ABSTRACT Rhizophora mangle L. (Rhizophoraceae) grows on aerial roots, which emerge above the water level, giving stands of this tree the characteristic “mangrove” appearance. To produce in situ and ex situ information for genetic conservation programs for this species, we investigated the genetic diversity and mating system of one R. mangle population. We sampled 30 adult trees and a total of 349 seeds in Northern Brazil. We genotyped all adult trees and seeds with four microsatellite loci. The average fixation index was -0.222 for adult trees and 0.030 for seeds. The multilocus outcrossing rate (t m =0.921) was significantly lower than unity (1.0). There was no substantial evidence of null alleles nor genotypic disequilibrium among the loci. The combined power to exclude the first parent probability was 0.921. The average coancestry coefficient (=0.180) was similar to that expected for half-sib progenies (=0.125). Thus, the number of adult trees necessary for seed collection to obtain progeny arrays with an effective size of 150 was estimated to be 62. In conclusion, this study produced important information for the management and conservation of R. mangle and will contribute to conservation and management programs for this species.

The potato (Solanum tuberosum L.) mitochondrial cox3/sdh4/pseudo-cox2 gene cluster has previously been identified by heterologous hybridization using a Marchantia polymorpha sdh4 probe. In our present study we used Southern blotting using sdh4 and cox2 probes to show that the sdh4 and cox2 genes are clustered in the mitochondria of potato, soybean and pea. Northern blotting revealed cotranscription of sdh4 and cox2 in potato but not in cauliflower, indicating that these genes are not clustered in cauliflower. A putative recombination point was detected downstream of the cox2 pseudogene (pseudo-cox2) in potato mitochondrial DNA (mtDNA). This sequence corresponds to a 32 bp sequence which appears to be well-conserved and is adjacent to the terminals of some mitochondrial genes in Citrullus lanatus, Beta vulgaris and Arabidopsis thaliana and is probably involved in the genic rearrangements. It is possible the potato mtDNA pseudo-cox2 gene was generated by recombination during evolution in the same way as that of several other mitochondrial genes and remains as an inactive partial copy of the functional cox2 which was also detected in potato mtDNA.

Foram comparadas metodologias para preservação de amostras de folhas de plantas de savanas neotropicais, visando a posterior obtenção de DNA de alta qualidade para estudos moleculares. Foram também testadas diferentes metodologias para extração de DNA das amostras investigadas. A qualidade do DNA extraído foi avaliada através de dois métodos: por digestão, utilizando-se três enzimas de restrição e através da amplificação do gene mitocondrial cox3 (subunidade III da citocromo oxidase) por PCR. Os resultados revelaram que, para as plantas de savanas investigadas, a metodologia de preservação de amostras mais comumente empregada pelos botânicos (desidratação rápida usando sílica-gel) não é a mais eficiente. Os resultados obtidos demonstram a importância de testar diferentes métodos de preservação de amostras vegetais para estudos moleculares, para garantir a obtenção de DNA de alta qualidade. Para as plantas de savanas neotropicais, o método de preservação em sílica gel pode ser menos eficiente do que outros métodos igualmente simples de preservação de amostras.

Methods for the preservation of leaf sample from plants occurring in neotropical savannas were compared, with the aim to get high quality DNA for molecular studies. Different methods for DNA isolation of the same samples were also compared. The DNA quality was evaluated by two empirical methods: digestion of the DNA with three restriction enzymes and by amplification of the mitochondrial cox3 gene by PCR. The results show that, for the savanna plants investigated, the most commonly employed method for plant sample preservation (quick dehydratation using silica gel) is not the most efficient. Our results demonstrate the importance of testing different methods of plant tissue preservation for molecular studies to guarantee high quality DNA. For plants from neotropical savannas, the silica gel method might be often less efficient than other equally simple methods of sample preservation.

In order to compare their relative efficiencies as markers and to find the most suitable marker for maize diversity studies we evaluated 18 inbred tropical maize lines using a number of different loci as markers. The loci used were: 774 amplified fragment length polymorphisms (AFLPs); 262 random amplified polymorphic DNAs (RAPDs); 185 restriction fragment length polymorphisms (RFLPs); and 68 simple sequence repeats (SSR). For estimating genetic distance the AFLP and RFLP markers gave the most correlated results, with a correlation coefficient of r = 0.87. Bootstrap analysis were used to evaluate the number of loci for the markers and the coefficients of variation (CV) revealed a skewed distribution. The dominant markers (AFLP and RAPD) had small CV values indicating a skewed distribution while the codominant markers gave high CV values. The use of maximum values of genetic distance CVs within each sample size was efficient in determining the number of loci needed to obtain a maximum CV of 10%. The number of RFLP and AFLP loci used was enough to give CV values of below 5%, while the SSRs and RAPD loci gave higher CV values. Except for the RAPD markers, all the markers correlated genetic distance with single cross performance and heterosis which showed that they could be useful in predicting single cross performance and heterosis in intrapopulation crosses for broad-based populations. Our results indicate that AFLP seemed to be the best-suited molecular assay for fingerprinting and assessing genetic relationships among tropical maize inbred lines with high accuracy.