Roots of Panax ginseng, one of the most famous medicinal plants, contain various phytosterols and bioactive triterpene saponins (ginsenosides). In P. ginseng, phytosterols and triterpenes share the common biosynthetic intermediate, squalene. Here, we investigate the regulatory role of Panax ginseng squalene synthase (PgSS1) on the biosynthesis of phytosterols and triterpene saponins. PgSS1 transcripts are expressed ubiquitously in the various plant tissues, but higher in shoot apex and root. The transcript levels of PgSS1 increased markedly in the adventitious roots during 12- to 96-h period after metyl jasmonate (MeJA) treatment; MeJA treatment induced the activation of the transcripts of squalene epoxidase (SE), beta-amyrin synthase (bAS), but not cycloartenol synthase (CAS). Unlike MeJA treatment, overexpression of PgSS1 in adventitious roots of transgenic P. ginseng was followed by the up-regulation of all the downstream genes tested, such as SE, bAS, and CAS. The enhanced activity of PgSS1 enzyme resulted in remarkable increase of phytosterols as well as ginsenoside contents. These results demonstrate that PgSS1 is a key regulatory enzyme not only for phytosterol but also for triterpene biosynthesis and overexpressing of PgSS1 confers the hyperproduction of triterpene saponins to P. ginseng.
Natural releasable attachment systems of insect legs, where attachment-detachment performances are often very fast, seem to be optimized to get a maximum of real contact to the substratum. Tarsi of Tettigonia viridissima bear flexible attachment pads with unusual ultrastructural architecture of the cuticle. The indentation of the attachment pads was measured under different loads using a force-tester. Since the mechanical properties are influenced by material structure, the freeze-substitution experiments were undertaken to investigate the influence of loads on material structure. Both profile changes of the surface and the orientation of cuticle microfibrils were visualized by means of scanning electron microscopy followed by fracturing of the frozen material. The results show that the flexible pad material deforms replicating the substrate profile down to the micrometer roughness. The pad material showed both elastic and viscous behavior under loads. Elastic deformation of the pad occurred under normal force applied for 4-6 s (elastic modulus 27.2 +/- 11.6 kPa). Two viscous relaxation processes were found, of time constants tau1 = 1.88+/-0.616 s and tau2 =41.2 +/- 9.95 s. Low stiffness of material studied here aids in surface replication and increase of area of real contact between the pad and the underlying substrate.
The suitability of three common atomic force microscope (AFM) imaging modes for quantitative height and volume measurements on soft samples was investigated. The height and volume of rehydrated human metaphase chromosomes in liquid were measured using the contact mode, the tapping mode, and the force mapping mode. In both the contact and tapping modes, the measured height and volume strongly depended on the imaging setpoint that sets the imaging force. Measurement deviations up to 50% were observed. The force mapping mode, on the other hand, yielded reproducible height and volume measurements independent of the imaging force. It is therefore suggested that the force mapping mode should be used whenever the height or volume of soft samples need to be accurately determined.
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