Jian-Jie Gao

Forced expression of Mdmyb10, a myb transcription factor gene from apple, enhances tolerance to osmotic stress in transgenic Arabidopsis

In plants, anthocyanins often appear at specific developmental stages, but are also induced by a number of environmental factors. The coordinated expression of genes encoding the anthocyanin biosynthetic pathway enzymes is controlled at the transcriptional level usually by an R2R3Myb transcription factor. However, little is known about the effects of R2R3-Myb on plant resistance to environmental stresses. In this study, we introduced an R2R3Myb transcription factor gene Mdmyb10, a regulatory gene of anthocyanin biosynthesis in apple fruit, into Arabidopsis and analyzed its function to osmotic stress in transgenic plants. Under high osmotic stress, the Mdmyb10 over-expressing plants exhibited growth better than wild-type plants. The elevated tolerance of the transgenic plants to osmotic stress was confirmed by the changes of flavonoids, chlorophyll, malondialdehyde and proline contents. These results preliminarily showed that the Mdmyb10 can possibly be used to enhance the high osmotic-tolerant ability of plants.

The myb transcription factor MdMYB6 suppresses anthocyanin biosynthesis in transgenic Arabidopsis

The R2R3-Myb proteins comprise a large family of plant transcription factors that regulate the expression of multiple drought-responsive and cold-responsive genes by binding special cis-elements at the promoter. There have been previous studies in transgenic plant over-expression of the myb protein. In this article, we analyzed the function of MdMYB6, an R2R3-type Myb transcription factor from apples (Malus × domestica), in transgenic Arabidopsis plants. In contrast to wild type plants, these transgenic lines accumulated less anthocyanin. Equimolar concentrations of sorbitol did not alter the anthocyanin accumulation. Real-time polymerase chain reaction (PCR) indicated that MdMYB6 over-expression suppressed enzymes involved in anthocyanin biosynthesis and some Arabidopsis basic helix-loop-helix (bHLH) genes. The MdMYB6 transcription faction may be an important repressor of anthocyanin biosynthesis in plants.

Over‐expression of AtGSTU19 provides tolerance to salt, drought and methyl viologen stresses in Arabidopsis

The plant-specific tau class of glutathione S-transferases (GSTs) is often highly stress-inducible and expressed in a tissue-specific manner, thereby suggesting its important protective roles. Although activities associated with the binding and transport of reactive metabolites have been proposed, little is known about the regulatory functions of GSTs. Expression of AtGSTU19 is induced by several stimuli, but the function of this GST remains unknown. In this study, we demonstrated that transgenic over-expressing (OE) plants showed enhanced tolerance to different abiotic stresses and increased percentage of seed germination and cotyledon emergence. Transgenic plants exhibited an increased level of proline and activities of antioxidant enzymes, along with decreased malonyldialdehyde level under stress conditions. Real-time polymerase chain reaction (PCR) analyses revealed that the expression levels of several stress-regulated genes were altered in AtGSTU19 OE plants. These results indicate that AtGSTU19 plays an important role in tolerance to salt/drought/methyl viologen stress in Arabidopsis.

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.

Transformation with a gene for myo-inositol O-methyltransferase enhances the cold tolerance of Arabidopsis thaliana

In this study, we report a function of myo-inositol-O-methyltransferase (Imt1) in response to low temperature stress using transgenic Arabidopsis thaliana. Imt1 gene was constructed identical to the Imt1 gene from a halophyte Mesembryanthemum crystallinum. After cold stress, the Imt1 transgenic plants exhibited stronger growth than the wild type plants. The elevated cold tolerance of the Imt1 over-expressing plants was confirmed by the lower electrolyte leakage and accumulation of malondialdehyde, but higher proline and soluble sugar contents in transgenic than wild type plants.

Transcription factor MdCBF1 gene increases freezing stress tolerance in transgenic Arabidopsis thaliana

Transcription factors play vital roles in stress signal transduction and gene expression modulation. The sequence analysis shows that MdCBF1 from Malus domestica Borkh. cv. Fuji contained an AP2 core domain of 56 amino acids. By comparison of deduced amino acid sequences of CBF related proteins, we deduced that MdCBF1 is a CBF transcription factor gene which belongs to AP2/EREBP family, DREB-A1 subfamily. Further, we reported that transgenic Arabidopsis thaliana plants expressing the MdCBF1 gene exhibited stronger growth than wild type plants under freezing stress. The analysis of RT-PCR for stress-responsive genes implied that MdCBF1 over-expressing plants had a higher expression of COR15a, RD29A, and RD29B genes than wild type plants. Collectively, our results indicate that MdCBF1 might play an important role in the response of transgenic Arabidopsis plants to freezing stress.

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.

Phytoremediation potential of Arabidopsis with reference to acrylamide and microarray analysis of acrylamide-response genes

Acrylamide (ACR) is a widely used industrial chemical. However, it is a dangerous compound because it showed neurotoxic effects in humans and act as reproductive toxicant and carcinogen in many animal species. In the environment, acrylamide has high soil mobility and may travel via groundwater. Phytoremediation is an effective method to remove the environmental pollutants, but the mechanism of plant response to acrylamide remains unknown. With the purpose of assessing remediation potentials of plants for acrylamide, we have examined acrylamide uptake by the model plant Arabidopsis grown on contaminated substrates with high performance liquid chromatography (HPLC) analysis. The result revealed that acrylamide could be absorbed and degraded by Arabidopsis. Further microarray analysis showed that 527 transcripts were up-regulated within 2-days under acrylamide exposure condition. We have found many potential acrylamide-induced genes playing a major role in plant metabolism and phytoremediation.

Phytoremediation and phytosensing of chemical contaminant, toluene: identification of the required target genes

As an industrial chemical produced worldwide in high volumes, toluene is commonly detected in ambient air and water. It can combine with oxygen and form compounds that are harmful to humans. In recent years, phytoremediation has been increasingly applied to repair the environmental damage caused by pollutants. However, insufficient knowledge is available regarding the response of plants to toluene. To detect the potential genes in plants that are related to the sensing mechanism and metabolism of toluene, a microarray analysis has been conducted on Arabidopsis thaliana seedlings grown on toluene-containing media. Following the validation of data and the application of appropriate selection criteria, the results show a coordinated induction and suppression of 202 and 67 toluene-responsive genes, respectively. Within the functional class “metabolism”, the genes encoding detoxification proteins represent the most strongly up-regulated group. These include genes encoding cytochrome P450s, glucosyl transferases, and transporters. Subsequently, the toluene-induced genes of Arabidopsis are analyzed in detail.

Disulfide isomerase-like protein AtPDIL1–2 is a good candidate for trichlorophenol phytodetoxification

Trichlorophenol (TCP) is a widely used and persistent environmentally toxic compound that poses a carcinogenic risk to humans. Phytoremediation is a proficient cleanup technology for organic pollutants. In this study, we found that the disulfide isomerase-like protein AtPDIL1–2 in plants is a good candidate for enhancing 2,4,6-TCP phytoremediation. The expression of AtPDIL1-2 in Arabidopsis was induced by 2,4,6-TCP. The heterologously expressed AtPDIL1-2 in Escherichia coli exhibited both oxidase and isomerase activities as protein disulfide isomerase and improved bacteria tolerance to 2,4,6-TCP. Further research revealed that transgenic tobacco overexpressing AtPDIL1-2 was more tolerant to high concentrations of 2,4,6-TCP and removed the toxic compound at far greater rates than the control plants. To elucidate the mechanism of action of AtPDIL1-2, we investigated the chemical interaction of AtPDIL1-2 with 2,4,6-TCP for the first time. HPLC analysis implied that AtPDIL1-2 exerts a TCP-binding activity. A suitable configuration of AtPDIL1-2-TCP binding was obtained by molecular docking studies using the AutoDock program. It predicted that the TCP binding site is located in the b-b′ domain of AtPDIL1-2 and that His254 of the protein is critical for the binding interaction. These findings imply that AtPDIL1-2 can be used for TCP detoxification by the way of overexpression in plants.