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Keywords = AMT1 family

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16 pages, 8050 KiB  
Article
Functional Characterization of Ammonium Transporter LjAMT2;4 During Lotus japonicus Symbiosis with Rhizobia and Arbuscular Mycorrhizal Fungi
by Kailing Xie, Ying Ni, Lijie Bai, Yuqian Zhai, Wenqing Zhou, Beijiu Cheng and Xiaoyu Li
J. Fungi 2025, 11(5), 340; https://doi.org/10.3390/jof11050340 - 27 Apr 2025
Viewed by 489
Abstract
Arbuscular mycorrhizal fungi (AMF) and rhizobia are important symbiotic microorganisms in soil, which can symbiose with legumes to form mycorrhizal symbionts and nodules, respectively. Once a stable symbiotic relationship is established, these microorganisms have been found to enhance nitrogen absorption by legumes. Although [...] Read more.
Arbuscular mycorrhizal fungi (AMF) and rhizobia are important symbiotic microorganisms in soil, which can symbiose with legumes to form mycorrhizal symbionts and nodules, respectively. Once a stable symbiotic relationship is established, these microorganisms have been found to enhance nitrogen absorption by legumes. Although plants can directly utilize ammonium through ammonium transporters (AMTs), there is limited research on the role of the AMT gene family in promoting ammonium transport in symbiotic relationships. Lotus japonicus, a common host of arbuscular mycorrhizal fungi and rhizobia, serves as a model legume plant. In this study, we examined the characteristics of the ammonium transporter LjAMT2;4 in L. japonicus and found that LjAMT2;4 is localized to the plasma membrane and is predominantly expressed in roots. The promoter region of LjAMT2;4 contains cis-acting elements induced by arbuscular mycorrhizal fungi and rhizomes, and the expression of LjAMT2;4 was induced by AM fungi and rhizobia. However, there was no significant difference in the mycorrhizal colonization rate of ljamt2;4 compared to the wild type, while the absence of LjAMT2;4 significantly increased the number of root nodules under nitrogen-starved conditions, enhancing nitrogen fixation and alleviating nitrogen stress in extremely nitrogen-starved environments, ultimately promoting plant growth. These findings suggest that manipulating the genes involved in symbiotic nitrogen fixation, such as LjAMT2;4, could offer new strategies for sustainable agricultural production. Given that AM and rhizobia symbiosis are critical for crop growth, our findings may inform strategies to improve agricultural management. Full article
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13 pages, 2282 KiB  
Article
Ammonium Transporter 1 (AMT1) Gene Family in Pomegranate: Genome-Wide Analysis and Expression Profiles in Response to Salt Stress
by Fatima Omari Alzahrani
Curr. Issues Mol. Biol. 2025, 47(1), 59; https://doi.org/10.3390/cimb47010059 - 16 Jan 2025
Cited by 1 | Viewed by 1095
Abstract
Understanding the ammonium (NH4+) uptake and transport systems, particularly AMT1 genes, is important for plant growth and defense. However, there is a lack of research on identifying and analyzing AMT1 genes in pomegranate, emphasizing the need for further investigation in [...] Read more.
Understanding the ammonium (NH4+) uptake and transport systems, particularly AMT1 genes, is important for plant growth and defense. However, there is a lack of research on identifying and analyzing AMT1 genes in pomegranate, emphasizing the need for further investigation in this area. Five AMT1 genes (PgAMT1-1 to PgAMT1-5) were identified, all of which contain the PF00909 domain, a feature of ammonium transporters. Various characteristics of these genes, including gene length, coding sequence length, and chromosomal locations, were examined. This study evaluated the isoelectric point, hydropathicity, conserved domains, motifs, and synteny of the PgAMT1 proteins. Phylogenetic analysis confirmed the homology of PgAMT1 genes with previously reported AMT in Arabidopsis and tomato. The tissue-specific expression analysis of PgAMT1 genes revealed distinct patterns: PgAMT1-1 and PgAMT1-2 were predominantly expressed in flowers, PgAMT1-3 exhibited notable expression in roots, leaves, and flowers, PgAMT1-4 was primarily expressed in leaf tissue, while the expression of PgAMT1-5 was detected in both leaves and roots. The impact of salt-induced stress on AMT1 gene expression was also examined, revealing that PgAMT1-1, PgAMT1-2, and PgAMT1-4 expression is reduced under increased salt stress. These expression modifications can help regulate NH4+ assimilation in conditions of elevated salinity, maintaining cellular homeostasis and ion balance. This study contributes to the comprehensive identification of the AMT1s gene family in pomegranate; however, further research on the functional characterization of the identified PgAMT1s is needed. Full article
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24 pages, 6615 KiB  
Article
The Identification of AMT Family Genes and Their Expression, Function, and Regulation in Chenopodium quinoa
by Xiangxiang Wang, He Wu, Nazer Manzoor, Wenhua Dongcheng, Youbo Su, Zhengjie Liu, Chun Lin and Zichao Mao
Plants 2024, 13(24), 3524; https://doi.org/10.3390/plants13243524 - 17 Dec 2024
Cited by 1 | Viewed by 1136
Abstract
Quinoa (Chenopodium quinoa) is an Andean allotetraploid pseudocereal crop with higher protein content and balanced amino acid composition in the seeds. Ammonium (NH4+), a direct source of organic nitrogen assimilation, mainly transported by specific transmembrane ammonium transporters ( [...] Read more.
Quinoa (Chenopodium quinoa) is an Andean allotetraploid pseudocereal crop with higher protein content and balanced amino acid composition in the seeds. Ammonium (NH4+), a direct source of organic nitrogen assimilation, mainly transported by specific transmembrane ammonium transporters (AMTs), plays important roles in the development, yield, and quality of crops. Many AMTs and their functions have been identified in major crops; however, no systematic analyses of AMTs and their regulatory networks, which is important to increase the yield and protein accumulation in the seeds of quinoa, have been performed to date. In this study, the CqAMTs were identified, followed by the quantification of the gene expression, while the regulatory networks were predicted based on weighted gene co-expression network analysis (WGCNA), with the putative transcriptional factors (TFs) having binding sites on the promoters of CqAMTs, nitrate transporters (CqNRTs), and glutamine-synthases (CqGSs), as well as the putative TF expression being correlated with the phenotypes and activities of GSs, glutamate synthase (GOGAT), nitrite reductase (NiR), and nitrate reductase (NR) of quinoa roots. The results showed a total of 12 members of the CqAMT family with varying expressions in different organs and in the same organs at different developmental stages. Complementation expression analyses in the triple mep1/2/3 mutant of yeast showed that except for CqAMT2.2b, 11/12 CqAMTs restored the uptake of NH4+ in the host yeast. CqAMT1.2a was found to mainly locate on the cell membrane, while TFs (e.g., CqNLPs, CqG2Ls, B3 TFs, CqbHLHs, CqZFs, CqMYBs, CqNF-YA/YB/YC, CqNACs, and CqWRKY) were predicted to be predominantly involved in the regulation, transportation, and assimilation of nitrogen. These results provide the functions of CqAMTs and their possible regulatory networks, which will lead to improved nitrogen use efficiency (NUE) in quinoa as well as other major crops. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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18 pages, 6059 KiB  
Article
Moso Bamboo’s Survival Strategy Against Chilling Stress in Signaling Dynamics
by Xiong Jing, Chunju Cai, Pengfei Geng and Yi Wang
Forests 2024, 15(12), 2132; https://doi.org/10.3390/f15122132 - 2 Dec 2024
Cited by 1 | Viewed by 1035
Abstract
Phyllostachys edulis, an economically and ecologically significant bamboo species, has substantial research value in applications as a bamboo substitute for plastic and in forest carbon sequestration. However, frequent seasonal low-temperature events due to global climate change affect the growth, development, and productivity [...] Read more.
Phyllostachys edulis, an economically and ecologically significant bamboo species, has substantial research value in applications as a bamboo substitute for plastic and in forest carbon sequestration. However, frequent seasonal low-temperature events due to global climate change affect the growth, development, and productivity of P. edulis. Calcium signaling, serving as a versatile second messenger, is involved in various stress responses and nitrogen metabolism. In this study, we analyzed the calcium signaling dynamics and regulatory strategies in P. edulis under chilling stress. Differentially expressed genes (DEGs) from the CBF families, AMT families, NRT families, and Ca2+ sensor families, including CaM, CDPK, and CBL, were identified using transcriptomics. Additionally, we explored the law of Ca2+ flux and distribution in the roots of P. edulis under chilling stress and validated these findings by assessing the content or activity of Ca2+ sensor proteins and nitrogen transport proteins in the roots. The results indicated that the Ca2+ sensor families of CaM, CDPK, and CBL in P. edulis exhibited significant transcriptional changes under chilling stress. Notably, PH02Gene03957, PH02Gene42787, and PH02Gene19300 were significantly upregulated, while the expressions of PH02Gene08456, PH02Gene01209, and PH02Gene37879 were suppressed. In particular, the expression levels of the CBF family gene PH02Gene14168, a downstream target gene of the calcium channels, increased significantly. P. edulis exhibited an influx of Ca2+ at the root, accompanied by oscillating negative peaks under chilling stress. Spatially, the cytosolic calcium concentration ([Ca2+]cyt) within the root cells increased. The CIPK family genes, interacting with Ca2+-CBL in downstream signaling pathways, showed significant differential expressions. In addition, the expressions of the NRT and AMT family genes changed correspondingly. Our study demonstrates that Ca2+ signaling is involved in the regulatory network of P. edulis under chilling stress. [Ca2+]cyt fluctuations in the roots of P. edulis are induced by chilling stress, reflecting an influx of extracellular Ca2+. Upon binding to Ca2+, downstream target genes from the CBF family are activated. Within the Ca2+–CBL–CIPK signaling network, the CIPK family plays a crucial role in nitrogen metabolism pathways. Full article
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16 pages, 4447 KiB  
Article
Molecular Characterization, Evolution and Expression Analysis of Ammonium Transporter from Four Closely Related Bactrocera Species (Tephritidae)
by Jie Zhang, Qi Wang, Chenhao Liu, Jiaying Liu, Qian Qian, Chuanjian Ru, Leyuan Liu, Shanchun Yan, Wei Liu and Guirong Wang
Life 2024, 14(9), 1114; https://doi.org/10.3390/life14091114 - 4 Sep 2024
Viewed by 1215
Abstract
Numerous insects are attracted to low levels of ammonia, utilizing it as a cue to locate food sources. The Ammonium Transporter (Amt), a highly conserved, atypical olfactory receptor, has been shown to mediate the detection of ammonia in insects. While the attraction of [...] Read more.
Numerous insects are attracted to low levels of ammonia, utilizing it as a cue to locate food sources. The Ammonium Transporter (Amt), a highly conserved, atypical olfactory receptor, has been shown to mediate the detection of ammonia in insects. While the attraction of Tephritidae to ammonia is well established, knowledge about the Amt in this family is limited. The species Bactrocera dorsalis (Hendel 1912), Bactrocera cucurbitae (Coquillett 1899), Bactrocera correcta Bezzi 1916 and Bactrocera tau (Walker 1849), which are common agricultural pests within Tephritidae, exhibit numerous ecological similarities, offering a solid foundation for studying Amt characteristics in this family. In this study, we elucidated the sequences, evolutionary relationships, and expression patterns of Amt in these four species. The results indicated that these Amts share the same open reading frame, containing 1770 bp that encode a protein of 589 amino acid residues. These Amt proteins exhibit the typical structural characteristics of Amts, including an 11-transmembrane domain with an extracellular N-terminus and an intracellular C-terminus. They also have the ability to form trimers in the membrane. Additionally, they contain three conserved amino acid residues essential for ammonia transport: A189, H195, and H352. Phylogenetic and expression pattern analyses showed that they are highly conserved in Diptera and are significantly expressed in antennae. This study is the first report characterizing the Amt gene in four Tephritidae species. These findings provide a foundation for further exploration into the roles of these genes in their particular biological contexts. Full article
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20 pages, 5637 KiB  
Article
Genome-Wide Identification and Characterization of Ammonium Transporter (AMT) Genes in Chlamydomonas reinhardtii
by Wenhui Hu, Dan Wang, Shuangshuang Zhao, Jiaqi Ji, Jing Yang, Yiqin Wan and Chao Yu
Genes 2024, 15(8), 1002; https://doi.org/10.3390/genes15081002 - 31 Jul 2024
Cited by 2 | Viewed by 1618
Abstract
Ammonium transporters (AMTs) are vital plasma membrane proteins facilitating NH4+ uptake and transport, crucial for plant growth. The identification of favorable AMT genes is the main goal of improving ammonium-tolerant algas. However, there have been no reports on the systematic identification [...] Read more.
Ammonium transporters (AMTs) are vital plasma membrane proteins facilitating NH4+ uptake and transport, crucial for plant growth. The identification of favorable AMT genes is the main goal of improving ammonium-tolerant algas. However, there have been no reports on the systematic identification and expression analysis of Chlamydomonas reinhardtii (C. reinhardtii) AMT genes. This study comprehensively identified eight CrAMT genes, distributed across eight chromosomes, all containing more than 10 transmembrane structures. Phylogenetic analysis revealed that all CrAMTs belonged to the AMT1 subfamily. The conserved motifs and domains of CrAMTs were similar to those of the AMT1 members of OsAMTs and AtAMTs. Notably, the gene fragments of CrAMTs are longer and contain more introns compared to those of AtAMTs and OsAMTs. And the promoter regions of CrAMTs are enriched with cis-elements associated with plant hormones and light response. Under NH4+ treatment, CrAMT1;1 and CrAMT1;3 were significantly upregulated, while CrAMT1;2, CrAMT1;4, and CrAMT1;6 saw a notable decrease. CrAMT1;7 and CrAMT1;8 also experienced a decline, albeit less pronounced. Transgenic algas with overexpressed CrAMT1;7 did not show a significant difference in growth compared to CC-125, while transgenic algas with CrAMT1;7 knockdown exhibited growth inhibition. Transgenic algas with overexpressed or knocked-down CrAMT1;8 displayed reduced growth compared to CC-125, which also resulted in the suppression of other CrAMT genes. None of the transgenic algas showed better growth than CC-125 at high ammonium levels. In summary, our study has unveiled the potential role of CrAMT genes in high-ammonium environments and can serve as a foundational research platform for investigating ammonium-tolerant algal species. Full article
(This article belongs to the Collection Feature Papers in Bioinformatics)
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14 pages, 3607 KiB  
Article
Identification and Expression Analysis of R2R3-MYB Transcription Factors Associated with Flavonoid Biosynthesis in Panax quinquefolius
by Guimei Song, Yan Yan, Chun Guo, Jiankang Chen, Yumeng Wang, Yingping Wang, Jiaxin Zhang, Chang Gao, Junmei Lian, Xiangmin Piao and Peng Di
Int. J. Mol. Sci. 2024, 25(7), 3709; https://doi.org/10.3390/ijms25073709 - 26 Mar 2024
Cited by 4 | Viewed by 1428
Abstract
Panax quinquefolius L. is an important medicinal plant, and flavonoids are among its main secondary metabolites. The R2R3-MYB transcription factor plays an irreplaceable role in plant growth, development, and secondary metabolism. In our study, we identified 159 R2R3-MYBs and analyzed their physical and [...] Read more.
Panax quinquefolius L. is an important medicinal plant, and flavonoids are among its main secondary metabolites. The R2R3-MYB transcription factor plays an irreplaceable role in plant growth, development, and secondary metabolism. In our study, we identified 159 R2R3-MYBs and analyzed their physical and chemical properties in P. quinquefolius. The protein length of 159 PqMYBs varied from 107 to 1050 amino acids. The molecular weight ranged from 12.21 to 116.44 kDa. The isoelectric point was between 4.57 and 10.34. We constructed a phylogenetic tree of P. quinquefolius and Arabidopsis thaliana R2R3-MYB family members, and PqMYB members were divided into 33 subgroups. Transcriptome data analysis showed that the expression patterns of PqMYBs in root, leaf, and flower were significantly different. Following the MeJA treatment of seedlings, five candidate PqMYB genes demonstrated a response. A correlation analysis of PqMYBs and candidate flavonoid pathway genes showed that PqMYB2, PqMYB46, and PqMYB72 had correlation coefficients that were higher than 0.8 with PqCHS, PqANS4, and PqCCoAMT10, respectively. Furthermore, a transient expression assay confirmed that the three PqMYBs were localized in the nucleus. We speculated that these three PqMYBs were related to flavonoid biosynthesis in P. quinquefolius. These results provided a theoretical basis and a new perspective for further understanding the R2R3-MYB gene family and the biosynthesis mechanism of secondary metabolites in P. quinquefolius. Full article
(This article belongs to the Special Issue Advances in Genetics and Phylogenomics of Tree)
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18 pages, 30283 KiB  
Article
Genome-Wide Analysis of Sweet Potato Ammonium Transporter (AMT): Influence on Nitrogen Utilization, Storage Root Development and Yield
by Ya-Yi Meng, Ning Wang, Hai-Yan Zhang, Ran Xu and Cheng-Cheng Si
Int. J. Mol. Sci. 2023, 24(24), 17424; https://doi.org/10.3390/ijms242417424 - 13 Dec 2023
Cited by 3 | Viewed by 1645
Abstract
Ammonium, as a major inorganic source of nitrogen (N) for sweet potato N utilization and growth, is specifically transported by ammonium transporters (AMTs). However, the activities of AMT family members in sweet potatoes have not been analyzed. In the present study, [...] Read more.
Ammonium, as a major inorganic source of nitrogen (N) for sweet potato N utilization and growth, is specifically transported by ammonium transporters (AMTs). However, the activities of AMT family members in sweet potatoes have not been analyzed. In the present study, the sweet potato cultivar ‘Pushu 32’, which is planted in a large area in China, was used in field experiments at the Agricultural Base of Hainan University (20°06′ N, 110°33′ E) in 2021, and Sanya Nanfan Research Institute of Hainan University (18°30′ N, 109°60′ E) in 2022. Four N levels were tested: 0, 60, 120, and 180 kg ha−1. The results are as follows. Twelve IbAMT genes were identified in the sweet potato genome, which were classified into three distinct subgroups based on phylogeny; the same subgroup genes had similar properties and structures. IbAMT1.3 and IbAMT1.5 were mostly expressed in the storage roots under N deficiency. Compared with the NN and HN groups, IbAMT1.3 and IbAMT1.5 expressions, N content in storage roots, N uptake efficiency at the canopy closure, N fertilization contribution rates, number of storage roots per plant, storage root weight, and yield were all increased in the MN group. Furthermore, there was a significant positive correlation between the expressions of IbAMT1.3 and IbAMT1.5 with N content in the storage roots of sweet potato. In a word, IbAMT1.3 and IbAMT1.5 may regulate N utilization, affect the development of the storage root. and determine the yield of sweet potato. The results provide valuable insights into the AMT gene family’s role in the use of N and effects on storage root development and yield in sweet potatoes. Full article
(This article belongs to the Special Issue Abiotic Stress in Plant: From Gene to the Fields 2.0)
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22 pages, 9472 KiB  
Article
Characterization of VvmiR166s-Target Modules and Their Interaction Pathways in Modulation of Gibberellic-Acid-Induced Grape Seedless Berries
by Yunhe Bai, Zhuangwei Wang, Linjia Luo, Xuxian Xuan, Wei Tang, Ziyang Qu, Tianyu Dong, Ziyang Qi, Mucheng Yu, Weimin Wu, Jinggui Fang and Chen Wang
Int. J. Mol. Sci. 2023, 24(22), 16279; https://doi.org/10.3390/ijms242216279 - 14 Nov 2023
Cited by 4 | Viewed by 1393
Abstract
Exogenous GA is widely used to efficiently induce grape seedless berry development for significantly improving berry quality. Recently, we found that VvmiR166s are important regulators of response to GA in grapes, but its roles in GA-induced seedless grape berry development remain elusive. Here, [...] Read more.
Exogenous GA is widely used to efficiently induce grape seedless berry development for significantly improving berry quality. Recently, we found that VvmiR166s are important regulators of response to GA in grapes, but its roles in GA-induced seedless grape berry development remain elusive. Here, the precise sequences of VvmiR166s and their targets VvREV, VvHB15 and VvHOX32 were determined in grape cv. ‘Rosario Bianco’, and the cleavage interactions of VvmiR166s-VvHB15/VvHOX32/VvREV modules and the variations in their cleavage roles were confirmed in grape berries. Exogenous GA treatment significantly induced a change in their expression correlations from positive to negative between VvmiR166s and their target genes at the seeds during the stone-hardening stages (32 DAF–46 DAF) in grape berries, indicating exogenous GA change action modes of VvmiR166s on their targets in this process, in which exogenous GA mainly enhanced the negative regulatory roles of VvmiR166s on VvHB15 among all three VvmiR166s-target pairs. The transient OE-VvmiR166a-h/OE-VvHB15 in tobacco confirmed that out of the VvmiR166 family, VvmiR166h/a/b might be the main factors in modulating lignin synthesis through inhibiting VvHB15, of which VvmiR166h-VvHB15-NtPAL4/NtCCR1/NtCCR2/NtCCoAMT5/NtCOMT1 and VvmiR166a/b-VvHB15-NtCAD1 are the potential key regulatory modules in lignin synthesis. Together with the GA-induced expression modes of VvmiR166s-VvHB15 and genes related to lignin synthesis in grape berries, we revealed that GA might repress lignin synthesis mainly by repressing VvCAD1/VvCCR2/VvPAL2/VvPAL3/Vv4CL/VvLac7 levels via mediating VvmiR166s-VvHB15 modules in GA-induced grape seedless berries. Our findings present a novel insight into the roles of VvmiR66s that are responsive to GA in repressing the lignin synthesis of grape seedless berries, with different lignin-synthesis-enzyme-dependent action pathways in diverse plants, which have important implications for the molecular breeding of high-quality seedless grape berries. Full article
(This article belongs to the Special Issue Molecular Research in Fruit Crop)
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11 pages, 2279 KiB  
Article
Structural Study of a New MbtI-Inhibitor Complex: Towards an Optimized Model for Structure-Based Drug Discovery
by Matteo Mori, Stefania Villa, Laurent R. Chiarelli, Fiorella Meneghetti and Marco Bellinzoni
Pharmaceuticals 2023, 16(11), 1559; https://doi.org/10.3390/ph16111559 - 3 Nov 2023
Cited by 3 | Viewed by 2124
Abstract
MbtI from Mycobacterium tuberculosis (Mtb) is a Mg2+-dependent salicylate synthase, belonging to the chorismate-utilizing enzyme (CUE) family. As a fundamental player in iron acquisition, MbtI promotes the survival and pathogenicity of Mtb in the infected host. Hence, it has [...] Read more.
MbtI from Mycobacterium tuberculosis (Mtb) is a Mg2+-dependent salicylate synthase, belonging to the chorismate-utilizing enzyme (CUE) family. As a fundamental player in iron acquisition, MbtI promotes the survival and pathogenicity of Mtb in the infected host. Hence, it has emerged in the last decade as an innovative, potential target for the anti-virulence therapy of tuberculosis. In this context, 5-phenylfuran-2-carboxylic acids have been identified as potent MbtI inhibitors. The first co-crystal structure of MbtI in complex with a member of this class was described in 2020, showing the enzyme adopting an open configuration. Due to the high mobility of the loop adjacent to the binding pocket, large portions of the amino acid chain were not defined in the electron density map, hindering computational efforts aimed at structure-driven ligand optimization. Herein, we report a new, high-resolution co-crystal structure of MbtI with a furan-based derivative, in which the closed configuration of the enzyme allowed tracing the entirety of the active site pocket in the presence of the bound inhibitor. Moreover, we describe a new crystal structure of MbtI in open conformation and in complex with the known inhibitor methyl-AMT, suggesting that in vitro potency is not related to the observed enzyme conformation. These findings will prove fundamental to enhance the potency of this series via rational structure-based drug-design approaches. Full article
(This article belongs to the Special Issue Novel Insights into Tuberculosis Research and Drug Discovery)
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21 pages, 11033 KiB  
Article
Functional Identification and Genetic Transformation of the Ammonium Transporter PtrAMT1;6 in Populus
by Chengjun Yang, Chunxi Huang, Luzheng Gou, Han Yang and Guanjun Liu
Int. J. Mol. Sci. 2023, 24(10), 8511; https://doi.org/10.3390/ijms24108511 - 10 May 2023
Cited by 2 | Viewed by 2767
Abstract
The ammonium transporter (AMT) family gene is an important transporter involved in ammonium uptake and transfer in plants and is mainly engaged in the uptake and transport of ammonium from the environment by roots and the reabsorption of ammonium in the aboveground parts. [...] Read more.
The ammonium transporter (AMT) family gene is an important transporter involved in ammonium uptake and transfer in plants and is mainly engaged in the uptake and transport of ammonium from the environment by roots and the reabsorption of ammonium in the aboveground parts. In this study, the expression pattern, functional identification, and genetic transformation of the PtrAMT1;6 gene, a member of the ammonium transporter protein family in P. trichocarpa, were investigated as follows: (1) Fluorescence quantitative PCR demonstrated that the PtrAMT1;6 gene was preferentially expressed in the leaves, with both dark-induced and light-inhibited expression patterns. (2) A functional restoration assay using the yeast ammonium transporter protein mutant strain indicated that the PtrAMT1;6 gene restored the ability of the mutant to transport ammonium with high affinity. (3) Arabidopsis was transformed with pCAMBIA-PtrAMT1;6P, and the transformed lines were stained with GUS, which showed that the rootstock junction, cotyledon petioles, and the leaf veins and pulp near the petioles of the transformed plants could be stained blue, indicating that the promoter of the PtrAMT1;6 gene had promoter activity. (4) The overexpression of the PtrAMT1;6 gene caused an imbalance in carbon and nitrogen metabolism and reduced nitrogen assimilation ability in ‘84K’ poplar and ultimately reduced biomass. The above results suggest that PtrAMT1;6 may be involved in ammonia recycling during nitrogen metabolism in aboveground parts, and overexpression of PtrAMT1;6 may affect the process of carbon and nitrogen metabolism, as well as nitrogen assimilation in plants, resulting in stunted growth of overexpression plants. Full article
(This article belongs to the Section Molecular Plant Sciences)
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21 pages, 8316 KiB  
Article
Genome-Wide Identification and Characterization of Ammonium Transporter (AMT) Genes in Rapeseed (Brassica napus L.)
by Jing Dai, Peipei Han, Thomas C. Walk, Ling Yang, Liyu Chen, Yinshui Li, Chiming Gu, Xing Liao and Lu Qin
Genes 2023, 14(3), 658; https://doi.org/10.3390/genes14030658 - 6 Mar 2023
Cited by 10 | Viewed by 2890
Abstract
Ammonium transporters (AMTs) are plasma membrane proteins mediating ammonium uptake and transport. As such, AMTs play vital roles in ammonium acquisition and mobilization, plant growth and development, and stress and pathogen defense responses. Identification of favorable AMT genotypes is a prime target for [...] Read more.
Ammonium transporters (AMTs) are plasma membrane proteins mediating ammonium uptake and transport. As such, AMTs play vital roles in ammonium acquisition and mobilization, plant growth and development, and stress and pathogen defense responses. Identification of favorable AMT genotypes is a prime target for crop improvement. However, to date, systematic identification and expression analysis of AMT gene family members has not yet been reported for rapeseed (Brassica napus L.). In this study, 20 AMT genes were identified in a comprehensive search of the B. napus genome, 14 members of AMT1 and 6 members of AMT2. Tissue expression analyses revealed that the 14 AMT genes were primarily expressed in vegetative organs, suggesting that different BnaAMT genes might function in specific tissues at the different development stages. Meanwhile, qRT-PCR analysis found that several BnaAMTs strongly respond to the exogenous N conditions, implying the functional roles of AMT genes in ammonium absorption in rapeseed. Moreover, the rapeseed AMT genes were found to be differentially regulated by N, P, and K deficiency, indicating that crosstalk might exist in response to different stresses. Additionally, the subcellular localization of several BnaAMT proteins was confirmed in Arabidopsis protoplasts, and their functions were studied in detail by heterologous expression in yeast. In summary, our studies revealed the potential roles of BnaAMT genes in N acquisition or transportation and abiotic stress response and could provide valuable resources for revealing the functionality of AMTs in rapeseed. Full article
(This article belongs to the Special Issue Genomics and Breeding of Oil Crops)
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23 pages, 6696 KiB  
Article
Genome-Wide Identification and Expression Analysis of the Ammonium Transporter Family Genes in Soybean
by Wei Yang, Xiaoxu Dong, Zhanxin Yuan, Yan Zhang, Xia Li and Youning Wang
Int. J. Mol. Sci. 2023, 24(4), 3991; https://doi.org/10.3390/ijms24043991 - 16 Feb 2023
Cited by 14 | Viewed by 3185
Abstract
Ammonium transporters (AMTs) are responsible for ammonium absorption and utilization in plants. As a high-nitrogen-demand crop and a legume, soybean can also obtain ammonium from symbiotic root nodules in which nitrogen-fixing rhizobia convert atmospheric nitrogen (N2) into ammonium. Although increasing evidence [...] Read more.
Ammonium transporters (AMTs) are responsible for ammonium absorption and utilization in plants. As a high-nitrogen-demand crop and a legume, soybean can also obtain ammonium from symbiotic root nodules in which nitrogen-fixing rhizobia convert atmospheric nitrogen (N2) into ammonium. Although increasing evidence implicates vital roles of ammonium transport in soybean, no systematic analyses of AMTs in soybean (named GmAMTs) or functional analyses of GmAMTs are available. In this study, we aimed to identify all GmAMT family genes and gain a better understanding of the characteristics of GmAMT genes in soybean. Here, due to the improved genome assembly and annotation of soybean, we tried to generate a phylogenetic tree of 16 GmAMTs based on new information. Consistent with reported data, GmAMT family members can be divided into two subfamilies of GmAMT1 (6 genes) and GmAMT2 (10 genes). Interestingly, unlike Arabidopsis, which has only one AMT2, soybean has substantially increased the number of GmAMT2s, suggesting enhanced demand for ammonium transport. These genes were distributed on nine chromosomes, of which GmAMT1.3, GmAMT1.4, and GmAMT1.5 were three tandem repeat genes. The gene structures and conserved protein motifs of the GmAMT1 and GmAMT2 subfamilies were different. All the GmAMTs were membrane proteins with varying numbers of transmembrane domains ranging from 4 to 11. Promoter analysis found that these GmAMT genes have phytohormone-, circadian control-, and organ expression-related cis-elements in their promoters, and notably, there were nodulation-specific and nitrogen-responsive elements in the promoters of the GmAMT1 and GmAMT2 genes. Further expression data showed that these GmAMT family genes exhibited different spatiotemporal expression patterns across tissues and organs. In addition, GmAMT1.1, GmAMT1.2, GmAMT2.2, and GmAMT2.3 were responsive to nitrogen treatment, while GmAMT1.2, GmAMT1.3, GmAMT1.4, GmAMT1.5, GmAMT1.6, GmAMT2.1, GmAMT2.2, GmAMT2.3, GmAMT3.1, and GmAMT4.6 showed circadian rhythms in transcription. RT-qPCR validated the expression patterns of GmAMTs in response to different forms of nitrogen and exogenous ABA treatments. Gene expression analysis also confirmed that GmAMTs are regulated by key nodulation gene GmNINa, indicating a role of GmAMTs in symbiosis. Together, these data indicate that GmAMTs may differentially and/or redundantly regulate ammonium transport during plant development and in response to environmental factors. These findings provide a basis for future research on the functions of GmAMTs and the mechanisms through which GmAMTs regulate ammonium metabolism and nodulation in soybean. Full article
(This article belongs to the Special Issue Genetics and Novel Techniques for Soybean Yield Enhancement)
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16 pages, 3452 KiB  
Article
The Relationship between Ectomycorrhizal Fungi, Nitrogen Deposition, and Pinus massoniana Seedling Nitrogen Transporter Gene Expression and Nitrogen Uptake Kinetics
by Pengfei Sun, Ruimei Cheng, Wenfa Xiao, Lixiong Zeng, Yafei Shen, Lijun Wang, Tian Chen and Meng Zhang
J. Fungi 2023, 9(1), 65; https://doi.org/10.3390/jof9010065 - 31 Dec 2022
Cited by 5 | Viewed by 2543
Abstract
Analyzing the molecular and physiological processes that govern the uptake and transport of nitrogen (N) in plants is central to efforts to fully understand the optimization of plant N use and the changes in the N-use efficiency in relation to changes in atmospheric [...] Read more.
Analyzing the molecular and physiological processes that govern the uptake and transport of nitrogen (N) in plants is central to efforts to fully understand the optimization of plant N use and the changes in the N-use efficiency in relation to changes in atmospheric N deposition changes. Here, a field experiment was conducted using the ectomycorrhizal fungi (EMF), Pisolithus tinctorius (Pt) and Suillus grevillei (Sg). The effects of N deposition were investigated using concentrations of 0 kg·N·hm−2a−1 (N0), a normal N deposition of 30 kg·N·hm−2a−1 (N30), a moderate N deposition of 60 kg·N·hm−2a−1 (N60), and a severe N deposition of 90 kg·N·hm−2a−1 (N90), with the goal of examining how these factors impacted root activity, root absorbing area, NH4+ and NO3 uptake kinetics, and the expression of ammonium and nitrate transporter genes in Pinus massoniana seedlings under different levels of N deposition. These data revealed that EMF inoculation led to increased root dry weight, activity, and absorbing area. The NH4+ and NO3 uptake kinetics in seedlings conformed to the Michaelis–Menten equation, and uptake rates declined with increasing levels of N addition, with NH4+ uptake rates remaining higher than NO3 uptake rates for all tested concentrations. EMF inoculation was associated with higher Vmax values than were observed for non-mycorrhizal plants. Nitrogen addition resulted in the upregulation of genes in the AMT1 family and the downregulation of genes in the NRT family. EMF inoculation under the N60 and N90 treatment conditions resulted in the increased expression of each of both these gene families. NH4+ and NO3 uptake kinetics were also positively correlated with associated transporter gene expression in P. massoniana roots. Together, these data offer a theoretical foundation for EMF inoculation under conditions of increased N deposition associated with climate change in an effort to improve N absorption and transport rates through the regulation of key nitrogen transporter genes, thereby enhancing N utilization efficiency and promoting plant growth. Synopsis: EMF could enhance the efficiency of N utilization and promote the growth of Pinus massoniana under conditions of increased N deposition. Full article
(This article belongs to the Section Environmental and Ecological Interactions of Fungi)
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20 pages, 4139 KiB  
Article
Genome-Wide Identification and Expression Analysis of AMT and NRT Gene Family in Pecan (Carya illinoinensis) Seedlings Revealed a Preference for NH4+-N
by Mengyun Chen, Kaikai Zhu, Junyi Xie, Junping Liu, Pengpeng Tan and Fangren Peng
Int. J. Mol. Sci. 2022, 23(21), 13314; https://doi.org/10.3390/ijms232113314 - 1 Nov 2022
Cited by 5 | Viewed by 2416
Abstract
Nitrogen (N) is a major limiting factor for plant growth and crop production. The use of N fertilizer in forestry production is increasing each year, but the loss is substantial. Mastering the regulatory mechanisms of N uptake and transport is a key way [...] Read more.
Nitrogen (N) is a major limiting factor for plant growth and crop production. The use of N fertilizer in forestry production is increasing each year, but the loss is substantial. Mastering the regulatory mechanisms of N uptake and transport is a key way to improve plant nitrogen use efficiency (NUE). However, this has rarely been studied in pecans. In this study, 10 AMT and 69 NRT gene family members were identified and systematically analyzed from the whole pecan genome using a bioinformatics approach, and the expression patterns of AMT and NRT genes and the uptake characteristics of NH4+ and NO3 in pecan were analyzed by aeroponic cultivation at varying NH4+/NO3 ratios (0/0, 0/100,25/75, 50/50, 75/25,100/0 as CK, T1, T2, T3, T4, and T5). The results showed that gene duplication was the main reason for the amplification of the AMT and NRT gene families in pecan, both of which experienced purifying selection. Based on qRT-PCR results, CiAMTs were primarily expressed in roots, and CiNRTs were majorly expressed in leaves, which were consistent with the distribution of pecan NH4+ and NO3 concentrations in the organs. The expression levels of CiAMTs and CiNRTs were mainly significantly upregulated under N deficiency and T4 treatment. Meanwhile, T4 treatment significantly increased the NH4+, NO3, and NO2 concentrations as well as the Vmax and Km values of NH4+ and NO3 in pecans, and Vmax/Km indicated that pecan seedlings preferred to absorb NH4+. In summary, considering the single N source of T5, we suggested that the NH4+/NO3 ratio of 75:25 was more beneficial to improve the NUE of pecan, thus increasing pecan yield, which provides a theoretical basis for promoting the scale development of pecan and provides a basis for further identification of the functions of AMT and NRT genes in the N uptake and transport process of pecan. Full article
(This article belongs to the Special Issue Nitrogen with Plant Growth and Development)
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