Genetic Diversity and Differentiation of Eleven Medicago Species from Campania Region Revealed by Nuclear and Chloroplast Microsatellites Markers

The species belonging to the genus Medicago are considered a very important genetic resource at global level both for planet’s food security and for sustainable rangelands management. The checklist of the Italian flora (2021) includes a total number of 40 Medicago species for Italy, and 27 for Campania region, with a number of doubtful records or related to species no more found in the wild. In this study, 10 Medicago species native to Campania region, and one archaeophyte (M. sativa), identified by means of morphological diagnostic characters, were analyzed in a blind test to assay the efficacy of nine microsatellite markers (five cp-SSRs and four n-SSRs). A total number of 33 individuals from 6 locations were sampled and genotyped. All markers were polymorphic, 40 alleles were obtained with n-SSRs ranging from 8–12 alleles per locus with an average of 10 alleles per marker, PIC values ranged from 0.672 to 0.847, and the most polymorphic SSR was MTIC 564. The cp-SSRs markers were highly polymorphic too; PIC values ranged from 0.644 to 0.891 with an average of 0.776, the most polymorphic cp-SSR was CCMP10. 56 alleles were obtained with cp-SSRs ranging from 7 to 17 alleles per locus with an average of 11. AMOVA analysis with n-SSR markers highlighted a great level of genetic differentiation among the 11 species, with a statistically significant fixation index (FST). UPGMA clustering and Bayesian-based population structure analysis assigned these 11 species to two main clusters, but the distribution of species within clusters was not the same for the two analyses. In conclusion, our results demonstrated that the combination of the used SSRs well distinguished the 11 Medicago species. Moreover, our results demonstrated that the use of a limited number of SSRs might be considered for further genetic studies on other Medicago species.

Impact of Sprouting under Potassium Nitrate Priming on Nitrogen Assimilation and Bioactivity of Three Medicago Species

Edible sprouts are rich in flavonoids and other polyphenols, as well as proteins, minerals, and vitamins. Increasing sprout consumption necessitates improving their quality, palatability, and bioactivity. The purpose of this study was to test how KNO3 priming affects the sprouting process species on three Medicago species (Medicago indicus, Medicago interexta, and Medicago polymorpha) and their nutritional values. Targeted species of Medicago were primed with KNO3, and the levels of different primary and secondary metabolites were determined. KNO3 induced biomass accumulation in the sprouts of the three species, accompanied by an increased content of total mineral nutrients, pigments, vitamins, and essential amino acids. Besides, our results showed that KNO3 enhanced the activity of nitrate reductase (NR), glutamate dehydrogenase (GDH), and glutamine synthetase (GS) enzymes, which are involved in the nitrogen metabolism and GOGAT cycle, which, in turn, increase the nitrogen and protein production. KNO3 treatment improved the bioactive compound activities of Medicago sprouts by increasing total phenolic and flavonoid contents and enhancing the antioxidant and antidiabetic activities. Furthermore, species-specific responses toward KNO3 priming were noticeable, where Medicago interexta showed the highest antioxidant and antidiabetic activities, followed by Medicago polymorpha. Overall, this study sheds the light on the physiological and biochemical bases of growth, metabolism, and tissue quality improvement impact of KNO3 on Medicago sprouts.

Biological Activities and Chemistry of Triterpene Saponins from Medicago Species: An Update Review

Plants are known to be a great source of phytochemicals for centuries. Medicago, belonging to the Family Fabaceae, is a large and well spread genus comprising about 83 cosmopolitan species, of which one-third are annuals and span diverse ecological niches. Medicago species are rich in saponins mainly classified into three classes, namely, steroid alkaloid glycosides, triterpene glycosides, and steroid glycosides. These saponins are important compounds having diverse pharmacological and biological activities. As a whole, 95 of saponins are reported to date occurring in Medicago species using various latest extraction/isolation techniques. Considering the multiple biological and pharmacological potential of Medicago species due to saponins along with structural diversity, we compiled this review article to sum up the recent reports for the pharmacological potential of the Medicago’s derived saponins in modern as well as traditional medication systems. The current manuscript produces data of chemical structures and molecular masses of all Medicago species saponins simultaneously. The toxicity of certain pure saponins (aglycones) has been reported in vitro; hederagenin appeared highly toxic in comparison to medicagenic acid and bayogenin against X. index, while soyasaponin I, containing soyasapogenol B as a glycone, appeared as the least toxic saponin. The diversity in the structural forms shows a close relationship for its biological and pharmacological actions. Moreover, saponins showed antioxidant properties and the mechanism behind antimicrobial potential also elaborated in this review article is mainly because of the side sugar groups on these compounds. The collected data presented herein include chemical structures and molecular masses of all saponins so far. Their biological activity and therapeutic potential are also discussed. This information can be the starting point for future research on this important genus.

Phoma medicaginis Isolate Differences Determine Disease Severity and Phytoestrogen Production in Annual Medicago spp.

Phoma black stem and leaf spot disease of annual Medicago spp., caused by Phoma medicaginis, not only can devastate forage and seed yield, but also reduces herbage quality by inducing production of phytoestrogens, particularly coumestrol and 4'-O-methylcoumestrol, that can reduce ovulation rates of animals grazing infected forage. We determined the consequent phytoestrogen levels on three different annual Medicago species/cultivars (M. truncatula cv. Cyprus, M. polymorpha var. brevispina cv. Serena and M. murex cv. Zodiac), following inoculation with 35 isolates of P. medicaginis. Across the isolate x cultivar combinations, leaf disease incidence (%LDI), petiole disease incidence (%PDI), leaf disease severity (%LDS), petiole disease severity (%PDS), and leaf yellowing severity (%LYS) ranged up to 100, 89.4, 100, 58.1 and 61.2%, respectively. Cultivars Cyprus and Serena were most susceptible and cv. Zodiac the most resistant to P. medicaginis. Isolates WAC3653, WAC3658 and WAC4252 produced the most severe disease. Levels of phytoestrogens in stems ranged from 25 to 1995 mg/kg for coumestrol and from 0 to 418 mg/kg for 4'-O-methylcoumestrol. There was a significant positive relationship of disease incidence and severity parameters with both coumestrol and 4'-O-methylcoumestrol contents, as noted across individual cultivars, and as across the three cultivars overall where r=0.39 and 0.37 for coumestrol and 4'-O-methylcoumestrol, respectively (P<0.05). Although, cv. Serena was most susceptible to P. medicaginis and produced the highest levels of phytoestrogens in the presence of P. medicaginis, cv. Zodiac contained the highest levels of phytoestrogens in comparison with other cultivars in the absence of P. medicaginis. There was a 15-fold increase in coumestrol in cv. Serena but only a 7-fold increase in cv. Zodiac from infection of P. medicaginis. The study highlights that the intrinsic ability of a particular cultivar to produce phytoestrogens in the absence of the pathogen, and its comparative ability to produce phytoestrogens in the presence of the P. medicaginis, are both important and highly relevant to developing new annual Medicago spp. cultivars that offer improved disease resistance and better animal reproductive outcomes.

Short-Time Impact of Soil Amendments with Medicago Plant Materials on Soil Nematofauna

Soil amendments with plant materials from Medicago species are widely acknowledged for a suppressive effect on plant-parasitic nematodes but their impact on beneficial components of soil nematofauna is still unknown. A study on potted tomato was carried out to investigate the short-time effects on the overall nematofauna of dry biomasses from six different Medicago species, i.e., M. sativa, M. heyniana, M. hybrida, M. lupulina, M. murex and M. truncatula, incorporated to natural soil at 10, 20, or 40 g kg−1 soil rates. All amendments resulted in a significant decrease of the total nematofauna biomass, whereas total abundance was significantly reduced only by M. heyniana, M. hybrida, and M. lupulina biomasses. Almost all the Medicago amendments significantly reduced the relative abundance of plant-parasites and root fungal feeders. All amendments significantly increased the abundance of bacterivores, whereas fungivores significantly increased only in soil amended with M. heyniana, M. lupulina and M. sativa plant materials. Mesorhabditis and Rhabditis were the most abundant genera of bacterivores, whereas Aphelenchoides and Aphelenchus prevailed among the fungivores. Predators were poorly influenced by all the tested Medicago biomasses, whereas the abundance of omnivores was negatively affected by M. heyniana and M. lupulina. Values of the Maturity Index and Sum Maturity Index were reduced by treatments with M. heyniana, M. hybrida, M. lupulina and M. sativa plant materials, whereas most of the tested amendments decreased values of the Channel Index while increasing those of the Enrichment Index. Enrichment and bacterivore footprints raised following soil addition with Medicago biomasses, whereas composite and fungivore footprints were significantly reduced. According to their overall positive effects on soil nematofauna, amendments with Medicago plant materials or their formulated derivatives could represent an additional tool for a sustainable management of plant-parasitic nematodes.

Activity of Saponins from Medicago Species against Phytoparasitic Nematodes

Content of bioactive saponins of Medicago species suggests that they may also exert, as previously demonstrated on M. sativa, nematicidal properties exploitable for the formulation of new products for sustainable phytoparasitic nematode management. This study was addressed to highlight the bioactivity of saponins from five different Medicago species still poorly known for their biological efficacy, i.e., M. heyniana, M. hybrida, M. lupulina, M. murex and M. truncatula, against the plant parasitic nematodes Meloidogyne incognita, Xiphinema index and Globodera rostochiensis. The bioactivity of the extracts from the five Medicago species was assessed by in vitro assays on the juveniles (J2) and eggs of M. incognita and G. rostochiensis and the adult females of X. index. The suppressiveness to M. incognita of soil treatments with the Medicago plant biomasses was also investigated in a tomato experiment. The nematicidal activity of the five Medicago species was reported and discussed in relation to their phytochemical profile.