Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze)
The microRNA156 (miR156) family, one of the first miRNA families discovered in plants, plays various important roles in plant growth and resistance to various abiotic stresses. Previously, miR156s were shown to respond to drought stress, but miR156s in tea plants (<i>Camellia sinensis</i>...
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| Format: | Article |
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MDPI AG
2024-01-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/13/2/201 |
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| author | Shengjing Wen Chengzhe Zhou Caiyun Tian Niannian Yang Cheng Zhang Anru Zheng Yixing Chen Zhongxiong Lai Yuqiong Guo |
| author_facet | Shengjing Wen Chengzhe Zhou Caiyun Tian Niannian Yang Cheng Zhang Anru Zheng Yixing Chen Zhongxiong Lai Yuqiong Guo |
| author_sort | Shengjing Wen |
| collection | DOAJ |
| description | The microRNA156 (miR156) family, one of the first miRNA families discovered in plants, plays various important roles in plant growth and resistance to various abiotic stresses. Previously, miR156s were shown to respond to drought stress, but miR156s in tea plants (<i>Camellia sinensis</i> (L.) O. Kuntze) have not been comprehensively identified and analyzed. Herein, we identify 47 mature sequences and 28 precursor sequences in tea plants. Our evolutionary analysis and multiple sequence alignment revealed that csn-miR156s were highly conserved during evolution and that the rates of the csn-miR156 members’ evolution were different. The precursor sequences formed typical and stable stem–loop structures. The prediction of <i>cis</i>-acting elements in the <i>CsMIR156s</i> promoter region showed that the <i>CsMIR156s</i> had diverse <i>cis</i>-acting elements; of these, 12 <i>CsMIR156s</i> were found to be drought-responsive elements. The results of reverse transcription quantitative PCR (RT-qPCR) testing showed that csn-miR156 family members respond to drought and demonstrate different expression patterns under the conditions of drought stress. This suggests that csn-miR156 family members may be significantly involved in the response of tea plants to drought stress. Csn-miR156f-2-5p knockdown significantly reduced the <i>Fv/Fm</i> value and chlorophyll content and led to the accumulation of more-reactive oxygen species and proline compared with the control. The results of target gene prediction showed that csn-miR156f-2-5p targeted SQUAMOSA promoter binding protein-like (<i>SPL</i>) genes. Further analyses showed that <i>CsSPL14</i> was targeted by csn-miR156f-2-5p, as confirmed through RT-qPCR, 5′ RLM-RACE, and antisense oligonucleotide validation. Our results demonstrate that csn-miR156f-2-5p and <i>CsSPL14</i> are involved in drought response and represent a new strategy for increasing drought tolerance via the breeding of tea plants. |
| first_indexed | 2024-03-08T09:48:57Z |
| format | Article |
| id | doaj.art-a129684208aa4e1c98ffd921a45e181b |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-08T09:48:57Z |
| publishDate | 2024-01-01 |
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| series | Plants |
| spelling | doaj.art-a129684208aa4e1c98ffd921a45e181b2024-01-29T14:10:54ZengMDPI AGPlants2223-77472024-01-0113220110.3390/plants13020201Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze)Shengjing Wen0Chengzhe Zhou1Caiyun Tian2Niannian Yang3Cheng Zhang4Anru Zheng5Yixing Chen6Zhongxiong Lai7Yuqiong Guo8College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaThe microRNA156 (miR156) family, one of the first miRNA families discovered in plants, plays various important roles in plant growth and resistance to various abiotic stresses. Previously, miR156s were shown to respond to drought stress, but miR156s in tea plants (<i>Camellia sinensis</i> (L.) O. Kuntze) have not been comprehensively identified and analyzed. Herein, we identify 47 mature sequences and 28 precursor sequences in tea plants. Our evolutionary analysis and multiple sequence alignment revealed that csn-miR156s were highly conserved during evolution and that the rates of the csn-miR156 members’ evolution were different. The precursor sequences formed typical and stable stem–loop structures. The prediction of <i>cis</i>-acting elements in the <i>CsMIR156s</i> promoter region showed that the <i>CsMIR156s</i> had diverse <i>cis</i>-acting elements; of these, 12 <i>CsMIR156s</i> were found to be drought-responsive elements. The results of reverse transcription quantitative PCR (RT-qPCR) testing showed that csn-miR156 family members respond to drought and demonstrate different expression patterns under the conditions of drought stress. This suggests that csn-miR156 family members may be significantly involved in the response of tea plants to drought stress. Csn-miR156f-2-5p knockdown significantly reduced the <i>Fv/Fm</i> value and chlorophyll content and led to the accumulation of more-reactive oxygen species and proline compared with the control. The results of target gene prediction showed that csn-miR156f-2-5p targeted SQUAMOSA promoter binding protein-like (<i>SPL</i>) genes. Further analyses showed that <i>CsSPL14</i> was targeted by csn-miR156f-2-5p, as confirmed through RT-qPCR, 5′ RLM-RACE, and antisense oligonucleotide validation. Our results demonstrate that csn-miR156f-2-5p and <i>CsSPL14</i> are involved in drought response and represent a new strategy for increasing drought tolerance via the breeding of tea plants.https://www.mdpi.com/2223-7747/13/2/201tea plantmiR156sdrought stressSQUAMOSA promoter binding protein-like |
| spellingShingle | Shengjing Wen Chengzhe Zhou Caiyun Tian Niannian Yang Cheng Zhang Anru Zheng Yixing Chen Zhongxiong Lai Yuqiong Guo Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) Plants tea plant miR156s drought stress SQUAMOSA promoter binding protein-like |
| title | Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) |
| title_full | Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) |
| title_fullStr | Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) |
| title_full_unstemmed | Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) |
| title_short | Identification and Validation of the miR156 Family Involved in Drought Responses and Tolerance in Tea Plants (<i>Camellia sinensis</i> (L.) O. Kuntze) |
| title_sort | identification and validation of the mir156 family involved in drought responses and tolerance in tea plants i camellia sinensis i l o kuntze |
| topic | tea plant miR156s drought stress SQUAMOSA promoter binding protein-like |
| url | https://www.mdpi.com/2223-7747/13/2/201 |
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