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Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica

Juglans mandshurica (Manchurian walnut) is a precious timber and woody grain and oil species in Northeast China. The heterodichogamous characteristic phenomenon resulted in the non-synchronous flowering and development of male and female flowers, which limited the mating and the yield and quality of...

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Autores principales: Zhang, Lijie, Fu, Jingqi, Dong, Tianyi, Zhang, Mengmeng, Wu, Jingwen, Liu, Chunping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10602732/
https://www.ncbi.nlm.nih.gov/pubmed/37900747
http://dx.doi.org/10.3389/fpls.2023.1243030
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author Zhang, Lijie
Fu, Jingqi
Dong, Tianyi
Zhang, Mengmeng
Wu, Jingwen
Liu, Chunping
author_facet Zhang, Lijie
Fu, Jingqi
Dong, Tianyi
Zhang, Mengmeng
Wu, Jingwen
Liu, Chunping
author_sort Zhang, Lijie
collection PubMed
description Juglans mandshurica (Manchurian walnut) is a precious timber and woody grain and oil species in Northeast China. The heterodichogamous characteristic phenomenon resulted in the non-synchronous flowering and development of male and female flowers, which limited the mating and the yield and quality of fruits. LFY is a core gene in the flowering regulatory networks, which has been cloned in J. mandshurica, and the function has also been verified preliminarily. In this study, the JmLFY promoter sequence with different lengths of 5′-deletion (pLFY1-pLFY6) were cloned and conducted bioinformatics analysis, the promoter activities were analyzed by detecting their driving activity to GUS gene in the tobacco plants that transformed with different promoter sequence stably or transiently. After that, the interaction between JmSOC1 and JmLFY gene promoter was also analyzed via yeast single-hybrid. The results showed that the promoter sequence contains core cis-acting elements essential for eukaryotic promoters, hormone response elements, defense- and stress-responsive elements, flowering-related elements, etc. Transgenic tobacco plants with pLFY1 were obtained by Agrobacterium infection using the pCAMBIA1301 expression vector, and the GUS gene driven by the JmLFY promoter was detected to express in the leaf, stem, flower, and root of the transformed tobacco plant, which indicated that the obtained JmLFY promoter had driving activity. GUS histochemical staining and enzyme activity detection showed that promoter fragments with different lengths had promoter activity and could respond to the induction of long photoperiod, low temperature, salicylic acid (SA), IAA, GA3, and methyl jasmonate (MeJA). The core regulatory region of JmLFY gene promoter in J. mandshurica was between −657 bp and −1,904 bp. Point-to-point validation of yeast single-hybrid confirmed the interaction between JmSOC1 and JmLFY gene promoter, which indicated that JmLFY gene is the downstream target of JmSOC1. These results reveal relevant factors affecting JmLFY gene expression and clarify the molecular mechanism of JmLFY gene regulation in the flower developmental partially, which will provide a theoretical basis for regulating the flowering time by regulating JmLFY gene expression in J. mandshurica.
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spelling pubmed-106027322023-10-27 Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica Zhang, Lijie Fu, Jingqi Dong, Tianyi Zhang, Mengmeng Wu, Jingwen Liu, Chunping Front Plant Sci Plant Science Juglans mandshurica (Manchurian walnut) is a precious timber and woody grain and oil species in Northeast China. The heterodichogamous characteristic phenomenon resulted in the non-synchronous flowering and development of male and female flowers, which limited the mating and the yield and quality of fruits. LFY is a core gene in the flowering regulatory networks, which has been cloned in J. mandshurica, and the function has also been verified preliminarily. In this study, the JmLFY promoter sequence with different lengths of 5′-deletion (pLFY1-pLFY6) were cloned and conducted bioinformatics analysis, the promoter activities were analyzed by detecting their driving activity to GUS gene in the tobacco plants that transformed with different promoter sequence stably or transiently. After that, the interaction between JmSOC1 and JmLFY gene promoter was also analyzed via yeast single-hybrid. The results showed that the promoter sequence contains core cis-acting elements essential for eukaryotic promoters, hormone response elements, defense- and stress-responsive elements, flowering-related elements, etc. Transgenic tobacco plants with pLFY1 were obtained by Agrobacterium infection using the pCAMBIA1301 expression vector, and the GUS gene driven by the JmLFY promoter was detected to express in the leaf, stem, flower, and root of the transformed tobacco plant, which indicated that the obtained JmLFY promoter had driving activity. GUS histochemical staining and enzyme activity detection showed that promoter fragments with different lengths had promoter activity and could respond to the induction of long photoperiod, low temperature, salicylic acid (SA), IAA, GA3, and methyl jasmonate (MeJA). The core regulatory region of JmLFY gene promoter in J. mandshurica was between −657 bp and −1,904 bp. Point-to-point validation of yeast single-hybrid confirmed the interaction between JmSOC1 and JmLFY gene promoter, which indicated that JmLFY gene is the downstream target of JmSOC1. These results reveal relevant factors affecting JmLFY gene expression and clarify the molecular mechanism of JmLFY gene regulation in the flower developmental partially, which will provide a theoretical basis for regulating the flowering time by regulating JmLFY gene expression in J. mandshurica. Frontiers Media S.A. 2023-10-12 /pmc/articles/PMC10602732/ /pubmed/37900747 http://dx.doi.org/10.3389/fpls.2023.1243030 Text en Copyright © 2023 Zhang, Fu, Dong, Zhang, Wu and Liu https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Zhang, Lijie
Fu, Jingqi
Dong, Tianyi
Zhang, Mengmeng
Wu, Jingwen
Liu, Chunping
Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title_full Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title_fullStr Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title_full_unstemmed Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title_short Promoter cloning and activities analysis of JmLFY, a key gene for flowering in Juglans mandshurica
title_sort promoter cloning and activities analysis of jmlfy, a key gene for flowering in juglans mandshurica
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10602732/
https://www.ncbi.nlm.nih.gov/pubmed/37900747
http://dx.doi.org/10.3389/fpls.2023.1243030
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