To develop transgenic cucumber tolerant to abiotic stress, a cotyledonary-node explants were co-cultivated with Agrobacterium tumefaciens (EHA101) carrying TPSP gene (pHC30-TPSP). After transfer to fresh medium every two week for eight weeks, putative transgenic plants were selected when shoots grown a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 5 mgl -1 phospinotricin as selectable agent. The confirmation of transgenic cucumber was based on the Northern blot analysis. Thirty four shoots (5.2%) with resistance to phospinotricin were obtained from 660 explants inoculated. Of them, transformants were only confirmed from 11 plants (1.7%). Transgenic cucumber expressing TPSP gene was more synthesized at 3.8 times amounts of trehalose (0.014 mg g fresh wt ). However, all of transgenic plants showed abnormal morphology, including stunted growth (< height 15 cm), shrunken leaves, and sterility as compared with non-transgenic plants (> height 150 cm) under the same growth environment. These results lead us to speculate that the overproduction of trehalose was toxic for cucumber, even though that had known for rice as non-toxic.
Colpomenia sinuosa is an annual brown algal species that occurs in temperate to tropical waters of the world. In order to examine the genetic diversity among populations of the species and to discuss its current distribution, we analyzed the nuclear ribosomal DNA internal transcribed spacer (ITS) region from 18 specimens of C. sinuosa, and, for comparison, plastid rbcL from the same specimens. The C. sinuosa ITS region (2141-2534 base pairs) is approximately 2.2 times the length of most other brown algae. We found a long repeated sequence of approximately 190 base pairs in the first half of the ITS1 region; five repeats in Northern Hemisphere collections and three in those from the Southern Hemisphere, which result in ITS length variation. The unequal occurrence of tandem repeats of C. sinuosa corresponds to the geographical distribution of the species. The rbcL sequences from all the specimens of C. sinuosa, except the Canary Island samples, were identical, indicating that they indeed belong to the same species.
Pyrolysis mass spectrometry (PyMS) is a rapid, simple, high-resolution analytical method based on thermal degradation of complex material in a vacuum and has been widely applied to the discrimination of closely related microbial strains. Leaf samples of six species and one variety of higher plants (Rosa multiflora, R. multiflora var. platyphylla, Sedum kamtschaticum, S. takesimense, S. sarmentosum, Hepatica insularis, and H. asiatica) were subjected to PyMS for spectral fingerprinting. Principal component analysis of PyMS data was not able to discriminate these plants in discrete clusters. However, canonical variate analysis of PyMS data separated these plants from one another. A hierarchical dendrogram based on canonical variate analysis was in agreement with the known taxonomy of the plants at the variety level. These results indicate that PyMS is able to discriminate higher plants based on taxonomic classification at the family, genus, species, and variety level.
Researchers have widely adopted the hairy root culture system as a means for producing secondary metabolites, including ginsenosides from ginseng. Although bacterial genes are involved, the aspects of plant gene expression are unclear. Using a cDNA microarray approach, we identified genes that are differentially expressed in ginseng hairy roots after Agrobacterium rhizogenes infection. Our goal was to gain an initial understanding of the correlation between hairy root morphology and ginsenoside produc'Uon. Among the 250 genes analyzed here, 63 (including 14 that are unclassified) were differentially expressed in a hairy root line containing a high level of ginsenosides. Of the genes that had been functionally categorized, 29% and 17% were active in metabolism and s~ress responses, respectively. Most were primarily associated with ribosomal proteins, thereby functioning in protein synthesis and destination. Their expression was down-regulated in hairy roots having less lateral branching. This phenotype may have resulted from the manipulation of metabolic activities by the translational machinery.
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