. The putative targets of vsiRNAs were predicted by the bioinformatics software psRNATarget. RT-qPCR and VIGS were employed to identify the function of selected target transcripts. To confirm the interaction between vsiRNA and the target, 5’ RACE was performed to analyze the specific cleavage sites.Results: From the sequencing data, we obtained a total of 11,384 detected vsiRNAs. The length distribution of these vsiRNAs was mostly 21 and 22 nt, and an A/U bias was observed at the 5' terminus. We also observed that the production region of vsiRNAs had no strand polarity. The vsiRNAs were predicted to target 23,719 wheat transcripts. GO and KEGG enrichment analysis demonstrated that these targets were mostly involved in cell components, catalytic activity and plant-pathogen interactions. The results of RT-qPCR analysis showed that most chloroplast-related genes were downregulated in BYDV-GAV-infected wheat plants. Silencing of a chlorophyll synthase gene caused leaf yellowing that was similar to the symptoms exhibited by BYDV-GAV-inoculated wheat plants. A vsiRNA from an overlapping region of BYDV-GAV MP and CP was observed to target chlorophyll synthase for gene silencing. Next, 5’ RACE validated that vsiRNA8856 could cleave the chlorophyll synthase transcript in a sequence-specific manner.Conclusion: This report is the first to demonstrate that BYDV-GAV-derived vsiRNAs can target wheat transcripts for symptom development, and the results of this study help to elucidate the molecular mechanisms underlying leaf yellowing after viral infection.

Tolerance to Barley Yellow Dwarf Virus in Triticale

Crop Science ◽  
1990 ◽  
Vol 30 (5) ◽  
pp. 1008 ◽  
Author(s):  
J. Collin ◽  
A. Comeau ◽  
C.A. St-Pierre
Keyword(s):  
Barley Yellow Dwarf Virus ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Barley Yellow Dwarf ◽  
Yellow Dwarf Virus
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