Groundnut (Apios americana Medik) Extract Enhances the Osteoblast Differentiation of MC3T3-E1 Cells

The effects of groundnut ( Apios americana Medik) extract on osteoblast differentiation were examined using MC3T3-E1 cells. MC3T3-E1 cells were treated with the crude extract along with other differentiating reagents. The alkaline phosphatase (ALP) activity of cells cultured in a differentiation medium supplemented with 0.01% crude groundnut extract was 1.5‐1.6 times higher than that of cells cultured in a differentiation medium without the extract. Crude groundnut extract was further separated into aqueous and methanol fractions. The methanol fraction enhanced ALP activity, osteocalcin, integrin-binding sialoprotein, and type I collagen expression, and calcium mineralization. Conversely, the aqueous fraction did not show such effects. Groundnut extract may enhance osteoblast differentiation, and this effect is likely conferred by water insoluble substance(s).

First Report of Post-Harvest Tuber Rot of American Groundnut (Apios americana) Caused by Fusarium acuminatum

Apios americana Medik, commonly known as American groundnut, is a leguminous perennial vine crop native to North America and is cultivated in Japan and Korea (Chu et al. 2019). Its tubers are edible and believed to be very nutritious, especially for women just after childbirth. The tubers also contain secondary metabolites, saponin and genistein, which is good for human health (Ichige et al. 2013). However, the storage of tubers at inappropriate temperatures and humidity levels can cause severe fungal infection, and adversely affect tuber quality. During March and April 2020, a white to pale-orange fungal mycelia were observed on stored American groundnut tubers, with 10 to 15% of seed tubers rotten. Infected tubers were collected, and fungal isolates were isolated on potato dextrose agar (PDA) using the single spore isolation method (Leslie and Summerell 2006). A pure culture (isolate JC20003) was obtained and stored at the Bioenergy Crop Research Institute, NICS, Muan, Republic of Korea. The fungus was cultured on PDA and V8 liquid media for 7 days at 25℃ to observe its morphological characteristics. The length and width of macroconidia ranged from 20.6 to 52.9 μm and 2.9 to 5.1 μm, respectively (n = 30). The microconidia were 8.5 to 14.9 μm and 2.3 to 4.2 μm in length and width, respectively (n = 30). Macroconidia were broadly falcate, strongly septate, 2 to 6 septations with dorsiventral curvature; chlamydospores were formed in chains; and microconidia were fusiform with 0 to 1 septation observed. Genomic DNA of the isolate was extracted using Solgent DNA extraction kit (Solgent, Daejeon, Korea), followed by PCR analysis using the internal transcribed spacer (ITS5/ITS4) and elongation factor (EF-1/EF2) genes (White et al. 1990; O’Donnel 2000). PCR products were sequenced and analyzed to confirm species identity (Yang et al. 2018). These sequences were deposited in GenBank (accession numbers MT703859/ITS and MT731939/EF). BLASTn search analysis showed 100% sequence similarity with Fusarium acuminatum (isolates N-51-1/ITS and WXWH24/EF). Based on morphological and molecular data analysis, the fungus was identified as F. acuminatum (Leslie and Summerell 2006; Marin et al. 2012). Pathogenicity tests were conducted on five tubers inoculated with 5 mm mycelial plugs with three replicates, while a non-mycelial plug served as the control. After 5 days of incubation in plastic containers at 25 °C with high humidity, typical symptoms developed. No symptoms were observed on the control tubers; F. acuminatum was re-isolated from artificially inoculated tubers to complete Koch’s postulates. This is the first report on post-harvest tuber rot caused by F. acuminatum in Apios americana.

Flavonoids from Apios americana Medikus Leaves Protect RAW264.7 Cells against Inflammation via Inhibition of MAPKs, Akt-mTOR Pathways, and Nfr2 Activation

Apios americana Medikus was once widely accepted as staple food in India for a long time, and the tuber of which possesses high nutrients. During the past decades, most of the research has focused on the biological activity in the tubers of Apios americana Medikus whereas the leaves were ignored. In this study, the Apios americana Medikus leaf extract (ALE) was obtained and seven compounds were identified. LPS-induced RAW264.7 cells were used to study the anti-inflammation activity of ALE. As expected, ALE reduced the secretion of nitric oxide (NO) and inflammatory cytokines via inhibition of NF-κB and MAPK signaling together with activation of Nrf2-Keap1 and FOXO pathways, as well as alleviating the oxidative stress and mitochondrial dysfunction. In addition, ALE could activate HMGB1-Beclin1 and Sirt1-FoxO1 pathways and inhibit the Akt-mTOR signaling pathway to activate autophagy, protecting RAW264.7 cells from inflammation. In summary, our results suggested that ALE might help activate the anti-inflammation system, resulting in the prevention of LPS-induced damage in RAW264.7 cells.