Screening and identification of cucumber germplasm and rootstock resistance against the root-knot nematode (Meloidogyne incognita).

Root-knot nematodes (Meloidogyne spp) are destructive agricultural pests that reduce the productivity of cultivated vegetables worldwide, especially when vegetables are cropped continuously in greenhouses. Cucumbers (Cucumis sativus L.), in particular, suffer extensive damage due to root-knot nematodes, and only a few wild species are known to be resistant. Grafting of cultivated plants to rootstocks of known resistant germplasms could be an effective method to resolve this problem. In this study, 21 cucumber germplasms and seven rootstocks were evaluated for resistance based on the growth of cucumber seedlings and resistance indexes to Meloidogyne incognita, which were surveyed 25 days after inoculation with M. incognita. Cluster analysis and principal component analysis (PCA) were used to investigate the resistance of 21 cucumber germplasms and seven rootstocks based on their growth and resistance indexes after inoculation with M. incognita. These analyses showed that the 21 germplasms and seven rootstocks could be divided into three groups based upon their resistance levels: moderately resistant, susceptible, and highly susceptible to M. incognita. All 21 cucumber germplasms exhibited susceptibility or high susceptibility to M. incognita and most rootstocks exhibited moderate resistance. The PCA results were consistent with those of the clustering analysis. The Jinyou No.1 cultivar had the highest resistance to M. incognita among the 21 cucumber germplasms, and Huangzhen No.1 cultivar had the highest resistance among the seven rootstock cultivars.

Heterozygosities and genetic relationship of tea cultivars revealed by simple sequence repeat markers and implications for breeding and genetic mapping programs.

Genetic maps are essential tools for quantitative trait locus analysis and marker-assisted selection breeding. In order to select parents that are highly heterozygous for genetic mapping, the heterozygosity (HS) of 24 tea cultivars (Camellia sinensis) was analyzed with 72 simple sequence repeat markers. In total, 359 alleles were obtained with an average of 4.99 per marker. The HS varied greatly from 37.5 to 71.0% with an average of 51.3%. On average, tea cultivars from Fujian Province showed a higher level of heterozygosity (59.8%) than those from Zhejiang (48.5%) and Yunnan (44.5%), and the 12 national tea cultivars were generally more heterozygous than the 12 provincial cultivars. Unweighted pair-group analysis using the arithmetic average grouping divided the 24 cultivars into 2 groups that are consistent with the morphological classification. All dual combinations of the 24 cultivars were studied to calculate the percentage of mappable markers when using pseudo-testcross mapping strategy, and results showed that this value also varied greatly from 51.4 to 90.3%. The genetic relationships and HS differences among different cultivars were discussed, and tea cultivars with high HS were recommended as cross parents for genetic mapping programs.

Assessment of genetic diversity of cucumber cultivars in China based on simple sequence repeats and fruit traits.

The cucumber (Cucumis sativus L.) is an important crop grown worldwide. In this study, the genetic diversity of 42 cucumber cultivars in China was analyzed using 51 pairs of simple sequence repeat (SSR) primers. These primers identified 129 polymorphic loci, 95.6% of which were polymorphic. The mean effective number of alleles, mean Nei's gene diversity, and mean Shannon's information index were 0.36, 0.16, and 0.21, respectively. A cluster analysis demonstrated that the 42 cultivars could be divided into three groups, a result that was largely consistent with those of a principal component analysis (PCA). The PCA indicated that the three groups displayed significant variation in fruit traits. The cultivars of group 1 tended to have longer fruits (>30 cm), longer fruit ends (>4 cm), larger fruit diameters (>5 cm), a sharp strigose fruit spine, and the same fruit end shape. The basal color of the fruit in group 2 was dark green. Group 3 cultivars have no wax or mottling on the fruit surface. Our study demonstrates the value of our SSR primers for assessing genetic diversity in cucumber.

Cell origins and significance of IL-17 in malignant pleural effusion.

PURPOSE: T cells are dominant in the immune regulation of malignant pleural effusion (MPE). However, it is unclear about the role of IL-17+ T cells, particularly for IL-17+CD8+ Tc17 cells in antitumor immunity. This retrospective study is aimed at evaluating the prognostic significance of IL-17+ T cells in patients with MPE. METHODS: The frequency of IL-17+CD4+ Th17 and IL-17+CD8+ Tc17 cells in peripheral blood (PB), pleural fluids (PF), and tumor tissues in 24 patients undergoing thoracoscopy was determined by flow cytometry, immunohistochemistry, and ELISA. The association among the different measures was analyzed by Spearman's correlation tests. RESULTS: The percentages of PF Th17 and Tc17 cells were significantly higher than those in the PB of MPE patients and healthy controls (p < 0.01). Analysis of Th17 and Tc17 cells in the tumor tissues indicated that the percentages of Th17 and Tc17 cells in the invading tumor edge were significantly higher than those in the non-tumor tissues and intra-tumor regions (p < 0.05). More importantly, the percentages of IL-17+ T cells were associated with prolonged survival of patients with MPE. CONCLUSIONS: Both Th17 and Tc17 cells were involved in the tumor immunity against MPE. Increased frequency of Tc17 cells may serve as a biomarker for the prognosis of patients with MPE.

RNAi-mediated knockdown of pituitary tumor- transforming gene-1 (PTTG1) suppresses the proliferation and invasive potential of PC3 human prostate cancer cells

Pituitary tumor-transforming gene-1 (PTTG1) is a proto-oncogene that promotes tumorigenesis and metastasis in numerous cell types and is overexpressed in a variety of human tumors. We have demonstrated that PTTG1 expression was up-regulated in both human prostate cancer specimens and prostate cancer cell lines. For a more direct assessment of the function of PTTG1 in prostate tumorigenesis, RNAi-mediated knockdown was used to selectively decrease PTTG1 expression in PC3 human prostate tumor cells. After three weeks of selection, colonies stably transfected with PTTG1-targeted RNAi (the knockdown PC3 cell line) or empty vector (the control PC3 cell line) were selected and expanded to investigate the role of PTTG1 expression in PC3 cell growth and invasion. Cell proliferation rate was significantly slower (28%) in the PTTG1 knockdown line after 6 days of growth as indicated by an MTT cell viability assay (P < 0.05). Similarly, a soft agar colony formation assay revealed significantly fewer (66.7%) PTTG1 knockdown PC3 cell colonies than control colonies after three weeks of growth. In addition, PTTG1 knockdown resulted in cell cycle arrest at G1 as indicated by fluorescence-activated cell sorting. The PTTG1 knockdown PC3 cell line also exhibited significantly reduced migration through Matrigel in a transwell assay of invasive potential, and down-regulation of PTTG1 could lead to increased sensitivity of these prostate cancer cells to a commonly used anticancer drug, taxol. Thus, PTTG1 expression is crucial for PC3 cell proliferation and invasion, and could be a promising new target for prostate cancer therapy.

Identification of markers tightly linked to tomato yellow leaf curl disease and root-knot nematode resistance by multiplex PCR.

Seven different commercial F1 hybrids and two F2 populations were evaluated by multiplex PCR to identify plants that are homozygous or heterozygous for Ty-1 and Mi, which confer resistance to tomato yellow leaf curl disease and root-knot nematode, respectively. The Ty-1 and Mi markers were amplified by PCR and identified by digestion of the amplicons with the TaqI enzyme. The hybrids E13 and 288 were found to be Ty/ty heterozygous plants with 398-, 303-, and 95-bp bands, and B08, 314, 198, and A10 were found to be ty/ty homozygous plants with a 398-bp band; whereas 098 did not give any PCR products. The hybrids E13 and 198 were found to be Mi/Mi homozygous plants with 570- and 180-bp bands, and 288 and A10 were found to be Mi/mi heterozygous plants, with 750-, 570- and 180-bp bands, and B08, 109 and 314 were found to be mi/mi homozygous plants with only a 750-bp band. We additionally developed a multiplex PCR technique for JB-1 and Mi, which confer resistance to tomato yellow leaf curl disease and root-knot nematode. The JB-1 marker identified the genotype of the Ty gene, and the plants that produced the 400-bp band were ty/ty homozygous plants, whereas the plants that produced 400- and 500-bp bands were resistant to tomato yellow leaf curl disease. We conclude that multiplex PCRs can be used to reproducibly and efficiently detect these resistance genes.

RNAi-mediated knockdown of pituitary tumor- transforming gene-1 (PTTG1) suppresses the proliferation and invasive potential of PC3 human prostate cancer cells.

Pituitary tumor-transforming gene-1 (PTTG1) is a proto-oncogene that promotes tumorigenesis and metastasis in numerous cell types and is overexpressed in a variety of human tumors. We have demonstrated that PTTG1 expression was up-regulated in both human prostate cancer specimens and prostate cancer cell lines. For a more direct assessment of the function of PTTG1 in prostate tumorigenesis, RNAi-mediated knockdown was used to selectively decrease PTTG1 expression in PC3 human prostate tumor cells. After three weeks of selection, colonies stably transfected with PTTG1-targeted RNAi (the knockdown PC3 cell line) or empty vector (the control PC3 cell line) were selected and expanded to investigate the role of PTTG1 expression in PC3 cell growth and invasion. Cell proliferation rate was significantly slower (28%) in the PTTG1 knockdown line after 6 days of growth as indicated by an MTT cell viability assay (P < 0.05). Similarly, a soft agar colony formation assay revealed significantly fewer (66.7%) PTTG1 knockdown PC3 cell colonies than control colonies after three weeks of growth. In addition, PTTG1 knockdown resulted in cell cycle arrest at G1 as indicated by fluorescence-activated cell sorting. The PTTG1 knockdown PC3 cell line also exhibited significantly reduced migration through Matrigel in a transwell assay of invasive potential, and down-regulation of PTTG1 could lead to increased sensitivity of these prostate cancer cells to a commonly used anticancer drug, taxol. Thus, PTTG1 expression is crucial for PC3 cell proliferation and invasion, and could be a promising new target for prostate cancer therapy.