Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.)
Rapeseed (<i>Brassica napus</i> L.) is an important crop for edible oil, vegetables, and biofuel. Rapeseed growth and development require a minimum temperature of ~1–3 °C. Notably, frost damage occurs during overwintering, posing a serious threat to the productivity and yield of rapeseed...
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MDPI AG
2023-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/11/9514 |
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| author | Dan Luo Ali Raza Yong Cheng Xiling Zou Yan Lv |
| author_facet | Dan Luo Ali Raza Yong Cheng Xiling Zou Yan Lv |
| author_sort | Dan Luo |
| collection | DOAJ |
| description | Rapeseed (<i>Brassica napus</i> L.) is an important crop for edible oil, vegetables, and biofuel. Rapeseed growth and development require a minimum temperature of ~1–3 °C. Notably, frost damage occurs during overwintering, posing a serious threat to the productivity and yield of rapeseed. MYB proteins are important transcription factors (TFs) in plants, and have been proven to be involved in the regulation of stress responses. However, the roles of the MYB TFs in rapeseed under cold stress conditions are yet to be fully elucidated. To better understand the molecular mechanisms of one MYB-like 17 gene, <i>BnaMYBL17</i>, in response to low temperature, the present study found that the transcript level of <i>BnaMYBL17</i> is induced by cold stress. To characterize the gene’s function, the 591 bp coding sequence (CDS) from rapeseed was isolated and stably transformed into rapeseed. The further functional analysis revealed significant sensitivity in <i>BnaMYBL17</i> overexpression lines (<i>BnaMYBL17</i>-OE) after freezing stress, suggesting its involvement in freezing response. A total of 14,298 differentially expressed genes relative to freezing response were found based on transcriptomic analysis of <i>BnaMYBL17</i>-OE. Overall, 1321 candidate target genes were identified based on differential expression, including Phospholipases C1 (<i>PLC1</i>), FCS-like zinc finger 8 (<i>FLZ8</i>), and Kinase on the inside (<i>KOIN</i>). The qPCR results confirmed that the expression levels of certain genes showed fold changes ranging from two to six when compared between <i>BnaMYBL17</i>-OE and WT lines after exposure to freezing stress. Furthermore, verification indicated that <i>BnaMYBL17</i> affects the promoter of <i>BnaPLC1</i>, <i>BnaFLZ8,</i> and <i>BnaKOIN</i> genes. In summary, the results suggest that <i>BnaMYBL17</i> acts as a transcriptional repressor in regulating certain genes related to growth and development during freezing stress. These findings provide valuable genetic and theoretical targets for molecular breeding to enhance freezing tolerance in rapeseed. |
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| issn | 1661-6596 1422-0067 |
| language | English |
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| spelling | doaj.art-d3106c94e4724e8486bcbf27d58b38cd2023-11-18T08:00:02ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-05-012411951410.3390/ijms24119514Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.)Dan Luo0Ali Raza1Yong Cheng2Xiling Zou3Yan Lv4Key Laboratory of Biology and Genetic Improvement of Oil Crops, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Ministry of Agriculture, Wuhan 430062, ChinaKey Laboratory of Biology and Genetic Improvement of Oil Crops, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Ministry of Agriculture, Wuhan 430062, ChinaKey Laboratory of Biology and Genetic Improvement of Oil Crops, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Ministry of Agriculture, Wuhan 430062, ChinaKey Laboratory of Biology and Genetic Improvement of Oil Crops, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Ministry of Agriculture, Wuhan 430062, ChinaKey Laboratory of Biology and Genetic Improvement of Oil Crops, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Ministry of Agriculture, Wuhan 430062, ChinaRapeseed (<i>Brassica napus</i> L.) is an important crop for edible oil, vegetables, and biofuel. Rapeseed growth and development require a minimum temperature of ~1–3 °C. Notably, frost damage occurs during overwintering, posing a serious threat to the productivity and yield of rapeseed. MYB proteins are important transcription factors (TFs) in plants, and have been proven to be involved in the regulation of stress responses. However, the roles of the MYB TFs in rapeseed under cold stress conditions are yet to be fully elucidated. To better understand the molecular mechanisms of one MYB-like 17 gene, <i>BnaMYBL17</i>, in response to low temperature, the present study found that the transcript level of <i>BnaMYBL17</i> is induced by cold stress. To characterize the gene’s function, the 591 bp coding sequence (CDS) from rapeseed was isolated and stably transformed into rapeseed. The further functional analysis revealed significant sensitivity in <i>BnaMYBL17</i> overexpression lines (<i>BnaMYBL17</i>-OE) after freezing stress, suggesting its involvement in freezing response. A total of 14,298 differentially expressed genes relative to freezing response were found based on transcriptomic analysis of <i>BnaMYBL17</i>-OE. Overall, 1321 candidate target genes were identified based on differential expression, including Phospholipases C1 (<i>PLC1</i>), FCS-like zinc finger 8 (<i>FLZ8</i>), and Kinase on the inside (<i>KOIN</i>). The qPCR results confirmed that the expression levels of certain genes showed fold changes ranging from two to six when compared between <i>BnaMYBL17</i>-OE and WT lines after exposure to freezing stress. Furthermore, verification indicated that <i>BnaMYBL17</i> affects the promoter of <i>BnaPLC1</i>, <i>BnaFLZ8,</i> and <i>BnaKOIN</i> genes. In summary, the results suggest that <i>BnaMYBL17</i> acts as a transcriptional repressor in regulating certain genes related to growth and development during freezing stress. These findings provide valuable genetic and theoretical targets for molecular breeding to enhance freezing tolerance in rapeseed.https://www.mdpi.com/1422-0067/24/11/9514abiotic stresscanolagene overexpressionlow temperatureosmoprotectantstranscriptome analysis |
| spellingShingle | Dan Luo Ali Raza Yong Cheng Xiling Zou Yan Lv Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) International Journal of Molecular Sciences abiotic stress canola gene overexpression low temperature osmoprotectants transcriptome analysis |
| title | Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) |
| title_full | Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) |
| title_fullStr | Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) |
| title_full_unstemmed | Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) |
| title_short | Cloning and Functional Characterization of Cold-Inducible MYB-like 17 Transcription Factor in Rapeseed (<i>Brassica napus</i> L.) |
| title_sort | cloning and functional characterization of cold inducible myb like 17 transcription factor in rapeseed i brassica napus i l |
| topic | abiotic stress canola gene overexpression low temperature osmoprotectants transcriptome analysis |
| url | https://www.mdpi.com/1422-0067/24/11/9514 |
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