Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance
Trichomes are specialized hair-like organs found on epidermal cells of many terrestrial plants, which protect plant from excessive transpiration and numerous abiotic and biotic stresses. However, the genetic basis and underlying mechanisms are largely unknown in Nicotiana tabacum (common tobacco), a...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-10-01
|
| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2022.1005811/full |
| _version_ | 1811228182030843904 |
|---|---|
| author | Jing Yu Jing Yu Bo Lei Huina Zhao Bing Wang Kaleem U. Kakar Yushuang Guo Xiaolian Zhang Xiaolian Zhang Mengao Jia Hui Yang Degang Zhao Degang Zhao |
| author_facet | Jing Yu Jing Yu Bo Lei Huina Zhao Bing Wang Kaleem U. Kakar Yushuang Guo Xiaolian Zhang Xiaolian Zhang Mengao Jia Hui Yang Degang Zhao Degang Zhao |
| author_sort | Jing Yu |
| collection | DOAJ |
| description | Trichomes are specialized hair-like organs found on epidermal cells of many terrestrial plants, which protect plant from excessive transpiration and numerous abiotic and biotic stresses. However, the genetic basis and underlying mechanisms are largely unknown in Nicotiana tabacum (common tobacco), an established model system for genetic engineering and plant breeding. In present study, we identified, cloned and characterized an unknown function transcription factor NtMYB306a from tobacco cultivar K326 trichomes. Results obtained from sequence phylogenetic tree analysis showed that NtMYB306a-encoded protein belonged to S1 subgroup of the plants’ R2R3-MYB transcription factors (TFs). Observation of the green fluorescent signals from NtMYB306a-GFP fusion protein construct exhibited that NtMYB306a was localized in nucleus. In yeast transactivation assays, the transformed yeast containing pGBKT7-NtMYB306a construct was able to grow on SD/-Trp-Ade+X-α-gal selection media, signifying that NtMYB306a exhibits transcriptional activation activity. Results from qRT-PCR, in-situ hybridization and GUS staining of transgenic tobacco plants revealed that NtMYB306a is primarily expressed in tobacco trichomes, especially tall glandular trichomes (TGTs) and short glandular trichomes (SGTs). RNA sequencing (RNA-seq) and qRT-PCR analysis of the NtMYB306a-overexpressing transgenic tobacco line revealed that NtMYB306a activated the expression of a set of key target genes which were associated with wax alkane biosynthesis. Gas Chromatography–Mass Spectrometry (GC-MS) exhibited that the total alkane contents and the contents of n-C28, n-C29, n-C31, and ai-C31 alkanes in leaf exudates of NtMYB306a-OE lines (OE-3, OE-13, and OE-20) were significantly greater when compared to WT. Besides, the promoter region of NtMYB306a contained numerous stress-responsive cis-acting elements, and their differential expression towards salicylic acid and cold stress treatments reflected their roles in signal transduction and cold-stress tolerance. Together, these results suggest that NtMYB306a is necessarily a positive regulator of alkane metabolism in tobacco trichomes that does not affect the number and morphology of tobacco trichomes, and that it can be used as a candidate gene for improving stress resistance and the quality of tobacco. |
| first_indexed | 2024-04-12T09:53:58Z |
| format | Article |
| id | doaj.art-3331d268fe324c0e81c5bf55b65c69d6 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-12T09:53:58Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-3331d268fe324c0e81c5bf55b65c69d62022-12-22T03:37:45ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2022-10-011310.3389/fpls.2022.10058111005811Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress toleranceJing Yu0Jing Yu1Bo Lei2Huina Zhao3Bing Wang4Kaleem U. Kakar5Yushuang Guo6Xiaolian Zhang7Xiaolian Zhang8Mengao Jia9Hui Yang10Degang Zhao11Degang Zhao12Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-bioengineering, Guizhou University, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaDepartment of Microbiology, Baluchistan University of Information Technology and Managemnet Sciences, Quetta, PakistanGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaKey Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-bioengineering, Guizhou University, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaGuizhou Academy of Tobacco Science, Molecular Genetics Key Laboratory of China Tobacco, Guiyang, ChinaKey Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-bioengineering, Guizhou University, Guiyang, ChinaPlant Conservation Technology Center, Guizhou Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, ChinaTrichomes are specialized hair-like organs found on epidermal cells of many terrestrial plants, which protect plant from excessive transpiration and numerous abiotic and biotic stresses. However, the genetic basis and underlying mechanisms are largely unknown in Nicotiana tabacum (common tobacco), an established model system for genetic engineering and plant breeding. In present study, we identified, cloned and characterized an unknown function transcription factor NtMYB306a from tobacco cultivar K326 trichomes. Results obtained from sequence phylogenetic tree analysis showed that NtMYB306a-encoded protein belonged to S1 subgroup of the plants’ R2R3-MYB transcription factors (TFs). Observation of the green fluorescent signals from NtMYB306a-GFP fusion protein construct exhibited that NtMYB306a was localized in nucleus. In yeast transactivation assays, the transformed yeast containing pGBKT7-NtMYB306a construct was able to grow on SD/-Trp-Ade+X-α-gal selection media, signifying that NtMYB306a exhibits transcriptional activation activity. Results from qRT-PCR, in-situ hybridization and GUS staining of transgenic tobacco plants revealed that NtMYB306a is primarily expressed in tobacco trichomes, especially tall glandular trichomes (TGTs) and short glandular trichomes (SGTs). RNA sequencing (RNA-seq) and qRT-PCR analysis of the NtMYB306a-overexpressing transgenic tobacco line revealed that NtMYB306a activated the expression of a set of key target genes which were associated with wax alkane biosynthesis. Gas Chromatography–Mass Spectrometry (GC-MS) exhibited that the total alkane contents and the contents of n-C28, n-C29, n-C31, and ai-C31 alkanes in leaf exudates of NtMYB306a-OE lines (OE-3, OE-13, and OE-20) were significantly greater when compared to WT. Besides, the promoter region of NtMYB306a contained numerous stress-responsive cis-acting elements, and their differential expression towards salicylic acid and cold stress treatments reflected their roles in signal transduction and cold-stress tolerance. Together, these results suggest that NtMYB306a is necessarily a positive regulator of alkane metabolism in tobacco trichomes that does not affect the number and morphology of tobacco trichomes, and that it can be used as a candidate gene for improving stress resistance and the quality of tobacco.https://www.frontiersin.org/articles/10.3389/fpls.2022.1005811/fullNtMYB306atranscription factorstrichomesNicotiana tabacumalkane biosynthesis |
| spellingShingle | Jing Yu Jing Yu Bo Lei Huina Zhao Bing Wang Kaleem U. Kakar Yushuang Guo Xiaolian Zhang Xiaolian Zhang Mengao Jia Hui Yang Degang Zhao Degang Zhao Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance Frontiers in Plant Science NtMYB306a transcription factors trichomes Nicotiana tabacum alkane biosynthesis |
| title | Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| title_full | Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| title_fullStr | Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| title_full_unstemmed | Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| title_short | Cloning, characterization and functional analysis of NtMYB306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| title_sort | cloning characterization and functional analysis of ntmyb306a gene reveals its role in wax alkane biosynthesis of tobacco trichomes and stress tolerance |
| topic | NtMYB306a transcription factors trichomes Nicotiana tabacum alkane biosynthesis |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2022.1005811/full |
| work_keys_str_mv | AT jingyu cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT jingyu cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT bolei cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT huinazhao cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT bingwang cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT kaleemukakar cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT yushuangguo cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT xiaolianzhang cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT xiaolianzhang cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT mengaojia cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT huiyang cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT degangzhao cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance AT degangzhao cloningcharacterizationandfunctionalanalysisofntmyb306agenerevealsitsroleinwaxalkanebiosynthesisoftobaccotrichomesandstresstolerance |