Analysis of genetic diversity of Trypanosoma cruzi: an application of riboprinting and gradient gel electrophoresis methods

Analysis of restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S from Trypanosoma cruzi yields a typical `riboprint' profile that can vary intraspecifically. A selection of 21 stocks of T. cruzi and three outgroup taxa: T. rangeli, T. conorhini and Leishmania braziliensis were analysed by riboprinting to assess divergence within and between taxa. T. rangeli, T. conorhini and L. braziliensis could be easily differentiated from each other and from T. cruzi. Phenetic analysis of PCR-RFLP profiles indicated that, with one or two exceptions, stocks of T. cruzi could be broadly partitioned into two groups that formally corresponded to T. cruzi I and T. cruzi II respectively. To test if ribosomal 18S sequences were homogeneous within each taxon, gradient gel electrophoresis methods were employed utilising either chemical or temperature gradients. Upon interpretation of the melting profiles of riboprints and a section of the 18S independently amplified by PCR, there would appear to be at least two divergent 18S types present within T. cruzi. Heterogeneity within copies of the ribosomal 18S within a single genome has therefore been demonstrated and interestingly, this dimorphic arrangement was also present in the outgroup taxa. Presumably the ancestral duplicative event that led to the divergent 18S types preceded that of speciation within this group. These divergent 18S paralogues may have, or had, different functional pressures or rates of molecular evolution. Whether or not these divergent types are equally transcriptionally active throughout the life cycle, remain to be assessed.

Water use efficiency of cassava plants under competition conditions

The objective of this work was to evaluate characteristics associated with the photosynthetic activity of cassava plants in competition with weeds or not. The trial was performed on open environment conditions, with experimental units consisting of fiber glass vases with 150 dm³ filled with Red Yellow Latosol, previously fertilized. Treatments consisted in the cultivation of cassava plants isolated and associated to three weed species (Bidens pilosa, Commelina benghalensis and Brachiaria plantaginea). After cassava shooting, 15 days after planting, a removal of the weeds excess was performed, sown at the time of cassava planting, leaving six plants m-2 of B. pilosa and four plants m-2 of C. benghalensis and B. plantaginea. At 60 days after emergence (DAE), stomatal conductance (Gs), vapor pressure in the substomatal cavity (Ean), temperature gradient between leaf and air (ΔT), transpiration rate (E) and water use efficiency (WUE) were evaluated. B. pilosa showed greater capacity to affect growth of cassava plants. B. plantaginea is very efficient in using water, especially by presenting C4 metabolism, and remains competitive with cassava even under temporarily low water status. C. benghalensis, in turn, is not a good competitor for light and apparently is not the primary cause of water depletion in the soil. The effects of weeds, in this case, were more associated with the competition. However, they were found between moderate to low. This implies that the competition established at experimental level was low.