Brown juice is a byproduct of fractionated green biomass during leaf protein isolation. It represents approximately 45%–50% of the total pressed fresh biomass. Disposal of brown juice is a serious issue in leaf protein production due to its high biological oxygen demand and carbohydrates content. The current study aimed to find a possible potential use of brown juice. Therefore, chemical and biochemical properties of brown juice—derived from alfalfa green biomass—were determined before and after fermentation by lactic acid bacteria. Additionally, the growth stimulation potential of fermented brown juice on plumed cockscomb (Celosia argantea var. plumose ‘Arrabona’) plants were tested. Celosia seedlings were sprayed at different rates of fermented brown juice (i.e., 0.5%, 1%, 2.5%, 5%, and 10%) and tap water was applied as control. The results revealed that lactic acid bacteria successfully enhanced the stabilization of brown juice via reducing sugars content and increasing organic acids content. After fermentation, contents of glucose monomers were 15 times lower; while concentrations of lactic and acetic acids increased by 7- and 10-fold, respectively. This caused a reduction in the pH of fermented brown juice by 13.9%. Treating Celosia plants at lower rates of fermented brown juice (up to 1.0%) significantly induced their growth dynamics and antioxidant capacity. Higher values of vegetative parameters were measured in treated plants compared to control. The brown juice treatments caused significant changes in histological parameters as well. The activity of catalase and peroxidase increased in plants that received fermented brown juice especially at low rates. Moreover, an increase in water-soluble protein and phenol was measured in different tissues of plants sprayed with fermented brown juice. Malondialdehyde content was lowered in treated plants compared to control. Fermented brown juice at high rates slightly reduced the amount of photosynthetic pigments; however, this reduction was not reported for low rates of fermented brown juice. These results surely illustrate the potential use of fermented alfalfa brown juice as a growth stimulator for crops particularly at rates below 2.5%.

Fertilization management is a key issue in plant nutrition to produce plants with good quality and quantity. Deproteinized leaf juice or brown juice (BJ) is a by-product during the isolation of leaf protein from biomass crops such as alfalfa. The idea of using BJ as a biostimulant fits well in the aspect of circular economy since BJ is currently a problematic issue of the leaf protein production approach. Fractionation of one-kilogram fresh biomass results in approximately 500 cm³ BJ. Due to fast spoil of fresh BJ, if left on room temperature, it is found that fermentation of fresh BJ using lactic acid bacteria and reducing its pH increases its stability and storage on room temperature. In the present study, we examined the effect of fermented alfalfa BJ on vegetative, physiological, and anatomical properties of the versatile sweet basil (Ocimum basilicum L. ‘Bíborfelhő’) plants. Sweet basil seedlings were sprayed at different doses of fermented alfalfa BJ (i.e., 0.5%, 1.0%, and 2.5%) and tap water served as a control (0.0% BJ). The results revealed that foliar application of fermented alfalfa BJ significantly improved the biometrical features of sweet basil plants. Plants treated with fermented BJ showed significantly higher values of all the measured parameters compared to the control (0.0%), except for the number of leaves per plants where control plants (0.0%) had more leaves. However, the leaves of control plants (0.0%) were smaller than treated plants as data of leaf area showed. Fermented alfalfa BJ significantly increased the content of photosynthetic pigments (chl a and chl b), relative chlorophyll (SPAD value), lengths of stem and root, fresh masses of stem, root, and leaves, volumes of stem and root, and leaf area. Despite all rates of fermented BJ displayed higher values over control plants (0.0%), the rate of 0.5% was the best one supported by results. Application of fermented alfalfa BJ influenced the anatomical parameters as well. These findings demonstrate the possible use of fermented alfalfa BJ as a promising novel plant biostimulant.

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.

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. ’Csemő’) 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.