Plant growth and development influenced by transgenic insertion of bacterial chitinolytic enzymes

The role of the chitinolytic enzymes in plants is not necessarilyrestricted to plant defense. Tomato plants transformed with an endochitinaseand a chitobiosidase gene from Streptomyces albidoflavus andgrowth under greenhouse conditions showed a significant reduction in plantheight, and reduced time to flowering compared with the control(non-transformed) plants. The levels of chitobiosidase and endochitinaseactivity in the transgenic tomato plants were positively correlated with earlyflowering, and negatively correlated with plant height. We have not determinedwhether these effects are exclusively due to the expression of the transgenesof endochitinase and chitobiosidase from S. albidoflavus orthe additive effect of these 2 enzymes combined with the endogenouschitinolytic enzymes produced by the plants. However, when control plants were trimmed,early flowering was observed compared with the controls that were not trimmed, whichindicates that wound induced proteins such as chitinolytic enzymes affect thetime of flowering. In addition, the expression of the endochitinase andchitobiosidase genes significantly increased the number of flowers and fruit onthe plants, resulting in an increase in yield of fruit. One of the primarygoals of crop breeding programs is to increase the productivity of plants. These twogenes were directly associated with plant productivity, and should be studied further.     

Rearrangement of chromatin structure induced by increasing ionic strength and temperature.

Native rat liver chromatin fragments exposed to 600 mM NaCl at 37 degrees C for 45 min exhibit substantial modification of their original (approximately 200 base pairs) repeating subunit structure: a new repeat of 140 base pairs, superimposed on a high background, is observed after micrococcal nuclease digestion. The same material appears, in the electron microscope, as clusters of tightly packed beads connected by stretches of 'free' DNA. These modifications are not observed when the native chromatin is incubated at 37 degrees C at NaCl concentrations up to 400 mM. When native rat liver chromatin depleted of histone H1 by tRNA extraction is exposed to ionic strengths up to 600 mM NaCl at 4 degrees C, almost no modifications of the original native repeating structure are observed. However, when the incubation is carried out at 37 degrees C in 150, 300 or 400 mM NaCl, rearrangements of the native structure occur as indicated by micrococcal nuclease digestion and electron microscopic studies. Incubation of H1-depleted chromatin at 600 mM NaCl for 45 min at 37 degrees C induces, as for the native chromatin, a complete rearrangement characterized by the appearance of a 140-base-pair repeat superimposed on a high background upon digestion by micrococcal nuclease. It is suggested that these rearrangements are mediated by hydrophobic interactions between the histone cores and are prevented at ionic strengths lower than 500 mM by the presence of histone H1.     

Seagrass light acclimation: 2-DE protein analysis in Posidonia leaves grown in chronic low light conditions

Posidonia oceanica meadows are among the most valuable coastal systems in the Mediterranean basin. They provide nursery and forage areas for many commercially important species, including juvenile mollusc, finfish, and crustaceans. In the Mediterranean Sea, P. oceanica beds have recently suffered from progressive die-offs attributed to lower light availability from elevated water turbidity. In order to understand adaptive low-light responses of this seagrass, we compared the protein expression in plants collected from turbid waters (low-light) with plants collected from pristine-clear waters (high-light). More than 2600 proteins were detected in leaves from both sites. Among them, 26 proteins were differentially expressed in low-light conditions, 12 of which were identified through MASCOT analyses. The remaining 14 proteins, did not receive significant identity scores due to a lack of genomic and proteomic information in available databases. Nevertheless, we observed a 30% down-regulation of RuBisCo large subunit in low-light acclimated leaves. Whereas, enzymes involved in carbohydrate cleavage (1-fructose-bisphosphate aldolase, nucleoside diphosphate kinase, and beta-amylase) were upregulated in low-light conditions. Electron microscopy studies also revealed substantial changes in the stroma lamellae/grana ratios in chloroplasts receiving low-light, possibly as a mechanism for re-establishing optimal PSI/PSII ratios. Furthermore, under low-light conditions, four components of the ubiquitin/mediated proteolysis pathway (26 S proteasome regulatory, proteasome beta type 1, proteasome 7 D beta type, and proteasome alpha 7), and the perchloric acid soluble translation inhibitor protein, were upregulated. This suggests that, in P. oceanica leaves, enhanced protein turnover mediates acclimation to low-light conditions. Also, enzymes involved in defending against cellular stress (superoxide dismutase, pyridoxine, and 2-caffeic-acido-methyl transferase) were differentially expressed in low-light regime. Subsequent aquaria studies involving P. oceanica transplants maintained in low- and high-light conditions, also demonstrate RuBisCo down-regulation and proteasomes upregulation in low-light acclimated plants.