Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana
Effects of the empty pYL156 vector (pYL156∷00), the most commonly used control vector for Tobacco rattle virus (TRV)-induced gene silencing analysis on viral symptom development and growth of VIGS-treated tomato and Nicotiana benthamiana plants were investigated. It was shown that VIGS-treatment for...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2012-01-01
|
| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.002 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849412734994612224 |
|---|---|
| author | CHENG Wei-shun XU Qiu-fang LI Fei XU You-ping CAI Xin-zhong |
| author_facet | CHENG Wei-shun XU Qiu-fang LI Fei XU You-ping CAI Xin-zhong |
| author_sort | CHENG Wei-shun |
| collection | DOAJ |
| description | Effects of the empty pYL156 vector (pYL156∷00), the most commonly used control vector for Tobacco rattle virus (TRV)-induced gene silencing analysis on viral symptom development and growth of VIGS-treated tomato and Nicotiana benthamiana plants were investigated. It was shown that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and obvious growth repression in treated plants. Therefore, pYL156∷00 is not a good control vector for TRV-induced gene silencing analysis in these plant species. To set a better control for this analysis, two new constructs, pYL156∷NIRi and pYL156∷eGFP, were made. They were released by inserting a 253 bp fragment of a Phaseolus vulgaris nitrite reductase gene intron and a 400 bp fragment of the jellyfish GFP coding sequence in pYL156∷00, respectively. Effects of the two new control constructs on viral symptom development and growth of VIGS-treated N. benthamiana plants were compared with pYL156∷00. The results showed that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and significant growth repression in treated plants, resulting in death of 26.7% plants. VIGS-treatment for pYL156∷NIRi also led to obvious viral symptoms and growth repression, but in a less severe extent than that for pYL156∷00, resulting in death of 13.3% plants. However, VIGS-treatment for pYL156∷eGFP did not cause any obvious viral symptom and growth repression. Severity of viral symptom and growth repression was well correlated with the accumulation level of TRV virus. These results demonstrate that pYL156∷eGFP is an excellent control vector for TRV-induced gene silencing analysis, and provide some insights into the direction to establish an excellent control for VIGS analysis. |
| format | Article |
| id | doaj-art-0ba70725c3a74daab44e60b7daecb0ba |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-0ba70725c3a74daab44e60b7daecb0ba2025-08-20T03:34:21ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552012-01-0138102010.3785/j.issn.1008-9209.2012.01.00210089209Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamianaCHENG Wei-shunXU Qiu-fangLI FeiXU You-pingCAI Xin-zhongEffects of the empty pYL156 vector (pYL156∷00), the most commonly used control vector for Tobacco rattle virus (TRV)-induced gene silencing analysis on viral symptom development and growth of VIGS-treated tomato and Nicotiana benthamiana plants were investigated. It was shown that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and obvious growth repression in treated plants. Therefore, pYL156∷00 is not a good control vector for TRV-induced gene silencing analysis in these plant species. To set a better control for this analysis, two new constructs, pYL156∷NIRi and pYL156∷eGFP, were made. They were released by inserting a 253 bp fragment of a Phaseolus vulgaris nitrite reductase gene intron and a 400 bp fragment of the jellyfish GFP coding sequence in pYL156∷00, respectively. Effects of the two new control constructs on viral symptom development and growth of VIGS-treated N. benthamiana plants were compared with pYL156∷00. The results showed that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and significant growth repression in treated plants, resulting in death of 26.7% plants. VIGS-treatment for pYL156∷NIRi also led to obvious viral symptoms and growth repression, but in a less severe extent than that for pYL156∷00, resulting in death of 13.3% plants. However, VIGS-treatment for pYL156∷eGFP did not cause any obvious viral symptom and growth repression. Severity of viral symptom and growth repression was well correlated with the accumulation level of TRV virus. These results demonstrate that pYL156∷eGFP is an excellent control vector for TRV-induced gene silencing analysis, and provide some insights into the direction to establish an excellent control for VIGS analysis.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.002<italic>Tobacco rattle virus</italic>gene silencingcontrol vectorpYL156∷<italic>eGFP</italic>pYL156∷<italic>NIRi</italic> |
| spellingShingle | CHENG Wei-shun XU Qiu-fang LI Fei XU You-ping CAI Xin-zhong Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana 浙江大学学报. 农业与生命科学版 <italic>Tobacco rattle virus</italic> gene silencing control vector pYL156∷<italic>eGFP</italic> pYL156∷<italic>NIRi</italic> |
| title | Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana |
| title_full | Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana |
| title_fullStr | Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana |
| title_full_unstemmed | Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana |
| title_short | Establishment of a suitable control vector for Tobacco rattle virus-induced gene silencing analysis in Nicotiana benthamiana |
| title_sort | establishment of a suitable control vector for tobacco rattle virus induced gene silencing analysis in nicotiana benthamiana |
| topic | <italic>Tobacco rattle virus</italic> gene silencing control vector pYL156∷<italic>eGFP</italic> pYL156∷<italic>NIRi</italic> |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.01.002 |
| work_keys_str_mv | AT chengweishun establishmentofasuitablecontrolvectorfortobaccorattlevirusinducedgenesilencinganalysisinnicotianabenthamiana AT xuqiufang establishmentofasuitablecontrolvectorfortobaccorattlevirusinducedgenesilencinganalysisinnicotianabenthamiana AT lifei establishmentofasuitablecontrolvectorfortobaccorattlevirusinducedgenesilencinganalysisinnicotianabenthamiana AT xuyouping establishmentofasuitablecontrolvectorfortobaccorattlevirusinducedgenesilencinganalysisinnicotianabenthamiana AT caixinzhong establishmentofasuitablecontrolvectorfortobaccorattlevirusinducedgenesilencinganalysisinnicotianabenthamiana |