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1. Detection of plant genes using a rapid, nonorganic DNA purification method.

Author

Lin JJ, Fleming R, Kuo J, Matthews BF, and Saunders JA

Subjects
Base Sequence, Biotechnology, DNA Primers genetics, Gene Amplification, Paper, Plant Leaves chemistry, Plants, Genetically Modified, DNA, Plant genetics, DNA, Plant isolation & purification, Genes, Plant, Genetic Techniques
Abstract

We have developed a simple procedure for the preparation of plant genomic DNA using FTA paper. Plant leaves were crushed against FTA paper, and the genomic DNA was purified using simple, nonorganic reagents. The 18S rRNA gene and the gene encoding the ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (rbcL) from the chloroplast genome were detected by PCR amplification of DNA on FTA paper. DNA amplification was successful using extracts from 16 dicot and monocot plants. Studies of specific plant extracts revealed that extracts of leaf samples could be collected and stored at room temperature on FTA paper without a decrease in the DNA amplification success rate for more than a month. Both the 18S RNA gene and the rbcL gene were detected in the genomic DNA isolated from various soybean cultivars stored in this manner. Furthermore, by modestly increasing the number of cycles of DNA amplification, we were able to detect the uidA gene in transgenic tobacco and rice leaves as well as a single copy gene linked to the resistance gene of cyst nematode race 3 using genomic DNA isolated on FTA paper. These results demonstrate that genomic DNA isolated using FTA paper can be used for the detection of plant genes, from a wide range of plants with either high or low gene copy number and of either nuclear or cytoplasmic origin.

Published
2000
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2. Improvement of paper mulberry tolerance to abiotic stresses by ectopic expression of tall fescue FaDREB1

Author

Hongqing Li, Yan Li, Meiru Li, and Guojiang Wu

Subjects
Festuca, Paper, Physiology, Drought tolerance, Plant Science, Sodium Chloride, Genes, Plant, chemistry.chemical_compound, Gene Expression Regulation, Plant, Stress, Physiological, Botany, Plant Proteins, Abiotic component, biology, Abiotic stress, fungi, Paper mulberry, Reproducibility of Results, food and beverages, Broussonetia, Blotting, Northern, Plants, Genetically Modified, biology.organism_classification, Adaptation, Physiological, Blotting, Southern, Mutagenesis, Insertional, chemistry, Chlorophyll, Ectopic expression, Festuca arundinacea
Abstract

Dehydration-responsive element binding/C-repeat-binding factors (DREB/CBF) control the activity of multiple stress response genes and therefore represent attractive targets for genetic improvement of abiotic stress tolerance. Paper mulberry (Broussonetia papyrifera L. Vent) is well known for its bark fibers and high levels of chalcone and flavonoid derivatives. Transgenic paper mulberry plants expressing a tall fescue (Festuca arundinacea Schreb.) FaDREB1 gene under the control of CaMV 35S were produced to examine the potential utility of FaDREB1 to increase the tolerance of paper mulberry plants to abiotic stress. The overexpressing FaDREB1 plants showed higher salt and drought tolerance than the wild-type plants (WT). After 13 days of withholding water, or 15 days in the presence of 250 mM NaCl, all the WT plants died, while the over-expressing FaDREB1 plants survived. The FaDREB1 plants had higher leaf water and leaf chlorophyll contents, accumulated more proline and soluble sugars, and had less ion leakage (which reflects membrane damage) than the WT plants had under high salt- and water-deficient conditions. The 35S promoter-driven expression of FaDREB1 did not cause growth retardation under normal growth conditions. Therefore, improved tolerance to multiple environmental stresses in paper mulberry might be achieved via genetic engineering through the ectopic expression of an FaDREB1 gene.

Published
2011