Showing papers by "Dabing Zhang published in 2007"

Genome-Wide Comparative Analysis and Expression Pattern of TCP Gene Families in Arabidopsis thaliana and Oryza sativa

Abstract: Several TCP genes have been reported to play important roles in plant development; the TCP homologs encode a plant-specific family of putative transcription factors. To understand the evolutionary relationship of TCP genes of Arabidopsis thaliana and Oryza sativa L. (hereafter called rice), we have identified 23 and 22 TCP genes in the Arabidopsis and rice genomes, respectively. Using phylogenetic analysis, we grouped these TCP genes into three classes. In addition, the motifs outside the TCP domain further support the evolutionary relationships among these genes. The genome distribution of the TCP genes strongly supports the hypothesis that genome-wide and tandem duplication contributed to the expansion of the TCP gene family. The expression pattern of the TCP genes was analyzed further, providing useful clues about the function of these genes.

Event-specific quantitative detection of nine genetically modified maizes using one novel standard reference molecule.

Abstract: With the development of genetically modified organism (GMO) detection techniques, the Polymerase Chain Reaction (PCR) technique has been the mainstay for GMO detection, and real-time PCR is the most effective and important method for GMO quantification. An event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity. This study establishes the event-specific detection methods for TC1507 and CBH351 maizes. In addition, the event-specific TaqMan real-time PCR detection methods for another seven GM maize events (Bt11, Bt176, GA21, MON810, MON863, NK603, and T25) were systematically optimized and developed. In these PCR assays, the fluorescent quencher, TAMRA, was dyed on the T-base of the probe at the internal position to improve the intensity of the fluorescent signal. To overcome the difficulties in obtaining the certified reference materials of these GM maizes, one novel standard reference molecule containing all nine specific integration junction sequences of these GM maizes and the maize endogenous reference gene, zSSIIb, was constructed and used for quantitative analysis. The limits of detection of these methods were 20 copies for these different GM maizes, the limits of quantitation were about 20 copies, and the dynamic ranges for quantification were from 0.05 to 100% in 100 ng of DNA template. Furthermore, nine groups of the mixed maize samples of these nine GM maize events were quantitatively analyzed to evaluate the accuracy and precision. The accuracy expressed as bias varied from 0.67 to 28.00% for the nine tested groups of GM maize samples, and the precision expressed as relative standard deviations was from 0.83 to 26.20%. All of these indicated that the established event-specific real-time PCR detection systems and the reference molecule in this study are suitable for the identification and quantification of these GM maizes.

Cloning and characterization of squalene synthase (SQS) gene from Ganoderma lucidum

Abstract: This report provides the complete nucleotide sequences of the full-length cDNA encoding squalene synthase (SQS) and its genomic DNA sequence from a triterpene-producing fungus, Ganoderma lucidum. The cDNA of the squalene synthase (SQS) (GenBank Accession Number: DQ494674) was found to contain an open reading frame (ORF) of 1,404 bp encoding a 468-amino-acid polypeptide, whereas the SQS genomic DNA sequence (GenBank Accession Number: DQ494675) consisted of 1,984 bp and contained four exons and three introns. Only one gene copy was present in the G lucidum genome. The deduced amino acid sequence of Ganoderma lucidum squalene synthase (Gl-SQS) exhibited a high homology with other fungal squalene synthase genes and contained six conserved domains. A phylogenetic analysis revealed that G. lucidum SQS belonged to the fungi SQS group, and was more closely related to the SQS of U. maydis than to those of other fungi. A gene expression analysis showed that the expression level was relatively low in mycelia incubated for 12 days, increased after 14 to 20 days of incubation, and reached a relatively high level in the mushroom primordia. Functional complementation of Gl-SQS in a SQS-deficient strain of Saccharomyces cerevisiae confirmed that the cloned cDNA encoded a squalene synthase.

Varied Transcriptional Efficiencies of Multiple Arabidopsis U6 Small Nuclear RNA Genes

Abstract: Arabidopsis U6 small nuclear RNA (snRNA) promoters are those transcribed by RNA polymerase III, but all the core elements for transcriptional initiation are located in the 5’promoter region Previously, three Arabidopsis U6 snRNA genes (U6-1, U6-26, and U6-29) were identified Herein, we have further identified three new U6 loci (U6-4, U6-5, and U6-6) in the Arabidopsis genome Alignment of these revealed that the upstream sequence element and TATA elements were contained in six U6 promoters In addition, a unique, highly conserved element named the “CAT” element was observed in the promoter region To understand the expression patterns of these U6 genes in Arabidopsis, we fused these promoters to the DNA segment of β-glucuronidase and then transferred these six constructs into Arabidopsis Real-time reverse transcription-polymerase chain reaction analysis of these fused transcripts indicated that the newly identified U6 genes are active in Arabidopsis and that the U6-26 promoter seems to have higher transcriptional activity in leaf, stem, flower and silique These results help to understand the function of these U6 snRNAs in Arabidopsis

The Shoot Apical Meristem Size Regulated by FON4 in Rice.

Abstract: CLAVATA pathway is one of best-characterized signaling pathway involves in the regulation of meristem development in Arabidopsis. Increasing evidence indicated that this pathway also exist in the monocots as well as in the dicots. We have recently identified FON4 in rice as an ortholog of CLV3 in Arabidopsis. FON4 is putative ligand of FON1, which play a role in restricting the meristem size in rice. FON4 and CLV3 are the members of CLE gene family, which encode small functional secreted peptide with a conserved 14-amino acid motif (CLE motif) near or at the C termini.

Interlaboratory trial validation of an event-specific qualitative polymerase chain reaction-based detection method for genetically modified RT73 rapeseed.

Abstract: The qualitative event-specific polymerase chain reaction detection method of genetically modified (GM) RT73 rapeseed was developed based on the cloned 3' end flanking sequence of RT73 rapeseed integration. The specificity of the method for GM RT73 rapeseed was validated using several different GM rapeseed lines, GM maize lines, GM soybean line, non-GM rapeseed, and other non-GM crops. In this study, the developed method was validated through an interlaboratory study by 12 laboratories from 6 countries. The sensitivity of this method was evaluated using several mixed rapeseed meals with different GM RT73 rapeseed contents from 5.0 to 0.01% prepared by our laboratory. The evaluated results showed that all of the rapeseed endogenous reference high mobility group protein gene (HMG I/Y), figwort mosaic virus 35S (FMV 35S) promoter, and RT73 event-specific fragment could be detected from rapeseed samples at 0.1% (w/w) with a confidence level of more than 95%. All results from the 12 laboratories indicated that the developed method could be considered fit for the detection and identification of GM RT73 rapeseed.

Genome-wide Expression Profiling in Seedlings of the Arabidopsis Mutant uro that is Defective in the Secondary Cell Wall Formation

Abstract: Plant secondary growth is of tremendous importance, not only for plant growth and development but also for economic usefulness. Secondary tissues such as xylem and phloem are the conducting tissues in plant vascular systems, essentially for water and nutrient transport, respectively. On the other hand, products of plant secondary growth are important raw materials and renewable sources of energy. Although advances have been recently made towards describing molecular mechanisms that regulate secondary growth, the genetic control for this process is not yet fully understood. Secondary cell wall formation in plants shares some common mechanisms with other plant secondary growth processes. Thus, studies on the secondary cell wall formation using Arabidopsis may help to understand the regulatory mechanisms for plant secondary growth. We previously reported phenotypic characterizations of an Arabidopsis semi-dominant mutant, upright rosette (uro), which is defective in secondary cell wall growth and has an unusually soft stem. Here, we show that lignification in the secondary cell wall in uro is aberrant by analyzing hypocotyl and stem. We also show genome-wide expression profiles of uro seedlings, using the Affymetrix GeneChip that contains approximately 24 000 Arabidopsis genes. Genes identified with altered expression levels include those that function in plant hormone biosynthesis and signaling, cell division and plant secondary tissue growth. These results provide useful information for further characterizations of the regulatory network in plant secondary cell wall formation.

Phenotypic characterization of a rice mutant Oryza sativa extraordinary glume 1 (Oseg 1) and its genetic analysis

Abstract: A rice mutant with Japonica 9522 cultivar background Oryza sativa extraordinary glume 1 (Oseg 1) was identified from the M 2 mutant pool mutagenized by 60Co γ-ray. Compared with wild type plants, Oseg 1 developed longer empty glumes and rudimentary glumes. In some Oseg 1 mutants, the number of stamens of flowers was reduced and leaf-like lodicules occurred, and excessive lemma/palea-like organ could be observed in some mutant spikelets. This indicated that OsEG1 could regulate the edevelopment of rudimentary glumes, empty glumes, lemma/palea, lodicules, and stamens. Genetic analysis indicated that Oseg 1 came from a single recessive genetic locus. To clone OsEG1 gene, F 2 population was constructed by a cross between Oseg 1 (Japonica) and Guangluai4 (Indica). Using map-based cloning approach, OsEG1 was mapped on chromosome 4, between INDEL marker OS407 and WHM0466 with genetic distance of 2.0 cm and 1.0 cm, respectively. These results are useful for further cloning and functional analysis of the OsEG1 gene.