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Selection and functional identification of Dof genes expressed in response to nitrogen in Populus simonii × Populus nigra Полный текст
2022
Wang Shenmeng | Wang Ruoning | Yang Chengjun
Selection and functional identification of Dof genes expressed in response to nitrogen in Populus simonii × Populus nigra Полный текст
2022
Wang Shenmeng | Wang Ruoning | Yang Chengjun
In plants, Dof transcription factors are involved in regulating the expression of a series of genes related to N uptake and utilization. Therefore, the present study investigated how DNA-binding with one finger (Dof) genes are expressed in response to nitrogen (N) form and concentration to clarify the role of Dof genes and their functions in promoting N assimilation and utilization in poplar. The basic characteristics and expression patterns of Dof genes in poplar were analyzed by the use of bioinformatics methods. Dof genes expressed in response to N were screened, after which the related genes were cloned and transformed into Arabidopsis thaliana; the physiological indexes and the expression of related genes were subsequently determined. The function of Dof genes was then verified in Arabidopsis thaliana plants grown in the presence of different N forms and concentrations. Forty-four Dof genes were identified, most of which were expressed in the roots and young leaves, and some of the Dof genes were expressed under ammonia- and nitrate-N treatments. Three genes related to N induction were cloned, their proteins were found to localize in the nucleus, and PnDof30 was successfully transformed into Arabidopsis thaliana for functional verification. On comparing Arabidopsis thaliana with WT Arabidopsis thaliana plants, Arabidopsis thaliana plants overexpressing the Dof gene grew better under low N levels; the contents of soluble proteins and chlorophyll significantly increased, while the soluble sugar content significantly decreased. The expressions of several AMT, NRT, and GS genes were upregulated, while the expressions of several others were downregulated, and the expression of PEPC and PK genes significantly increased. In addition, the activity of PEPC, PK, GS, and NR enzymes significantly increased. The results showed that overexpression of PnDof30 significantly increased the level of carbon and N metabolism and improved the growth of transgenic Arabidopsis thaliana plants under low-N conditions. The study revealed the biological significance of poplar Dof transcription factors in N response and regulation of related downstream gene expression and provided some meaningful clues to explain the huge difference between poplar and Arabidopsis thaliana transformed by exogenous Dof gene, which could promote the comprehensive understanding of the molecular mechanism of efficient N uptake and utilization in trees.
Показать больше [+] Меньше [-]Selection and functional identification of Dof genes expressed in response to nitrogen in Populus simonii × Populus nigra Полный текст
2022
Wang, Shenmeng | Wang, Ruoning | Yang, Chengjun
In plants, Dof transcription factors are involved in regulating the expression of a series of genes related to N uptake and utilization. Therefore, the present study investigated how DNA-binding with one finger (Dof) genes are expressed in response to nitrogen (N) form and concentration to clarify the role of Dof genes and their functions in promoting N assimilation and utilization in poplar. The basic characteristics and expression patterns of Dof genes in poplar were analyzed by the use of bioinformatics methods. Dof genes expressed in response to N were screened, after which the related genes were cloned and transformed into Arabidopsis thaliana; the physiological indexes and the expression of related genes were subsequently determined. The function of Dof genes was then verified in Arabidopsis thaliana plants grown in the presence of different N forms and concentrations. Forty-four Dof genes were identified, most of which were expressed in the roots and young leaves, and some of the Dof genes were expressed under ammonia- and nitrate-N treatments. Three genes related to N induction were cloned, their proteins were found to localize in the nucleus, and PnDof30 was successfully transformed into Arabidopsis thaliana for functional verification. On comparing Arabidopsis thaliana with WT Arabidopsis thaliana plants, Arabidopsis thaliana plants overexpressing the Dof gene grew better under low N levels; the contents of soluble proteins and chlorophyll significantly increased, while the soluble sugar content significantly decreased. The expressions of several AMT, NRT, and GS genes were upregulated, while the expressions of several others were downregulated, and the expression of PEPC and PK genes significantly increased. In addition, the activity of PEPC, PK, GS, and NR enzymes significantly increased. The results showed that overexpression of PnDof30 significantly increased the level of carbon and N metabolism and improved the growth of transgenic Arabidopsis thaliana plants under low-N conditions. The study revealed the biological significance of poplar Dof transcription factors in N response and regulation of related downstream gene expression and provided some meaningful clues to explain the huge difference between poplar and Arabidopsis thaliana transformed by exogenous Dof gene, which could promote the comprehensive understanding of the molecular mechanism of efficient N uptake and utilization in trees.
Показать больше [+] Меньше [-]PseAraUbi: predicting arabidopsis ubiquitination sites by incorporating the physico-chemical and structural features Полный текст
2022
Wang, Wei | Zhang, Yu | Liu, Dong | Zhang, Hongjun | Wang, XianFang | Zhou, Yun
KEY MESSAGE: We makes three kinds of important features from Arabidopsis thaliana: protein secondary structure based on the Chou-Fasman parameter, amino acids hydrophobicity and polarity information, and analyze their properties. Ubiquitination modification is an important post-translational modification of proteins, which participates in the regulation of many important life activities in cells. At present, ubiquitination proteomics research is mostly concentrated in animals and yeasts, while relatively few studies have been carried out in plants. It can be said that the calculation and prediction of Arabidopsis thaliana ubiquitination sites is still in its infancy. Based on this, we describe a calculation method, PseAraUbi (Prediction of Arabidopsis thaliana ubiquitination sites using pseudo amino acid composition), that can effectively detect ubiquitination sites on Arabidopsis thaliana using support vector machine learning classifiers. Based on protein sequence information, extract features from the Chou-Fasman parameter, amino acids hydrophobicity features, polarity information and selected for classification with the Boruta algorithm. PseAraUbi achieves promising performances with an AUC score of 0.953 with fivefold cross-validation on the training dataset, which are significantly better than that of the pioneer Arabidopsis thaliana ubiquitination sites method. We also proved the ability of our proposed method on independent test sets, thus gaining a competitive advantage. In addition, we also in-depth analyzed the physicochemical properties of amino acids in the region adjacent to the ubiquitination site. To facilitate the community, the source code, optimal feature subset, ubiquitination sites dataset in the Arbidopsis proteome are available at GitHub (https://github.com/HNUBioinformatics/PseAraUbi.git) for interest users.
Показать больше [+] Меньше [-]Dual-transcriptomic datasets evaluating the effect of the necrotrophic fungus Alternaria brassicicola on Arabidopsis germinating seeds Полный текст
2022
Ortega-Cuadros, Mailen | Chir, Laurine | Aligon, Sophie | Arias, Tatiana | Verdier, Jerome | Grappin, Philippe
Dual-transcriptomic datasets evaluating the effect of the necrotrophic fungus Alternaria brassicicola on Arabidopsis germinating seeds Полный текст
2022
Ortega-Cuadros, Mailen | Chir, Laurine | Aligon, Sophie | Arias, Tatiana | Verdier, Jerome | Grappin, Philippe
Many fungal pathogens are carried and transmitted by seeds. These pathogens affect germination and seed quality. Their transmission from the germinating seed to seedling causes many diseases in crops. Seed defense mechanisms during germination are poorly documented. RNA-seq experiments were used to describe the molecular mechanisms involved in seed interaction with a necrotrophic fungus. Here the Arabidopsis thaliana/Alternaria brassicicola pathosystem was used to perform dual-transcriptomic approach. Arabidopsis thaliana seeds and necrotrophic fungus transcripts were identified at critical germination and seedling establishment stages. Total RNA was extracted from healthy and infected germinating seeds and seedlings at 3, 6 and 10 days after sowing. Transcript libraries were made and sequenced, then fungal and plant short reads were mapped and quantified respectively against Arabidopsis thaliana and Alternaria brassicicola reference transcriptomes. This dual-transcriptomic approach revealed that 3409, 7506 and 8589 Arabidopsis thaliana genes showed a differential expression at respectevely 3, 6 and 10 days after sowing between healthy and infected seeds, including 1192 genes differentially expressed at the three studied stages. Moreover, in this experiement, we also identified the dynamic of the transcript changes occurring at the same stages in the necrotrophic fungus concomitantly during germination and seedling establishment.
Показать больше [+] Меньше [-]Development of bi-oligo (ethylene glycol)-functionalized fluorescent probe for two-photon and noninvasive imaging in Arabidopsis thaliana Полный текст
2022
Jingzhi Du | Keming Chen | Zhenyang Yu | Yuhao Qiao | Jiaxin Liu | Qianqian Zhai | Zhe Hu | Sheng-Gang Yang | Jun Li | Huailong Teng
Fluorescent probe is an important tool for investigation of biological events in plant tissues. However, the research on small-molecule based fluorescent probe for plant imaging still stays at the initial stage. In order to noninvasively achieve the subcellular information of Arabidopsis thaliana, a two-photon and fluorene based fluorophore with a bi-oligo (ethylene glycol) functional group was synthesized. Under the laser's irradiation at 750 nm, the subcellular structures of Arabidopsis thaliana including stomata and root tips were clearly observed without slicing, which displayed significant superiorities over traditional single-photon excitation microscopy. Subsequently, the above functionalized fluorophore was further modified with a recognition group (dipicolylamine) to form a smart fluorescent probe (LJTP1). As results, LJTP1 not only can sensitively detect Cu2+/S2− with fluorescence “on-off-on” response in vitro and in vivo, but also can achieve noninvasive imaging in Arabidopsis thaliana tissues under two-photon microscopy. We believe this study will open up a new mind for noninvasive imaging in plant tissues.
Показать больше [+] Меньше [-]Dual-transcriptomic datasets evaluating the effect of the necrotrophic fungus Alternaria brassicicola on Arabidopsis germinating seeds Полный текст
2022
Ortega-Cuadros, Mailen | Chir, Laurine | Aligon, Sophie | Arias, Tatiana | Verdier, Jerome | Grappin, Philippe | Universidad de Antioquia = University of Antioquia [Medellín, Colombia] | Institut de Recherche en Horticulture et Semences (IRHS) ; Université d'Angers (UA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Rennes Angers ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro) | Marie Selby Bot Gardens, | Université Nice Sophia Antipolis - Faculté de Médecine (UNS UFR Médecine) ; Université Nice Sophia Antipolis (1965 - 2019) (UNS) | French Region Pays de la Loire, Angers Loire Métropole and the European Regional Development Fund ("Objectif Végétal, Research, Education and Innovation in Pays de la Loire“).
International audience | Many fungal pathogens are carried and transmitted by seeds. These pathogens affect germination and seed quality. Their transmission from the germinating seed to seedling causes many diseases in crops. Seed defense mechanisms during germination are poorly documented. RNA-seq experiments were used to describe the molecular mechanisms involved in seed interaction with a necrotrophic fungus. Here the Arabidopsis thaliana/Alternaria brassicicola pathosystem was used to perform dual-transcriptomic approach. Arabidopsis thaliana seeds and necrotrophic fungus transcripts were identified at critical germination and seedling establishment stages. Total RNA was extracted from healthy and infected germinating seeds and seedlings, sampled at 3, 6 and 10 days after sowing. Transcript libraries were made and sequenced, then fungal and plant short reads were mapped and quantified respectively against Arabidopsis thaliana and Alternaria brassicicola reference transcriptomes. This dual-transcriptomic approach revealed that 3409, 7506 and 8589 Arabidopsis thaliana genes showed a differential expression at respectevely 3, 6 and 10 days after sowing between healthy and infected seeds, including 1192 genes differentially expressed at the three studied stages. Moreover, in this experiement, we also identified the dynamic of the transcript changes occurring at the same stages in the necrotrophic fungus concomitantly during germination and seedling establishment.
Показать больше [+] Меньше [-]Dual-transcriptomic datasets evaluating the effect of the necrotrophic fungus Alternaria brassicicola on Arabidopsis germinating seeds Полный текст
2022
Ortega-Cuadros, Mailen | Chir, Laurine | Aligon, Sophie | Arias, Tatiana | Verdier, Jerome | Grappin, Philippe | Universidad de Antioquia = University of Antioquia [Medellín, Colombia] | Institut de Recherche en Horticulture et Semences (IRHS) ; Université d'Angers (UA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Rennes Angers ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro) | Marie Selby Bot Gardens, | Université Nice Sophia Antipolis - Faculté de Médecine (UNS UFR Médecine) ; Université Nice Sophia Antipolis (1965 - 2019) (UNS) | French Region Pays de la Loire, Angers Loire Métropole and the European Regional Development Fund ("Objectif Végétal, Research, Education and Innovation in Pays de la Loire“).
International audience | Many fungal pathogens are carried and transmitted by seeds. These pathogens affect germination and seed quality. Their transmission from the germinating seed to seedling causes many diseases in crops. Seed defense mechanisms during germination are poorly documented. RNA-seq experiments were used to describe the molecular mechanisms involved in seed interaction with a necrotrophic fungus. Here the Arabidopsis thaliana/Alternaria brassicicola pathosystem was used to perform dual-transcriptomic approach. Arabidopsis thaliana seeds and necrotrophic fungus transcripts were identified at critical germination and seedling establishment stages. Total RNA was extracted from healthy and infected germinating seeds and seedlings, sampled at 3, 6 and 10 days after sowing. Transcript libraries were made and sequenced, then fungal and plant short reads were mapped and quantified respectively against Arabidopsis thaliana and Alternaria brassicicola reference transcriptomes. This dual-transcriptomic approach revealed that 3409, 7506 and 8589 Arabidopsis thaliana genes showed a differential expression at respectevely 3, 6 and 10 days after sowing between healthy and infected seeds, including 1192 genes differentially expressed at the three studied stages. Moreover, in this experiement, we also identified the dynamic of the transcript changes occurring at the same stages in the necrotrophic fungus concomitantly during germination and seedling establishment.
Показать больше [+] Меньше [-]Genome-Wide Identification of Brassica napus PEN1-LIKE Genes and Their Expression Profiling in Insect-Susceptible and Resistant Cultivars Полный текст
2022
Lei Sheng | Zengbei Feng | Zhongping Hao | Shumin Hou
Genome-Wide Identification of Brassica napus PEN1-LIKE Genes and Their Expression Profiling in Insect-Susceptible and Resistant Cultivars Полный текст
2022
Lei Sheng | Zengbei Feng | Zhongping Hao | Shumin Hou
Recently, it has been reported that a gene (PEN1) in Arabidopsis thaliana is highly resistant to Plutella xylostella. We screened all the homologous genes of PEN1 in Arabidopsis thaliana and found that the motif of these genes was very conserved. At present, few insect resistance genes have been identified and characterized in Brassica napus. Therefore, we screened all the homologous genes containing this motif in the Brassica napus genome and systematically analyzed the basic information, conserved domain, evolutionary relationship, chromosomal localization and expression analysis of these genes. In this study, 12 PEN1 homologous genes were identified in the Brassica napus genome, which is more than the number in Arabidopsis thaliana. These genes are unevenly distributed on the 12 chromosomes in Brassica napus. Furthermore, all the PEN1 homologous genes contained light responsiveness elements, and most of the genes contained gibberellin-responsive elements, meJA-responsive elements and abscisic-acid-responsive elements. The results will provide a theoretical basis for screening insect resistance genes from the genome of Brassica napus and analyzing the molecular mechanism of insect resistance in Brassica napus.
Показать больше [+] Меньше [-]Genome-Wide Identification of <i>Brassica napus</i> PEN1-LIKE Genes and Their Expression Profiling in Insect-Susceptible and Resistant Cultivars Полный текст
2022
Lei Sheng | Zengbei Feng | Zhongping Hao | Shumin Hou
Recently, it has been reported that a gene (PEN1) in <i>Arabidopsis thaliana</i> is highly resistant to <i>Plutella xylostella</i>. We screened all the homologous genes of PEN1 in <i>Arabidopsis thaliana</i> and found that the motif of these genes was very conserved. At present, few insect resistance genes have been identified and characterized in <i>Brassica napus</i>. Therefore, we screened all the homologous genes containing this motif in the <i>Brassica napus</i> genome and systematically analyzed the basic information, conserved domain, evolutionary relationship, chromosomal localization and expression analysis of these genes. In this study, 12 PEN1 homologous genes were identified in the <i>Brassica napus</i> genome, which is more than the number in <i>Arabidopsis thaliana</i>. These genes are unevenly distributed on the 12 chromosomes in <i>Brassica napus</i>. Furthermore, all the PEN1 homologous genes contained light responsiveness elements, and most of the genes contained gibberellin-responsive elements, meJA-responsive elements and abscisic-acid-responsive elements. The results will provide a theoretical basis for screening insect resistance genes from the genome of <i>Brassica napus</i> and analyzing the molecular mechanism of insect resistance in <i>Brassica napus</i>.
Показать больше [+] Меньше [-]Expression and Functional Study of BcWRKY1 in Baphicacanthus cusia (Nees) Bremek Полный текст
2022
Meijuan Zeng | Meijuan Zeng | Yongjia Zhong | Zhiying Guo | Zhiying Guo | Huiyong Yang | Haisheng Zhu | Liling Zheng | Yong Diao
Baphicacanthus cusia (Nees) Bremek (B. cusia) is an important medicinal plant. Its effective substances including indigo and indirubin are metabolites in indoleacetate metabolic pathway. Based on a previous transcriptome sequencing analysis, a WRKY transcription factor, BcWRKY1, in B. cusia was identified, showing significant correlation with effective substances from B. cusia. In this study, BcWRKY1 was cloned by reverse transcription-polymerase chain reaction (RT-PCR). Further analysis showed that the BcWRKY1 gene was 916 bp in length, containing three exons and two introns. The open reading frame (ORF) of BcWRKY1 was 534 bp in length and encoded a WRKY domain-containing protein with 177 amino acids residues. Subcellular localization showed that BcWRKY1 protein was mainly localized in the nucleus. It could bind to the W-box motif and its role in transcriptional activation was confirmed in yeast. The function of BcWRKY1 was investigated by overexpressing BcWRKY1 in Arabidopsis thaliana. Metabolic profiles in wild type and BcWRKY1-OX1 transgenic Arabidopsis thaliana were analyzed with LC-MS. Results showed that the metabolic profile was significantly changed in BcWRKY1-OX1 transgenic Arabidopsis thaliana compared with wild type. Furthermore, indole-related metabolites were significantly increased in BcWRKY1-OX1 transgenic Arabidopsis thaliana, and the metabolic pathway analysis showed that flavonoid biosynthesis was significantly enriched. Overexpression of BcWRKY1 significantly changed flavonoid and indole metabolism and indole-related metabolites were significantly upregulated. We postulated that the BcWRKY1 transcription factor might be involved in the regulation of effective substances metabolism in B. cusia.
Показать больше [+] Меньше [-]Regulation of ammonium transport in Arabidopsis thaliana Полный текст
2022
Ganz, Pascal
The overarching question of this thesis deals with how plants ensure the selective uptake of ammonium while maintaining ion and pH homeostasis. A key component of this is ammonium transporters (AMTs) with high affinity towards their substrate, which are at the same time part of a multilayered protection system against uncontrolled ammonium influx. Conserved protein sequences inside the transporter were analyzed as well as the regulatory system based on post-translational modification of the transporter. | Die übergeordnete Fragestellung dieser Arbeit beschäftigt sich damit, wie Pflanzen die selektive Aufnahme von Ammonium, bei gleichzeitiger Erhaltung der Ionen- und pH-Homöostase, sicherstellen. Eine Schlüsselkomponente sind dabei Ammoniumtransporter (AMTs) mit hoher Affinität gegenüber ihrem Substrat die gleichzeitig Teil eines mehrschichtigen Schutzsystems gegen unkontrollierten Ammonium Zustrom sind. Konservierte Proteinsequenzen im Inneren des Transporters wurden ebenso analysiert wie das regulatorische System, das auf post-translationaler Modifikation des Transporters beruht.
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