Bioactive metabolite profile and antioxidant properties of brown juice, a processed alfalfa (Medicago sativa) by-product.

Recently, leaf protein concentrate (LPC) has gained increased attention in response to the constantly growing protein demand. Green biorefineries can become more economical by valorizing their by-products and reducing environmental risks. The current study describes the variations in the antioxidant capacity and phytochemical composition of a liquid by-product (referred to as brown juice (BJ)) obtained during the extraction of leaf protein concentrate (LPC) from the fresh biomass of alfalfa (Medicago sativa L.). Four varieties of alfalfa were investigated during three harvest times, i.e., August 2017 (first harvest), September 2017 (second harvest), and June 2018 (third harvest). Also, the fresh BJ was lacto-fermented to extend its preservation period but also modifying its composition. The results of different general phytochemical composition analyses and antioxidant assays revealed similar tendencies across different alfalfa varieties and harvest times. Most of the phytochemicals in the BJ identified by HPLC-MS/MS can be classified as flavonoids/flavonoid derivatives, e.g., apigenin, naringenin, luteolin, formononetin. Substantially, the lacto-fermentation process induced a switch into aglycones, e.g., apigenin content increased by an order of magnitude, while apigenin-7-O-glucuronide content was halved after lacto-fermentation. Additionally, several B vitamins were detected, including B2, B3, and B7. These results could provide a basis for various ways of industrial valorization but need to be strengthened by data generated from large-scale production.

Comparison of Wet Fractionation Methods for Processing Broccoli Agricultural Wastes and Evaluation of the Nutri-Chemical Values of Obtained Products.

The main objective of this study was to increase the economic value of broccoli green agro-waste using three wet fractionation methods in the shadow of green biorefinery and the circular economy. Product candidates were obtained directly by using a mechanical press, and indirectly by using microwave coagulation or via lactic acid fermentation of green juice. The leaf protein concentrates (LPC) fractions displayed significantly higher dry matter content and crude protein content (34-39 m/m% on average) than the green juice fraction (27.4 m/m% on average), without considerable changes in the amino acids composition ratio. UHPLC-ESI-ORBITRAP-MS/MS analysis showed that kaemferol and quercetin are the most abundant flavonols, forming complexes with glycosides and hydroxycinnamic acids in green juice. Lacto-ermentation induced a considerable increase in the quantity of quercetin (48.75 µg·g-1 dry weight) and kaempferol aglycons (895.26 µg·g-1 dry weight) of LPC. In contrast, chlorogenic acid isomers and sulforaphane disappeared from LPC after lactic acid fermentation, while microwave treatment did not cause significant differences. These results confirm that both microwave treatment and lacto-fermentation coagulate and concentrate most of the soluble proteins. Also, these two processes affect the amount of valuable phytochemicals differently, so it should be considered when setting the goals.

Uptake Dynamics of Ionic and Elemental Selenium Forms and Their Metabolism in Multiple-Harvested Alfalfa (Medicago sativa L.).

A pot experiment, under greenhouse conditions, was carried out aiming at investigating the agronomic biofortification of alfalfa (Medicago sativa L.) with Se and monitoring the Se uptake and accumulation dynamics within four consecutive harvests within the same growing season. Two ionic Se forms, i.e., sodium selenate (Se (VI)) and sodium selenite (Se (IV)), were applied once at a rate of 1, 10, and 50 mg kg-1 (added on Se basis), while 10 and 50 mg L-1 of a red elemental Se (red Se0) were used; all Se treatments were added as soil application. Application of Se (VI) at the rate of 50 mg kg-1 was toxic to alfalfa plants. The effect of Se forms on Se accumulation in alfalfa tissues, regardless of the applied Se concentration, follows: Se (VI) > Se (IV) > red Se0. The leaf, in general, possessed higher total Se content than the stem in all the treatments. The accumulation of Se in stem and leaf tissues showed a gradual decline between the harvests, especially for plants treated with either Se (VI) or Se (IV); however, the chemically synthesized red Se0 showed different results. The treatment of 10 mg kg-1 Se (VI) resulted in the highest total Se content in stem (202.5 and 98.0 µg g-1) and leaf (643.4 and 284.5 µg g-1) in the 1st and 2nd harvests, respectively. Similar tendency is reported for the Se (IV)-treated plants. Otherwise, the application of red Se0 resulted in a lower Se uptake; however, less fluctuation in total Se content between the four harvests was noticed compared to the ionic Se forms. The Se forms in stem and leaf of alfalfa extracted by water and subsequently by protease XIV enzyme were measured by strong anion exchange (SAX) HPLC-ICP-MS. The major Se forms in our samples were selenomethionine (SeMet) and Se (VI), while neither selenocysteine (SeCys) nor Se (IV) was detected. In water extract, however, Se (VI) was the major Se form, while SeMet was the predominant form in the enzyme extract. Yet, Se (VI) and SeMet contents declined within the harvests, except in stem of plants treated with 50 mg L-1 red Se0. The highest stem or leaf SeMet yield %, in all harvests, corresponded to the treatment of 50 mg L-1 red Se0. For instance, 63.6% (in stem) and 38.0% (in leaf) were calculated for SeMet yield % in the 4th harvest of plants treated with 50 mg L-1 red Se0. Our results provide information about uptake and accumulation dynamics of different ionic Se forms in case of multiple-harvested alfalfa, which, besides being a good model plant, is an important target plant species in green biorefining.

Raw and Fermented Alfalfa Brown Juice Induces Changes in the Germination and Development of French Marigold (Tagetes patula L.) Plants.

Organic and ecological farming programs require new and efficient biostimulants with beneficial properties for the sustainable and safe production of seedlings and ornamental plants. We examined the effect of non-fermented and lacto-fermented alfalfa brown juice (BJ) on seed germination and the vegetative, physiological, and anatomical properties of French marigold (Tagetes patula L. 'Csemo') plants which were treated with 0.5-10% fermented and non-fermented BJ, with tap water applied as a control. Applying 0.5% fermented BJ significantly improved seed germination compared with non-fermented BJ, resulting in an increase of 9.6, 11.2, 10.9, and 41.7% in the final germination percent, germination rate index, germination index, and vigor index, respectively. In addition, it increased the root and shoot length by 7.9 and 16.1%, respectively, root and shoot dry mass by 20 and 47.6%, respectively, and the number of leaves by 28.8% compared to the control. Furthermore, an increase in contents of water-soluble phenol, chlorophyll a and b, and carotenoid was reported upon the application of 0.5% fermented BJ, while peroxidase activity decreased. Our results prove that alfalfa BJ can be enrolled as a biostimulant as part of the circular farming approach which supports the sustainable horticultural practice.

Industrial side-products as possible soil-amendments.

The protection of our environment is a common task. All pollution that expose our soils, plants or in the narrower and wider sense environment will appear sooner or later in the food chain and in human beings who are at the top of the food-chain pyramid. The aim of our work is to give a brief overview about the effects of some industrial wastes on the physiological parameters of plants. Compost, black soot, sewage sludge and lime sludge dust was examined. Sunflower seeds were used in the experiments. The filtrates of examined materials were added to the nutrient solution in different quantities because of different solubility. The contents of sample elements and uptake of the element were measured by ICP, the relative chlorophyll contents by SPAD 502. Disadvantageous and advantageous physiological effects of compost, black soot, sewage sludge and lime sludge were proved. Larger concentrations of aluminium were measured in the roots than in the shoots. The concentrations of chrome were below the control value in the shoots when black soot, compost, lime sludge and sewage sludge were added to the nutrient solution. The concentrations of zinc, phosphorous, magnesium and copper were very low when black soot was used, and it was lower than the control. The dry matter of shoots increased when compost and sewage sludge was used, but the growth of roots remained under the control level.