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1.
Sci Rep ; 12(1): 12693, 2022 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-35882960

RESUMO

The objectives of this study were to determine drought tolerant novel mutant of alfalfa (Medicago sativa L.) genotypes by screening EMS mutagenized 340675 M3 seeds at germination stages in the presence of osmotic stress of 35% PEG6000. Root growth assay provided several drought tolerant candidate mutants. Of those, 4 mutants were further evaluated at water deficit conditions applied for 24 days after the first cutting at flowering bud stage. The results revealed that mutants determined as drought tolerant at germination stage were also tolerant to water deficit conditions. Protein content and superoxide dismutase values were found to be higher in all mutants than controls. Ascorbate peroxides, glutton reductase and lipid peroxidase values varied based on the mutant genotype and duration of drought stress. Drought stress significantly changed transcriptional levels of MtP5CS, MtDehyd, MseIF-2, MtRD2 and MsNAC genes. These results indicated that in vitro screening of alfalfa mutant seeds for osmatic tolerance at germination and early seedling growth stages was successfully able to determine the drought tolerant alfalfa mutants which were also tolerant to water deficit conditions after the first cutting at flowering bud stage.


Assuntos
Germinação , Medicago sativa , Secas , Germinação/genética , Medicago sativa/metabolismo , Plântula , Água/metabolismo
2.
Int J Mol Sci ; 23(14)2022 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-35887111

RESUMO

Cinnamoyl-CoA reductase (CCR) is a pivotal enzyme in plant lignin synthesis, which has a role in plant secondary cell wall development and environmental stress defense. Alfalfa is a predominant legume forage with excellent quality, but the lignin content negatively affects fodder digestibility. Currently, there is limited information on CCR characteristics, gene expression, and its role in lignin metabolism in alfalfa. In this study, we identified 30 members in the CCR gene family of Medicago sativa. In addition, gene structure, conserved motif, and evolution analysis suggested MsCCR1-7 presumably functioned as CCR, while the 23 MsCCR-likes fell into three categories. The expression patterns of MsCCRs/MsCCR-likes suggested their role in plant development, response to environmental stresses, and phytohormone treatment. These results were consistent with the cis-elements in their promoters. Histochemical staining showed that lignin accumulation gradually deepened with the development, which was consistent with gene expression results. Furthermore, recombinant MsCCR1 and MsCCR-like1 were purified and the kinetic parameters were tested under four substrates. In addition, three-dimensional structure models of MsCCR1 and MsCCR-like1 proteins showed the difference in the substrate-binding motif H212(X)2K215R263. These results will be useful for further application for legume forage quality modification and biofuels industry engineering in the future.


Assuntos
Lignina , Medicago sativa , Aldeído Oxirredutases/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Lignina/metabolismo , Medicago sativa/genética , Medicago sativa/metabolismo
3.
BMC Plant Biol ; 22(1): 323, 2022 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-35790925

RESUMO

The application of nanotechnology in agriculture can remarkably improve the cultivation and growth of crop plants. Many studies showed that nanoparticles (NPs) made plants grow more vigorously. Light can make NPs aggregated, leading to the reduction of the NPs toxicity. In addition, treatment with NPs had a "hormesis effect" on plants. In this study, light-induced silver nanoparticles (AgNPs) were synthesized by using the alfalfa (Medicago sativa L.) extracts, and then the optimal synthetic condition was determined. Light-induced AgNPs were aggregated, spherical and pink, and they were coated with esters, phenols, acids, terpenes, amino acids and sugars, which were the compositions of alfalfa extracts. The concentration of free Ag+ was less than 2 % of the AgNPs concentration. Through nanopriming, Ag+ got into the seedlings and caused the impact of AgNPs on alfalfa. Compared with the control group, low concentration of light-induced AgNPs had a positive effect on the photosynthesis. It was also harmless to the leaf cells, and there was no elongation effect on shoots. Although high concentration of AgNPs was especially beneficial to root elongation, it had a slight toxic effect on seedlings due to the accumulation of silver. With the increase of AgNPs concentration, the content of silver in the seedlings increased and the silver enriched in plants was at the mg/kg level. Just as available research reported the toxicity of NPs can be reduced by using suitable synthesis and application methods, the present light induction, active material encapsulation and nanopriming minimized the toxicity of AgNPs to plants, enhancing the antioxidant enzyme system.


Assuntos
Nanopartículas Metálicas , Prata , Medicago sativa/metabolismo , Nanopartículas Metálicas/química , Plântula/metabolismo , Prata/química , Nitrato de Prata/farmacologia
4.
Int J Mol Sci ; 23(13)2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35805915

RESUMO

Nuclear factor YB (NF-YB) are plant-specific transcription factors that play a critical regulatory role in plant growth and development as well as in plant resistance against various stresses. In this study, a total of 49 NF-YB genes were identified from the genomes of Medicago truncatula and Medicago sativa. Multiple sequence alignment analysis showed that all of these NF-YB members contain DNA binding domain, NF-YA interaction domain and NF-YC interaction domain. Phylogenetic analysis suggested that these NF-YB proteins could be classified into five distinct clusters. We also analyzed the exon-intron organizations and conserved motifs of these NF-YB genes and their deduced proteins. We also found many stress-related cis-acting elements in their promoter region. In addition, analyses on genechip for M. truncatula and transcriptome data for M. sativa indicated that these NF-YB genes exhibited a distinct expression pattern in various tissues; many of these could be induced by drought and/or salt treatments. In particular, RT-qPCR analysis revealed that the expression levels of gene pairs MsNF-YB27/MtNF-YB15 and MsNF-YB28/MtNF-YB16 were significantly up-regulated under NaCl and mannitol treatments, indicating that they are most likely involved in salt and drought stress response. Taken together, our study on NF-YB family genes in Medicago is valuable for their functional characterization, as well as for the application of NF-YB genes in genetic breeding for high-yield and high-resistance alfalfa.


Assuntos
Regulação da Expressão Gênica de Plantas , Medicago truncatula , Medicago sativa/genética , Medicago sativa/metabolismo , Medicago truncatula/genética , Medicago truncatula/metabolismo , Família Multigênica , Filogenia , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Cloreto de Sódio/farmacologia , Estresse Fisiológico/genética , Fatores de Transcrição/metabolismo
5.
Int J Mol Sci ; 23(13)2022 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-35806246

RESUMO

Drought stress is one of the major constraints that decreases global crop productivity. Alfalfa, planted mainly in arid and semi-arid areas, is of crucial importance in sustaining the agricultural system. The family 1 UDP-glycosyltransferases (UGT) is indispensable because it takes part in the regulation of plant growth and stress resistance. However, a comprehensive insight into the participation of the UGT family in adaptation of alfalfa to drought environments is lacking. In the present study, a genome-wide analysis and profiling of the UGT in alfalfa were carried out. A total of 409 UGT genes in alfalfa (MsUGT) were identified and they are clustered into 13 groups. The expression pattern of MsUGT genes were analyzed by RNA-seq data in six tissues and under different stresses. The quantitative real-time PCR verification genes suggested the distinct role of the MsUGT genes under different drought stresses and abscisic acid (ABA) treatment. Furthermore, the function of MsUGT003 and MsUGT024, which were upregulated under drought stress and ABA treatment, were characterized by heterologous expression in yeast. Taken together, this study comprehensively analyzed the UGT gene family in alfalfa for the first time and provided useful information for improving drought tolerance and in molecular breeding of alfalfa.


Assuntos
Secas , Medicago sativa , Ácido Abscísico/metabolismo , Regulação da Expressão Gênica de Plantas , Glicosiltransferases/metabolismo , Medicago sativa/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética , Difosfato de Uridina/metabolismo
6.
J Hazard Mater ; 435: 129077, 2022 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-35650732

RESUMO

Cadmium (Cd) pollution in croplands is a global environmental problem. Measures to improve the tolerance of sensitive crops and reduce pollutant absorption and accumulation are needed in contaminated agricultural areas, and inoculation with rhizosphere microorganisms to regulate plant resistance and heavy metal transport can provide an effective solution. A pot experiment was conducted to analyse the impact of arbuscular mycorrhizal fungi (AMF) on alfalfa oxidase activity, heavy metal resistance genes and transport proteins, metabolism, and other biochemical regulation mechanisms that lead to complexation, compartmentalisation, efflux, enrichment, and antioxidant detoxification pathways. The AMF reduced shoot and protoplasm Cd inflow, and promoted organic compound production (e.g., by upregulating HM-Res4 for 1.2 times), to complex with Cd, reducing its biological toxicity. The AMF increased the ROS scavenging efficiency and osmotic regulatory substance content of the alfalfa plants, reduced oxidative stress (ROS dereased), and maintained homeostasis. It also alleviated Cd inhibition of photosynthetic electron transport, tricarboxylic acid circulation, and nitrogen assimilation. These AMF effects improved leaf and root biomass by 43.87% and 59.71% and facilitated recovery of a conservative root economic strategy. It is speculated that AMF induces the resistance signal switch by regulating the negative feedback regulation mode of indole acetic acid upward transport and methyl jasmonate downward transmission in plants.


Assuntos
Metais Pesados , Micorrizas , Poluentes do Solo , Cádmio/metabolismo , Medicago sativa/metabolismo , Metais Pesados/metabolismo , Micorrizas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Poluentes do Solo/metabolismo
7.
BMC Plant Biol ; 22(1): 295, 2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35705909

RESUMO

BACKGROUND: SQUAMOSA promoter-binding protein-like (SPL) transcription factors are widely present in plants and are involved in signal transduction, the stress response and development. The SPL gene family has been characterized in several model species, such as A. thaliana and G. max. However, there is no in-depth analysis of the SPL gene family in forage, especially alfalfa (Medicago sativa L.), one of the most important forage crops worldwide. RESULT: In total, 76 putative MsSPL genes were identified in the alfalfa genome with an uneven distribution. Based on their identity and gene structure, these MsSPLs were divided into eight phylogenetic groups. Seventy-three MsSPL gene pairs arose from segmental duplication events, and the MsSPLs on the four subgenomes of individual chromosomes displayed high collinearity with the corresponding M. truncatula genome. The prediction of the cis-elements in the promoter regions of the MsSPLs detected two copies of ABA (abscisic acid)-responsive elements (ABREs) on average, implying their potential involvement in alfalfa adaptation to adverse environments. The transcriptome sequencing of MsSPLs in roots and leaves revealed that 54 MsSPLs were expressed in both tissues. Upon salt treatment, three MsSPLs (MsSPL17, MsSPL23 and MsSPL36) were significantly regulated, and the transcription level of MsSPL36 in leaves was repressed to 46.6% of the control level. CONCLUSION: In this study, based on sequence homology, we identified 76 SPL genes in the alfalfa. The SPLs with high identity shared similar gene structures and motifs. In total, 71.1% (54 of 76) of the MsSPLs were expressed in both roots and leaves, and the majority (74.1%) preferred underground tissues to aerial tissues. MsSPL36 in leaves was significantly repressed under salt stress. These findings provide comprehensive information regarding the SPB-box gene family for improve alfalfa tolerance to high salinity.


Assuntos
Regulação da Expressão Gênica de Plantas , Medicago sativa , Ácido Abscísico/metabolismo , Medicago sativa/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Estresse Salino/genética , Estresse Fisiológico/genética
8.
Int J Mol Sci ; 23(12)2022 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-35742869

RESUMO

Nuclear factor Y (NF-Y) is a heterotrimeric transcription factor that plays an important role in various biological processes in plants, such as flowering regulation, drought resistance, and salt stress. However, few in-depth studies investigated the alfalfa NF-Y gene family. In this study, in total, 60 MsNF-Y genes, including 9 MsNF-YAs, 26 MsNF-YBs, and 25 MsNF-YCs, were identified in the alfalfa genome. The genomic locations, gene structures, protein molecular weights, conserved domains, phylogenetic relationships, and gene expression patterns in different tissues and under different stresses (cold stress, drought stress, and salt stress) of these NF-Y genes were analyzed. The illustration of the conserved domains and specific domains of the different subfamilies of the MsNF-Y genes implicates the conservation and diversity of their functions in alfalfa growth, development, and stress resistance. The gene expression analysis showed that 48 MsNF-Y genes (7 MsNF-YAs, 22 MsNF-YBs, and 19 MsNF-YCs) were expressed in all tissues at different expression levels, indicating that these genes have tissue expression specificity and different biological functions. In total, seven, seven, six, and eight MsNF-Y genes responded to cold stress, the ABA treatment, drought stress, and salt stress in alfalfa, respectively. According to the WGCNA, molecular regulatory networks related to salt stress were constructed for MsNF-YB2, MsNF-YB5, MsNF-YB7, MsNF-YB15, MsNF-YC5, and MsNF-YC6. This study could provide valuable information for further elucidating the biological functions of MsNF-Ys and improving salt tolerance and other abiotic stress resistance in alfalfa.


Assuntos
Medicago sativa , Fatores de Transcrição , Fator de Ligação a CCAAT , Secas , Regulação da Expressão Gênica de Plantas , Medicago sativa/genética , Medicago sativa/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tolerância ao Sal/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
9.
Plant Signal Behav ; 17(1): 2081420, 2022 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-35642507

RESUMO

Dehydration-responsive element-binding proteins (DREBs) belong to members of the AP2/ERF transcription factor superfamily, which has been reported to involve various abiotic-stress responses and tolerance in plants. However, research on the DREB-family is still limited in alfalfa (Medicago sativa L.), a forage legume cultivated worldwide. The recent genome-sequence release of the alfalfa cultivar "XinJiangDaYe" allowed us to identify 172 DREBs by a multi-step homolog search. The phylogenetic analysis indicated that such MsDREBs could be classified into 5 groups, namely A-1 (56 members), A-2 (39), A-3 (3), A-4 (61) and 13 (A-5 (13), thus adding substantial new members to the DREB-family in alfalfa. Furthermore, a comprehensive survey in silico of conserved motif, gene structure, molecular weight, and isoelectric point (pI) as well as gene expression was conducted. The resulting data showed that, for cold-stress response, 33 differentially expressed MsDREBs were identified with a threshold of Log2-fold > 1, and most of which were transcriptionally upregulated within 48 h during a cold treatment(s). Moreover, the expression profiling of MsDREBs from two ecotypes of alfalfa subspecies i.e. M. sativa ssp. falcata (F56, from a colder region of Central Asia) and M. sativa ssp. sativa (B47, from Near East) revealed that most of the cold-stress responsive MsDREBs exhibited a significantly lower expression in F56, leading to a proposal of the existence of a distinct mechanism(s) for cold tolerance regulated by DREB-related action, which would have been evolved in alfalfa with a genotypic specificity. Additionally, by examining the transcriptome of a freezing-tolerance species (M. sativa cv. Zhaodong), eight DREBs were found to be implicated in a long-term freezing-stress adaptation with a great potential. Taken together, the current genome-wide identification in alfalfa points to the importance of some MsDREBs in the cold-stress response, providing some promising molecular targets to be functionally characterized for the improvement of cold tolerance in crops including alfalfa.


Assuntos
Resposta ao Choque Frio , Medicago sativa , Resposta ao Choque Frio/genética , Regulação da Expressão Gênica de Plantas/genética , Medicago sativa/genética , Medicago sativa/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
10.
Ecotoxicol Environ Saf ; 241: 113780, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35738100

RESUMO

Although listed as endocrine disruptor compounds, atrazine (ATZ) is still used in large quantities in agricultural production. Here, alfalfa seedling was cultivated in hydroponic media to investigate the toxic effects of ATZ on alfalfa and accumulation of ATZ in tissues of different plant parts. Alfalfa had a strong upward translocation ability to ATZ. The stress response of alfalfa under ATZ stress was studied using metabolomic and transcriptomic techniques. S-adenosylmethionine, glutathione, 3-mercaptopyruvic acid, ornithine, and aminopropylcadaverine were significantly increased by ATZ in pathways mtr00270 and mtr00480. Several genes of cysteine synthase and spermidine synthase were significantly up-regulated by ATZ induction. They may be markers and genes with potential physiological functions of alfalfa in response to ATZ stress. In addition, using high resolution mass spectrometry, a total of five ATZ metabolites secreted from alfalfa roots were detected. Among them, acetylated deisopropylated ATZ was discovered for the first time. Hydroxylated ATZ and acetylated deethylated ATZ were more readily excreted by the root system. This study not only provides potential genes for the construction of engineering plants to remediate ATZ-contaminated soil, but also provides monitoring objects for the ecological research of ATZ metabolites.


Assuntos
Atrazina , Disruptores Endócrinos , Herbicidas , Atrazina/metabolismo , Atrazina/toxicidade , Disruptores Endócrinos/metabolismo , Poluição Ambiental , Herbicidas/metabolismo , Medicago sativa/metabolismo
12.
Sci Rep ; 12(1): 5952, 2022 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-35396458

RESUMO

Comprehensive climate change mitigation necessitates soil carbon (C) storage in cultivated terrestrial ecosystems. Deep-rooted perennial crops may help to turn agricultural soils into efficient C sinks, especially in deeper soil layers. Here, we compared C allocation and potential stabilization to 150 cm depth from two functionally distinct deep-rooted perennials, i.e., lucerne (Medicago sativa L.) and intermediate wheatgrass (kernza; Thinopyrum intermedium), representing legume and non-legume crops, respectively. Belowground C input and stabilization was decoupled from nitrogen (N) fertilizer rate in kernza (100 and 200 kg mineral N ha-1), with no direct link between increasing mineral N fertilization, rhizodeposited C, and microbial C stabilization. Further, both crops displayed a high ability to bring C to deeper soil layers and remarkably, the N2-fixing lucerne showed greater potential to induce microbial C stabilization than the non-legume kernza. Lucerne stimulated greater microbial biomass and abundance of N cycling genes in rhizosphere soil, likely linked to greater amino acid rhizodeposition, hence underlining the importance of coupled C and N for microbial C stabilization efficiency. Inclusion of legumes in perennial cropping systems is not only key for improved productivity at low fertilizer N inputs, but also appears critical for enhancing soil C stabilization, in particular in N limited deep subsoils.


Assuntos
Fertilizantes , Solo , Agricultura , Carbono/metabolismo , Produtos Agrícolas/metabolismo , Ecossistema , Medicago sativa/metabolismo , Nitrogênio , Solo/química
13.
BMC Plant Biol ; 22(1): 199, 2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35428186

RESUMO

BACKGROUND: The use of heterosis to produce hybrid seeds is a challenge to breeding for improved crop yield. In previous studies, we isolated a male sterile alfalfa hybrid and successfully obtained a genetically stable alfalfa male sterile line through backcrossing, henceforth named MS-4. In this study, we used RNA-seq technology to analyze the transcriptome profiles of the male sterile line (MS-4) and the male fertile line (MF) of alfalfa to elucidate the mechanism of male sterility. RESULTS: We screened a total of 11,812 differentially expressed genes (DEGs) from both MS-4 and MF lines at three different stages of anther development. Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that these DEGs are mainly involved in processes such as energy metabolism, lipid and amino acid metabolism, carbohydrate metabolism, in addition to cell synthesis and aging. The results from protein-protein interaction (PPI) network analysis showed that the ribosomal protein (MS.Gene25178) was the core gene in the network. We also found that transcriptional regulation was an influential factor in the development of anthers. CONCLUSIONS: Our findings provide new insights into understanding of the fertility changes in the male sterile (MS-4) of alfalfa.


Assuntos
Infertilidade Masculina , Infertilidade das Plantas , Flores/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Humanos , Infertilidade Masculina/metabolismo , Masculino , Medicago sativa/genética , Medicago sativa/metabolismo , Melhoramento Vegetal , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transcriptoma
14.
Oxid Med Cell Longev ; 2022: 2691577, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35378828

RESUMO

Medicago sativa Linn or alfalfa is a tonic plant rich in proteins, vitamins, and minerals that is used to treat many diseases due to its pharmacological properties such as anti-inflammatory and antioxidant activities. So, the aim of this study was to evaluate the efficacy of alfalfa methanolic extract (AME) on the prevention of liver damage caused by nicotine. The total phenols, flavonoids levels, and the free radical scavenging activity of its extract (IC50) were measured. In this study, 30 Wistar rats were randomly divided into 5 groups as control (untreated), N (nicotine only), T1, T2, and T3 (nicotine + AME 100, 250, and 500 mg/kg/day, respectively). AME (orally) and nicotine (intraperitoneal injection, 0.5 mg/kg/day) were then administered for 21 days. Weight gain, the liver-to-body weight ratio, liver functional enzymes, and the lipid profile were measured. Moreover, we evaluated oxidative stress, proinflammatory parameters, and histopathological changes in the liver. Total phenols, flavonoids, and IC50 were determined as 51.68 ± 0.62 mg GAE/g, 18.55 ± 1.01 mg QE/g, and 350.91 ± 16.46 µg/ml, respectively. Nicotine changed the measured parameters to abnormal. AME increased weight gain, the liver-to-body weight ratio, and enzymatic antioxidant levels and decreased malondialdehyde, liver functional enzymes, and proinflammatory cytokine levels. The lipid profile and histopathological changes have also been improved by AME in a dose-dependent manner. The results showed that AME in a dose-dependent manner by improving the inflammation and oxidative damage could improve the liver damage caused by nicotine.


Assuntos
Antioxidantes , Medicago sativa , Animais , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacologia , Antioxidantes/metabolismo , Fígado/metabolismo , Medicago sativa/metabolismo , Nicotina/farmacologia , Estresse Oxidativo , Ratos , Ratos Wistar
15.
Genome ; 65(6): 315-330, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35298891

RESUMO

Heat is one of the major environmental stressors that negatively affects alfalfa production. Previously, we reported the role of microRNA156 (miR156) in heat tolerance; however, the mechanism and downstream genes involved in this process were not fully studied. To provide further insight, we compared an empty vector control and miR156-overexpressing alfalfa plants (miR156+) after exposing them to heat stress (40 °C) for 24 h. We collected leaf samples for transcriptome analysis to illustrate the miR156-regulated molecular mechanisms underlying the heat stress response. A total of 3579 differentially expressed genes (DEGs) were detected exclusively in miR156+ plants under heat stress using the Medicago sativa genome as a reference. GO and KEGG analysis indicated that these DEGs were mainly involved in "polysaccharide metabolism", "response to chemical", "secondary metabolism", "carbon metabolism", and "cell cycle". Transcription factors predicted in miR156+ plants belonged to the TCP family, MYB, ABA response element-binding factor, WRKY, and heat shock transcription factor. We also identified two new SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) family gene members (SPL8a and SPL12a), putatively regulated by miR156. The present study provided a comprehensive transcriptome profile of alfalfa, identified a number of genes and pathways, and revealed an miR156-regulated network of mechanisms at the gene expression level to modulate heat responses in alfalfa.


Assuntos
MicroRNAs , Termotolerância , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Resposta ao Choque Térmico/genética , Medicago sativa/genética , Medicago sativa/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Transcriptoma
16.
Plant Cell Rep ; 41(5): 1273-1284, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35305132

RESUMO

KEY MESSAGE: Alfalfa has the ability to degrade TNT. TNT exposure caused root disruption of mineral nutrient metabolism. The exposure of TNT imbalanced basal cell energy metabolism. The mechanism of 2,4,6-trinitrotoluene (TNT) toxicity effects was analyzed in alfalfa (Medicago sativa L.) seedlings by examining the mineral nutrition and secondary metabolism of the plant roots. Exposure to 25-100 mg·L-1 TNT in a hydroponic solution for 72 h resulted in a TNT absorption rate of 26.8-63.0%. The contents of S, K, and B in root mineral nutrition metabolism increased significantly by 1.70-5.46 times, 1.38-4.01 times, and 1.40-4.03 times, respectively, after TNT exposure. Non-targeted metabolomics analysis of the roots identified 189 significantly upregulated metabolites and 420 significantly downregulated metabolites. The altered metabolites were primarily lipids and lipid-like molecules, and the most significant enrichment pathways were alanine, aspartate, and glutamate metabolism and glycerophospholipid metabolism. TNT itself was transformed in the root system into several intermediate products, including 4-hydroxylamino-2,6-dinitrotoluene, 4-amino-2,6-dinitrotoluene, 2-hydroxylamino-4,6-dinitrotoluene, 2,4',6,6'-tetranitro-2',4-azoxytoluene, 4,4',6,6'-tetranitro-2,2'-azoxytoluene, and 2,4-dinitrotoluene. Overall, TNT exposure disturbed the mineral metabolism balance, and significantly interfered with basic plant metabolism.


Assuntos
Trinitrotolueno , Medicago sativa/metabolismo , Minerais , Metabolismo Secundário , Trinitrotolueno/metabolismo , Trinitrotolueno/toxicidade
17.
Food Chem ; 386: 132716, 2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-35358860

RESUMO

Phytoferritin was explored as an attractive nanocarrier to encapsulate bioactive compounds due to its excellent stability and biocompatibility. In the present study, a novel phytoferritin derived from alfalfa (Medicago sativa) was successfully expressed, purified and characterized. Results confirmed that alfalfa ferritin, self-assembled by 24 subunits, formed a spherical hollow structure. Baicalein exhibits superior antioxidant properties and nutritious values, but low bioavailability and solubility limit its application. Herein, we fabricated water-soluble chitosan-ferritin-baicalein nanoparticles to overcome its drawbacks. It was calculated that one apoferritin cage could encapsulate 52 molecules of baicalein. Moreover, chitosan-ferritin-baicalein nanoparticles prolonged the release of baicalein in simulated gastrointestinal tract digestion. Caco-2 cell monolayer absorption analysis demonstrated that baicalein encapsulated within ferritin-chitosan double layers was more efficient in cellular transportation. These results indicated that alfalfa ferritin, as a novel cage-like protein, has potential application in improving the bioavailability of insoluble bioactive molecules.


Assuntos
Quitosana , Nanopartículas , Células CACO-2 , Quitosana/química , Digestão , Portadores de Fármacos/química , Ferritinas/metabolismo , Flavanonas , Humanos , Medicago sativa/metabolismo
18.
J Sci Food Agric ; 102(11): 4577-4588, 2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-35170039

RESUMO

BACKGROUND: Selenium (Se) is an increasing concern for investigators predominantly because of its consumption in the human body mainly from crops. As the fourth largest plant crop globally, alfalfa is one of the most important forages. Alfalfa was fertilized with selenium(IV) (Se(IV)) under field conditions to study the accumulation and assimilation of Se(IV) and to assess the impact of Se fertilization. RESULTS: It was analyzed that the physio-biochemistry, Se species, combined with transcriptome after spraying Se(IV) at different times (0, 12, and 48 h). 9402 and 12 607 differentially expressed genes (DEGs) were identified at 12 h (versus 0 h) and 48 h (versus 12 h). DEG functional enrichments proposed two time-specific biological processes: Se(IV) accumulation was the primary process at 0-12 h, and its assimilation mainly occurred during 12-48 h. This was further proved by the separation of various Se speciation at different times. It showed that Se-supplementation also affected the soluble protein, soluble sugar, pigment contents and antioxidant capacity. Selenium-biofortification could improve the stress resistance of alfalfa by enhancing antioxidant system to scavenge reactive oxygen species (e.g. hydrogen peroxide) and boosting carbohydrate metabolism. CONCLUSION: By integrating physio-biochemistry, Se-related metabolites, and transcriptome under Se(IV) treatment, this study provides data to guide further work on Se-fortification in alfalfa. © 2022 Society of Chemical Industry.


Assuntos
Medicago sativa , Selênio , Antioxidantes/metabolismo , Perfilação da Expressão Gênica , Humanos , Medicago sativa/genética , Medicago sativa/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Selênio/metabolismo , Transcriptoma
19.
J Agric Food Chem ; 70(7): 2221-2230, 2022 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-35157803

RESUMO

The aim of this study was to characterize the effects of tannins on plant protein during sheep digestion using a digestomic approach combining in vivo (rumen) conditions and an in vitro digestive system (abomasum and small intestine). Ruminal fluid from wethers infused with a tannin solution or water (control) was introduced into the digester, and protein degradation was followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Tannin infusion in the rumen led to a clear decrease in protein degradation-related fermentation end-products, whereas ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) protein was more abundant than in control wethers. In the simulated abomasum, peptidomic analysis showed more degradation products of RuBisCo in the presence of tannins. The effect of RuBisCo protection by tannins continued to impact Rubisco digestion into early-stage intestinal digestion but was no longer detectable in late-stage intestinal digestion. The peptidomics approach proved a potent tool for identifying and quantifying the type of protein hydrolyzed throughout the gastrointestinal tract.


Assuntos
Medicago sativa , Taninos , Ração Animal/análise , Animais , Cromatografia Líquida , Suplementos Nutricionais/análise , Digestão , Fermentação , Medicago sativa/metabolismo , Proteólise , Rúmen/metabolismo , Ovinos , Espectrometria de Massas em Tandem , Taninos/metabolismo
20.
PLoS One ; 17(1): e0260918, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34982779

RESUMO

Desmanthus (Desmanthus spp.), a tropically adapted pasture legume, is highly productive and has the potential to reduce methane emissions in beef cattle. However, liveweight gain response to desmanthus supplementation has been inconclusive in ruminants. This study aimed to evaluate weight gain, rumen fermentation and plasma metabolites of Australian tropical beef cattle in response to supplementation with incremental levels of desmanthus forage legume in isonitrogenous diets. Forty-eight Brahman, Charbray and Droughtmaster crossbred beef steers were pen-housed and fed a basal diet of Rhodes grass (Chloris gayana) hay supplemented with 0, 15, 30 or 45% freshly chopped desmanthus forage on dry matter basis, for 140 days. Varying levels of lucerne (Medicago sativa) hay were added in the 0, 15 and 30% diets to ensure that all diets were isonitrogenous with the 45% desmanthus diet. Data were analyzed using the Mixed Model procedures of SAS software. Results showed that the proportion of desmanthus in the diet had no significant effect on steer liveweight, rumen volatile fatty acids molar proportions and plasma metabolites (P ≥ 0.067). Total bilirubin ranged between 3.0 and 3.6 µmol/L for all the diet treatments (P = 0.67). All plasma metabolites measured were within the expected normal range reported for beef cattle. Rumen ammonia nitrogen content was above the 10 mg/dl threshold required to maintain effective rumen microbial activity and maximize voluntary feed intake in cattle fed low-quality tropical forages. The average daily weight gains averaged 0.5 to 0.6 kg/day (P = 0.13) and were within the range required to meet the target slaughter weight for prime beef markets within 2.5 years of age. These results indicate that desmanthus alone or mixed with other high-quality legume forages can be used to supplement grass-based diets to improve tropical beef cattle production in northern Australia with no adverse effect on cattle health.


Assuntos
Dieta/veterinária , Rúmen/metabolismo , Vicia/química , Amônia/química , Ração Animal/análise , Animais , Austrália , Bilirrubina/sangue , Bovinos , Creatinina/sangue , Suplementos Nutricionais , Ácidos Graxos Voláteis/sangue , Ácidos Graxos Voláteis/metabolismo , Concentração de Íons de Hidrogênio , Hidroxibutiratos/sangue , Masculino , Medicago sativa/química , Medicago sativa/metabolismo , Rúmen/química , Rúmen/microbiologia , Vicia/metabolismo , Aumento de Peso
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