Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i>
Previous studies have identified numerous transcription factors involved in drought response, each of which play different roles in plants. The objective of the present study was to evaluate the effectiveness of two transcription factors on drought response in <i>Jatropha curcas</i> L.,...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/12/17/3029 |
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| author | Xianfei Niu Zhiping Lai Linghui Wang Rui Ma Yingying Ren Xueying Wang Cheng Cheng Ting Wang Fang Chen Ying Xu |
| author_facet | Xianfei Niu Zhiping Lai Linghui Wang Rui Ma Yingying Ren Xueying Wang Cheng Cheng Ting Wang Fang Chen Ying Xu |
| author_sort | Xianfei Niu |
| collection | DOAJ |
| description | Previous studies have identified numerous transcription factors involved in drought response, each of which play different roles in plants. The objective of the present study was to evaluate the effectiveness of two transcription factors on drought response in <i>Jatropha curcas</i> L., <i>JcNAC1</i> and <i>JcZFP8</i>. The overexpression of these transcription factors in tobacco (<i>Nicotiana benthamiana</i> L.) improved drought resistance, but <i>JcZFP8</i> delayed germination and <i>JcNAC1</i> reduced biomass and yield. By constitutively co-expressing these two genes in tobacco, drought resistance was improved, and the negative effects of each of them were overcome. The transgenic plants with double-gene co-expression showed stronger drought tolerance with 1.76-fold greater accumulation of proline and lower H<sub>2</sub>O<sub>2</sub> and malondialdehyde (MDA) content to 43 and 65% of wildtype (WT) levels, respectively. The expression levels of <i>NbbHLH1</i> and <i>NbbHLH2</i> genes upregulated linearly with the increased drought tolerance of double genes co-expression plants. In drought conditions, the leaf water contents of <i>bhlh1</i>, <i>bhlh2</i>, and <i>bhlh1bhlh2</i> deletion mutants obtained by CRISPR-CAS9 knockout technique were maintained at 99%, 97%, and 97% of WT. The <i>bhlh1bhlh2</i> was found with lower germination rate but with higher reactive oxygen levels (1.64-fold H<sub>2</sub>O<sub>2</sub> and 1.41-fold MDA levels). Thus, the co-expression of two transcription factors with different functions overcame the adverse traits brought by a single gene and enhanced the shared drought-tolerant traits, which can provide guidance on theory and selection of gene combinations for the application of multi-gene co-expression in agriculture in the future. |
| first_indexed | 2024-03-10T23:15:29Z |
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| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T23:15:29Z |
| publishDate | 2023-08-01 |
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| record_format | Article |
| series | Plants |
| spelling | doaj.art-7a0c36ed9e53475aab4027af87666b9d2023-11-19T08:40:19ZengMDPI AGPlants2223-77472023-08-011217302910.3390/plants12173029Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i>Xianfei Niu0Zhiping Lai1Linghui Wang2Rui Ma3Yingying Ren4Xueying Wang5Cheng Cheng6Ting Wang7Fang Chen8Ying Xu9Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaCollege of Life Science and Food Engineering, Yibin University, Yibin 644000, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaKey Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, ChinaPrevious studies have identified numerous transcription factors involved in drought response, each of which play different roles in plants. The objective of the present study was to evaluate the effectiveness of two transcription factors on drought response in <i>Jatropha curcas</i> L., <i>JcNAC1</i> and <i>JcZFP8</i>. The overexpression of these transcription factors in tobacco (<i>Nicotiana benthamiana</i> L.) improved drought resistance, but <i>JcZFP8</i> delayed germination and <i>JcNAC1</i> reduced biomass and yield. By constitutively co-expressing these two genes in tobacco, drought resistance was improved, and the negative effects of each of them were overcome. The transgenic plants with double-gene co-expression showed stronger drought tolerance with 1.76-fold greater accumulation of proline and lower H<sub>2</sub>O<sub>2</sub> and malondialdehyde (MDA) content to 43 and 65% of wildtype (WT) levels, respectively. The expression levels of <i>NbbHLH1</i> and <i>NbbHLH2</i> genes upregulated linearly with the increased drought tolerance of double genes co-expression plants. In drought conditions, the leaf water contents of <i>bhlh1</i>, <i>bhlh2</i>, and <i>bhlh1bhlh2</i> deletion mutants obtained by CRISPR-CAS9 knockout technique were maintained at 99%, 97%, and 97% of WT. The <i>bhlh1bhlh2</i> was found with lower germination rate but with higher reactive oxygen levels (1.64-fold H<sub>2</sub>O<sub>2</sub> and 1.41-fold MDA levels). Thus, the co-expression of two transcription factors with different functions overcame the adverse traits brought by a single gene and enhanced the shared drought-tolerant traits, which can provide guidance on theory and selection of gene combinations for the application of multi-gene co-expression in agriculture in the future.https://www.mdpi.com/2223-7747/12/17/3029<i>JcNAC1</i><i>JcZFP8</i>double gene co-expressiongrowth traitsdrought stress |
| spellingShingle | Xianfei Niu Zhiping Lai Linghui Wang Rui Ma Yingying Ren Xueying Wang Cheng Cheng Ting Wang Fang Chen Ying Xu Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> Plants <i>JcNAC1</i> <i>JcZFP8</i> double gene co-expression growth traits drought stress |
| title | Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> |
| title_full | Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> |
| title_fullStr | Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> |
| title_full_unstemmed | Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> |
| title_short | Co-Expression of <i>JcNAC1</i>- and <i>JcZFP8</i>-Improved Agronomic Traits of Tobacco and Enhanced Drought Resistance through <i>NbbHLH1</i> and <i>NbbHLH2</i> |
| title_sort | co expression of i jcnac1 i and i jczfp8 i improved agronomic traits of tobacco and enhanced drought resistance through i nbbhlh1 i and i nbbhlh2 i |
| topic | <i>JcNAC1</i> <i>JcZFP8</i> double gene co-expression growth traits drought stress |
| url | https://www.mdpi.com/2223-7747/12/17/3029 |
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