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1.
PLoS One ; 19(4): e0300864, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38635849

RESUMO

Chia (Salvia hispanica L.) seed (CS) and Pumpkin (Cucurbita moschata) seed (PS) are used in ruminant diets as energy sources. The current experiment studied the impact of dietary inclusion of CS and PS on nutrient intake and digestibility, milk yield, and milk composition of dairy sheep. Twelve primiparous Texel × Suffolk ewes [70 ± 5 days in milk (DIM); 0.320 ± 0.029 kg milk yield] were distributed in a 4 × 3 Latin square design and fed either a butter-based control diet [CON; 13 g/kg dry matter] or two diets with 61 g/kg DM of either CS or PS. Dietary inclusion of CS and PS did not alter live weight (p >0.1) and DM intake (p >0.1). However, compared to the CON, dietary inclusion of both CS and PS increased the digestibility of neutral detergent fiber (p <0.001) and acid detergent lignin (p < 0.001). Milk production (p = 0.001), fat-corrected milk (p < 0.001), and feed efficiency (p < 0.001) were enhanced with PS, while the highest milk protein yield (p < 0.05) and lactose yield (p < 0.001) were for CS-fed ewes. Compared to the CON diet, the ingestion of either CS and/or PS decreased (p < 0.001) the C16:0 in milk. Moreover, both CS and PS tended to enhance the content of C18:3n6 (p > 0.05) and C18:3n3 (p > 0.05). Overall short-term feeding of CS and/or PS (up to 6.1% DM of diet) not only maintains the production performance and digestibility of nutrients but also positively modifies the milk FA composition.


Assuntos
Cucurbita , Animais , Feminino , Ovinos , Cucurbita/metabolismo , Lactação , Salvia hispanica , Detergentes , Fibras na Dieta/metabolismo , Dieta/veterinária , Sementes/metabolismo , Digestão , Ração Animal/análise , Zea mays/metabolismo , Suplementos Nutricionais/análise , Rúmen/metabolismo
2.
J Sci Food Agric ; 104(7): 4189-4200, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38349054

RESUMO

BACKGROUND: We investigated the impact of using canola meal (CM) or corn distillers dried grain soluble (cDDGS) in place of soybean meal (SBM) in low-crude-protein diets supplemented with amino acids (AA) on AA digestibility, gut morphometrics, and AA transporter genes in broiler chicken. On day 0, 540 Cobb 500 male broilers were allocated to six diets in 36-floor pens. The positive control (PC) was a corn-SBM diet with adequate crude protein (CP). The CP level of negative control (NC) was decreased by 45 and 40 g kg-1 relative to PC for grower and finisher phases, respectively. The subsequent two diets had the same CP levels as NC but with cDDGS added at 50 or 125 g kg-1. The last two diets had the same CP as NC but with CM added at 50 or 100 g kg-1. RESULTS: Dietary CP reduction in corn-SBM diets increased (P < 0.05) the digestibility of Lys (88.5%), Met (90.7%), Thr (77.4%), Cys (80.7%), and Gly (84.7%). Increasing levels of cDDGS linearly decreased (P < 0.05) the digestibility of Asp, Cys, Glu, and Ser, whereas increasing CM level linearly decreased (P < 0.05) the digestibility of Cys, Pro, and Ser. The CP reduction in corn-SBM diets produced downward expression of peptide transporter1 and decreased (P < 0.05) absolute pancreas and ileum weight and length of jejunum and ileum. CONCLUSIONS: Partial replacement of SBM with alternative protein feedstuffs (cDDGS or CM) in low-CP diets had minimal effects on AA digestibility and mRNA levels of peptides and AA transporters. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Aminoácidos , Brassica napus , Animais , Masculino , Aminoácidos/metabolismo , Galinhas/metabolismo , Zea mays/genética , Zea mays/metabolismo , Farinha , Digestão , Ração Animal/análise , Dieta/veterinária , Dieta com Restrição de Proteínas , Íleo/metabolismo , Brassica napus/genética , Brassica napus/metabolismo , Peptídeos/metabolismo , Glycine max , Expressão Gênica , Fenômenos Fisiológicos da Nutrição Animal
3.
Biochem Biophys Res Commun ; 703: 149637, 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38354464

RESUMO

The normal development of pollen grains and the completion of double fertilization in embryos are crucial for both the sexual reproduction of angiosperms and grain production. Actin depolymerizing factor (ADF) regulates growth, development, and responses to biotic and abiotic stress by binding to actin in plants. In this study, the function of the ZmADF1 gene was validated through bioinformatic analysis, subcellular localization, overexpression in maize and Arabidopsis, and knockout via CRISPR/Cas9. The amino acid sequence of ZmADF1 exhibited high conservation and a similar tertiary structure to that of ADF homologs. Subcellular localization analysis revealed that ZmADF1 is localized mainly to the nucleus and cytoplasm. The ZmADF1 gene was specifically expressed in maize pollen, and overexpression of the ZmADF1 gene decreased the number of pollen grains in the anthers of transgenic Arabidopsis plants. The germination rate of pollen and the empty seed shell rate in the fruit pods of the overexpressing plants were significantly greater than those in the wild-type (WT) plants. In maize, the pollen viability of the knockout lines was significantly greater than that of both the WT and the overexpressing lines. Our results confirmed that the ZmADF1 gene was specifically expressed in pollen and negatively regulated pollen quantity, vigor, germination rate, and seed setting rate. This study provides insights into ADF gene function and possible pathways for improving high-yield maize breeding.


Assuntos
Arabidopsis , Destrina , Pólen , Zea mays , Sequência de Aminoácidos , Arabidopsis/metabolismo , Destrina/genética , Destrina/metabolismo , Gelsolina/metabolismo , Regulação da Expressão Gênica de Plantas , Pólen/genética , Pólen/crescimento & desenvolvimento , Zea mays/metabolismo
4.
Plant Physiol Biochem ; 207: 108396, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38310727

RESUMO

Drought stress poses a substantial threat to global plant productivity amid increasing population and rising agricultural demand. To overcome this problem, the utilization of organic plant growth ingredients aligns with the emphasis on eco-friendly farming practices. Therefore, the present study aimed to assess the influence of 30 botanical extracts on seed germination, seedling vigor, and subsequent maize plant growth under normal and water deficit conditions. Specifically, eight extracts showed significant enhancement in agronomical parameters (ranging from ∼2 % to ∼ 183 %) and photosynthetic pigments (ranging from ∼21 % to âˆ¼ 195 %) of seedlings under drought conditions. Extended tests on maize in a greenhouse setting confirmed that the application of six extracts viz Moringa oleifera leaf (MLE), bark (MBE), Terminalia arjuna leaf (ALE), bark (ABE), Aegel marmelos leaf (BLE), and Phyllanthus niruri leaf (AmLE) improved plant growth and drought tolerance, as evident in improved physio-biochemical parameters. GC-MS analysis of the selected extracts unveiled a total of 51 bioactive compounds, including sugars, sugar alcohols, organic acids, and amino acids, and might be playing pivotal roles in plant acclimatization to drought stress. In conclusion, MLE, MBE, BLE, and ABE extracts exhibit significant potential for enhancing seedling establishment and growth in maize under both normal and water deficit conditions.


Assuntos
Antioxidantes , Zea mays , Antioxidantes/metabolismo , Zea mays/metabolismo , Secas , Plântula/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/metabolismo , Água/metabolismo , Estresse Fisiológico
5.
Gene ; 895: 148001, 2024 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-37977314

RESUMO

Demand for maize oil is progressively increasing due to its diverse industrial applications, aside from its primary role in human nutrition and animal feed. Oil content and composition are two crucial determinants of maize oil in the international market. As kernel oil in maize is a complex quantitative trait, improving this trait presents a challenge for plant breeders and biotechnologists. Here, we characterized a set of 292 diverse maize inbreds of both indigenous and exotic origin by exploiting functional polymorphism of the dgat1-2, fatb, ge2, and wri1a genes governing kernel oil in maize. Genotyping using gene-based functional markers revealed a lower frequencies of dgat1-2 (0.15) and fatb (0.12) mutant alleles and a higher frequencies of wild-type alleles (Dgat1-2: 0.85; fatB: 0.88). The favorable wri1a allele was conserved across genotypes, while its wild-type allele (WRI1a) was not detected. In contrast, none of the genotypes possessed the ge2 favorable allele. The frequency of favorable alleles of both dgat1-2 and fatb decreased to 0.03 when considered together. Furthermore, pairwise protein-protein interactions among target gene products were conducted to understand the effect of one protein on another and their responses to kernel oil through functional enrichments. Thus, the identified maize genotypes with dgat1-2, fatb, and wri1a favourable alleles, along with insights gained through the protein-protein association network, serve as prominent and unique genetic resources for high-oil maize breeding programs. This is the first comprehensive report on the functional characterization of diverse genotypes at the molecular and protein levels.


Assuntos
Óleo de Milho , Zea mays , Humanos , Zea mays/genética , Zea mays/metabolismo , Óleo de Milho/genética , Óleo de Milho/metabolismo , Ácidos Graxos/genética , Ácidos Graxos/metabolismo , Melhoramento Vegetal , Marcadores Genéticos , Alelos
6.
J Anim Physiol Anim Nutr (Berl) ; 108(1): 111-125, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37602531

RESUMO

Hydrolysable tannins (HT) show potential as silage additive for autumn herbage silages, high in (rumen degradable) protein, as they may reduce proteolysis. Additionally, they have abilities to form pH-reversible tannin-protein complexes, non-degradable in the rumen but degradable in the abomasum and intestines of ruminants. Therefore they can improve milk N efficiency and shift N excretions from urine to faeces, possibly mitigating the environmental impact of ruminants. In this study, two small bunker silos were filled with autumn grass. One was treated with 20 g/kg DM HT extract (TAN) (TannoSan-L), the other with 8 mg/kg DM inoculant containing lactic acid bacteria (INO) (Bonsilage Fit G). Secondly, micro-silos (2.75 L) were filled with four treatments; (1) grass without additive (CON) (n = 5); (2) TAN (n = 5); (3) INO (n = 5); and (4) TAN + INO (n = 5). The bunker silos were used in a cross-over feeding experiment with periods of 4 weeks involving 22 lactating Holstein cows (average ± SD: 183 ± 36.3 days in milk, 665 ± 71.0 kg body weight, and 33.8 ± 3.91 kg/day milk yield). The HT dose was insufficient to reduce proteolysis or alter chemical composition and nutritional value in the micro- and bunker silages. Including grass silage added with TAN (3.2 g HT/kg DM) in the diet, did not affect feed intake nor fat and protein corrected milk yield in comparison to feeding the grass silage added with INO in a similar diet. The TAN-fed cows had an increased faecal N excretion and decreased apparent total-tract N and organic matter digestibility, but no improvement in the cows' N utilization could be confirmed in milk and blood urea levels. Overall, feeding an autumn grass silage treated with 20 g/kg chestnut HT extract did not affect the performance of dairy cows in comparison to feeding an autumn grass silage treated with a lactic acid bacteria inoculant.


Assuntos
Inoculantes Agrícolas , Lactobacillales , Feminino , Bovinos , Animais , Poaceae/metabolismo , Silagem/análise , Taninos/farmacologia , Lactação , Inoculantes Agrícolas/metabolismo , Fermentação , Ácido Láctico/metabolismo , Digestão , Leite/química , Dieta/veterinária , Taninos Hidrolisáveis/análise , Taninos Hidrolisáveis/metabolismo , Taninos Hidrolisáveis/farmacologia , Rúmen/metabolismo , Extratos Vegetais/farmacologia , Ruminantes , Valor Nutritivo , Zea mays/metabolismo
7.
Plant Biotechnol J ; 22(1): 216-232, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37792967

RESUMO

Lipid biosynthesis and transport are essential for plant male reproduction. Compared with Arabidopsis and rice, relatively fewer maize lipid metabolic genic male-sterility (GMS) genes have been identified, and the sporopollenin metabolon in maize anther remains unknown. Here, we identified two maize GMS genes, ZmTKPR1-1 and ZmTKPR1-2, by CRISPR/Cas9 mutagenesis of 14 lipid metabolic genes with anther stage-specific expression patterns. Among them, tkpr1-1/-2 double mutants displayed complete male sterility with delayed tapetum degradation and abortive pollen. ZmTKPR1-1 and ZmTKPR1-2 encode tetraketide α-pyrone reductases and have catalytic activities in reducing tetraketide α-pyrone produced by ZmPKSB (polyketide synthase B). Several conserved catalytic sites (S128/130, Y164/166 and K168/170 in ZmTKPR1-1/-2) are essential for their enzymatic activities. Both ZmTKPR1-1 and ZmTKPR1-2 are directly activated by ZmMYB84, and their encoded proteins are localized in both the endoplasmic reticulum and nuclei. Based on protein structure prediction, molecular docking, site-directed mutagenesis and biochemical assays, the sporopollenin biosynthetic metabolon ZmPKSB-ZmTKPR1-1/-2 was identified to control pollen exine formation in maize anther. Although ZmTKPR1-1/-2 and ZmPKSB formed a protein complex, their mutants showed different, even opposite, defective phenotypes of anther cuticle and pollen exine. Our findings discover new maize GMS genes that can contribute to male-sterility line-assisted maize breeding and also provide new insights into the metabolon-regulated sporopollenin biosynthesis in maize anther.


Assuntos
Arabidopsis , Infertilidade , Zea mays/genética , Zea mays/metabolismo , Edição de Genes , Sistemas CRISPR-Cas/genética , Simulação de Acoplamento Molecular , Pironas/metabolismo , Melhoramento Vegetal , Arabidopsis/genética , Lipídeos , Pólen/genética , Pólen/metabolismo , Infertilidade/genética , Infertilidade/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
8.
Plant Physiol Biochem ; 206: 108280, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38103337

RESUMO

Soil calcium (Ca) and magnesium (Mg) mineral states in rain-fed arid regions of Northwest China are inefficient, and their levels of substitution and water-soluble states are far below the lowest threshold required for maize growth, resulting in frequent physiological diseases, restricting synthesis of kernel protein (CRP). Our study set up different levels of foliar spraying of Ca and Mg fertilizers before maize pollination to examine the response characteristics of physiological and biochemical indicators in kernel, and the driving process of CRP synthesis. The main findings were: (1) Ca and Mg significantly increased the levels of CRP and endogenous hormones, and the activities of defense enzymes and CRP synthesis enzymes, which decreased significantly and stabilized at the maturity stage of maize. (2) The synthesis and accumulation of CRP were synergistically regulated by endogenous hormones, defense enzymes, and CRP synthase enzymes, with the degree of regulation varying with the level of Ca and Mg supplementation. Indole-3-acetic acid (IAA), gibberellin (GA), zeatin riboside (ZR), catalase (CAT), malondialdehyde (MDA), and glutamate dehydrogenase (GDH) were the primary physiological driving indicators of CRP synthesis, with CRP having a significant synergistic relationship with CAT and a remarkable trade-off with other driving indicators. (3) The dominant driving pathway of CRP synthesis was "Ca, Mg-IAA or GA or ZR-CAT-GDH-CRP". Ca and Mg positively affected IAA and GA levels, and IAA and GA positively regulated CAT activity. However, CAT negatively regulated GDH levels, causing GDH to negatively influence the synthesis and accumulation of CRP and its components. The findings provide theoretical support for further study of inter-root endogenous hormones and soil microbe-driven processes in the regulation of maize quality by Ca and Mg.


Assuntos
Reguladores de Crescimento de Plantas , Zea mays , Reguladores de Crescimento de Plantas/metabolismo , Zea mays/metabolismo , Magnésio/metabolismo , Hormônios/metabolismo , Solo
9.
Poult Sci ; 103(2): 103297, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38104413

RESUMO

A study was conducted to evaluate the efficiency of guanidinoacetic acid (GAA) in diets containing varying levels of corn replacement with low-tannin sorghum (LTS) for laying hens in the later stage of production. In a 12-wk study, a total of 288 laying hens at 52 wk of age were divided into 6 treatment groups. Each treatment group had 8 replicates, each of which consisted of 6 hens. A 2 × 3 factorial design was used to investigate the impact of substituting corn with LTS at 3 levels (100% corn, 50% LTS, and 100% LTS) with 2 doses of GAA supplementation (0 and 0.6 g/kg). The results indicate that there were interaction effects (P < 0.05) between diet type and GAA supplementation on protein digestibility and AMEn, with the GAA supplement being more effective in the 100% LTS group. Replacing corn with LTS at both levels had no negative effects on performance and metabolic profile. In contrast, the 100% LTS diet increased monounsaturated fatty acids in the yolk (P < 0.05), but decreased the yolk color index, the ratio of polyunsaturated fatty acids (PUFA) to saturated fatty acids (SFA) in the yolk, ileal digestibility of energy, and AMEn when compared to the 100% corn diet (P < 0.05). Regardless of the diet, dietary supplementation with GAA resulted in increases (P < 0.05) in shell-breaking strength, the PUFA to SFA ratio in egg yolk, and concentrations of creatine and nitric oxide in serum. There was also a decrease (P < 0.05) in serum malondialdehyde concentration with GAA supplementation. In conclusion, the positive effects of GAA on protein digestibility and AMEn were found to be more pronounced when corn was completely replaced with LTS. However, the positive effects of GAA on egg-laying performance, eggshell quality, antioxidant status, and yolk fatty acid composition remained consistent regardless of the extent to which corn was substituted with LTS.


Assuntos
Ácidos Graxos , Glicina/análogos & derivados , Sorghum , Animais , Feminino , Ácidos Graxos/metabolismo , Zea mays/metabolismo , Taninos , Galinhas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Gema de Ovo/metabolismo , Nutrientes , Ração Animal/análise
10.
Huan Jing Ke Xue ; 44(12): 7014-7023, 2023 Dec 08.
Artigo em Chinês | MEDLINE | ID: mdl-38098424

RESUMO

Fertilizer reduction and efficiency improvement is an important basis for ensuring the safety of the agricultural ecological environment. Microorganisms are the key driving force for regulating the soil nitrogen and phosphorus cycle. Studying the nitrogen and phosphorus transformation function of rhizosphere microorganisms can provide a microbiological regulation approach for further improving the use efficiency of soil nitrogen and phosphorus. Based on the field micro-plot experiments of three typical farmland soils(phaeozem, cambisol, and acrisol), metagenomic sequencing technology was used to study the differences in functional genes and regulatory factors of maize rhizosphere microorganisms during soil nitrogen and phosphorus transformation. The results showed that the functional diversity of maize rhizosphere microorganisms was affected by soil type. The functional diversity of rhizosphere microorganisms in phaeozem and cambisol was mainly affected by water content and nutrient content, and that in acrisol was affected by total phosphorus(TP) and available phosphorus(AP). For soil nitrogen transformation, the gene abundance of related enzymes in the pathway of nitrogen transformation was the highest in the urease gene(ureC) and glucose dehydrogenase gene(gdh), which were 7.25×10-5-12.88×10-5 and 4.47×10-5-7.49×10-5, respectively. The total abundance of assimilatory nitrate reduction functional genes in acrisol was higher than that in phaeozem and cambisol, and the total abundance of functional genes related to other processes was the highest in cambisol. The abundance of functional genes encoding enzymes related to nitrogen metabolism was mainly driven by soil bacterial richness, total potassium(TK), and TP. For soil phosphorus transformation, the number of alkaline phosphatase genes(phoD) catalyzing organic phosphorus mineralization was 1093, and the number of acid phosphatase genes(PHO) was 42. The abundance of phoD was two orders of magnitude higher than that of PHO. In addition, fertilization had no significant effect on the abundance of phoD and PHO in the same soil type. Random forest analysis showed that the abundances of phoD and PHO were significantly affected by soil moisture, organic matter(OM), and total nitrogen(TN), but AP content had the greatest impact on PHO abundance. These results clarified the nitrogen and phosphorus transformation characteristics of maize rhizosphere microorganisms at the functional genomic level and enriched the molecular biological mechanism of the microbial nitrogen and phosphorus transformation function.


Assuntos
Rizosfera , Zea mays , Zea mays/metabolismo , Fósforo/metabolismo , Nitrogênio/análise , Solo , Genômica , Microbiologia do Solo , Fertilizantes/análise
11.
Water Sci Technol ; 88(12): 3129-3141, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38154799

RESUMO

To comprehensively assess the efficacy of employing the internal circulation (IC) anaerobic reactor for corn alcohol wastewater treatment and investigate its feasibility, this study focused on anaerobic digestion parameters, energy balance, and the composition of the prokaryotic microbial community. During the operation of the reactor, the hydraulic retention time was progressively reduced from 4.8 to 1.6 days while achieving an average organic loading rate of 12.46 kg chemical oxygen demand (COD)/(m3·d). Moreover, the removal rate of COD exceeded 98%, and the energy balance (ΔE) reached 10.29 kJ/g fed COD. The initial manifestation of organic acidosis in the reactor was a decline in gas production, which is primarily caused by propionic acid accumulation. The subsequent analysis revealed a high diversity of prokaryotes in granular sludge, with the predominant archaea primarily involved in methane production through the acetic acid pathway. The IC anaerobic reactor shows exceptional performance in treating corn alcohol wastewater by optimizing its operating conditions. Energy balance analysis confirmed the feasibility of the process. The findings of this study may offer valuable insights for optimizing control strategies and engineering applications.


Assuntos
Águas Residuárias , Zea mays , Zea mays/metabolismo , Eliminação de Resíduos Líquidos , Anaerobiose , Reatores Biológicos , Esgotos/química , Etanol , Metano/metabolismo
12.
PeerJ ; 11: e16386, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38025755

RESUMO

This study aimed to examine the impact of nitrogen (N) fertilization on phyllosphere microorganisms in silage maize (Zea mays) to enhance the production of high-quality silage. The effects of different N application rates (160, 240, and 320 kg ha-1) and maturity stages (flowering and dough stages) on microbial diversity, abundance and physiochemical properties of the leaf surfaces were evaluated in a field experiment. The results showed that N application rates did not significantly impact the abundance of lactic acid bacteria (LAB), aerobic bacteria (AB), yeasts, or molds on the leaf surfaces. However, these microbes were more abundant during the flowering stage compared to the dough stage. Furthermore, the N application rate had no significant impact on inorganic phosphorus, soluble sugar, free amino acids, total phenolic content, and soluble protein concentrations, or pH levels on the leaf surfaces. Notably, these chemical indices were lower during the dough stage. The abundance of Pantoea decreased with higher N application rates, while that of other microorganisms did not changes significantly. The abundance of AB, LAB, yeasts, and molds were positively correlated with soluble sugar, soluble protein, inorganic phosphorus, free amino acids, and total phenolic concentrations on leaf surfaces. Moreover, water loss was negatively correlated with the abundance of AB, LAB, yeasts, and molds, whereas water retention capacity and stomatal density were positively correlated with microbial abundance. We recommend applying an optimal N rate of 160 kg ha-1 to silage maize and harvesting at the flowering stage is recommended.


Assuntos
Microbiota , Silagem , Silagem/análise , Zea mays/metabolismo , Nitrogênio/farmacologia , Folhas de Planta , Carboidratos , Fungos , Leveduras , Açúcares/metabolismo , Aminoácidos/metabolismo , Fósforo/metabolismo , Água/metabolismo
13.
Nat Protoc ; 18(11): 3512-3533, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37783945

RESUMO

RNA-sequencing (RNA-seq) provides invaluable knowledge on developmental pathways and the effects of mutant phenotypes. Plant reproductive cells have traditionally been difficult to isolate for genomics because they are rare and often deeply embedded within somatic tissues. Here, we present a protocol to isolate single maize meiocytes and pollen grains for RNA-seq. We discuss how to identify and isolate each sample type under a microscope, prepare RNA-seq libraries and analyze the data. This technique has several advantages over alternative methods, combining the ability to target specific rare cell types while resolving cell-to-cell heterogeneity with single-cell RNA-seq. The technique is compatible with minute amounts of starting material (e.g., a single anther), making it possible to collect dense time courses. Furthermore, developmentally synchronized anthers are saved for microscopy, allowing staging to be performed in parallel with expression analysis. Up to 200 cells can be collected in 4-5 h by someone proficient in tissue dissection, and library preparation can be completed in 2 d by researchers experienced in molecular biology and genomics. This protocol will facilitate research on plant reproduction, providing insights critical to plant breeding, genetics and agriculture.


Assuntos
Análise da Expressão Gênica de Célula Única , Zea mays , Zea mays/genética , Zea mays/metabolismo , Genômica , Pólen/genética , Pólen/metabolismo , Fenótipo
14.
Int J Mol Sci ; 24(19)2023 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-37834187

RESUMO

Common smut caused by Ustilago maydis is one of the dominant fungal diseases in plants. The resistance mechanism to U. maydis infection involving alterations in the cell wall is poorly studied. In this study, the resistant single segment substitution line (SSSL) R445 and its susceptible recurrent parent line Ye478 of maize were infected with U. maydis, and the changes in cell wall components and structure were studied at 0, 2, 4, 8, and 12 days postinfection. In R445 and Ye478, the contents of cellulose, hemicellulose, pectin, and lignin increased by varying degrees, and pectin methylesterase (PME) activity increased. The changes in hemicellulose and pectin in the cell wall after U. maydis infection were analyzed via immunolabeling using monoclonal antibodies against hemicellulsic xylans and high/low-methylated pectin. U. maydis infection altered methyl esterification of pectin, and the degree of methyl esterification was correlated with the resistance of maize to U. maydis. Furthermore, the relationship between methyl esterification of pectin and host resistance was validated using 15 maize inbred lines with different resistance levels. The results revealed that cell wall components, particularly pectin, were important factors affecting the colonization and propagation of U. maydis in maize, and methyl esterification of pectin played a role in the resistance of maize to U. maydis infection.


Assuntos
Doenças das Plantas , Ustilago , Doenças das Plantas/microbiologia , Esterificação , Zea mays/metabolismo , Pectinas/metabolismo , Ustilago/metabolismo , Parede Celular/metabolismo
15.
Molecules ; 28(19)2023 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-37836621

RESUMO

A theoretical analysis of the potential inhibition of human sucrase-isomaltase (SI) by flavonoids was carried out with the aim of identifying potential candidates for an alternative treatment of type 2 diabetes. Two compounds from maize silks, maysin and luteolin, were selected to be studied with the structure-based density functional theory (DFT), molecular docking (MDock), and molecular dynamics (MD) approaches. The docking score and MD simulations suggested that the compounds maysin and luteolin presented higher binding affinities in N-terminal sucrase-isomaltase (NtSI) than in C-terminal sucrase-isomaltase (CtSI). The reactivity parameters, such as chemical hardness (η) and chemical potential (µ), of the ligands, as well as of the active site amino acids of the NtSI, were calculated by the meta-GGA M06 functional in combination with the 6-31G(d) basis set. The lower value of chemical hardness calculated for the maysin molecule indicated that this might interact more easily with the active site of NtSI, in comparison with the values of the acarbose and luteolin structures. Additionally, a possible oxidative process was proposed through the quantum chemical calculations of the electronic charge transfer values (∆N) between the active site amino acids of the NtSI and the ligands. In addition, maysin displayed a higher ability to generate more oxidative damage in the NtSI active site. Our results suggest that maysin and luteolin can be used to develop novel α-glucosidase inhibitors via NtSI inhibition.


Assuntos
Diabetes Mellitus Tipo 2 , Humanos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Zea mays/metabolismo , Simulação de Acoplamento Molecular , Luteolina/farmacologia , Oligo-1,6-Glucosidase/química , Sacarase/metabolismo , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/uso terapêutico , Aminoácidos
16.
Plant Commun ; 4(6): 100682, 2023 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-37691288

RESUMO

Sporopollenin in the pollen cell wall protects male gametophytes from stresses. Phenylpropanoid derivatives, including guaiacyl (G) lignin units, are known to be structural components of sporopollenin, but the exact composition of sporopollenin remains to be fully resolved. We analyzed the phenylpropanoid derivatives in sporopollenin from maize and Arabidopsis by thioacidolysis coupled with nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS). The NMR and GC-MS results confirmed the presence of p-hydroxyphenyl (H), G, and syringyl (S) lignin units in sporopollenin from maize and Arabidopsis. Strikingly, H units account for the majority of lignin monomers in sporopollenin from these species. We next performed a genome-wide association study to explore the genetic basis of maize sporopollenin composition and identified a vesicle-associated membrane protein (ZmVAMP726) that is strongly associated with lignin monomer composition of maize sporopollenin. Genetic manipulation of VAMP726 affected not only lignin monomer composition in sporopollenin but also pollen resistance to heat and UV radiation in maize and Arabidopsis, indicating that VAMP726 is functionally conserved in monocot and dicot plants. Our work provides new insight into the lignin monomers that serve as structural components of sporopollenin and characterizes VAMP726, which affects sporopollenin composition and stress resistance in pollen.


Assuntos
Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Estudo de Associação Genômica Ampla , Temperatura Alta , Lignina/química , Lignina/genética , Lignina/metabolismo , Pólen/genética , Pólen/metabolismo , Raios Ultravioleta , Zea mays/genética , Zea mays/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
17.
Braz J Biol ; 83: e274601, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37585933

RESUMO

An experiment was performed to investigate the effect of mycorrhizal symbiosis and foliar application of salicylic acid on quantitative and qualitative traits of maize during 2018 and 2019 in the research farm of Islamic Azad University, Chalous Branch. Split plot in a randomized complete block design with three replications was used. Experimental factors included mycorrhiza species of (G. mosseae), (G. geosporum) and (G. intraradices) at two levels (no consumption and consumption of mycorrhiza) and salicylic acid at two levels (no consumption and consumption of 1 mµ of salicylic acid). Results of interaction effects of mycorrhiza and salicylic acid on the measured traits revealed that the maximum 1000-grain weight, grain yield, biological yield, phosphorus, potassium, nitrogen percentage and yield of maize grain protein were observed in G. mosseae treatment under foliar application of salicylic acid. Foliar application of salicylic acid increases the root length and provides the necessary conditions for increasing water and nutrient uptake alongwith increase in photosynthesis and thus allocates more photosynthetic substance for development of reproductive organs. Hence, it increases maize grain weight and accordingly grain yield. In general, the results revealed that mycorrhiza and foliar application of salicylic acid increase growth indicators, yield and yield components. It also improved the quality traits of the maize plant. Based on results, the interaction effect of G. mosseae treatment and foliar application of salicylic acid yielded better results than other treatments. Mycorrhiza increases the number of grain in the ear, the number of rows in the ear, increases the plant's ability to absorb phosphorus, and the increase of mycorrhiza along with salicylic acid shows the maximum grain yield in maize. Finally, it can be concluded that the use of mycorrhiza and salicylic acid can be effective in increasing grain in the plant.


Assuntos
Micorrizas , Zea mays , Humanos , Grão Comestível , Micorrizas/metabolismo , Fósforo/metabolismo , Fotossíntese , Ácido Salicílico/farmacologia , Zea mays/genética , Zea mays/metabolismo , Locos de Características Quantitativas
18.
Int J Mol Sci ; 24(15)2023 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-37569319

RESUMO

Talaromyces purpurogenus, an endophytic fungus, exhibits beneficial effects on plants during plant-fungus interactions. However, the molecular mechanisms underlying plants' responses to T. purpurogenus under low-phosphorous (P) stress are not fully understood. In this study, we investigated the transcriptomic changes in maize with low-P-sensitive (31778) and -tolerant (CCM454) genotypes under low-P stress and its symbiotic interaction with T. purpurogenus. Its colonization enhanced plant growth and facilitated P uptake, particularly in 31778. Transcriptome sequencing revealed that 135 DEGs from CCM454 and 389 from 31778 were identified, and that only 6 DEGs were common. This suggested that CCM454 and 31778 exhibited distinct molecular responses to T. purpurogenus inoculation. GO and KEGG analysis revealed that DEGs in 31778 were associated with nicotianamine biosynthesis, organic acid metabolic process, inorganic anion transport, biosynthesis of various secondary metabolites and nitrogen metabolism. In CCM454, DEGs were associated with anthocyanin biosynthesis, diterpenoid biosynthesis and metabolic process. After T. purpurogenus inoculation, the genes associated with phosphate transporter, phosphatase, peroxidase and high-affinity nitrate transporter were upregulated in 31778, whereas AP2-EREBP-transcription factors were detected at significantly higher levels in CCM454. This study provided insights on the molecular mechanisms underlying plant-endophytic fungus symbiosis and low-P stress in maize with low-P-sensitive and -tolerant genotypes.


Assuntos
Simbiose , Transcriptoma , Simbiose/genética , Zea mays/metabolismo , Perfilação da Expressão Gênica , Genótipo , Fósforo/metabolismo , Regulação da Expressão Gênica de Plantas
19.
J Dairy Sci ; 106(12): 8627-8641, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37641245

RESUMO

This study aimed to quantify the effects of dietary inclusion of tannin-rich pomegranate peel (PP) on intake, methane and nitrogen (N) losses, and metabolic and health indicators in dairy cows. Four multiparous, late-lactating Brown Swiss dairy cows (796 kg body weight; 29 kg/d of energy corrected milk yield) were randomly allocated to 3 treatments in a randomized cyclic change-over design with 3 periods, each comprising 14 d of adaptation, 7 d of milk, urine, and feces collection, and 2 d of methane measurements. Treatments were formulated using PP that replaced on a dry matter (DM) basis 0% (control), 5%, and 10% of the basal mixed ration (BMR) consisting of corn and grass silage, alfalfa, and concentrate. Gaseous exchange of the cows was determined in open-circuit respiration chambers. Blood samples were collected on d 15 of each period. Individual feed intake as well as feces and urine excretion were quantified, and representative samples were collected for analyses of nutrients and phenol composition. Milk was analyzed for concentrations of fat, protein, lactose, milk urea N, and fatty acids. Total phenols and antioxidant capacity in milk and plasma were determined. In serum, the concentrations of urea and bilirubin as well as the activities of alanine aminotransferase (ALT), aspartate aminotransferase, glutamate dehydrogenase, alkaline phosphatase, and γ-glutamyl transferase were measured. The data were subjected to ANOVA with the Mixed procedure of SAS, with treatment and period as fixed and animal as random effects. The PP and BMR contained 218 and 3.5 g of total extractable tannins per kg DM, respectively, and thereof 203 and 3.3 g of hydrolyzable tannins. Total DM intake, energy corrected milk, and methane emission (total, yield, and intensity) were not affected by PP supplementation. The proportions of C18:2n-6 and C18:3n-3 in milk increased linearly as the amount of PP was increased in the diet. Milk urea N, blood urea N, and urinary N excretion decreased linearly with the increase in dietary PP content. Total phenols and antioxidant capacity in milk and plasma were not affected by the inclusion of PP. The activity of ALT increased in a linear manner with the inclusion of PP. In conclusion, replacing up to 10% of BMR with PP improved milk fatty acid composition and alleviated metabolic and environmental N load. However, the elevated serum ALT activity indicates an onset of liver stress even at 5% PP, requiring the development of adaptation protocols for safe inclusion of PP in ruminant diets.


Assuntos
Lactação , Punica granatum , Feminino , Bovinos , Animais , Nitrogênio/metabolismo , Metano/metabolismo , Antioxidantes/metabolismo , Dieta/veterinária , Leite/química , Zea mays/metabolismo , Ácidos Graxos/análise , Silagem/análise , Taninos , Ureia/metabolismo , Rúmen/metabolismo
20.
Pest Manag Sci ; 79(12): 4868-4878, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37506299

RESUMO

BACKGROUND: The fall armyworm Spodoptera frugiperda (J.E. Smith), is an important pest of agronomical crops. It is interesting to discover secondary metabolites in plants that are environmentally safer than synthetic pesticides. For this purpose, Combretum trifoliatum crude extract and its isolated compounds were investigated for their insecticidal activities against S. frugiperda. RESULTS: The median lethal dose (LD50 ) was evaluated in the second-instar larvae using the topical application method. The isolated compounds, apigenin and camphor, demonstrated a highly toxic effect on larvae at a lower LD50 dose than crude extract. Moreover, when the larvae were exposed to crude extract concentrations, the development to pupa and adult stages was reduced by more than 50%. The ovicidal toxicity was examined using a hand sprayer. The extract concentration 5, 10, and 20 µg/egg significantly decreased the egg hatchability. In addition, crude extract showed a significant difference in inhibiting acetylcholinesterase (AChE) activity while crude extract and camphor showed significant inhibitory effects on carboxylesterase (CE) and glutathione-S-transferase (GST) activities. CONCLUSION: The crude ethanol extract of Combretum trifoliatum was toxic to S. frugiperda in terms of larval mortality, negatively affecting biological parameters, and decreasing egg hatchability. Additionally, the activities of cholinergic and detoxifying enzymes were affected by crude extract and its isolated compounds. These results highlight that Combretum trifoliatum might be efficient as a bioinsecticide to control S. frugiperda. © 2023 Society of Chemical Industry.


Assuntos
Combretaceae , Combretum , Inseticidas , Myrtales , Animais , Inseticidas/farmacologia , Spodoptera , Combretum/metabolismo , Combretaceae/metabolismo , Myrtales/metabolismo , Cânfora/toxicidade , Acetilcolinesterase/metabolismo , Larva , Extratos Vegetais/farmacologia , Zea mays/metabolismo
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