Gene expression analysis using reverse transcription quantitative real-time PCR (RT-qPCR) requires the use of reference gene(s) in the target species. The long yellow daylily, Hemerocallis citrina Baroni. is rich in beneficial secondary metabolites and is considered as a functional vegetable. It is widely cultivated and consumed in East Asian countries. However, reference genes for use in RT-qPCR in H. citrina are not available. In the present study, six potential reference genes, actin (ACT), AP-4 complex subunit (AP4), tubulin (TUB), ubiquitin (UBQ), 18S and 60S ribosomal RNA, were selected and their expression stability in different developmental stages, organs and accessions was evaluated using four statistical software packages (geNorm, NormFinder, BestKeeper, and RefFinder). For commercial flower buds of different landraces, the combination of 60S, TUB, and AP4 was appropriate whereas ACT and 60S was suitable for normalization of different organs. In addition, AP4 exhibited the most stable expression in flower buds among different developmental stages. UBQ was less stable than the other reference genes under the experimental conditions except under different organs was 18S. The relative expression levels of two genes, primary-amine oxidase (HcAOC3) and tyrosine aminotransferase (HcTAT) which play important roles in alkaloid biosynthesis were also examined in different organs of the 'Datong' landrace, which further confirmed the results of selected reference genes. This is the first report to evaluate the stability of reference genes in the long yellow daylily that can serve as a foundation for RT-qPCR analysis of gene expression in this species.
The N-myc downstream-regulated gene 2 (NDRG2) is involved in tumor cell differentiation and apoptosis, but its function in tumor angiogenesis remains to be established. Here, we employed adenovirus overexpressing NDRG2 (Ad-NDRG2) to efficiently up-regulate target gene expression in the NDRG2-low-expressing, breast cancer cell line MCF-7. Moreover, VEGF secretion was decreased in MCF-7 cells infected by Ad-NDRG2, and medium conditioned by these infected cells could significantly inhibit the proliferation, tube formation and invasion of human umbilical vein endothelial cells (HUVECs). Further study indicated that the angiogenesis promoting factors VEGF and HIF-1α were down-regulated, whereas the angiogenesis suppressing factors p53 and VHL were up-regulated in MCF-7 cells infected by Ad-NDRG2. Finally, in a nude mouse model, intratumoral injections of Ad-NDRG2 every 3 days for 20 days significantly inhibited the growth and angiogenesis of xenografted MCF-7 tumors. In summary, these data indicate that NDRG2 may be involved in angiogenesis by impacting the expression of angiogenesis related factors. Thus, specific overexpression of NDRG2 by adenovirus represents a promising approach for the treatment of tumor angiogenesis.
Abstract Blue carbon in mangrove ecosystems contributes significantly to the global carbon cycle. However, large uncertainties maintain in the soil organic carbon (SOC) storage throughout the tide-induced salinity and alkalinity transect in the mangrove restoration region in Southern China. Total 125 soil samples were obtained to detect the SOC content and physicochemical properties. The mean SOC content of each layer ranged from 6.82 to 7.86 g kg −1 , while the SOC density ranged from 2.99 to 11.41 kg m −2 , increasing with soil depths. From different land covers in the study region, the SOC content varied from 4.63 to 9.71 g kg −1 , increasing across the salinity and alkalinity transect, while the SOC density fluctuated from 3.01 kg m −2 in mudflats to 10.05 kg m −2 in mangrove forests. SOC concentration was favorably linked with total nitrogen (r = 0.95), and total phosphorus (r = 0.74), and negatively correlated with Cl − (r = − 0.95), electrical conductivity (r = − 0.24), and total dissolved solids (r = − 0.08). There were significant logarithmic relationships between SOC content and the concentrations of clay (r = 0.76), fine silt (r = 0.81), medium silt (r = − 0.82), and coarse silt (r = − 0.78). The spatial patterns of SOC concentration were notably affected by soil texture, physicochemical properties, and land-cover type, providing essential reference for future investigations of blue carbon budget in restored mangrove forests.
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