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PubMed | Luzhou Medical College, Guangdong Academy of Agriculture science and Sichuan Agricultural University
Type: Journal Article | Journal: Biochemical and biophysical research communications | Year: 2015

Lead (Pb), as a heavy metal element, has become the most important metal pollutant of the environment. With allocating a relatively higher proportion of its biomass in roots, maize could be a potential important model to study the phytoremediation of Pb-contaminated soil. Here we analyzed the maize root transcriptome of inbred lines 9782 under heavy metal lead (Pb) pollution, which was identified as a non-hyperaccumulator for Pb in roots. In the present study, more than 98 millions reads were mapped to define gene structure and detect polymorphism, thereby to qualify transcript abundance along roots development under Pb treatment. A total of 17,707, 17,440, 16,998 and 16,586 genes were identified in maize roots at four developmental stages (0, 12 h, 24 h and 48 h) respectively and 2,825, 2,626, 2161 and 2260 stage-specifically expressed genes were also identified respectively. In addition, based on our RNA-Seq data, transcriptomic changes during maize root development responsive to Pb were investigated. A total of 384 differentially expressed genes (DEGs) (log2Ratio 1, FDR 0.001) were identified, of which, 36 genes with significant alteration in expression were detected in four developmental stages; 12 DEGs were randomly selected and successful validated by qRT-PCR. Additionally, many transcription factor families might act as the important regulators at different developmental stages, such as bZIP, ERF and GARP et al. These results will expand our understanding of the complex molecular and cellular events in maize root development and provide a foundation for future study on root development in maize under heavy metal pollution and other cereal crops.

PubMed | University of South China and Guangdong Academy of Agriculture science
Type: Journal Article | Journal: PloS one | Year: 2016

Staphylococcus hyicus has caused great losses in the swine industry by inducing piglet exudative epidermitis (EE), sow mastitis, metritis, and other diseases and is a threat to human health. The pathogenesis of EE, sow mastitis, and metritis involves the interaction between the host and virulent protein factors of S. hyicus, however, the proteins that interact with the host, especially the host immune system, are unclear. In the present study, immunoproteomics was used to screen the immunogenic proteins of S. hyicus strain ZC-4. The cellular and secreted proteins of S. hyicus strain ZC-4 were obtained, separated by 2D gel electrophoresis, and further analyzed by western blot with S. hyicus strain ZC-4-infected swine serum. Finally, 28 specific immunogenic proteins including 15 cellular proteins and 13 secreted proteins, 26 of which were novel immunogenic proteins from S. hyicus, were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. To further verify their immunogenicity, two representative proteins (acetate kinase [cellular] and enolase [secreted]) were chosen for expression, and the resultant recombinant proteins could react with S. hyicus ZC-4-infected swine serum. In mice, both acetate kinase and enolase activated the immune response by increasing G-CSF and MCP-5 expression, and acetate kinase further activated the immune response by increasing IL-12 expression. Enolase can confer better protection against S.hycius than acetate kinase in mice. For the first time to our knowledge, our results provide detailed descriptions of the cellular and secreted proteins of S. hyicus strain ZC-4. These immunogenic proteins may contribute to investigation and elucidation of the pathogenesis of S. hyicus and provide new candidates for subunit vaccines in the future.

Jiang B.,Nanjing Agricultural University | Jiang B.,Guangdong Academy of Agriculture Science | Lou Q.,Nanjing Agricultural University | Wu Z.,Nanjing Agricultural University | And 6 more authors.
Plant Molecular Biology | Year: 2011

Allopolyploidization is considered an essential evolutionary process in plants that could trigger genomic shock in allopolyploid genome through activation of transcription of retrotransposons, which may be important in plant evolution. Two retrotransposon-based markers, inter-retrotransposon amplified polymorphism and retrotransposon-microsatellite amplified polymorphism and a microsatellite-based marker, inter simple sequence repeat were employed to investigate genomic changes in early generations of a newly synthesized allotetraploid Cucumis × hytivus Chen & Kirkbride (2n = 4x = 38) which was derived from crossing between cultivated cucumber C. sativus L. (2n = 2x = 14) and its wild relative C. hystrix Chakr. (2n = 2x = 24). Extensive genomic changes were observed, most of which involved the loss of parental DNA fragments and gain of novel fragments in the allotetraploid. Among the 28 fragments examined, 24 were lost while four were novel, suggesting that DNA sequence elimination is a relatively frequent event during polyploidization in Cucumis. Interestingly, of the 24 lost fragments, 18 were of C. hystrix origin, four were C. sativus-specific, and the remaining two were shared by both species, implying that fragment loss may be correlated with haploid DNA content (genome size) of diploid parents. Most changes were observed in the first generation after polyploidization (S 1) and stably inherited in the subsequent three generations (S 2-S 4), indicating that genomic changes might be a rapid driving force for the stabilization of allotetraploids. Sequence analysis of 11 of the 28 altered DNA fragments showed that genomic changes in the allotetraploid occurred in both coding and non-coding regions, which might suggest that retrotransposons inserted into genome randomly and had a genome-wide effect on the allotetraploid evolution. Fluorescence in situ hybridization (FISH) analysis revealed a unique distribution of retrotransposon and/or microsatellite flanking sequences in mitotic and meiotic chromosomes, where the preferential FISH signals occurred in the centromeric and telomeric regions, implying that these regions were the possible hotspots for genomic changes. © 2011 Springer Science+Business Media B.V.

Feng K.,South China Agricultural University | Feng K.,Key Laboratory of Chicken Genetics | Xue Y.,Guangdong Wen's Food Co. | Wang F.,Guangdong Wen's Food Co. | And 5 more authors.
Virus Genes | Year: 2014

Sixty-two strains of avian infectious bronchitis virus (IBV) were isolated from diseased chickens at different farms in southern China during 2011–2012, and 66.1 % of the isolated strains were associated with typical nephritis. Analysis of the S1 gene sequences amplified from the 62 isolated strains together with 40 reference strains published in Genbank showed nucleotide homologies ranging from 63.5 to 99.9 % and amino acid homologies ranging from 57.9 to 100 %. Phylogenetic analysis revealed that all Chinese IBV strains were clustered into six distinct genetic groups (I–VI). Most of the isolated strains belonged to group I, and the isolation of group V strains was increased compared with an earlier period of surveillance. Current vaccine strains used in China (H120, H52, W93, and Ma5) formed the group Mass which is evolutionarily distant from Chinese isolates. Alignment of S1 amino acid sequences revealed polymorphic and diverse substitutions, insertions, and deletions, and the S1 protein of major pandemic strains contained 540 amino acids with a cleavage site sequence of HRRRR or RRF(L/S)RR. Further analysis showed that recombination events formed a new subgroup. Taken together, these findings suggest that various IBV variants were co-circulating and undergoing genetic evolution in southern China during the observation period. Therefore, long-term continuing surveillance is significantly important for prevention and control of IBV infection. © 2014, Springer Science+Business Media New York.

Li X.,Huazhong Agricultural University | Chen L.,Guizhou Academy of Agricultural science | Hong M.,Huazhong Agricultural University | Zhang Y.,Guangdong Academy of Agriculture science | And 7 more authors.
PLoS ONE | Year: 2012

Yellow seed is a desirable quality trait of the Brassica oilseed species. Previously, several seed coat color genes have been mapped in the Brassica species, but the molecular mechanism is still unknown. In the present investigation, map-based cloning method was used to identify a seed coat color gene, located on A9 in B. rapa. Blast analysis with the Arabidopsis genome showed that there were 22 Arabidopsis genes in this region including at4g09820 to at4g10620. Functional complementation test exhibited a phenotype reversion in the Arabidopsis thaliana tt8-1 mutant and yellow-seeded plant. These results suggested that the candidate gene was a homolog of TRANSPARENT TESTA8 (TT8) locus. BrTT8 regulated the accumulation of proanthocyanidins (PAs) in the seed coat. Sequence analysis of two alleles revealed a large insertion of a new class of transposable elements, Helitron in yellow sarson. In addition, no mRNA expression of BrTT8 was detected in the yellow-seeded line. It indicated that the natural transposon might have caused the loss in function of BrTT8. BrTT8 encodes a basic/helix-loop-helix (bHLH) protein that shares a high degree of similarity with other bHLH proteins in the Brassica. Further expression analysis also revealed that BrTT8 was involved in controlling the late biosynthetic genes (LBGs) of the flavonoid pathway. Our present findings provided with further studies could assist in understanding the molecular mechanism involved in seed coat color formation in Brassica species, which is an important oil yielding quality trait. © 2012 Li et al.

Guo D.,South China Normal University | Guo D.,Guangdong Academy of Agriculture Science | Liang J.,South China Normal University | Qiao Y.,Guangzhou Academy of Agriculture Science | And 3 more authors.
Journal of Plant Physiology | Year: 2012

Previous study indicated that increasing endogenous abscisic acid (ABA) level could inhibit the lateral root (LR) formation of peanuts. In this study, we investigated the mechanisms by which ABA regulated lateral root primordia (LRP) initiation in peanuts (Arachis hypogaea L). Results suggested that ABA inhibited LRP initiation through blocking G1-to-S transition in seedlings and mature roots: e.g. 5.8% increase in the proportion of G1 phase and 18% decrease in the proportion of S phase after ABA treatment for 6 days. Further study of the expression of the cell cycle marker gene for G2-to-M transition in peanut roots suggested that AhCYCB1 expression was regulated by ABA. We also investigated the cooperative regulation of LRP initiation by ABA and indole-3-acetic acid (IAA). ABA treatment greatly reduced the effects of endogenous IAA on mature roots. The expression of the IAA polar transport gene AhAUX1 appeared to be regulated by ABA since ABA inhibited auxin-mediated LRP initiation by suppressing AhAUX-dependent auxin transport in peanut roots. We further examined the effect of ABA on the expression of DR5::GUS and AtAUX1 in the model plant Arabidopsis. The results of Arabidopsis were consistent with that of the peanut. © 2012 Elsevier GmbH.

Jia L.,Nanjing Agricultural University | Lou Q.,Nanjing Agricultural University | Jiang B.,Guangdong Academy of Agriculture Science | Wang D.,Nanjing Agricultural University | Chen J.,Nanjing Agricultural University
Scientia Horticulturae | Year: 2014

Cucumis hytivus is a newly synthesized allotetraploid in which retrotransposons was strongly activated owing to allopolyploidization. In present study, the chromosomal distribution of LTR retrotransposons and their effects on expression of adjacent genes were observed by investigating the first four generations of C. hytivus. Fluorescent in situ hybridization (FISH) analysis revealed that LTR retrotransposons are distributed throughout all the chromosomes. The clusters on terminal regions indicated the high copy number of retrotransposons in C. hytivus. The extensive epigenetic changes (gene silencing and gene activation) were detected by cDNA-SSAP analysis. Totally, twenty transcripts subjected to gene expression alterations were sequenced, including 12 gene silencing and 8 gene activation. However, the silenced/activated transcripts consisted of known genes or putative proteins as well as new sequences that had no similarity to any known genes. Both gene silencing and activation mainly occurred in the early generations of allotetraploid. The interlaced distribution of retrotransposons and genes might lead to the transcriptional interference of retrotransposons on expression of their adjacent genes, which may contribute to the rapid genetic stabilization of newly formed allotetraploid. © 2014 Elsevier B.V.

PubMed | U.S. Department of Agriculture, South China Agricultural University, Shandong Agricultural University and Guangdong Academy of Agriculture science
Type: Journal Article | Journal: Archives of virology | Year: 2016

Avian leukosis virus (ALV) causes high mortality associated with tumor formation and decreased fertility, and results in major economic losses in the poultry industry worldwide. Recently, a putative novel ALV subgroup virus named ALV-K was observed in Chinese local chickens. In this study, a novel ALV strain named GD14LZ was isolated from a Chinese local yellow broiler in 2014. The proviral genome was sequenced and phylogenetically analyzed. The replication ability and pathogenicity of this virus were also evaluated. The complete proviral genome sequence of GD14LZ was 7482 nt in length, with a genetic organization typical of replication-competent type C retroviruses lacking viral oncogenes. Sequence analysis showed that the gag, pol and gp37 genes of GD14LZ have high sequence similarity to those of other ALV strains (A-E subgroups), especially to those of ALV-E. The gp85 gene of the GD14LZ isolate showed a low sequence similarity to those other ALV strains (A-E subgroups) but showed high similarity to strains previously described as ALV-K. Phylogenetic analysis of gp85 also suggested that the GD14LZ isolate was related to ALV-K strains. Further study showed that this isolate replicated more slowly and was less pathogenic than other ALV strains. These results indicate that the GD14LZ isolate belongs to the novel subgroup ALV-K and probably arose by recombination of ALV-K with endogenous viruses with low replication and pathogenicity. This virus might have existed in local Chinese chickens for a long time.

High-density map is a valuable tool for genetic and genomic analysis. Although wax gourd is a widely distributed vegetable of Cucurbitaceae and has important medicinal and health value, no genetic map has been constructed because of the lack of efficient markers. Specific-locus amplified fragment sequencing (SLAF-seq) is a newly developed high-throughput strategy for large-scale single nucleotide polymorphism (SNP) discovery and genotyping.In our present study, we constructed a high-density genetic map by using SLAF-seq and identified a locus controlling pericarp color in wax gourd. An F2 population of 140 individuals and their two parents were subjected to SLAF-seq. A total of 143.38 M pair-end reads were generated. The average sequencing depth was 26.51 in the maternal line (B214), 27.01 in the parental line (B227), and 5.11 in each F2 individual. When filtering low-depth SLAF tags, a total of 142,653 high-quality SLAFs were detected, and 22,151 of them were polymorphic, with a polymorphism rate of 15.42 %. And finally, 4,607 of the polymorphic markers were selected for genetic map construction, and 12 linkage groups (LGs) were generated. The map spanned 2,172.86 cM with an average distance between adjacent markers for 0.49 cM. The inheritance of pericarp color was also studied, which showed that the pericarp color was controlled by one single gene. And based on the newly constructed high-density map, a single locus locating on chromosome 5 was identified for controlling the pericarp color of wax gourd.This is the first report of high-density genetic map construction and gene mapping in wax gourd, which will be served as an invaluable tool for gene mapping, marker assisted breeding, map-based gene cloning, comparative mapping and draft genome assembling of wax gourd.

PubMed | U.S. Department of Agriculture, South China Agricultural University, Shandong Agricultural University and Guangdong Academy of Agriculture science
Type: | Journal: Scientific reports | Year: 2015

The group of avian sarcoma and leukosis virus (ASLV) in chickens contains six highly related subgroups, A to E and J. Four genetic loci, tva, tvb, tvc and tvj, encode for corresponding receptors that determine the susceptibility to the ASLV subgroups. The prevalence of ASLV in hosts may have imposed strong selection pressure toward resistance to ASLV infection, and the resistant alleles in all four receptor genes have been identified. In this study, two new alleles of the tva receptor gene, tva(r5) and tva(r6), with similar intronic deletions were identified in Chinese commercial broilers. These natural mutations delete the deduced branch point signal within the first intron, disrupting mRNA splicing of the tva receptor gene and leading to the retention of intron 1 and introduction of premature TGA stop codons in both the longer and shorter tva isoforms. As a result, decreased susceptibility to subgroup A ASLV in vitro and in vivo was observed in the subsequent analysis. In addition, we identified two groups of heterozygous allele pairs which exhibited quantitative differences in host susceptibility to ASLV-A. This study demonstrated that defective splicing of the tva receptor gene can confer genetic resistance to ASLV subgroup A in the host.

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