Heat stress-induced expression of Px-pdrg and Px-aspp2 in insecticide-resistant and -susceptible Plutella xylostella.

p53, DNA damage regulated gene (PDRG) and apoptosis-stimulating p53 protein 2 (ASPP2) are p53-related genes which can promote apoptosis. The full-length cDNA sequence of the Px-pdrg and Px-aspp2 genes were characterized and their mRNA expression dynamics under heat stress were studied in diamondback moth (DBM) Plutella xylostella collected from Fuzhou, China. The full-length cDNA of Px-pdrg and Px-aspp2 spans 721 and 4201 bp, containing 395 and 3216 bp of the open reading frame, which encode a putative protein comprising 130 and 1072 amino acids with a calculated molecular weight of 14.58 and 118.91 kDa, respectively. As compared to 25°C, both Px-pdrg and Px-aspp2 were upregulated in chlorpyrifos-resistant (Rc) and -susceptible (Sm) strains of DBM adults and pupae under heat stress. In addition, Rc DBM showed a significantly higher expression level of Px-pdrg and Px-aspp2 in contrast to Sm DBM. The results indicate that high temperature can significantly promote apoptosis process, especially in Rc-DBM. Significant fitness cost in Rc-DBM might be associated with drastically higher transcript abundance of Px-pdrg and Px-aspp2 under the heat stress.

Vitality and Stability of Insecticide Resistance in Adult Propylaea japonica (Coleoptera: Coccinellidae).

Propylaea japonica (Thunberg) was a dominant species among the predacious ladybirds in the fields and active from March to November during a year in Fuzhou, China. Stability of insecticide resistance and vitality in adult P. japonica were investigated. The field ladybird P. japonica in Fuzhou, China, showed 9- to 16-fold resistance ratios to chlorpyrifos, 13- to 2,083-fold to methamidophos, 32- to 230-fold to fenvalerate, and 4- to 49-fold to avermectins, respectively, based on the field monitoring during 2004, 2009, and 2012, as compared with insecticide-susceptible F39 progenies. The resistance levels in the field P. japonica were high during May and November but low during summer. The population growth tendency index in field P. japonica was 0.8-fold as high as that in insecticide-susceptible P. japonica. The field P. japonica also showed high tolerance to the insecticide as compared with pest Lipaphis erysimi Kaltenbach and two parasitoids Diaeretiella rapae and Pachyneuron aphidis. Stable insecticide resistance levels and high vitality were found first in adult P. japonica with 1-, 30-, or 60-d-old adults, or among the adults developed form the eggs produced by newly emerged adults or by 60-d-old adults. Increased activity of glutathione S-transferases, carboxylesterases, and cytochrome P450 monooxygenases might be involved in the resistance of P. japonica. The results indicated that, in certain areas, inclusion of P. japonica for pest control in the integrated pest management would be highly recommended.

Cloning of Wing-Development-Related Genes and mRNA Expression Under Heat Stress in Chlorpyrifos-Resistant and -Susceptible Plutella xylostella.

Chlorpyrifos-resistant (Rc) Plutella xylostella (DBM) shows higher wing-vein injury than chlorpyrifos-susceptible (Sm) DBM under heat stress in our previous study. To investigate the toxicological mechanisms of the differences in injury of wing vein between Rc- and Sm-DBM collected from Fuzhou, China, total ten cDNA sequences of wing-development-related genes were isolated and characterized in DBM, including seven open reading frame (ORF) (ash1, ah2, ash3, ase, dpp, srf and dll encoded 187 amino acids, 231 aa, 223aa, 397aa, 423aa, 229aa and 299aa, respectively), and three partly sequences (salm, ser and wnt-1 encoded 614aa, 369aa and 388aa, respectively). The mRNA expression of the genes was inhibited in Rc- and Sm-DBM under heat stress, as compared with that an average temperature (25 °C). And, in general, significantly higher down-regulated expressions of the mRNA expression of the wing development-related genes were found in Rc-DBM as compared to those in Sm-DBM under heat stress. The results indicated that Sm-DBM displayed higher adaptability at high temperature because of significantly lower inhibition the mRNA expressions of wing-development-related genes. We suggest that significantly higher injury of wing vein showed in Rc-DBM under heat stress might be associated with the strong down-regulation of wing-development-related genes.

An "up, no change, or down" system: Time-dependent expression of mRNAs in contused skeletal muscle of rats used for wound age estimation.

The combined use of multiple markers is considered a promising strategy in estimating the age of wounds. We sought to develop an "up, no change, or down" system and to explore how to combine and use various parameters. In total, 78 Sprague Dawley rats were divided randomly into a control group and contusion groups of 4-, 8-, 12-, 16-, 20-, 24-, 28-, 32-, 36-, 40-, 44-, and 48-h post-injury (n=6 per group). A contusion was produced in the right limb of the rats under diethyl ether anesthesia by a drop-ball technique; the animals were sacrificed at certain time points thereafter, using a lethal dose of pentobarbital. Levels of PUM2, TAB2, GJC1, and CHRNA1 mRNAs were detected in contused muscle using real-time PCR. An up, no change, or down system was developed with the relative quantities of the four mRNAs recorded as black, dark gray, or light gray boxes, representing up-, no change, or down-regulation of the gene of interest during wound repair. The four transcripts were combined and used as a marker cluster for color model analysis of each contusion group. Levels of PUM2, TAB2, and GJC1 mRNAs decreased, whereas that of CHRNA1 increased in wound repair (P<0.05). The up, no change, or down system was adequate to distinguish most time groups with the color model. Thus, the proposed up, no change, or down system provide the means to determine the minimal periods of early wounds.