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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Yuwei Cao, Mengmeng Bi, Panpan Yang, Meng Song, Guoren He, Jing Wang, Yue Yang, Leifeng Xu, Jun Ming
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Lilium Anthocyanins Transcriptional factor Yeast one-hybrid library LhMYBSPLATTER Botany QK1-989
Acceso en línea:	https://doaj.org/article/9ac8bfdcc01349248a36a000598f8024
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9ac8bfdcc01349248a36a000598f8024
record_format	dspace
spelling	oai:doaj.org-article:9ac8bfdcc01349248a36a000598f80242021-12-05T12:06:23ZConstruction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.)10.1186/s12870-021-03347-11471-2229https://doaj.org/article/9ac8bfdcc01349248a36a000598f80242021-11-01T00:00:00Zhttps://doi.org/10.1186/s12870-021-03347-1https://doaj.org/toc/1471-2229Abstract 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.Yuwei CaoMengmeng BiPanpan YangMeng SongGuoren HeJing WangYue YangLeifeng XuJun MingBMCarticleLiliumAnthocyaninsTranscriptional factorYeast one-hybrid libraryLhMYBSPLATTERBotanyQK1-989ENBMC Plant Biology, Vol 21, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Lilium Anthocyanins Transcriptional factor Yeast one-hybrid library LhMYBSPLATTER Botany QK1-989
spellingShingle	Lilium Anthocyanins Transcriptional factor Yeast one-hybrid library LhMYBSPLATTER Botany QK1-989 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.)
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.
format	article
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
title	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_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_sort	construction of yeast one-hybrid library and screening of transcription factors regulating lhmybsplatter expression in asiatic hybrid lilies (lilium spp.)
publisher	BMC
publishDate	2021
url	https://doaj.org/article/9ac8bfdcc01349248a36a000598f8024
work_keys_str_mv	AT yuweicao constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT mengmengbi constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT panpanyang constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT mengsong constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT guorenhe constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT jingwang constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT yueyang constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT leifengxu constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp AT junming constructionofyeastonehybridlibraryandscreeningoftranscriptionfactorsregulatinglhmybsplatterexpressioninasiatichybridliliesliliumspp
_version_	1718372272771694592

Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.)

Ejemplares similares