The influence of vegetal extracts derived from red grape, blueberry fruits and hawthorn leaves on Zea mays L. plant growth and the activity of phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid pathway, was investigated in laboratory experiments. The extracts were characterized using FT-IR and Raman spectroscopies in order to obtain a pattern of the main functional groups. In addition, phenols content was determined by HPLC, whereas the content of indoleacetic acid and isopentenyladenosine hormones was determined by ELISA test and the auxin and gibberellin-like activities by plant-bioassays. The treated maize revealed increased root and leaf biomass, chlorophyll and sugars content with respect to untreated plants. Hawthorn, red grape skin and blueberry at 1.0 mL/L induced high p-coumaric content values, whilst hawthorn also showed high amounts of gallic and p-hydroxybenzoic acids. PAL activity induced by hawthorn at 1.0 mL/L had the highest values (11.1-fold UNT) and was strongly and linearly related with the sum of leaf phenols. Our results suggest that these vegetal extracts contain more than one group of plant-promoting substances.

Humic substances (HS) are powerful natural plant biostimulants. However, there is still a lack of knowledge about the relationship between their structure and bioactivity in plants. We extracted HS (THE1-2) from two forest soils covered with Pinus mugo (1) or Pinus sylvestris (2). The extracts were subjected to weak acid treatment to produce size-fractionated HS (high molecular size, HMS1-2; low molecular size, LMS1-2). HS were characterized for total acidity, functional groups, element and auxin (IAA) contents, and hormone-like activity. HS concentrations ranging from 0 to 5 mg C L-1 were applied to garlic (Allium sativum L.) plantlets in hydroponics to ascertain differences between unfractionated and size-fractionated HS in the capacity to promote mineral nutrition, root growth and cell differentiation, activity of enzymes related to plant development (invertase, peroxidase, and esterase), and N (nitrate reductase, glutamine synthetase) and S (O-acetylserine sulphydrylase) assimilation into amino acids. A positive linear dose-response relationship was determined for all HS in the range 0-1 mg C L-1, while higher HS doses were less effective or ineffective in promoting physiological-biochemical attributes of garlic. Bioactivity was higher for size-fractionated HS according to the trend LMS1-2>HMS1-2>THE1-2, with LMS2 and HMS2 being overall more bioactive than LMS1 and HMS1, respectively. LMS1-2 contained more N, oxygenated functional groups and IAA compared to THE1-2 and HMS1-2. Also, they exhibited higher hormone-like activities. Such chemical properties likely accounted for the greater biostimulant action of LMS1-2. Beside plant growth, nutrition and N metabolism, HS stimulated S assimilation by promoting the enrichment of garlic plantlets with the S amino acid alliin, which has recognized beneficial properties in human health. Concluding, this study endorses that i) treating THE with a weak acid produced sized-fractionated HS with higher bioactivity and differing in properties, perhaps because of novel molecular arrangements of HS components that better interacted with garlic roots; ii) LMS from forest soils covered with P. mugo or P. sylvestris were the most bioactive; iii) the cover vegetation affected HS bioactivity iv); HS stimulated N and S metabolism with relevant benefits to crop nutritional quality.

The main aim of this study is to identify and investigate specific humates (Hs) as potential biostimulants. Five specialty lignosulfonates (LS1-5), one commercial leonardite-humate (PH), and one commercial lignosulfonate (LH), were analyzed for their carbon, nitrogen, and sulfur contents, and the distribution of functional groups using Fourier transform infrared (FTIR) and Raman spectroscopies. Hs were further supplied for two days to Zea mays L. in hydroponics to test their capacity to trigger changes in physiological target-responses. LS1, LS2, LS3, and LS5 determined the most pronounced effects on plant growth and accumulation of proteins and phenolics, perhaps because of their chemical and spectroscopic features. Root growth was more increased (+51–140%) than leaf growth (+5–35%). This effect was ascribed to higher stimulation of N metabolism in roots according to the increased activity of N-assimilation enzymes (GS and GOGAT) and high consumption of sugars for energy-dependent processes. Increased values of RuBisCO, SPAD (Soil Plant Analysis Development values), and leaf sugar accumulation refer to enhanced photosynthesis attributed to Hs. We conclude that Hs tested in this study functioned as biostimulants, but the specialty lignosulfonates were more efficient in this role, possibly because of the type of starting material and process used for their production, which may have influenced their chemical properties.

Significant benefit of soil organic matter (SOM) to crop productivity is scientifically well documented. The main constituents and active fractions of SOM are humic substances (HS) and phenolic compounds. Since both these two components strongly impact plant–soil relationship, it is importantly from an ecological point of view to discriminate their biological effects and relating them to their composition. In this study we compared the biological effects of HS, and the soil water soluble phenols (SWSP) on growth, antioxidant activities, carbohydrates, proteins, phenols, and vitamins of Pinus laricio callus. Each extract was assessed for the content of low molecular weight organic acids, soluble carbohydrates, fatty acids, and phenolic acids. Moreover, Fourier transform infrared (FT-IR) and surface-enhanced Raman scattering (SERS) spectroscopies were applied to study their molecular structure. The results showed that HS produced better callus growth compared to the control and SWSP. Carbohydrates decreased in presence of HS while proteins, vitamin C and E increased. In contrast, in callus treated with SWSP the amount of glucose and fructose increased as well as all the antioxidant activities. The data evidenced that HS rich in tartaric and fatty acids had beneficial effects on callus growth contrary to soil water-soluble phenols rich in aldehydes, and syringic, ferulic, and benzoic acids.