Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit
Auxin response factors (ARFs) play important roles in various plant physiological processes; however, knowledge of the exact role of ARFs in plant responses to water deficit is limited. In this study, SlARF4, a member of the ARF family, was functionally characterized under water deficit. Real-time f...
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Format: | Article |
Language: | English |
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MDPI AG
2021-03-01
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Series: | International Journal of Molecular Sciences |
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Online Access: | https://www.mdpi.com/1422-0067/22/7/3347 |
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doaj-c4bc1f77e1694b819559eb3b3fe96cd6 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mengyi Chen Xiaoyang Zhu Xiaojuan Liu Caiyu Wu Canye Yu Guojian Hu Lin Chen Riyuan Chen Mondher Bouzayen Mohammed Zouine Yanwei Hao |
spellingShingle |
Mengyi Chen Xiaoyang Zhu Xiaojuan Liu Caiyu Wu Canye Yu Guojian Hu Lin Chen Riyuan Chen Mondher Bouzayen Mohammed Zouine Yanwei Hao Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit International Journal of Molecular Sciences SlARF4 tomato water deficit drought ABA auxin |
author_facet |
Mengyi Chen Xiaoyang Zhu Xiaojuan Liu Caiyu Wu Canye Yu Guojian Hu Lin Chen Riyuan Chen Mondher Bouzayen Mohammed Zouine Yanwei Hao |
author_sort |
Mengyi Chen |
title |
Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit |
title_short |
Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit |
title_full |
Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit |
title_fullStr |
Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit |
title_full_unstemmed |
Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water Deficit |
title_sort |
knockout of auxin response factor slarf4 improves tomato resistance to water deficit |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-03-01 |
description |
Auxin response factors (ARFs) play important roles in various plant physiological processes; however, knowledge of the exact role of ARFs in plant responses to water deficit is limited. In this study, SlARF4, a member of the ARF family, was functionally characterized under water deficit. Real-time fluorescence quantitative polymerase chain reaction (PCR) and <i>β</i>-glucuronidase (GUS) staining showed that water deficit and abscisic acid (ABA) treatment reduced the expression of <i>SlARF4</i>. <i>SlARF4</i> was expressed in the vascular bundles and guard cells of tomato stomata. Loss of function of SlARF4 (<i>arf4</i>) by using Clustered Regularly Interspaced Short Palindromic Repeats/Cas 9 (CRISPR/Cas 9) technology enhanced plant resistance to water stress and rehydration ability. The <i>arf4</i> mutant plants exhibited curly leaves and a thick stem. Malondialdehyde content was significantly lower in <i>arf4</i> mutants than in wildtype plants under water stress; furthermore, <i>arf4</i> mutants showed higher content of antioxidant substances, superoxide dismutase, actual photochemical efficiency of photosystem II (PSII), and catalase activities. Stomatal and vascular bundle morphology was changed in <i>arf4</i> mutants. We identified 628 differentially expressed genes specifically expressed under water deficit in <i>arf4</i> mutants; six of these genes, including ABA signaling pathway-related genes, were differentially expressed between the wildtype and <i>arf4</i> mutants under water deficit and unlimited water supply. Auxin responsive element (AuxRE) elements were found in these genes’ promoters indicating that SlARF4 participates in ABA signaling pathways by regulating the expression of <i>SlABI5/ABF</i> and <i>SCL3</i>, thereby influencing stomatal morphology and vascular bundle development and ultimately improving plant resistance to water deficit. |
topic |
SlARF4 tomato water deficit drought ABA auxin |
url |
https://www.mdpi.com/1422-0067/22/7/3347 |
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doaj-c4bc1f77e1694b819559eb3b3fe96cd62021-03-26T00:00:36ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-03-01223347334710.3390/ijms22073347Knockout of Auxin Response Factor SlARF4 Improves Tomato Resistance to Water DeficitMengyi Chen0Xiaoyang Zhu1Xiaojuan Liu2Caiyu Wu3Canye Yu4Guojian Hu5Lin Chen6Riyuan Chen7Mondher Bouzayen8Mohammed Zouine9Yanwei Hao10Key Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaLaboratory Genomics and Biotechnology of Fruits, INRA, Toulouse INP, University of Toulouse, 31320 Castanet Tolosan, FranceLaboratory Genomics and Biotechnology of Fruits, INRA, Toulouse INP, University of Toulouse, 31320 Castanet Tolosan, FranceKey Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, ChinaAuxin response factors (ARFs) play important roles in various plant physiological processes; however, knowledge of the exact role of ARFs in plant responses to water deficit is limited. In this study, SlARF4, a member of the ARF family, was functionally characterized under water deficit. Real-time fluorescence quantitative polymerase chain reaction (PCR) and <i>β</i>-glucuronidase (GUS) staining showed that water deficit and abscisic acid (ABA) treatment reduced the expression of <i>SlARF4</i>. <i>SlARF4</i> was expressed in the vascular bundles and guard cells of tomato stomata. Loss of function of SlARF4 (<i>arf4</i>) by using Clustered Regularly Interspaced Short Palindromic Repeats/Cas 9 (CRISPR/Cas 9) technology enhanced plant resistance to water stress and rehydration ability. The <i>arf4</i> mutant plants exhibited curly leaves and a thick stem. Malondialdehyde content was significantly lower in <i>arf4</i> mutants than in wildtype plants under water stress; furthermore, <i>arf4</i> mutants showed higher content of antioxidant substances, superoxide dismutase, actual photochemical efficiency of photosystem II (PSII), and catalase activities. Stomatal and vascular bundle morphology was changed in <i>arf4</i> mutants. We identified 628 differentially expressed genes specifically expressed under water deficit in <i>arf4</i> mutants; six of these genes, including ABA signaling pathway-related genes, were differentially expressed between the wildtype and <i>arf4</i> mutants under water deficit and unlimited water supply. Auxin responsive element (AuxRE) elements were found in these genes’ promoters indicating that SlARF4 participates in ABA signaling pathways by regulating the expression of <i>SlABI5/ABF</i> and <i>SCL3</i>, thereby influencing stomatal morphology and vascular bundle development and ultimately improving plant resistance to water deficit.https://www.mdpi.com/1422-0067/22/7/3347SlARF4tomatowater deficitdroughtABAauxin |