Sunflower Verticillium wilt is a widespread and destructive disease caused by the soilborne pathogen Verticillium dahliae. To better understand the process of infection and seed transmission of the fungus, sunflower roots were inoculated with a V. dahliae strain (VdBM9-6) labeled with green fluorescent protein (GFP) and monitored microscopically. After 24 to 96 h postinoculation (hpi), conidia germinated and developed into mycelium on root hairs, elongation zones, and caps of lateral roots. Mycelium colonized vascular bundles of lateral roots and taproots at 7 days postinoculation (dpi). At 10 weeks postinoculation (wpi), the epidermal cells, cortical tissues, and vascular elements of stem, petiole, and leaf veins were colonized by mycelium. By 12 wpi, strong GFP signals were detected not only on different tissues of inflorescence but also on testa of seed and a small fraction of pollen grains. A GFP signal was not observed on cotyledon tissues in the seed. Additionally, the colonization of V. dahliae on testa was also confirmed with MNP-10 selection medium, indicating that the testa of seed is the main carrier for the long distance transmission of sunflower yellow wilt.

The Mongolian nationality has developed their unique lifestyle and dietary habit for thousands of years. However, by now, little research has been focused on Mongolian gut microbiota and how it is related to different dietary habits. In this study, denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR) methods were applied to reveal the diversity of predominant gut bacteria of 48 healthy Mongolians recruited from Hohhot city and the Xilin Gol pasturing area in Inner Mongolia. Compared to similar studies of other nationalities, results from the present study have confirmed that the composition of Mongolian gut microbiota is highly similar at the phylum level (Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria) but variable at the genus level. Especially, the numbers of Phascolarctobacterium, Lactobacillus and Bifidobacterium are rather high. DGGE profiles of Lactobacillus and Bifidobacterium revealed that Lactobacillus casei, Bifidobacterium longum and Bifidobacterium animalis subsp. lactis were predominant in the gut of the Mongolian subjects studied. On the contrary, Lactobacillus helveticus was detected in every pasturing area Mongolian, but not in any of the Hohhot city Mongolians. qPCR results revealed that the numbers of Lactobacillus and Bifidobacterium of Xilin Gol Mongolians were significantly higher (P<0.05) than that of Hohhot Mongolians, whereas the numbers of Enterobacterium were significantly lower (P<0.05). In addition, by partial least squares discriminate analysis and cluster analysis of data generated from DGGE and qPCR experiments, a striking difference in the composition of intestinal microbiota of Mongolians living in Hohhot city and the Xilin Gol pasturing area has been found. This study clearly shows that diet affects the microbiota composition of Mongolians living in different circumstances, i.e. urban versus rural.

We investigated the effects of leptin on the in vitro maturation (IVM) and development of calf oocytes. Cumulus-oocyte complexes were matured in IVM medium containing 0-100 ng/ml leptin. Experiment 1 showed that exposure of calf oocytes to IVM medium containing 1 or 10 ng/ml leptin significantly increased rates of development to the metaphase II stage compared with the control (81.7 ± 3.0% and 83.3 ± 2.1% for 1 and 10 ng/ml leptin, respectively, vs 64.1 ± 5.1% for control; p < 0.05). Experiment 2 showed that 1 or 10 ng/ml leptin significantly improved cleavage rates after in vitro fertilization when compared to control (58.6 ± 3.3% and 59.3 ± 2.9% for 1 and 10 ng/ml leptin, respectively, vs 48.5 ± 2.6% for control; p < 0.05); in addition, when compared to control medium, the addition of 10 ng/ml leptin to the IVM medium resulted in more presumptive zygotes reaching the 4- to 8-cell stage after 48 h of in vitro culture (30.3 ± 2.3% vs 20.1 ± 2.3%; p < 0.05) and developing into blastocysts after 8 days of culture (20.4 ± 1.6% vs 11.7 ± 1.7%; p < 0.05). Experiment 3 showed that the addition of 1 or 10 ng/ml leptin significantly increased the total number of blastocyst cells on day 8 of culture (114.6 ± 7.8 and 117.4 ± 5.9 for 1 and 10 ng/ml leptin, respectively, vs 92.7 ± 8.3 for control; p < 0.05) and trophectoderm (TE) cells (88.5 ± 5.5 and 90.6 ± 3.7 for 1 and 10 ng/ml leptin, respectively, vs 70.1 ± 5.9 for control; p < 0.05). In summary, these results indicate that the addition of leptin to IVM medium enhances meiotic maturation and embryo development from calf oocytes and improves the quality of embryos derived from these oocytes.