Pokemon (FBI-1) interacts with Smad4 to repress TGF-ß-induced transcriptional responses.

Pokemon, an important proto-oncoprotein, is a transcriptional repressor that belongs to the POK (POZ and Krüppel) family. Smad4, a key component of TGF-ß pathway, plays an essential role in TGF-ß-induced transcriptional responses. In this study, we show that Pokemon can interact directly with Smad4 both in vitro and in vivo. Overexpression of Pokemon decreases TGF-ß-induced transcriptional activities, whereas knockdown of Pokemon increases these activities. Interestingly, Pokemon does not affect activation of Smad2/3, formation of Smads complex, or DNA binding activity of Smad4. TGF-ß1 treatment increases the interaction between Pokemon and Smad4, and also enhances the recruitment of Pokemon to Smad4-DNA complex. In addition, we also find that Pokemon recruits HDAC1 to Smad4 complex but decreases the interaction between Smad4 and p300/CBP. Taken together, all these data suggest that Pokemon is a new partner of Smad4 and plays a negative role in TGF-ß pathway.

Indigenous oil-degrading bacteria in crude oil-contaminated seawater of the Yellow sea, China.

Indigenous oil-degrading bacteria play an important role in efficient remediation of polluted marine environments. In this study, we investigated the diversity and abundance of indigenous oil-degrading bacteria and functional genes in crude oil-contaminated seawater of the Dalian coast. The gene copy number bacterial 16S rRNA in total were determined to be about 10(10) copies L(-1) in contaminated seawater and 10(9) copies L(-1) in uncontaminated seawater. Bacteria of Alcanivorax, Marinobacter, Novosphingobium, Rhodococcus, and Pseudoalteromonas were found to be predominant oil-degrading bacteria in the polluted seawater in situ. In addition, bacteria belonging to Algoriphagus, Aestuariibacter, Celeribacter, Fabibacter, Zobellia, Tenacibaculum, Citreicella, Roseivirga, Winogradskyella, Thioclava, Polaribacter, and Pelagibaca were confirmed to be the first time as an oil-degrading bacterium. The indigenous functional enzymes, including AlkB or polycyclic aromatic hydrocarbons ring-hydroxylating dioxygenases α (PAH-RHDα) coding genes from Gram-positive (GP) and Gram-negative bacteria (GN), were revealed and quite diverse. About 10(10) to 10(11) copies L(-1) for the expression of alkB genes were recovered and showed that the two-thirds of all the AlkB sequences were closely related to widely distributed Alcanivorax and Marinobacter isolates. About 10(9) copies L(-1) seawater for the expression of RHDαGN genes in contaminated seawater and showed that almost all RHDαGN sequences were closely related to an uncultured bacterium; however, RHDαGP genes represented only about 10(5) copies L(-1) seawater for the expression of genes in contaminated seawater, and the naphthalene dioxygenase sequences from Rhodococcus and Mycobacterium species were most abundant. Together, their data provide evidence that there exists an active aerobic microbial community indigenous to the coastal area of the Yellow sea that is capable of degrading petroleum hydrocarbons.

Foliar spraying of exogenous uniconazole (S3307) at the flowering stage as an effective method to resist low-temperature stress on mung bean [Vigna radiata (L.) Wilczek].

Low temperature is one of the major constraints on agricultural productivity worldwide and is likely to further increase. Several adaptations and mitigation strategies are required to cope with low-temperature stress. Uniconazole (S3307) could play a significant role in the alleviation of abiotic stress in plants. In this study, the effects of S3307 on the reactive oxygen species (ROS) and antioxidant metabolism were studied in the leaves of mung bean [Vigna radiata (L.) Wilczek]. The experimental results showed that the low-temperature induced accumulation of superoxide anion (O2-) production rate, and malonaldehyde (MDA) contents. Increased proline content and enzymatic antioxidants, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), were found to alleviate oxidative damage under low temperatures. While, S3307 could reduce O2- production rate and MDA contents and increase the activities of SOD, POD, and CAT, slowed the decrease in ascorbic acid (AsA), dehydroascorbic acid (DHA), glutathione (GSH), and oxidized glutathione (GSSG), and promoted increase in soluble sugars (SS), soluble proteins (SP), and proline (Pro) content under low-temperature. At the same time, low temperature leads to lower 100 grain weight and number of grains per plant, which eventually causes yield reduction decreased. Foliar spraying of S3307 could alleviate the yield loss caused by low temperature, and the increase of S3307 treatment was 5.1%-12.5% and 6.3%-32.9% for the two varieties, respectively, compared with CK. In summary, exogenous S3307 pretreatment enhances plant tolerance to low-temperature by improving the antioxidant enzyme activities, increased non-enzymatic antioxidants content, and decreased O2- production rate and MDA contents and inducing alterations in endogenous S3307, and reduce the decrease in mung bean yield.

Identification of Finely Mapped Quantitative Trait Locus and Candidate Gene Mining for the Three-Seeded Pod Trait in Soybean.

The three-seeded pod number is an important trait that positively influences soybean yield. Soybean variety with increased three-seeded pod number contributes to the seed number/plant and higher yield. The candidate genes of the three-seeded pod may be the key for improving soybean yield. In this study, identification and validation of candidate genes for three-seeded pod has been carried out. First, a total of 36 quantitative trait locus (QTL) were detected from the investigation of recombinant inbred lines including 147 individuals derived from a cross between Charleston and Dongning 594 cultivars. Five consensus QTLs were integrated. Second, an introgressed line CSSL-182 carrying the target segment for the trait from the donor parent was selected to verify the consensus QTL based on its phenotype. Third, a secondary group was constructed by backcrossing with CSSL-182, and two QTLs were confirmed. There were a total of 162 genes in the two QTLs. The mining of candidate genes resulted in the annotation of eight genes with functions related to pod and seed sets. Finally, haplotype analysis and quantitative reverse transcriptase real-time PCR were carried to verify the candidate genes. Four of these genes had different haplotypes in the resource group, and the differences in the phenotype were highly significant. Moreover, the differences in the expression of the four genes during pod and seed development were also significant. These four genes were probably related to the development process underlying the three-seeded pod in soybean. Herein, we discuss the past and present studies related to the three-seeded pod trait in soybean.

An Alternative Hot Start PCR Method Using a Nuclease-Deficient ExoIII from Escherichia coli.

The Hot Start polymerase chain reaction (Hot Start PCR) is designed to reduce off-target amplification by blocking DNA polymerase extension at room temperature until the desired temperature is reached. In this study, we investigated a new method of Hot Start PCR that uses a modified Escherichia coli Exonuclease III (EcoExoIIIM) by substituting residues in the DNA-binding pocket and catalytic center. The results showed that PCR amplification yield and specificity were significantly promoted by the addition of EcoExoIIIM. We hypothesize that non-specific binding of primers at room temperature is prevented by binding of the primed template by EcoExoIIIM, which is then released from the DNA by heat denaturation before the first PCR cycle. Through this mechanism, PCR would be enhanced by reducing off-target extension at room temperature.

Resveratrol Represses Pokemon Expression in Human Glioma Cells.

POK erythroid myeloid ontogenic factor (Pokemon), an important proto-oncoprotein, is a transcriptional repressor that regulates the expression of many genes and plays an important role in tumorigenesis. Resveratrol (RSV), a natural polyphenolic compound, has many beneficial biological effects on health. In this study, we investigated the role of Pokemon in RSV-induced biological effects and the effect of RSV on the expression of Pokemon in glioma cells. We found that overexpression of Pokemon decreased RSV-induced cell apoptosis, senescence, and anti-proliferative effects. Moreover, we showed that RSV could efficiently decrease the activity of the Pokemon promoter and the expression of Pokemon. Meanwhile, RSV also inhibited Sp1 DNA binding activity to the Pokemon promoter; whereas, it did not influence the expression and nuclear translocation of Sp1. In addition, we found that RSV could increase the recruitment of HDAC1, but decreased p300 to the Pokemon promoter. Taken together, all these results extended our understanding on the anti-cancer mechanism of RSV in glioma cells.

Distribution and diversity of bacterioplankton communities in subtropical seawater around Xiamen Island, China.

Marine bacterioplankton communities have profound impact on global biogeochemical cycles and ecological balances. However, relatively little is known about the bacterioplankton communities and the factors shaping their spatial distribution in subtropical island. Here, the bacterioplankton communities around a typical subtropical island, Xiamen Island, were revealed by analyzing bacterial 16S rRNA gene through quantitative PCR (qPCR) and 454 pyrosequencing methods. The qPCR results indicated that the abundance of 16S rRNA gene ranged from 2.07 × 10(7) to 2.13 × 10(8)copies mL(-1) in surface seawater among eight sampling sites (S1-S8) around Xiamen Island, and the nitrogen and phosphorus-rich sites (S5 and S8) were detected with higher 16S rRNA gene abundance. Pyrosequencing evidenced that a total of 267 genera of 47 classes in 26 different phyla (or candidate phyla) and some unclassified bacteria were obtained from seawater around Xiamen Island. The most dominant phylum was Proteobacteria (49.62-76.84% among sites), followed by Bacteroidetes (6.64-20.88%), Actinobacteria (2.58-9.20%), Firmicutes (0.03-13.30%), Verrucomicrobia (0.23-2.67%) and Planctomycetes (0.14-2.20%). Among eight sites, the nitrogen and phosphorus-rich sites (S5 and S8) exhibited higher proportions of Gammaproteobacteria, Epsilonproteobacteria, Firmicutes and lower proportions of Alphaproteobacteria and Planctomycetes than other sites. S5 and S8 also had more similar ß-diversity, and sampling site near the estuary (S8) showed the highest bacterial diversity. Redundancy analysis (RDA) confirmed that total nitrogen and total phosphorus significantly (P<0.05 and P<0.01, respectively) influenced the bacterioplankton communities around Xiamen Island. These results will provide insights into bacterial abundance, diversity and distribution patterns, as well as their controlling factors, in subtropical marine ecosystems.

Draft Genome Sequence of the Agar-Degrading Bacterium Catenovulum sp. Strain DS-2, Isolated from Intestines of Haliotis diversicolor.

Catenovulum sp. strain DS-2, isolated from intestines of Haliotis diversicolor, is able to degrade agar and produce agaro-oligosaccharides. Here, we report the draft genome sequence of Catenovulum sp. strain DS-2.

Draft Genome Sequence of the Carrageenan-Degrading Bacterium Cellulophaga sp. Strain KL-A, Isolated from Decaying Marine Algae.

Cellulophaga sp. strain KL-A, isolated from decaying marine algae, is able to degrade iota-carrageenan. Here, we report the draft genome sequence of Cellulophaga sp. strain KL-A.

Cotton GhDREB1 increases plant tolerance to low temperature and is negatively regulated by gibberellic acid.

The transcription factors C-repeat binding factors/dehydration-responsive element binding proteins (CBFs/DREBs) control the expression of many stress-inducible genes in Arabidopsis. A cDNA clone, designated GhDREB1, was isolated from cotton (Gossypium hirsutum) by cDNA library screening. Northern blot analysis indicated that mRNA accumulation of GhDREB1 was induced by low temperatures and salt stress, but was not induced by abscisic acid (ABA) or drought stress in cotton seedlings. Transgenic tobacco (Nicotiana tabacum) plants overexpressing GhDREB1 displayed stronger chilling tolerance than wild-type plants. Their leaf chlorophyll fluorescence, net photosynthetic rate and proline concentrations were higher than those of control plants during low-temperature treatment. However, under normal growth conditions, the transgenic tobacco plants exhibited retarded growth and delayed flowering. Interestingly, GhDREB1 transcripts in cotton seedlings were negatively regulated by gibberellic acid (GA(3)) treatment. Analysis of the promoter of the GhDREB1 gene revealed the presence of one low-temperature and four gibberellin-responsive elements. Green fluorescent protein (GFP) signal intensity or beta-glucuronidase (GUS) activity driven by the GhDREB1 promoter was clearly enhanced by low temperature but repressed by GA(3). These results suggest that GhDREB1 functions as a transcription factor and plays an important role in improving cold tolerance, and also affects plant growth and development via GA(3).