Genome-wide characterization, transcriptome profiling, and functional analysis of the ALMT gene family in Medicago for aluminum resistance.

Aluminum (Al) is the major limiting factor affecting plant productivity in acidic soils. Al3+ ions exhibit increased solubility at a pH below 5, leading to plant root tip toxicity. Alternatively, plants can perceive very low concentrations of Al3+, and Al triggers downstream signaling even at pH 5.7 without causing Al toxicity. The ALUMINUM-ACTIVATED-MALATE-TRANSPORTER (ALMT) family members act as anion channels, with some regulating the secretion of malate from root apices to chelate Al, which is a crucial mechanism for plant Al resistance. To date, the role of the ALMT gene family within the legume Medicago species has not been fully characterized. In this study, we investigated the ALMT gene family in M. sativa and M. truncatula and identified 68 MsALMTs and 18 MtALMTs, respectively. Phylogenetic analysis classified these genes into five clades, and synteny analysis uncovered genuine paralogs and orthologs. The real-time quantitative reverse transcription PCR (qRT-PCR) analysis revealed that MtALMT8, MtALMT9, and MtALMT15 in clade 2-2b are expressed in both roots and root nodules, and MtALMT8 and MtALMT9 are significantly upregulated by Al in root tips. We also observed that MtALMT8 and MtALMT9 can partially restore the Al sensitivity of Atalmt1 in Arabidopsis. Moreover, transcriptome analysis examined the expression patterns of these genes in M. sativa in response to Al at both pH 5.7 and pH 4.6, as well as to protons, and found that Al and protons can independently induce some Al-resistance genes. Overall, our findings indicate that MtALMT8 and MtALMT9 may play a role in Al resistance, and highlight the resemblance between the ALMT genes in Medicago species and those in Arabidopsis.

MsWRKY44 regulates Mg-K homeostasis of shoots and promotes alfalfa sensitivities to acid and Al stresses.

Mg-K homeostasis is essential for plant response to abiotic stress, but its regulation remains largely unknown. MsWRKY44 cloned from alfalfa was highly expressed in leaves and petioles. Overexpression of it inhibited alfalfa growth, and promoted leaf senescence and alfalfa sensitivities to acid and Al stresses. The leaf tips, margins and interveins of old leaves occurred yellow spots in MsWRKY44-OE plants under pH4.5 and pH4.5 +Al conditions. Meanwhile, Mg-K homeostasis was substantially changed with reduction of K accumulation and increases of Mg as well as Al accumulation in shoots of MsWRKY44-OE plants. Further, MsWRKY44 was found to directly bind to the promoters of MsMGT7 and MsCIPK23, and positively activated their expression. Transiently overexpressed MsMGT7 and MsCIPK23 in tobacco leaves increased the Mg and Al accumulations but decreased K accumulation. These results revealed a novel regulatory module MsWRKY44-MsMGT7/MsCIPK23, which affects the transport and accumulation of Mg and K in shoots, and promotes alfalfa sensitivities to acid and Al stresses.

Transcriptomic analysis provides insights into the molecular mechanism of melatonin-mediated cadmium tolerance in Medicago sativa L.

Cadmium (Cd), a toxic element, often makes a serious threat to plant growth and development. Previous studies found that melatonin (Mel) reduced Cd accumulation and reestablished the redox balance to alleviate Cd stress in Medicago sativa L., however, the complex molecular mechanisms are still elusive. Here, comparative transcriptome analysis and biochemical experiments were conducted to explore the molecular mechanisms of Mel in enhancing Cd tolerance. Results showed that 7237 differentially expressed genes (DEGs) were regulated by Mel pretreatment to Cd stress compared to the control condition in roots of Medicago sativa L. Besides, in comparison with Cd stress alone, Mel upregulated 1081 DEGs, and downregulated 1085 DEGs. These DEGs were mainly involved in the transcription and translation of genes and folding, sorting and degradation of proteins, carbohydrate metabolism, and hormone signal network. Application of Mel regulated the expression of several genes encoding ribosomal protein and E3 ubiquitin-protein ligase involved in folding, sorting and degradation of proteins. Moreover, transcriptomic analyse suggested that Mel might regulate the expression of genes encoding pectin lyase, UDP-glucose dehydrogenase, sucrose-phosphate synthase, hexokinase-1, and protein phosphorylation in the sugar metabolism. Therefore, these could promote sucrose accumulation and subsequently alleviate the Cd damage. In conclusion, above findings provided the mining of important genes and molecular basis of Mel in mitigating Cd tolerance and genetic cultivation of Medicago sativa L.

Dehydrin MsDHN1 improves aluminum tolerance of alfalfa (Medicago sativa L.) by affecting oxalate exudation from root tips.

A SK3 -type dehydrin MsDHN1 was cloned from alfalfa (Medicago sativa L.). Its function and gene regulatory pathways were studied via overexpression and suppression of MsDHN1 in alfalfa seedlings or hairy roots. The results showed that MsDHN1 is a typical intrinsically disordered protein that exists in the form of monomers and homodimers in alfalfa. The plant growth rates increased as a result of MsDHN1 overexpression (MsDHN1-OE) and decreased upon MsDHN1 suppression (MsDHN1-RNAi) in seedlings or hairy roots of alfalfa compared with the wild-type or the vector line under Al stress. MsDHN1 interacting with aquaporin (AQP) MsPIP2;1 and MsTIP1;1 positively affected oxalate secretion from root tips and Al accumulation in root tips. MsABF2 was proven to be an upstream transcription factor of MsDHN1 and activated MsDHN1 expression by binding to the ABRE element of the MsDHN1 promoter. The transcriptional regulation of MsABF2 on MsDHN1 was dependent on the abscisic acid signaling pathway. These results indicate that MsDHN1 can increase alfalfa tolerance to Al stress via increasing oxalate secretion from root tips, which may involve in the interaction of MsDHN1 with two AQP.

Effect of Elicitation with Iron Chelate and Sodium Metasilicate on Phenolic Compounds in Legume Sprouts.

Seven-day-old sprouts of fenugreek (Trigonella foenum-graecum L.), lentil (Lens culinaris L.), and alfalfa (Medicagosativa L.) were studied. The legume seeds and then sprouts were soaked each day for 30 min during 6 days with water (control) or mixture of Fe-EDTA and sodium silicate (Optysil), or sodium silicate (Na-Sil) alone. Germination and sprout growing was carried out at temperature 20 ± 2 °C in 16/8 h (day/night) conditions. Phenolic compounds (free, ester, and glycosides) content were determined by HPLC-ESI-MS/MS using a multiple reaction monitoring of selected ions. Flavonoids and phenolic acids were released from their esters after acid hydrolysis and from glycosides by alkaline hydrolysis. The presence and high content of (-)-epicatechin (EC) in fenugreek sprouts was demonstrated for the first time. Applied elicitors decreased the level of free EC in fenugreek and alfalfa sprouts but enhanced the content of its esters. Besides, elicitors decreased the content of quercetin glycosides in lentil and fenugreek sprouts but increased the content of quercetin and apigenin glycosides in alfalfa sprouts. The applied elicitors decreased the glycoside levels of most phenolic acids in lentil and p-hydroxybenzoic acid in fenugreek, while they increased the content of this acid in alfalfa. The mixture of iron chelate and sodium silicate had less effect on changes in flavonoid and phenolic acid content in legume sprouts than silicate alone. In general, the used elicitors increased the content of total phenolic compounds in fenugreek and alfalfa sprouts and decreased the content in lentil sprouts. Among the evaluated elicitors, Optysil seems to be worth recommending due to the presence of iron chelate, which can be used to enrich sprouts with this element.

Chemical Profile and Phytotoxic Action of Hibiscus trionum Essential Oil.

The chemical profile and phytotoxic action of Hibiscus trionum essential oil (EO) was studied. In total 17 compounds were identified via GC/MS, representing 94.18 % of the entire oil, with phytol (40.37 %) being the dominant constituent. Bioassay revealed that the EO inhibited root elongation of Medicago sativa and Amaranthus retroflexus by 32.66 % and 61.86 % at 5 mg/mL, respectively; meanwhile, the major component phytol also exhibited significant phytotoxic activity, suppressing radical elongation of Pennisetum alopecuroides, M. sativa and A. retroflexus by 26.08 %, 27.55 % and 43.96 % at 1 mg/mL, respectively. The fact that the EO showed weaker activity than phytol implied that some constituents might trigger antagonistic action to decrease the oil's activity. Our study is the first on the chemical profile and phytotoxic effect of H. trionum EO.

Long-Term Cd Exposure Alters the Metabolite Profile in Stem Tissue of Medicago sativa.

As a common pollutant, cadmium (Cd) is one of the most toxic heavy metals accumulating in agricultural soils through anthropogenic activities. The uptake of Cd by plants is the main entry route into the human food chain, whilst in plants it elicits oxidative stress by unbalancing the cellular redox status. Medicago sativa was subjected to chronic Cd stress for five months. Targeted and untargeted metabolic analyses were performed. Long-term Cd exposure altered the amino acid composition with levels of asparagine, histidine and proline decreasing in stems but increasing in leaves. This suggests tissue-specific metabolic stress responses, which are often not considered in environmental studies focused on leaves. In stem tissue, profiles of secondary metabolites were clearly separated between control and Cd-exposed plants. Fifty-one secondary metabolites were identified that changed significantly upon Cd exposure, of which the majority are (iso)flavonoid conjugates. Cadmium exposure stimulated the phenylpropanoid pathway that led to the accumulation of secondary metabolites in stems rather than cell wall lignification. Those metabolites are antioxidants mitigating oxidative stress and preventing cellular damage. By an adequate adjustment of its metabolic composition, M. sativa reaches a new steady state, which enables the plant to acclimate under chronic Cd stress.

Interaction of zinc and IAA alleviate aluminum-induced damage on photosystems via promoting proton motive force and reducing proton gradient in alfalfa.

BACKGROUND: In acidic soils, aluminum (Al) competing with Zn results in Zn deficiency in plants. Zn is essential for auxin biosynthesis. Zn-mediated alleviation of Al toxicity has been rarely studied, the mechanism of Zn alleviation on Al-induced photoinhibition in photosystems remains unclear. The objective of this study was to investigate the effects of Zn and IAA on photosystems of Al-stressed alfalfa. Alfalfa seedlings with or without apical buds were exposed to 0 or100 µM AlCl3 combined with 0 or 50 µM ZnCl2, and then foliar spray with water or 6 mg L- 1 IAA. RESULTS: Our results showed that Al stress significantly decreased plant growth rate, net photosynthetic rate (Pn), quantum yields and electron transfer rates of PSI and PSII. Exogenous application of Zn and IAA significantly alleviated the Al-induced negative effects on photosynthetic machinery, and an interaction of Zn and IAA played an important role in the alleviative effects. After removing apical buds of Al-stressed alfalfa seedlings, the values of pmf, gH+ and Y(II) under exogenous spraying IAA were significantly higher, and ΔpHpmf was significantly lower in Zn addition than Al treatment alone, but the changes did not occur under none spraying IAA. The interaction of Zn and IAA directly increased Y(I), Y(II), ETRI and ETRII, and decreased O2- content of Al-stressed seedlings. In addition, the transcriptome analysis showed that fourteen functionally noted genes classified into functional category of energy production and conversion were differentially expressed in leaves of alfalfa seedlings with and without apical buds. CONCLUSION: Our results suggest that the interaction of zinc and IAA alleviate aluminum-induced damage on photosystems via increasing pmf and decreasing ΔpHpmf between lumen and stroma.

Protective roles of salicylic acid in maintaining integrity and functions of photosynthetic photosystems for alfalfa (Medicago sativa L.) tolerance to aluminum toxicity.

Aluminum (Al) can be detrimental to plant growth in areas with Al contamination. The objective of this study was to determine whether salicylic acid (SA) can improve plant tolerance to Al stress by mitigating Al toxicity for chloroplasts and photosynthetic systems in alfalfa (Medicago sativa L.). Plants were treated with Al (100 µM) for 3 d in a hydroponic system. The content of Al increased in leaves treated with Al, resulting in damage and deformation of chloroplasts. In Al-damaged leaves, chloroplast envelopes and starch granules disappeared; the lamellae and stroma lamella were loosely arranged and indistinguishable, and the number of grana was reduced; a large number of small plastoglobules appeared. Foliar spraying of 15 µM SA reduced Al content in roots and leaves and alleviated Al damages in chloroplasts. With 15 µM SA treatments, the chloroplast shape returned to a flat ellipsoid, thylakoids were arranged closely and regularly, chloroplasts had intact starch granules, and small plastoglobules disappeared. SA-treated plants had significantly higher aboveground biomass than the untreated control exposed to Al stress. Photosynthetic index and gene expression analyses demonstrated that SA could alleviate adverse effects of Al toxicity by increasing light capture efficiency, promoting electron transport in the photosynthetic electron transport chain and thylakoid lumen deacidification, and promoting synthesis of aenosine triphosphate (ATP) and nicotinamide adenine dinucleotide phosphate (NADPH). SA played protective roles in maintaining integrity and functions of photosystems in photosynthesis for plant tolerance to Al stress.

Influence of processing and pH on amino acid profile, morphology, electrophoretic pattern, bioactive potential and functional characteristics of alfalfa protein isolates.

Protein isolates were prepared from wet heat processed (APIp) and unprocessed alfalfa seeds (APIc) and characterized for composition and functionality at different pH. APIc and APIp exhibited high content of all the essential amino acids. Antinutrient content of APIp was lower in comparison to APIc and marked reduction in the trypsin inhibitor (85.97%) and lectin activity (100%) was observed. Processing did not cause much reduction of bioactive constituents and antioxidant activity of APIp. Alfalfa protein isolates exhibited complex polypeptide banding ranging from molecular weight of 11-75 kDa. APIp exhibited change in the conformation of protein discerned as alteration in interrelated nuances of ATR-FTIR spectra, XRD-pattern, morphology, charge on proteins and reduced solubility in comparison to APIc due to processing. APIp exhibited marked improvement in the functional properties in comparison to APIc discerned as improved hydration, surface active and gelation properties. Highest hydration and surface active properties were exhibited at pH 9.0, even though APIp at pH 7.0 showed fairly similar functional properties as APIc and APIp at pH 9.0. APIp exhibited reduced least gelation concentration in comparison to APIc at pH 7.0 and also engendered gelation at pH 4.0 and 9.0 contrary to APIc.

Influences of arbuscular mycorrhizae, phosphorus fertiliser and biochar on alfalfa growth, nutrient status and cadmium uptake.

The objective of the study was to explore the influences of arbuscular mycorrhizae (AM), phosphorus (P) fertiliser, biochar application (BC) and their interactions on Medicago sativa growth, nutrient, Cd content and AM fungi-plant symbioses. Applications of both P fertiliser and BC significantly increased total biomass and P and potassium (K) uptake, regardless of AM. When no P fertiliser or BC was used, the shoot biomass and nitrogen (N), P, and K contents in the +AM treatments were 1.39, 1.54, 4.53 and 2.06 times higher than those in the -AM treatments, respectively. AM fungi only elevated the total P uptake by 44.03% when P fertiliser was applied at a rate of 30 mg P kg-1 in the absence of BC addition. With BC application or high-P fertiliser input (100 mg P kg-1), the soil available P was significantly higher than that in the other treatments, and AM fungi significantly reduced the shoot biomass. The minimum Cd concentration occurred in the shoots of alfalfas treated with BC and high-P fertiliser inputs; this concentration was lower than the maximum permitted concentration in China. Although the BC and high-P inputs could eliminate the positive mycorrhizal response, the results suggested that BC application in combination with high-P fertiliser input could not only increase forage yields but also lower Cd concentrations to meet the forage safety standards by the dilution effect.

Physiological, biochemical and transcriptomic responses of Medicago sativa to nickel exposure.

Metal accumulation in soil could lead to severe damage to plants, animals, and humans. The present work aims to evaluate the effects of nickel (Ni) exposure on Medicago sativa at physiological, biochemical, and transcriptomic levels. Plants were exposed to five increasing concentrations of Ni (0, 50, 150, 250, and 500 mg/kg) for 60 days. Agronomic parameters (fresh and dry matter) and chlorophyll content (Chl) were determined in an alfalfa plant. Chemical analyses were conducted, involving the determination of Ni loads in plants (roots and shoots). Moreover, malondialdehyde accumulation (MDA), glutathione-S-transferase (GST), and peroxidase activities, termed as oxidative stress biomarkers, were measured. The gene expression levels of Prx1C, GST, and phytochelatins (PCs) were determined at different nickel concentrations. Our results showed that Ni concentration in plants increased significantly along with Ni concentration in the soil. Regarding oxidative stress biomarkers, Ni contamination caused an increase in peroxidase and GST activities, with a remarkable accumulation of MDA, especially for the highest Ni concentration (500 mg/kg of Ni). Our data showed also a significant upregulation of Prx1C and GST genes in shoots and roots. The PCs' gene expression was significantly enhanced in response to the different nickel concentrations, suggesting their important role in Ni detoxification in alfalfa plants. Our data provided evidence about the clear toxicity of Ni, an often-underestimated trace element.

Terrestrial Toxicity of Synthetic Gas-to-Liquid versus Crude Oil-Derived Drilling Fluids in Soil.

Unlike most other conventional petroleum products that are derived from crude oil, gas-to-liquids (GTLs) are petroleum products that are synthesized from natural gas (methane). This process results in GTL products having no sulfur and low aromatic content, so they should have less impact on human health and the environment compared with crude oil-derived products. The GTLs have been registered for use as nonaqueous base fluids (NABFs) in drilling muds, which aid in the process of drilling wells for oil and gas extraction; it is through these uses and others that they enter terrestrial environments. The primary objective of the present study was to determine whether GTLs were less toxic to terrestrial soil biota than conventional NABFs used for land-based drilling, such as diesel and low-toxicity mineral oil (LTMO). A second objective was to understand the fate and impact of these fluids under more realistic soil and aging conditions of a common west Texas (USA) oil-producing region (i.e., sandy loam soil with low organic matter and a hot arid climate). Acute terrestrial toxicity studies were conducted on the soft-bodied terrestrial invertebrate earthworm (Eisenia fetida) along with 3 plant species-alfalfa (Medicago stavia), thickspike wheatgrass (Elymus lanceolatus), and fourwing saltbrush (Atriplex canescens). We also assessed changes in microbial community structure of the soils following additions of NABF. Overall, the GTL NABFs had lower toxicity compared with conventional NABFs like diesel and LTMO, as measured by invertebrate toxicity, plant seed germination, and impact on the microbial community. Environ Toxicol Chem 2020;39:721-730. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Evaluation of seven chemical pesticides by mixed microbial culture (PCS-1): Degradation ability, microbial community, and Medicago sativa phytotoxicity.

Environmental problems caused by the large-scale use of chemical pesticides are becoming more and more serious, and the removal of chemical pesticides from the ecological environment by microbial degradation has attracted wide attention. In this study, using enrichment screening with seven chemical pesticides as the sole carbon source, a mixed microbial culture (PCS-1) was obtained from the continuous cropping of strawberry fields. The microbial community composition, degradation ability, and detoxification effect of PCS-1 was determined for the seven pesticides. Inoculation with PCS-1 showed significant degradation of and tolerance to the seven pesticides. Microbial community composition analysis indicated that Pseudomonas, Enterobacter, Aspergillus, and Rhodotorula were the dominant genera for the degradation of the seven pesticides by PCS-1. The concentration of the seven pesticides was 10 mg L-1 in hydroponic and soil culture experiments. The fresh weight, plant height, and root length of PCS-1-inoculated alfalfa (Medicago sativa) significantly increased compared with those of non-PCS-1-inoculated M. sativa. PCS-1 not only effectively degraded the residual content of the seven pesticides in water and soil but also reduced the pesticide residues in the roots, stems, and leaves of M. sativa. This study shows that PCS-1 may be important in environmental remediation involving the seven pesticides.

Physiological effects of zero-valent iron nanoparticles in rhizosphere on edible crop, Medicago sativa (Alfalfa), grown in soil.

We investigated the effects of nanoscale zero-valent iron (nZVI) that has been widely used for groundwater remediation on a terrestrial crop, Medicago sativa (Alfalfa), and comprehensively addressed its development and growth in soil culture. Root lengths, chlorophyll, carbohydrate and lignin contents were compared, and no physiological phytotoxicity was observed in the plants. In the roots, using an omics-based analytical, we found evidence of OH radical-induced cell wall loosening from exposure to nZVI, resulting in increased root lengths that were approximately 1.5 times greater than those of the control. Moreover, germination index (GI) was employed to physiologically evaluate the impact of nZVI on germination and root length. In regard to chlorophyll concentration, nZVI-treated alfalfa exhibited a higher value in 20-day-old seedlings, whereas the carbohydrate and lignin contents were slightly decreased in nZVI-treated alfalfa. Additionally, evidence for translocation of nZVI into plant tissues was also found. Vibrating sample magnetometry on shoots revealed the translocation of nZVI from the root to shoot. In this study, using an edible crop as a representative model, the potential impact of reactive engineered nanomaterials that can be exposed to the ecosystem on plant is discussed.

Application of signaling molecules in reducing metal accumulation in alfalfa and alleviating metal-induced phytotoxicity in Pb/Cd-contaminated soil.

The utilization of forages grown on metal-contaminated soil can increase the risk of heavy metals entering the food chain and affecting human health because of elevated toxic metal concentrations. Meanwhile, hydrogen sulfide (H2S) and nitric oxide (NO) as signaling molecules are known to promote plant growth in metal-contaminated soils. However, the regulatory mechanisms of such molecules in plant physiology and soil biochemistry have not been well-documented. Hence, we investigate the role of the exogenous application of H2S and NO on alfalfa growth in lead/cadmium (Pb/Cd)-contaminated soil. Our results indicate that the signaling molecules increase the alfalfa chlorophyll and biomass content and improve alfalfa growth. Further, H2S and NO reduce the translocation and bioconcentration factors of Pb and Cd, potentially reducing the risk of heavy metals entering the food chain. These signaling molecules reduce metal-induced oxidative damage to alfalfa by mitigating reactive oxygen species accumulation and increasing antioxidant enzyme activities. Their exogenous application increases soil enzymatic activities, particularly of catalase and polyphenol oxidase, without significantly changing the composition and structure of rhizosphere bacterial communities. Interestingly, H2S addition enriches the abundance of plant-growth-promoting rhizobacteria in soil, including Nocardioides, Rhizobium, and Glycomyces. H2S is more effective than NO in improving alfalfa growth and reducing heavy-metal contamination of the food chain. These results provide new insights into the exogenous application of signaling molecules in alleviating metal-induced phytotoxicity, including an efficient strategy for the safe use of forages.

Does long-term cadmium exposure influence the composition of pectic polysaccharides in the cell wall of Medicago sativa stems?

BACKGROUND: The heavy metal cadmium (Cd) accumulates in the environment due to anthropogenic influences. It is unessential and harmful to all life forms. The plant cell wall forms a physical barrier against environmental stress and changes in the cell wall structure have been observed upon Cd exposure. In the current study, changes in the cell wall composition and structure of Medicago sativa stems were investigated after long-term exposure to Cd. Liquid chromatography coupled to mass spectrometry (LC-MS) for quantitative protein analysis was complemented with targeted gene expression analysis and combined with analyses of the cell wall composition. RESULTS: Several proteins determining for the cell wall structure changed in abundance. Structural changes mainly appeared in the composition of pectic polysaccharides and data indicate an increased presence of xylogalacturonan in response to Cd. Although a higher abundance and enzymatic activity of pectin methylesterase was detected, the total pectin methylation was not affected. CONCLUSIONS: An increased abundance of xylogalacturonan might hinder Cd binding in the cell wall due to the methylation of its galacturonic acid backbone. Probably, the exclusion of Cd from the cell wall and apoplast limits the entry of the heavy metal into the symplast and is an important factor during tolerance acquisition.

Response and intraspecific differences in nitrogen metabolism of alfalfa (Medicago sativa L.) under cadmium stress.

Pot experiments were carried out to evaluate the response and intraspecific differences in nitrogen metabolism of 20 alfalfa cultivars under cadmium stress. To the aim, exogenous cadmium was added into soil with concentration of 0 (control) and 50 mg kg-1. Results showed that 20 alfalfa were ranked as following according to response index: Guochan (550.93) > Deqin (372.50) > Caoyuan No.1 (350.26) > Queen (345.45) > Xinmu No.2 (344.43) > Longzhong (274.85) > Victoria (233.13) > Emperor (233.13) > Giant (192.29) > Qianjing (101.21) > Xinjiangdaye (75.72) > Algonuin (-32.55) > Duoye (-62.44) > Altay (-102.77) > Sandeli (-155.02) > Turist (-193.24) > Gannong No.1 (-199.22) > Sijiwang (-245.14) > Zhongmu No.1 (-245.48) > WL525HQ (-268.26). Guochan was identified as cadmium tolerant cultivar. Compared with the control group, its plant height increased by 40.96%, shoot and root biomass respectively increased by 18.10% and 70.19%, total nitrogen content in shoot and root respectively increased by 26.69% and decreased by 12.59%, nitrate content decreased by 7.05%, content of ammonium, proline, free amino acid and soluble protein respectively increased by 13.67%, 89.63%, 28.09% and 14.86%, activity of nitrate reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase increased respectively 58.52%, 36.63%, 97.79% and 75.44%. WL525HQ, its above indicators appeared significant differences with those of Guochan, was identified as cadmium sensitive cultivar. In conclusion that the nitrogen metabolism process played an important role for alfalfa to adapt cadmium stress, and the response of nitrogen metabolism to cadmium stress varied with different alfalfa cultivars.

Efecto del consumo del extracto de alfalfa (medicago sativa) en anemia ferropénica inducida, en ratones (mus musculus) / Effect of consumption of alfalfa extract (Medicago sativa) on induced iron deficiency anemia in mice (Mus musculus)

RESUMEN Objetivos Determinar el efecto del consumo del extracto de alfalfa en anemia ferropé-nica inducida, en ratones. Materiales y Métodos Se utilizaron treinta ratones albinos M. musculus de la cepa Balb/c, machos de peso promedio 23±33,7 g. Se formaron tres grupos de diez ratones cada uno: a) grupo control negativo hierro suficiente (HS), recibió 40g/d de alimento balanceado durante siete semanas; b) grupo control positivo hierro deficiente (HD), recibió 40g/d de dieta ferropénica durante siete semanas y; c) grupo experimental hierro deficiente (HD), recibió 40g/d de dieta ferropénica durante siete semanas y a partir de la semana cinco se agregó 20g/d de extracto de alfalfa (EA). Resultados Al finalizar el tratamiento se observó diferencia significativa en los niveles de hemoglobina entre los grupos control positivo (8.41±3.9 g/dL) y experimental (13.4±3.3 g/dL) (t student, p<0,05). No se encontró diferencia significativa en los niveles de hemoglobina, al término del periodo de inducción entre los grupos control positivo (8.76±3.9 g/dL) y experimental (8.59± 3.1 g/dL) (t student, p>0,05). Conclusiones En condiciones experimentales, la alfalfa presenta efecto antianémico, sustentado en los resultados de los niveles de hemoglobina.(AU)

ABSTRACT Objective To determine the effect of alfalfa extract consumption on induced iron deficiency anemia in mice. Materials and Methods Thirty M. musculus albino mice of the BALB/c strain were used for this study. All of them were males, with a mean weight of 23±33.7 g. Three groups of 10 mice each were formed: a) negative control group with sufficient iron (HS), which received 40g/d of balanced feed for seven weeks; b) positive control group with iron deficiency (HD), which received 40g/d of a diet plan for anemia for seven weeks; and c) experimental group with iron deficiency (HD), which received 40g/d of a diet plan for anemia for seven weeks and 20g/d of alfalfa extract (EA) from week five. Hemoglobin levels were measured. Results At the end of the treatment, a significant difference was observed in hemoglobin levels between the positive (8.41±3.9 g/dL) and experimental (13.4±3.3 g/dL) control groups (T student, p<0.05). There was no significant difference in hemoglobin levels at the end of the induction period between the positive (8.76±3.9 g /dL) and experimental (8.59±3.1 g/dL) groups (T student, p>0.05). Conclusions Under experimental conditions, alfalfa has an antianemic effect based on the results of hemoglobin levels.(AU)

Changes in the Proteome of Medicago sativa Leaves in Response to Long-Term Cadmium Exposure Using a Cell-Wall Targeted Approach.

Accumulation of cadmium (Cd) shows a serious problem for the environment and poses a threat to plants. Plants employing various cellular and molecular mechanisms to limit Cd toxicity and alterations of the cell wall structure were observed upon Cd exposure. This study focuses on changes in the cell wall protein-enriched subproteome of alfalfa (Medicago sativa) leaves during long-term Cd exposure. Plants grew on Cd-contaminated soil (10 mg/kg dry weight (DW)) for an entire season. A targeted approach was used to sequentially extract cell wall protein-enriched fractions from the leaves and quantitative analyses were conducted with two-dimensional difference gel electrophoresis (2D DIGE) followed by protein identification with matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time of flight (TOF/TOF) mass spectrometry. In 212 spots that showed a significant change in intensity upon Cd exposure a single protein was identified. Of these, 163 proteins are predicted to be secreted and involved in various physiological processes. Proteins of other subcellular localization were mainly chloroplastic and decreased in response to Cd, which confirms the Cd-induced disturbance of the photosynthesis. The observed changes indicate an active defence response against a Cd-induced oxidative burst and a restructuring of the cell wall, which is, however, different to what is observed in M. sativa stems and will be discussed.