DNA binding specificity and cleavage activity of Pacmmar transposase.

Mariner-like elements (MLEs) are members of the Tc1/mariner superfamily of transposable elements which transpose by a "cut and paste" mechanism. Most of the MLEs characterized to date are transpositionally inactive due to the accumulation of mutations in their transposase gene. Here, we report the biochemical study of two copies of the Pacmmar element (Pacmmar1.1 and Pacmmar1.2), isolated from the coastal crab Pachygrapsus marmoratus. These two copies present an open reading frame encoding a putative active transposase. Using an in vitro transposition assay, we show that Pacmmar transposases are unable to perform by themselves the transposition reaction. However, we demonstrate by an electrophoretic mobility shift assay that both transposases bind specifically to the inverted terminal repeat of the Pacmmar element. Moreover, an in vitro cleavage assay showed that both transposases have the capacity to cleave the transposon. The in vitro cleavage activity of Pacmmar transposases appears imprecise, suggesting the requirement of specific host factors or the presence of mutations which have modified the cleavage specificity of the enzyme.

Osmotic adjustment, gas exchanges and chlorophyll fluorescence of a hexaploid triticale and its parental species under salt stress.

The effect of salt stress (NaCl 85.7 or 110 mmol/L) was investigated in the triticale T300 and its parental species, Triticum dicoccum farrum (Triticum df) and Secale cereale cv. Petkus. Triticum df and T300 were more salt-tolerant than the rye (110 mmol/L NaCl was the highest concentration allowing rye growth to the three-leaf stage). Na+, K+ and Cl- ions accounted for almost half of the osmotic adjustment in Triticum df and T300, and up to 90% in rye. Salinity decreased the net photosynthesis and transpiration rates of the three cereals as compared to control plants, but induced no significant change in chlorophyll a fluorescence parameters. Water-use efficiency (WUE) increased with salinity. In the presence of 110 mmol/L NaCl, the K+/Na+ ratio decreased markedly in rye as compared to the other two cereals. Proline concentration, which increased in Triticum df and T300, could have protected membrane selectivity in favour of K+. Proline content remained low in rye, and increasing soluble sugar content did not appear to prevent competition between Na+ and K+. The salt sensitivity of rye could be due to low K+ uptake in the presence of a high NaCl concentration.