The Expression of CAT Gene in Transgenic Tomato Plant

Bacterial CAT gene .with the 35S promoter of CaMV was transferred into leaf discs of cultivar Lycopersicon esculentum 462 with the help of Ti plasmid of Agrobacterium turnefaciens. These leaf discs were placed on MS salt and B5 vitamine medium containing kanamycin and carbenicillin. Several kinds of phytohormones were chosen in the medium, and it was found that zeatin and NAA have great effects on shooting and rooting. Transgenic tomato plants showed their normal flowering and fruiting. Leaves of these transgenic tomato plants were used for assaying the gene expression. Resuits of Southern Blot showed that the CAT gene was stably integrated into the genome of transgenic plants, The bacterial CAT, proteins were also detected, in transgenic tomato leaves with immunoreaction of CAT antibody. The methods presented here for culturing transformed tomato ceils will be a great help for transfering economically important genes into cultivars of tomato plants in China.     

An Arabidopsis mutant of inositol pentakisphosphate 2‐kinase AtIPK1 displays reduced arsenate tolerance

Arsenate [As(V)] toxicity is considered to be derived from similarities in the chemical properties of As(V) and phosphate (Pi). An Arabidopsis thaliana mutant of inositol pentakisphosphate 2‐kinase (AtIPK1), atipk1‐1, has previously exhibited lower level of phytate and higher level of Pi, relative to wild‐type (WT). Here, atipk1‐1 displayed hypersensitivity to As(V) stress and less As(V) uptake when compared to WT. Overexpression of AtIPK1 controlled by the CaMV 35S promoter partially rescued the As(V)‐sensitive phenotype of atipk1‐1. When compared to control Pi status, addition of Pi enhanced As(V) tolerance of both WT and atipk1‐1 plants, while the arsenic concentration was less reduced in the latter genotype. Despite the higher Pi level in atipk1‐1 than did WT plants, the mutant suffered more severe Pi starvation under Pi limitation stress, indicating that Pi homeostasis was altered in the mutant. Gene expression analysis of WT and atipk1‐1 plants showed the diverse effect of As(V) stress on Pi starvation‐dependent regulation of Pi‐responsive genes. Our study suggested that a particular mechanism of As(V) toxicity existed in atipk1‐1 mutant, and may offer new insights into the interactions between Pi homeostasis and As(V) detoxification in plants.     

Novel Histopathological and Molecular Effects of Natural Compound Pellitorine on Larval Midgut Epithelium and Anal Gills of Aedes aegypti

The yellow fever mosquito, Aedes aegypti, is a vector for transmitting dengue fever and yellow fever. In this study, we assessed the histopathological and molecular effects of pellitorine, an isobutylamide alkaloid, on the third instar of Ae. aegypti larvae. At 5 mg/l concentration of pellitorine, the whole body of the treated larvae became dark in color, particularly damaged thorax and abdominal regions. Pellitorine was targeted mainly on midgut epithelium and anal gills, indicating variably dramatic degenerative responses of the midgut through a sequential epithelial disorganization. The anterior and posterior midgut was entirely necrosed, bearing only gut lumen residues inside the peritrophic membranes. Pellitorine caused comprehensive damage of anal gill cells and branches of tracheole and debris was found in hemolymph of the anal gills. RT-PCR analysis indicates that the compound inhibited gene expression encoding V-type H⁺-ATPase and aquaporine 4 after treatment with 2.21 mg/l pellitorine. These results verify that pellitorine merits further study as a potential larvicide with a specific target site and a lead molecule for the control of mosquito populations.     

Real-time quantitative assay of telomerase activity using the duplex scorpion primer

A novel real-time quantitative method for measuring telomerase activity is described in which the duplex scorpion primer is used to provide an intramolecular probing mechanism for specific detection of telomerase activity. Using this method, linearity from 10 to 10(4) cells expressing telomerase activity could be obtained (R2 = 0.994). The requirement of post-PCR steps is thus obviated.     

Influence of p53 expression on sensitivity of cancer cells to bleomycin

In this study, we determined whether p53 expression affected the sensitivity of non–small cell lung cancer (NSCLC) and colon cancer cells to bleomycin (BLM). Two human NSCLC cell lines (A549 expressing wild‐type p53 and p53‐null H1299) and two colon cancer cell lines (HCT116 p53+/+ and its p53 deficient subline HCT116 p53?/?) were subjected to treatment with BLM. Cells were treated with various concentrations of BLM, and cellular viability was assessed by formazan assay. To investigate the role of p53 in BLM sensitivity, we transduced cells with adenovirus with wild‐type p53, dominant‐negative p53, and GFP control, and analyzed the effect on cellular viability. Cells expressing wild‐type p53 were more sensitive to BLM than p53‐null cells in both NSCLC and colon cancer cells. Sensitivity to BLM of the cells with wild‐type p53 was reduced by overexpression of dominant‐negative p53, while BLM sensitivity of p53‐null cells was increased by wild‐type p53 in both NSCLC cells and colon cancer cells. In conclusion, our data show that p53 sensitizes all four cancer cells lines tested to BLM toxicity and overexpression of p53 confers sensitivity to the cytotoxic activity of the anticancer agent in p53‐null cells. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:260–269, 2010; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.20334     

Lysophosphatidylcholine aggravates contact hypersensitivity by promoting neutrophil infiltration and IL17 expression

Lysophosphatidylcholine (LPC) is a bioactive lysolipid known to contribute to the development of lung allergic diseases. However, it remains unknown whether LPC possesses proinflammatory properties in the skin as well. Here, we investigated this issue by injection of LPC into the murine contact hypersensiti-vity (CHS) model induced by 2,4-dinitrofluorobenzene (DNFB). LPC increased the expression of IL17, recruited more neutrophils, and eventually aggravated the CHS in the skins. Moreover, the effects of LPC diminished after neutralizing IL17 or depleting neutrophils. Mechanistically, LPC upregulated not only IL17 but also CXCL1 and CXCL2 in a G2A-dependent manner. Taken together, our study demonstrated that the upregulation of LPC could contribute to allergic skin inflammation by increasing IL17 expression and neutrophil recruitment via G2A receptor.     

Isolation of hydrogen-producing bacteria from granular sludge of an upflow anaerobic sludge blanket reactor

H₂-producing bacteria were isolated from anaerobic granular sludge. Out of 72 colonies (36 grown under aerobic conditions and 36 under anaerobic conditions) arbitrarily chosen from the agar plate cultures of a suspended sludge, 34 colonies (15 under aerobic conditions and 19 under anaerobic conditions) produced H₂ under anaerobic conditions. Based on various biochemical tests and microscopic observations, they were classified into 13 groups and tentatively identified as follows: From aerobic isolates,Aeromonas spp. (7 strains),Pseudomonas spp. (3 strains), andVibrio spp. (5 strains); from anaerobic isolates,Actinomyces spp. (11 strains),Clostridium spp. (7 strains), andPorphyromonas sp. When glucose was used as the carbon substrate, all isolates showed a similar cell density and a H₂ production yield in the batch cultivations after 12h (2.24–2.74 OD at 600 nm and 1.02–1.22 mol H₂/mol glucose, respectively). The major fermentation by-products were ethanol and acetate for the aerobic isolates, and ethanol, acetate and propionate for the anaerobic isolates. This study demonstrated that several H₂ producers in an anaerobic granular sludge exist in large proportions and their performance in terms of H₂ production is quite similar.     

The Janzen-Connell effect on the population dynamics of a Fagus engleriana- Cyclobalanopsis oxyodon community in a subtropical zone of China

The Janzen-Connell (J-C) hypothesis provides a mechanism explaining the high species diversity in tropical rainforests. It postulates that predation could cause greater mortality on seeds and seedlings near their parental trees. In this study, we tested the hypothesis in a subtropical zone, a mixed evergreen-deciduous broad-leaved forest dominated by the Fagus engleriana and Cyclobalanopsis oxyodon. The study area was in the Shennongjia region, a key area of biodiversity conserva-tion in both China and the world. The recruitment probability index was used to detect the J-C effect on nine species of the community, which were more than 50 individuals. Six large adults of each species were selected, and the numbers of saplings and adults were counted at the distance intervals of 0-5, 5-10, 10-15, 15-20, and 20-25 m from each focal tree. Two species in saplings stage and six in adult stage supported the J-C hypothesis, but their χ2 was not significant. Three species, the F. engleri-ana, Rhododendron hypoglaucum, and Toona sinensis, showed a strong Hubbell pattern in the adult stage. Because of these results, we reject the J-C hypothesis and conclude that species could recruit near the conspecific trees in subtropical forest. The reasons why the J-C hypothesis fails to explain the species diversity in this community are the shortage of seed-consuming agents of subtropical forest and the influence of microsite topo-graphic variation.