Sweetpotato virus disease (SPVD), the most important disease affecting sweetpotato (Ipomoea batatas (L.) Lam), is caused by the synergistic interaction of the aphid-transmitted Sweetpotato feathery mottle virus (SPFMV) and whitefly-transmitted Sweetpotato chlorotic stunt virus (SPCSV). In this study, SPVD was the main disease in the Cañete Valley, the major sweetpotato-producing area in Peru. Studies on virus incidence showed that SPCSV and SPFMV were the most frequently identified viruses in Cañete Valley. Symptoms of different severity were associated with isolates of both viruses involved in the SPVD. Over 80% of plants infected with both SPFMV and SPCSV showed the symptoms (leaf reduction and deformation, vein clearing or mosaic, and stunting) typically attributed to SPVD elsewhere. SPFMV did not significantly affect the yield of the sweetpotato cultivars Jonathan and Costanero, but infection of these cultivars by SPCSV was associated with significant yield reduction. Double infection by the two viruses resulted in SPVD and greater yield reduction than for either alone. These results demonstrate that SPFMV and SPCSV interact synergistically and that the severity of SPVD symptoms also depends on the particular isolate of each virus.

Elevations in the intracellular Ca2+ concentration are a phenomena commonly observed during stem cell differentiation but cease after the process is complete. The transient receptor potential melastatin 4 (TRPM4) is an ion channel that controls Ca2+ signals in excitable and nonexcitable cells. However, its role in stem cells remains unknown. The aim of this study was to characterize TRPM4 in rat dental follicle stem cells (DFSCs) and to determine its impact on Ca2+ signaling and the differentiation process. We identified TRPM4 gene expression in DFSCs, but not TRPM5, a closely related channel with similar function. Perfusion of cells with increasing buffered Ca2+ resulted in a concentration-dependent activation of currents typical for TRPM4, which were also voltage-dependent and had Na+ conductivity. Molecular suppression with shRNA decreased channel activity and cell proliferation during osteogenesis but not adipogenesis. As a result, enhanced mineralization and phosphatase enzyme activity were observed during osteoblast formation, although DFSCs failed to differentiate into adipocytes. Furthermore, the normal agonist-induced first and secondary phases of Ca2+ signals were transformed into a gradual and sustained increase which confirmed the channels' ability to control Ca2+ signaling. Using whole genome microarray analysis, we identified several genes impacted by TRPM4 during DFSC differentiation. These findings suggest an inhibitory role for TRPM4 on osteogenesis while it appears to be required for adipogenesis. The data also provide a potential link between the Ca2+ signaling pattern and gene expression during stem cell differentiation.

The dental follicle (DF) plays an essential role in tooth eruption via regulation of bone resorption and bone formation. Bone morphogenetic protein-6 (BMP6) expression in the DF is coincident with bone growth in the tooth crypt. DF stem cells (DFSCs) have been shown to possess strong osteogenic capability. This study aims to determine the expression of BMP6 in DFSCs and to elucidate the role of BMP6 in the osteogenesis of DFSCs. DFSCs and their non-stem cell counterpart, DF cells (DFCs), were obtained from the DFs of rat pups. We showed that expression of BMP6 was significantly higher in the DFSCs than in the DFCs. DFSCs lost osteogenic capability during in vitro expansion, and DFSCs in late passages had reduced BMP6 expression as compared to early passages of DFSCs when they were subjected to osteogenic induction. Addition of exogenous human recombinant BMP6 (hrBMP6) to the osteogenic medium dramatically enhanced the osteogenesis of the late-passage DFSCs. Knockdown of BMP6 by short interfering RNA in the DFSCs in early passages resulted in a decrease in osteogenesis, which could be restored by addition of hrBMP6. We concluded that DFSCs need to express high levels of BMP6 to maintain their osteogenesis capability. Increased BMP6 expression seen in vivo in the DF may reflect the activation of DFSCs for osteogenic differentiation for bone growth during tooth eruption.