Exploring the Effects of an Alfalfa Leaf-Derived Adsorbent on Microbial Community, Ileal Morphology, Barrier Function, and Immunity in Turkey Poults during Chronic Aflatoxin B1 Exposure.

This article follows-up on our recently published work, which evaluated the impact of the addition of an alfalfa leaf-derived adsorbent in the aflatoxin B1 (AFB1)-contaminated diet in regard to the production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology of turkey poults. This paper presents complementary results on microbial community, ileal morphology, barrier function, and immunity. For this purpose, 350 1-day-old female turkey poults were randomly distributed into five groups: (1) Control, AFB1-free diet; (2) AF, AFB1-contaminated diet at 250 ng/g; (3) alfalfa, AFB1-free diet + 0.5% (w/w) adsorbent; (4) alfalfa + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) adsorbent; and (5) YCW + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) commercial yeast cell wall-based adsorbent (reference group). In general, in the AF group, the growth of opportunistic pathogens was promoted, which lead to gut dysbacteriosis, mainly influenced by Streptococcus lutetiensis. Conversely, a significant increase in beneficial bacteria (Faecalibacterium and Coprococcus catus) was promoted by the addition of the plant-based adsorbent. Moreover, the AF group had the lowest villus height and a compromised barrier function, as evidenced by a significant (p < 0.05) increase in fluorescein isothiocyanate dextran (FITC-d), but these negative effects were almost reversed by the addition of the alfalfa adsorbent. Furthermore, the AF + YCW and alfalfa + AF groups exhibited a significant increase in the cutaneous basophil hypersensitivity response compared to the rest of the experimental groups. Taken together, these results pointed out that the alfalfa counteracts the adverse effects of AFB1 in poults, facilitating the colonization of beneficial bacteria and improving the barrier function of the turkey poults.

Efficacy of powdered alfalfa leaves to ameliorate the toxic effects of aflatoxin B1 in turkey poults.

This experiment was conducted to determine the effect of an adsorbent material based on powdered alfalfa leaves added in the aflatoxin B1 (AFB1)-contaminated diet of turkey poults on production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology. For this purpose, three hundred and fifty female Nicholas-700 poults were randomly assigned into five treatments: (1) Control, AFB1-free diet; (2) AF, diet contaminated with 250 ng AFB1/g; (3) Alfalfa, AFB1-free diet + 0.5% (w/w) adsorbent; (4) AF+alfalfa, diet contaminated with 250 ng AFB1/g + 0.5% (w/w) adsorbent, and (5) AF+ yeast cell wall (YCW), diet contaminated with 250 ng AFB1/g + 0.5% (w/w) of yeast cell wall (a commercial mycotoxin binder used as reference material). The in vivo efficacy of powdered alfalfa leaves was assessed during a 28-day period. In general, the addition of powdered alfalfa leaves in the AFB1-free diet gave the best performance results (body weight, body weight gain, and feed intake) and improved the values of total protein, glucose, calcium, creatinine, and blood urea nitrogen. Moreover, the addition of powdered alfalfa leaves in the AFB1-contaminated diet enhanced body weight and body weight gain and significantly reduced the feed intake, compared to the AF and AF+YCW groups. Additionally, significant alterations in serum parameters were observed in poults intoxicated with the AFB1, compared to the Control group. Furthermore, typical histopathological lesions were observed in the liver of the AF group, which were significantly ameliorated with the addition of powdered alfalfa leaves. Conclusively, these results pointed out that low inclusion of powdered alfalfa leaves in the contaminated feed counteracted the adverse effects of AFB1 in turkey poults.

Removal of Aflatoxin B1 Using Alfalfa Leaves as an Adsorbent Material: A Comparison between Two In Vitro Experimental Models.

An adsorbent material derived from alfalfa leaves was prepared and further characterized, and its efficacy for removing aflatoxin B1 (AFB1) was investigated. Characterization consisted of the use of attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), environmental scanning electron microscopy (ESEM), X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD), point of zero charge (pHpzc), zeta potential (ζ-potential), UV-Vis diffuse reflectance spectroscopy, and spectral analysis. To determine the adsorption capacity against AFB1 (250 ng AFB1/mL), pH-dependent and avian intestinal in vitro models were used. The adsorbent inclusion percentage was 0.5% (w/w). In general, the pH-dependent model gave adsorption percentages of 98.2%, 99.9%, and 98.2%, evaluated at pH values of 2, 5, and 7, respectively. However, when the avian intestinal model was used, it was observed that the adsorption percentage of AFB1 significantly decreased (88.8%). Based on the characterization results, it is proposed that electrostatic, non-electrostatic, and the formation of chlorophyll-AFB1 complexes were the main mechanisms for AFB1 adsorption. From these results, it can be concluded that the adsorbent derived from alfalfa leaves could be used as an effective material for removing AFB1 in in vitro digestion models that mimic the physiological reality.

Effect of Dietary Guanidinoacetic Acid Levels on the Mitigation of Greenhouse Gas Production and the Rumen Fermentation Profile of Alfalfa-Based Diets.

The objective of this study was to evaluate the effect of different percentages of alfalfa (Medicago sativa L.) hay (AH) and doses of guanidinoacetic acid (GAA) in the diet on the mitigation of greenhouse gas production, the in vitro rumen fermentation profile and methane (CH4) conversion efficiency. AH percentages were defined for the diets of beef and dairy cattle, as well as under grazing conditions (10 (AH10), 25 (AH25) and 100% (AH100)), while the GAA doses were 0 (control), 0.0005, 0.0010, 0.0015, 0.0020, 0.0025 and 0.0030 g g-1 DM diet. With an increased dose of GAA, the total gas production (GP) and methane (CH4) increased (p = 0.0439) in the AH10 diet, while in AH25 diet, no effect was observed (p = 0.1311), and in AH100, GP and CH4 levels decreased (p = 0.0113). In addition, the increase in GAA decreased (p = 0.0042) the proportion of CH4 in the AH25 diet, with no influence (p = 0.1050) on CH4 in the AH10 and AH100 diet groups. Carbon monoxide production decreased (p = 0.0227) in the AH100 diet with most GAA doses, and the other diets did not show an effect (p = 0.0617) on carbon monoxide, while the production of hydrogen sulfide decreased (p = 0.0441) in the AH10 and AH100 diets with the addition of GAA, with no effect observed in association with the AH25 diet (p = 0.3162). The pH level increased (p < 0.0001) and dry matter degradation (DMD) decreased (p < 0.0001) when AH was increased from 10 to 25%, while 25 to 100% AH contents had the opposite effect. In addition, with an increased GAA dose, only the pH in the AH100 diet increased (p = 0.0142 and p = 0.0023) the DMD in the AH10 diet group. Similarly, GAA influenced (p = 0.0002) SCFA, ME and CH4 conversion efficiency but only in the AH10 diet group. In this diet group, it was observed that with an increased dose of GAA, SCFA and ME increased (p = 0.0002), while CH4 per unit of OM decreased (p = 0.0002) only with doses of 0.0010, 0.0015 and 0.0020 g, with no effect on CH4 per unit of SCFA and ME (p = 0.1790 and p = 0.1343). In conclusion, the positive effects of GAA depend on the percentage of AH, and diets with 25 and 100% AH showed very little improvement with the addition of GAA, while the diet with 10% AH presented the best results.

Including 8 hours of access to alfalfa in 1 or 2 grazing sessions in dairy cows fed a partial mixed ration: Effects on intake, behavior, digestion, and milk production and composition.

The aim of this study was to evaluate the inclusion of alfalfa grazing during 8 h continuous or partitioned in 2 separated sessions of 4 h after each milking, on nutrient intake, digestibility, ruminal fermentation, feeding behavior, milk production, milk composition, and milk fatty acid profile, in late-lactation cows fed a partial mixed ration (PMR). Twelve dairy cows (193 ± 83 d in milk, 584 ± 71 kg of body weight) were housed in individual outdoor pens and assigned to treatments according to a 3 × 3 Latin square design replicated 4 times. The treatments were as follows: (1) control (T0), cows were fed a total mixed ration (TMR) provided ad libitum 20.0% crude protein (CP), 32.2% neutral detergent fiber (NDF); (2) fed a diet combining a PMR which had the same ingredient composition as the TMR (60% of ad libitum intake) + 1 session of 8 h of pasture (T8), continuous grazing alfalfa (Medicago sativa; 20.6% CP, 35.8% NDF) after the p.m. milking; and (3) PMR (60% of ad libitum intake) + 2 daily sessions of 4 h of access to pasture after each milking (T4+4). The experiment lasted 57 d and was divided into 3 periods of 19 d each. The first 12 d of each period was used for diet adaptation, and the last 7 d was used for data collection. No differences among treatments were observed for any of the productive variables, feeding efficiency, or purine derivatives excretion. Cows in T0 had greater intake and apparent digestibility of dry matter, organic matter, and nonfibrous carbohydrates compared with T4+4 and T8. Compared with T0, alfalfa grazing increased the concentration of C18:1 trans-11 and decreased those of C16:0 and C17:0 in milk fat. Cows in T4+4 consumed 1.1 more kg DM/d of alfalfa and N provided by alfalfa in the diet was 3 percentage points higher compared with T8 cows (266 vs. 229 g/d, respectively). In addition, T4+4 cows had a greater daily range of ruminal pH than T8 (0.73 vs. 0.93), and the highest concentrations of NH3-N were recorded during the a.m. grazing session while in T8 cows it occurred during the night. In conclusion, including 8 h of alfalfa grazing in T8 and T4+4 treatments allowed the substitution between 35.8 and 38.7% of the total dry matter intake (DMI) of a PMR (with a similar CP concentration to alfalfa) for pasture, maintaining milk solids production and increasing the C18:1 trans-11 of milk fat compared with a TMR in mid late-lactation cows. In an herbage plus PMR diet, splitting the 1 continuous grazing session of 8 h into 2 sessions of 4 h increased the proportion of energy and N provided by alfalfa pasture and reduced PMR intake, without modifying the total nutrient intake or productive performance of cows.

MsTFL1A delays flowering and regulates shoot architecture and root development in Medicago sativa.

KEY MESSAGE: MsTFL1A is an important gene involved in flowering repression in alfalfa (Medicago sativa) which conditions not only above-ground plant shoot architecture but also root development and growth. Delayed flowering is an important trait for forage species, as it allows harvesting of high-quality forage for a longer time before nutritional values decline due to plant architecture changes related to flowering onset. Despite the relevance of delayed flowering, this trait has not yet been thoroughly exploited in alfalfa. This is mainly due to its complex genetics, sensitivity to inbreeding and to the fact that delayed flowering would be only advantageous if it allowed increased forage quality without compromising seed production. To develop new delayed-flowering varieties, we have characterized the three TERMINAL FLOWERING 1 (TFL1) family of genes in alfalfa: MsTFL1A, MsTFL1B and MsTFL1C. Constitutive expression of MsTFL1A in Arabidopsis caused late flowering and changes in inflorescence architecture, indicating that MsTFL1A is the ortholog of Arabidopsis TFL1. Overexpression of MsTFL1A in alfalfa consistently led to delayed flowering in both controlled and natural field conditions, coupled to an increase in leaf/stem ratio, a common indicator of forage quality. Additionally, overexpression of MsTFL1A reduced root development, reinforcing the role of MsTFL1A not only as a flowering repressor but also as a regulator of root development.We conclude that the precise manipulation of MsTFL1A gene expression may represent a powerful tool to improve alfalfa forage quality.

CRISPR/Cas9-mediated knockout of a polyester synthase-like gene delays flowering time in alfalfa.

KEY MESSAGE: T-DNA and CRISPR/Cas9-mediated knockout of polyester synthase-like genes delays flowering time in Arabidopsis thaliana and Medicago sativa (alfalfa). Thus, we here present the first report of edited alfalfa with delayed flowering.

Agromorphological, yield and quality characteristics of two populations of alfalfa developed by mass selection / Características agromorfológicas, produtivas e de qualidade de duas populações de alfafa desenvolvidas por seleção massal

This study was designed to test the two alfalfa advanced populations (L-1737 and L-1738) with four control cultivars (Bilensoy-80, Gözlü, Savas and Plato) for agromorphological, yield and quality properties under irrigated conditions in two various locations in Ankara, Turkey in the years of 2013, 2014, and 2015. The experiments were arranged in a randomized block design with four replications. Results of the study showed that there were statistically significant differences in plant height and stem diameter as well as stem number among the study materials. Statistically significant differences were observed for green forage yield (P < 0.05), while not in dry matter yield among the study materials. The average green forage and dry matter yields were reported as 67.0 t ha-1 and 13. 0 t ha-1, respectively. Green forage and dry matter yields in the advanced populations of L-1737 and L-1738 were 69.2 and 67.4 t ha-1; 13.7 and 13.6 t ha-1, respectively. Those yield values were 6.5 and 3.7%; 4.3 and 3.4% higher in green forage and dry matter yields of the advanced populations of L-1737 and L-1738 compared to the control cultivar Bilensoy-80, respectively. These increases in green forage and dry matter yields were due to the effect of the mass selection method. Statistically significant differences were observed for acid detergent fiber, neutral detergent fiber, and relative feeding value among the study materials, except crude protein contents. It was concluded that the two advanced populations had good adaptation ability, high yield and good quality performance under irrigated conditions and could be used as commercial cultivars.

Este estudo foi desenhado para testar duas populações avançadas de alfafa (L-1737 e L-1738) com quatro cultivares controle (Bilensoy-80, Gözlü, Savas e Plato) para propriedades agromorfológicas, produtividade e qualidade sob condições irrigadas em dois locais diferentes em Ancara, Turquia, nos anos de 2013, 2014 e 2015. Os experimentos foram dispostos em delineamento de blocos casualizados com quatro repetições. Os resultados do estudo mostraram que houveram diferenças estatisticamente significativas na altura das plantas e diâmetro do colmo, bem como no número de colmos entre os materiais avaliados. Foram observadas diferenças estatisticamente significativas para a produtividade de forragem verde (P < 0.05), enquanto não foram observadas diferenças na produção de matéria seca entre os materiais estudados. As produtividades médias de forragem verde e matéria seca foram de 67.0 t ha-1 e 13.0 t ha-1, respectivamente. Os rendimentos de forragem verde e matéria seca nas populações avançadas de L-1737 e L-1738 foram 69.2 e 67.4 t ha-1; 13.7 e 13.6 t ha-1, respectivamente. Esses valores de rendimento foram 6.5 e 3.7%; 4.3 e 3.4% maiores em rendimentos de forragem verde e matéria seca das populações avançadas de L-1737 e L-1738 em relação à cultivar controle Bilensoy-80, respectivamente. Esses aumentos nos rendimentos de forragem verde e matéria seca foram devidos ao efeito do método de seleção massal. Diferenças, estatisticamente significativas, foram observadas para fibra em detergente ácido, fibra em detergente neutro e valor alimentar relativo entre os materiais estudados, exceto nos teores de proteína bruta. Concluiu-se que as duas populações avançadas apresentaram boa capacidade de adaptação, alta produtividade e bom desempenho em condições irrigadas, podendo ser utilizadas como cultivares comerciais.

Agromorphological, yield and quality characteristics of two populations of alfalfa developed by mass selection / Características agromorfológicas, produtivas e de qualidade de duas populações de alfafa desenvolvidas por seleção massal

ABSTRACT: This study was designed to test the two alfalfa advanced populations (L-1737 and L-1738) with four control cultivars (Bilensoy-80, Gözlü, Savaş and Plato) for agromorphological, yield and quality properties under irrigated conditions in two various locations in Ankara, Turkey in the years of 2013, 2014, and 2015. The experiments were arranged in a randomized block design with four replications. Results of the study showed that there were statistically significant differences in plant height and stem diameter as well as stem number among the study materials. Statistically significant differences were observed for green forage yield (P < 0.05), while not in dry matter yield among the study materials. The average green forage and dry matter yields were reported as 67.0 t ha-1 and 13. 0 t ha-1, respectively. Green forage and dry matter yields in the advanced populations of L-1737 and L-1738 were 69.2 and 67.4 t ha-1; 13.7 and 13.6 t ha-1, respectively. Those yield values were 6.5 and 3.7%; 4.3 and 3.4% higher in green forage and dry matter yields of the advanced populations of L-1737 and L-1738 compared to the control cultivar Bilensoy-80, respectively. These increases in green forage and dry matter yields were due to the effect of the mass selection method. Statistically significant differences were observed for acid detergent fiber, neutral detergent fiber, and relative feeding value among the study materials, except crude protein contents. It was concluded that the two advanced populations had good adaptation ability, high yield and good quality performance under irrigated conditions and could be used as commercial cultivars.

RESUMO: Este estudo foi desenhado para testar duas populações avançadas de alfafa (L-1737 e L-1738) com quatro cultivares controle (Bilensoy-80, Gözlü, Savaş e Plato) para propriedades agromorfológicas, produtividade e qualidade sob condições irrigadas em dois locais diferentes em Ancara, Turquia, nos anos de 2013, 2014 e 2015. Os experimentos foram dispostos em delineamento de blocos casualizados com quatro repetições. Os resultados do estudo mostraram que houveram diferenças estatisticamente significativas na altura das plantas e diâmetro do colmo, bem como no número de colmos entre os materiais avaliados. Foram observadas diferenças estatisticamente significativas para a produtividade de forragem verde (P < 0.05), enquanto não foram observadas diferenças na produção de matéria seca entre os materiais estudados. As produtividades médias de forragem verde e matéria seca foram de 67.0 t ha-1 e 13.0 t ha-1, respectivamente. Os rendimentos de forragem verde e matéria seca nas populações avançadas de L-1737 e L-1738 foram 69.2 e 67.4 t ha-1; 13.7 e 13.6 t ha-1, respectivamente. Esses valores de rendimento foram 6.5 e 3.7%; 4.3 e 3.4% maiores em rendimentos de forragem verde e matéria seca das populações avançadas de L-1737 e L-1738 em relação à cultivar controle Bilensoy-80, respectivamente. Esses aumentos nos rendimentos de forragem verde e matéria seca foram devidos ao efeito do método de seleção massal. Diferenças, estatisticamente significativas, foram observadas para fibra em detergente ácido, fibra em detergente neutro e valor alimentar relativo entre os materiais estudados, exceto nos teores de proteína bruta. Concluiu-se que as duas populações avançadas apresentaram boa capacidade de adaptação, alta produtividade e bom desempenho em condições irrigadas, podendo ser utilizadas como cultivares comerciais.

Monitoring and Identification of Agricultural Crops through Multitemporal Analysis of Optical Images and Machine Learning Algorithms.

The information about where crops are distributed is useful for agri-environmental assessments, but is chiefly important for food security and agricultural policy managers. The quickness with which this information becomes available, especially over large areas, is important for decision makers. Methodologies have been proposed for the study of crops. Most of them require field survey for ground truth data and a single crop map is generated for the whole season at the end of the crop cycle and for the next crop cycle a new field survey is necessary. Here, we present models for recognizing maize (Zea mays L.), beans (Phaseolus vulgaris L.), and alfalfa (Medicago sativa L.) before the crop cycle ends without current-year field survey for ground truth data. The models were trained with an exhaustive field survey at plot level in a previous crop cycle. The field surveys begin since days before the emergence of crops to maturity. The algorithms used for classification were support vector machine (SVM) and bagged tree (BT), and the spectral information captured in the visible, red-edge, near infrared, and shortwave infrared regions bands of Sentinel 2 images was used. The models were validated within the next crop cycle each fifteen days before the mid-season. The overall accuracies range from 71.9% (38 days after the begin of cycle) to 87.5% (81 days after the begin cycle) and a kappa coefficient ranging from 0.53 at the beginning to 0.74 at mid-season.

Generation of a multi-herbicide-tolerant alfalfa by using base editing.

KEYMESSAGE: We present the first report on base editing in alfalfa. Specifically, we showed edited alfalfa with tolerance to both sulfonylurea- and imidazolinone-type herbicides.

Spontaneous Mutation in the Movement Protein of Citrus Leprosis Virus C2, in a Heterologous Virus Infection Context, Increases Cell-to-Cell Transport and Generates Fitness Advantage.

Previous results using a movement defective alfalfa mosaic virus (AMV) vector revealed that citrus leprosis virus C (CiLV-C) movement protein (MP) generates a more efficient local movement, but not more systemic transport, than citrus leprosis virus C2 (CiLV-C2) MP, MPs belonging to two important viruses for the citrus industry. Here, competition experiment assays in transgenic tobacco plants (P12) between transcripts of AMV constructs expressing the cilevirus MPs, followed by several biological passages, showed the prevalence of the AMV construct carrying the CiLV-C2 MP. The analysis of AMV RNA 3 progeny recovered from P12 plant at the second viral passage revealed the presence of a mix of progeny encompassing the CiLV-C2 MP wild type (MPWT) and two variants carrying serines instead phenylalanines at positions 72 (MPS72F) or 259 (MPS259F), respectively. We evaluated the effects of each modified residue in virus replication, and cell-to-cell and long-distance movements. Results indicated that phenylalanine at position 259 favors viral cell-to-cell transport with an improvement in viral fitness, but has no effect on viral replication, whereas mutation at position 72 (MPS72F) has a penalty in the viral fitness. Our findings indicate that the prevalence of a viral population may be correlated with its greater efficiency in cell-to-cell and systemic movements.

Alfalfa Plants (Medicago sativa L.) Expressing the 85B (MAP1609c) Antigen of Mycobacterium avium subsp. paratuberculosis Elicit Long-Lasting Immunity in Mice.

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Paratuberculosis, a contagious, untreatable, and chronic granulomatous enteritis that results in diarrhea, emaciation, and death in farmed ruminants (i.e., cattle, sheep, and goats). In this study, the Ag85B antigen from MAP was expressed in transgenic alfalfa as an attractive vaccine candidate. Agrobacterium-mediated transformation allowed the rescue of 56 putative transformed plants and transgenesis was confirmed in 19 lines by detection of the Ag85B gene (MAP1609c) by PCR. Line number 20 showed the highest Ag85B expression [840 ng Ag85B per gram of dry weight leaf tissue, 0.062% Total Soluble Protein (TSP)]. Antigenicity of the plant-made Ag85B was evidenced by its reactivity with a panel of sera from naturally MAP-infected animals, whereas immunogenicity was assessed in mice immunized by either oral or subcutaneous routes. The plant-made Ag85B antigen elicited humoral responses by the oral route when co-administered with cholera toxin as adjuvant; significant levels of anti-85B antibodies were induced in serum (IgG) and feces (IgA). Long-lasting immunity was evidenced at day 180 days post-first oral immunization. The obtained alfalfa lines expressing Ag85B constitute the first model of a plant-based vaccine targeting MAP. The initial immunogenicity assessment conducted in this study opens the path for a detailed characterization of the properties of this vaccine candidate.

Exopolysaccharide Characterization of Rhizobium favelukesii LPU83 and Its Role in the Symbiosis With Alfalfa.

One of the greatest inputs of available nitrogen into the biosphere occurs through the biological N2-fixation to ammonium as result of the symbiosis between rhizobia and leguminous plants. These interactions allow increased crop yields on nitrogen-poor soils. Exopolysaccharides (EPS) are key components for the establishment of an effective symbiosis between alfalfa and Ensifer meliloti, as bacteria that lack EPS are unable to infect the host plants. Rhizobium favelukesii LPU83 is an acid-tolerant rhizobia strain capable of nodulating alfalfa but inefficient to fix nitrogen. Aiming to identify the molecular determinants that allow R. favelukesii to infect plants, we studied its EPS biosynthesis. LPU83 produces an EPS I identical to the one present in E. meliloti, but the organization of the genes involved in its synthesis is different. The main gene cluster needed for the synthesis of EPS I in E. meliloti, is split into three different sections in R. favelukesii, which probably arose by a recent event of horizontal gene transfer. A R. favelukesii strain devoided of all the genes needed for the synthesis of EPS I is still able to infect and nodulate alfalfa, suggesting that attention should be directed to other molecules involved in the development of the symbiosis.

Microwave Assisted Extraction of Bioactive Carbohydrates from Different Morphological Parts of Alfalfa (Medicago sativa L.).

Despite the nutritional properties of alfalfa, its production is mainly for animal feed and it is undervalued as a food source. In this study, the valorization of alfalfa as a potential source of bioactive carbohydrates [inositols, α-galactooligosaccharides (α-GOS)] is presented. A Box-Behnken experimental design was used to optimize the extraction of these carbohydrates from leaves, stems, and seeds of alfalfa by solid-liquid extraction (SLE) and microwave-assisted extraction (MAE). Optimal extraction temperatures were similar for both treatments (40 °C leaves, 80 °C seeds); however, SLE required longer times (32.5 and 60 min vs. 5 min). In general, under similar extraction conditions, MAE provided higher yields of inositols (up to twice) and α-GOS (up to 7 times); hence, MAE was selected for their extraction from 13 alfalfa samples. Pinitol was the most abundant inositol of leaves and stems (24.2-31.0 mg·g-1 and 15.5-22.5 mg·g-1, respectively) while seed extracts were rich in α-GOS, mainly in stachyose (48.8-84.7 mg·g-1). In addition, inositols and α-GOS concentrations of lyophilized MAE extracts were stable for up to 26 days at 50 °C. These findings demonstrate that alfalfa is a valuable source of bioactive carbohydrates and MAE a promising alternative technique to obtain functional extracts.

Incidencia de distintas frecuencias de corte en parámetros nutritivos de pasturas de alfalfa (Medicago sativa L.) de distinto grado de reposo invernal / Incidence of cutoff frequency in nutritional parameters of alfalfa (Medicago sativa L.) of different degrees of winter rest

Resumen El objetivo de este trabajo fue evaluar la calidad nutricional en 3 cultivares de alfalfa, pertenecientes a diferentes grupos de reposo invernal (GRI), sometidos a dos frecuencias de corte, durante un año del cultivo, fundamentado en que es abundante la información existente en estudios realizados en productividad, cobertura y persistencia, no así en la determinación de calidad de forraje sometidos a diferentes frecuencias de cortes y de GRI contrastantes. Se utilizaron tres cultivares (GR6-Verzy), (GR9-Mecha) y (GR10-Ruano). El diseño experimental fue de parcelas divididas con cuatro repeticiones. Se establecieron 2 tratamientos definidos por los intervalos entre cortes: T1: 25 días y T2: 35 días. Estos fueron precisados para las estaciones de primavera, verano y otoño. En invierno el intervalo fue de 45 y 55 días en T1 y T2, respectivamente. Las variables evaluadas fueron: porcentaje de materia seca (% MS), porcentaje de proteína bruta (% PB), porcentaje de fibra detergente neutro (% FDN) y porcentaje de fibra detergente ácido (% FDA). Se encontraron diferencias significativas para todas las variables nutricionales estudiadas, para ambos tratamientos. Se observó interacción entre cultivares y tratamiento para % PB. Los cultivares estudiados con diferentes GRI, no expresaron diferencias significativas en los parámetros de calidad evaluados para ninguna de las frecuencias de corte realizadas. La frecuencia de corte de 35 días arrojó los menores % PB y mayores % FDN y % FDA, por lo cual el forraje resulta para esta frecuencia de menor calidad, llevando a la disminución de la digestibilidad y aprovechamiento de la pastura para el ganado.

Abstract The objective of this work was to evaluate the nutritional quality in 3 cultivars of alfalfa, belonging to different winter dormancy (GRI), subjected to two cutting frequencies during one year of cultivation, based on the abundant information existing in studies carried out in productivity, coverage and persistence, not so in the determination of quality of forage subjected to different frequencies of cuts and contrasting GRI. Three cultivars (GR6-Verzy), (GR9-Mecha) and (GR10-Ruano) were tested. The experimental design was a complete split plot with four repetitions. Two treatments defined by the cutting intervals were established: T1: 25 days and T2: 35 days. These were specified for spring, summer and fall seasons. For winter the interval was 45 and 55 days in T1 and T2, respectively. The evaluated variables were: percentage of dry matter (% DM), percentage of crude protein (% PB), percentage of neutral detergent fiber (% FDN) and percentage of acid detergent fiber (% FDA). Significant differences were found for all the nutritional variables studied, for both treatment with p <0.05 with InfoStat program. Interaction between cultivars and treatment was observed for % PB. The 35 days cutoff frequency of yielded the lowest percentages of PB and the highest percentages of FDN and FDA. Hence the forage resulting from this frequency is of a lower quality, leading to decreased digestibility and lower performance of pasture for livestock.

Expanding the Benefits of Tnt1 for the Identification of Dominant Mutations in Polyploid Crops: A Single Allelic Mutation in the MsNAC39 Gene Produces Multifoliated Alfalfa.

Most major crops are polyploid species and the production of genetically engineered cultivars normally requires the introgression of transgenic or gene-edited traits into elite germplasm. Thus, a main goal of plant research is the search of systems to identify dominant mutations. In this article, we show that the Tnt1 element can be used to identify dominant mutations in allogamous tetraploid cultivated alfalfa. Specifically, we show that a single allelic mutation in the MsNAC39 gene produces multifoliate leaves (mfl) alfalfa plants, a pivot trait of breeding programs of this forage species. Finally, we discuss the potential application of a combination of preliminary screening of beneficial dominant mutants using Tnt1 mutant libraries and genome editing via the CRISPR/Cas9 system to identify target genes and to rapidly improve both autogamous and allogamous polyploid crops.

Effects of whipped cream addition on fermentation quality and relative feed value of alfalfa silage / Efeitos da adição de creme de leite na qualidade de fermentação e valor alimentar relativo da silagem de alfafa

This study aims to determine the effects of whipped cream addition to alfalfa on fermentation quality and relative feed value (RFV). Alfalfa harvested at the beginning of blooming (5th cutting) was withered under laboratory conditions for 48 hours. In the study, 4 different levels of whipped cream were used, and no additives were added to the control group. A total of 20 packages of silage were left for fermentation in a closed storage (16±2 °C) for 90 days, 4 repetitions for each group. As a results of this study dry matter (DM) contents of silages increased due to the increase in the level of whipped cream addition. pH of silages was decreased significantly in the groups that participated in 20 ve 30 g kg-1 of whipped cream compared to the control (P < 0.01). In these groups, crude protein (CP) amount was higher (P < 0,01), and ammonia nitrogen (NH3-N) was lower (P < 0.01) than control and other whipped cream groups. The water soluble carbohydrate (WSC) contents increased in parallel with the increase in the whipped cream level added to the alfalfa, and lactic acid (LA), as for that, was determined to be the highest (80.40 g kg-1 DM) level in the group with 30 g kg-1 whipped cream. In the same group, neutral detergent fiber (NDF: 383.1 g kg-1 DM) and acid detergent fiber (ADF: 245.4 g kg-1 DM) were observed to be at the lowest level, yeast numbers decreased similarly, enterobacter and mold did not develop, and RFV increased. According to the results obtained from the study, it was demonstrated that by adding 20 or 30 mg kg-1 of whipped cream to alfalfa, quality alfalfa silage could be obtained, and the waste whipped cream can be used as a silage additive and can be recycled to the economy.(AU)

Este estudo teve como objetivo determinar os efeitos da adição de creme de leite à alfafa sobre a qualidade da fermentação e valor relativo da ração (VRR). A alfafa colhida no início da floração (5º corte) foi seca em laboratório por 48 horas. No estudo, 4 níveis diferentes de creme de leite foram usados e nenhum aditivo foi adicionado ao grupo controle. No total, 20 embalagens de silagens foram deixadas para fermentação em local fechado (16 ± 2 ° C) por 90 dias, sendo 4 repetições para cada grupo.Os teores de matéria seca (MS) das silagens aumentaram devido ao aumento do nível de adição de creme de leite. O pH das silagens diminuiu significativamente nos grupos que participaram de 20 a 30 g kg-1 de creme de leite em relação ao controle (P < 0,01).Nestes grupos, o teor de proteína bruta (PB) foi maior (P < 0,01) e o nitrogênio amoniacal (NH3-N) foi menor (P < 0,01) do que o controle e os demais grupos com creme de leite. Os teores de carboidratos solúveis em água (CSA) aumentaram paralelamente ao aumento do teor de creme de leite adicionado à alfafa, e o ácido lático (AL), nesse sentido, foi determinado como o maior nível (80,40 g kg-1 MS) no grupo com 30 g kg-1 de creme.No mesmo grupo, a fibra em detergente neutro (FDN: 383.1 g kg-1 DM) e a fibra em detergente ácido (FDA: 245.4 g kg-1 DM) foram observados no nível mais baixo. O número de leveduras diminuiu de forma semelhante, enterobactéria e fungos não se desenvolveram e a VRR aumentou.De acordo com os resultados obtidos no estudo, foi demonstrado que adicionando 20 ou 30 mg kg-1 de creme de leite à alfafa, pode-se obter silagem de alfafa de qualidade, e o creme de leite residual pode ser utilizado como aditivo de silagem e pode ser reciclado para a economia.(AU)