Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.)
Abstract Background Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TF) LhMYBSPLATTER (formerly known as LhMYB12-Lat), identified as the major kernel protein, regulating the anthocyanin biosynthe...
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| Format: | Article |
| Language: | English |
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BMC
2021-11-01
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| Series: | BMC Plant Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12870-021-03347-1 |
| _version_ | 1818573021686792192 |
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| author | Yuwei Cao Mengmeng Bi Panpan Yang Meng Song Guoren He Jing Wang Yue Yang Leifeng Xu Jun Ming |
| author_facet | Yuwei Cao Mengmeng Bi Panpan Yang Meng Song Guoren He Jing Wang Yue Yang Leifeng Xu Jun Ming |
| author_sort | Yuwei Cao |
| collection | DOAJ |
| description | Abstract Background Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TF) LhMYBSPLATTER (formerly known as LhMYB12-Lat), identified as the major kernel protein, regulating the anthocyanin biosynthesis pathway in ‘Tiny Padhye’ of Tango Series cultivars, which the pigmentation density is high in the lower half of tepals and this patterning is of exceptional ornamental value. However, the research on mechanism of regulating the spatial and temporal expression differences of LhMYBSPLATTER, which belongs to the R2R3-MYB subfamily, is still not well established. To explore the molecular mechanism of directly related regulatory proteins of LhMYBSPLATTER in the anthocyanin pigmentation, the yeast one-hybrid (Y1H) cDNA library was constructed and characterized. Results In this study, we describe a yeast one-hybrid library to screen transcription factors that regulate LhMYBSPLATTER gene expression in Lilium, with the library recombinant efficiency of over 98%. The lengths of inserted fragments ranged from 400 to 2000 bp, and the library capacity reached 1.6 × 106 CFU of cDNA insert, which is suitable to fulfill subsequent screening. Finally, seven prey proteins, including BTF3, MYB4, IAA6-like, ERF4, ARR1, ERF WIN1-like, and ERF061 were screened by the recombinant bait plasmid and verified by interaction with the LhMYBSPLATTER promoter. Among them, ERFs, AUX/IAA, and BTF3 may participate in the negative regulation of the anthocyanin biosynthesis pathway in Lilium. Conclusion A yeast one-hybrid library of lily was successfully constructed in the tepals for the first time. Seven candidate TFs of LhMYBSPLATTER were screened, which may provide a theoretical basis for the study of floral pigmentation. |
| first_indexed | 2024-12-15T00:05:32Z |
| format | Article |
| id | doaj.art-9ac8bfdcc01349248a36a000598f8024 |
| institution | Directory Open Access Journal |
| issn | 1471-2229 |
| language | English |
| last_indexed | 2024-12-15T00:05:32Z |
| publishDate | 2021-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Plant Biology |
| spelling | doaj.art-9ac8bfdcc01349248a36a000598f80242022-12-21T22:42:44ZengBMCBMC Plant Biology1471-22292021-11-0121111010.1186/s12870-021-03347-1Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.)Yuwei Cao0Mengmeng Bi1Panpan Yang2Meng Song3Guoren He4Jing Wang5Yue Yang6Leifeng Xu7Jun Ming8Institute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesInstitute of Vegetables and Flowers, Chinese Academy of Agricultural SciencesAbstract Background Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TF) LhMYBSPLATTER (formerly known as LhMYB12-Lat), identified as the major kernel protein, regulating the anthocyanin biosynthesis pathway in ‘Tiny Padhye’ of Tango Series cultivars, which the pigmentation density is high in the lower half of tepals and this patterning is of exceptional ornamental value. However, the research on mechanism of regulating the spatial and temporal expression differences of LhMYBSPLATTER, which belongs to the R2R3-MYB subfamily, is still not well established. To explore the molecular mechanism of directly related regulatory proteins of LhMYBSPLATTER in the anthocyanin pigmentation, the yeast one-hybrid (Y1H) cDNA library was constructed and characterized. Results In this study, we describe a yeast one-hybrid library to screen transcription factors that regulate LhMYBSPLATTER gene expression in Lilium, with the library recombinant efficiency of over 98%. The lengths of inserted fragments ranged from 400 to 2000 bp, and the library capacity reached 1.6 × 106 CFU of cDNA insert, which is suitable to fulfill subsequent screening. Finally, seven prey proteins, including BTF3, MYB4, IAA6-like, ERF4, ARR1, ERF WIN1-like, and ERF061 were screened by the recombinant bait plasmid and verified by interaction with the LhMYBSPLATTER promoter. Among them, ERFs, AUX/IAA, and BTF3 may participate in the negative regulation of the anthocyanin biosynthesis pathway in Lilium. Conclusion A yeast one-hybrid library of lily was successfully constructed in the tepals for the first time. Seven candidate TFs of LhMYBSPLATTER were screened, which may provide a theoretical basis for the study of floral pigmentation.https://doi.org/10.1186/s12870-021-03347-1LiliumAnthocyaninsTranscriptional factorYeast one-hybrid libraryLhMYBSPLATTER |
| spellingShingle | Yuwei Cao Mengmeng Bi Panpan Yang Meng Song Guoren He Jing Wang Yue Yang Leifeng Xu Jun Ming Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) BMC Plant Biology Lilium Anthocyanins Transcriptional factor Yeast one-hybrid library LhMYBSPLATTER |
| title | Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) |
| title_full | Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) |
| title_fullStr | Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) |
| title_full_unstemmed | Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) |
| title_short | Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.) |
| title_sort | construction of yeast one hybrid library and screening of transcription factors regulating lhmybsplatter expression in asiatic hybrid lilies lilium spp |
| topic | Lilium Anthocyanins Transcriptional factor Yeast one-hybrid library LhMYBSPLATTER |
| url | https://doi.org/10.1186/s12870-021-03347-1 |
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