Xiaofen Jin

AtCPK6, a functionally redundant and positive regulator involved in salt/drought stress tolerance in Arabidopsis

The calcium-dependent protein kinase (CDPK) family is needed in plant signaling during various physiological pathways. The ArabidopsisAtCPK6 gene belongs to the subclass of stress-inducible CDPKs, which is stimulated by salt and osmotic stress. To elucidate the physiological function of AtCPK6, transgenic Arabidopsis plants under the control of double CaMV 35S promoter were obtained. AtCPK6 over-expressing plants showed enhanced tolerance to salt/drought stresses. The elevated tolerance of the AtCPK6 over-expressing plants was confirmed by the change of proline and malondialdehyde (MDA). Real-time PCR analyses revealed that the expression levels of several stress-regulated genes were altered in AtCPK6 over-expressing plants. However, cpk6 mutant displayed no obvious difference with control. These results are likely to indicate that AtCPK6 is functionally redundant and a positive regulator involved in the tolerance to salt/drought stress in Arabidopsis.

OsNAC52, a rice NAC transcription factor, potentially responds to ABA and confers drought tolerance in transgenic plants

A novel NAC (NAM, ATAF1/2, CUC2) homologous gene, OsNAC52, was isolated from Oryza sativa L using RACE (rapid amplification of cDNA ends). Analysis of the amino acid sequence of OsNAC52 revealed a highly conserved NAC domain and a potential nuclear localization sequence in its N-terminus, and the transcriptional activation motif in the C-terminal region. Transgenic plants over-expressing OsNAC52 were highly sensitive to ABA (Abscisic acid), and the growth of the 35S-OsNAC52 transgenic seedlings was significantly more restrained by ABA treatment than those of the wild-type seedlings. Furthermore, over-expression of OsNAC52 activated the expression of downstream genes in transgenic Arabidopsis, resulting in enhanced tolerance to drought stresses but not growth retardation. The results from this study indicate that this novel rice OsNAC52 gene functions as an important transcriptional activator in ABA-inducible gene expression and may be useful in improving plant tolerance to abiotic stress.

Expression and Function of a Modified AP2/ERF Transcription Factor from Brassica napus Enhances Cold Tolerance in Transgenic Arabidopsis

One of the most rapid and effective defensive mechanisms plants have for protecting themselves, from a variety of biotic and abiotic stresses, is the regulation of plant signal transcription factors. AP2/ERF factors play an important role in plant development as well as in hormonal regulation and cold response. Directed evolution is a powerful tool to modify proteins, improving their properties, and for studying their structure–function relations. Here, the transgenic Arabidopsis plants over-expressed a mutant gene, BnaERF-B3-hy15-mu3, which encoded for a factor that exhibited more binding activity with the GCC box element than the wild-type gene BnaERF-B3-hy15 encode factor, and exhibited more freezing tolerance than transgenic plants containing the original BnaERF-B3-hy15 gene. Real-time PCR analyses also revealed that the expression levels of several stress-regulated genes were altered in the over-expressed BnaERF-B3-hy15-mu3 transgenic lines. The BnaERF-B3-hy15 responded to exogenous ABA. Using RT-PCR analysis, the expression of BnaERF-B3-hy15 at different stages and stress treatments were also analyzed.

Over-expression of ThpI from Choristoneura fumiferana enhances tolerance to cold in Arabidopsis.

Thermal hysteresis proteins (Thps) known as antifreeze proteins for their antifreeze activity, depress the freezing point of water below the melting point in many polar marine fishes, terrestrial arthropods and plants. For the purpose of breeding cold-resistant plants, we designed to introduce the Thp gene into the plants. The physiological and biochemical effect of high-lever expression of the modified Choristoneura fumiferanaThp (ThpI) in Arabidopsis thaliana plants was analyzed. Under low temperature stress, the ThpI transgenic plants exhibited stronger growth than wild-type plants. The elevated cold tolerance of the ThpI over-expressing plants was confirmed by the changes of electrolyte leakage activity, malonyldialdehyde and proline contents. These results preliminarily showed that the Thp possibly be used to enhance the low temperature-tolerant ability of plants.

Microarray analysis of the phytoremediation and phytosensing of occupational toxicant naphthalene.

Naphthalene is of global environmental concern because it is assumed to contribute considerably to human cancer risk. Plants are important in removing naphthalene from the atmosphere and soil. However, there remains insufficient knowledge on plant response to this compound. To determine the mechanism of naphthalene uptake and transduction in plants, as well as plant response to this compound, a microarray system was used to analyze gene expression patterns in Arabidopsis thaliana after irrigation with 2.0mM naphthalene. A total of 247 differentially expressed genes were identified as upregulated by naphthalene. These genes might specifically contribute to naphthalene uptake, transformation, conjugation, and compartmentalization in the plant. The potential role of upregulated genes in plant defense to naphthalene and the use of phytosensing for naphthalene detection were also discussed.

A Novel 5-Enolpyruvylshikimate-3-Phosphate Synthase from Rahnella aquatilis with Significantly Reduced Glyphosate Sensitivity

The 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 2.5.1.19) is a key enzyme in the shikimate pathway for the production of aromatic amino acids and chorismate-derived secondary metabolites in plants, fungi, and microorganisms. It is also the target of the broad-spectrum herbicide glyphosate. Natural glyphosate resistance is generally thought to occur within microorganisms in a strong selective pressure condition. Rahnella aquatilis strain GR20, an antagonist against pathogenic agrobacterial strains of grape crown gall, was isolated from the rhizosphere of grape in glyphosate-contaminated vineyards. A novel gene encoding EPSPS was identified from the isolated bacterium by complementation of an Escherichia coli auxotrophic aroA mutant. The EPSPS, named AroAR.aquatilis, was expressed and purified from E. coli, and key kinetic values were determined. The full-length enzyme exhibited higher tolerance to glyphosate than the E. coli EPSPS (AroAE.coli), while retaining high affinity for the substrate phosphoenolpyruvate. Transgenic plants of AroAR.aquatilis were also observed to be more resistant to glyphosate at a concentration of 5 mM than that of AroAE.coli. To probe the sites contributing to increased tolerance to glyphosate, mutant R.aquatilis EPSPS enzymes were produced with the c-strand of subdomain 3 and the f-strand of subdomain 5 (Thr38Lys, Arg40Val, Arg222Gln, Ser224Val, Ile225Val, and Gln226Lys) substituted by the corresponding region of the E. coli EPSPS. The mutant enzyme exhibited greater sensitivity to glyphosate than the wild type R.aquatilis EPSPS with little change of affinity for its first substrate, shikimate-3-phosphate (S3P) and phosphoenolpyruvate (PEP). The effect of the residues on subdomain 5 on glyphosate resistance was more obvious.

Stress responses to phenol in Arabidopsis and transcriptional changes revealed by microarray analysis

Phenols are toxic, environmentally persistent products of the chemical industry that are capable of bioaccumulation and biomagnifications in the food chain. Little is known of how plants respond to this compound. To understand the transcriptional changes under phenol, microarray experiments on Arabidopsis thaliana were performed. Microarray results revealed numerous perturbations in signaling and metabolic pathways. The results indicated that the phenol response was related to reactive oxygen species (ROS) accumulation and oxidative conditions, including ROS generated for pathogen defense.

Gene duplication and transfer events in plant mitochondria genome.

Gene or genome duplication events increase the amount of genetic material available to increase the genomic, and thereby phenotypic, complexity of organisms during evolution. Gene duplication and transfer events have been important to molecular evolution in all three domains of life, and may be the first step in the emergence of new gene functions. Gene transfer events have been proposed as another accelerator of evolution. The duplicated gene or genome, mainly nuclear, has been the subject of several recent reviews. In addition to the nuclear genome, organisms have organelle genomes, including mitochondrial genome. In this review, we briefly summarize gene duplication and transfer events in the plant mitochondrial genome.

Enhanced Transformation of TNT by Arabidopsis Plants Expressing an Old Yellow Enzyme

2,4,6-Trinitrotoluene (TNT) is released in nature from manufacturing or demilitarization facilities, as well as after the firing or detonation of munitions or leakage from explosive remnants of war. Environmental contamination by TNT is associated with human health risks, necessitating the development of cost-effective remediation techniques. The lack of affordable and effective cleanup technologies for explosives contamination requires the development of better processes. In this study, we present a system for TNT phytoremediation by overexpressing the old yellow enzyme (OYE3) gene from Saccharomyces cerevisiae. The resulting transgenic Arabidopsis plants demonstrated significantly enhanced TNT tolerances and a strikingly higher capacity to remove TNT from their media. The current work indicates that S. cerevisiae OYE3 overexpression in Arabidopsis is an efficient method for the phytoremoval and degradation of TNT. Our findings have the potential to provide a suitable remediation strategy for sites contaminated by TNT.

Analysis of gene expression profile of Arabidopsis genes under trichloroethylene stresses with the use of a full-length cDNA microarray

Trichloroethylene (TCE) is a widespread and persistent environmental contaminant. Plants are able to take up a range of harmful organic compounds, including some of the most abundant environmental pollutants like TCE. In this study, complementary DNA microarrays were constructed to have a better view of transcript expression in Arabidopsis thaliana during TCE-induced stress. The microarray analysis demonstrated the complexity of gene expression patterns resulting from TCE. A total of 1,020 transcripts were differentially up-regulated by TCE. Those genes might specifically contribute to the TCE transformation, conjugation, and compartmentation in plant. This study provides informative preliminary data for more in-depth analyses of TCE tolerance in Arabidopsis thaliana.