Estimation of outcrossing rate in a breeding population of Eucalyptus urophylla with dominant RAPD and AFLP markers

Eucalyptus breeding is typically conducted by selection in open-pollinated progenies. As mating is controlled only on the female side of the cross, knowledge of outcrossing versus selling rates is essential for maintaining adequate levels of genetic variability for continuous gains. Outcrossing rate in an open-pollinated breeding population of Eucalyptus urophylla was estimated by two PCR-based dominant marker technologies, RAPD and AFLP, using 11 open-pollinated progeny arrays of 24 individuals. Estimated outcrossing rates indicate predominant outcrossing and suggest maintenance of adequate genetic variability within families. The multilcous outcrossing rate (t(m)) estimated from RAPD markers (0.93 +/- 0.027), although in the same range, was higher (alpha > 0.01) than the estimate based on AFLP (0.89 +/- 0.033). Both estimates were of similar magnitude to those estimated for natural populations using isozymes. The estimated Wright's fixation index was lower than expected based on t, possibly resulting from selection against selfed seedlings when sampling plants for the study. An empirical analysis suggests that 18 is the minimum number of dominant marker loci necessary to achieve robust estimates of t,. This study demonstrates the usefulness of dominant markers, both RAPD and AFLP, for estimating the outcrossing rate in breeding and natural populations of forest trees. We anticipate an increasing use of such PCR-based technologies in mating-system studies, in view of their high throughput and universality of the reagents, particularly for species where isozyme systems have not yet been optimized.

THE USE OF AMPPD AS AN ALTERNATIVE SUBSTRATE FOR AP-MEDIATED DETECTION OF NONRADIOLABELED DNA PROBES IN EUCALYPTUS-SALIGNA

We present a non-radioactive alternative to Southern's (J. Mol. Biol. 98: 503-517, 1975) DNA-DNA hybridization technique. The use of AMPPD - Disodium 3-(4-Methoxyspiro {1,2-dioxetane-3,2'tricyclo[3.3.1.1(3,7)]decan}-4-yl)phyenyl phosphate as an alternative substrate for AP-mediated detection of digoxigenin-11 dUTP-labeled probes made possible the simple and nonhazardous reuse of blots. We used 0.8 % agarose gels containing 30 mug per lane of Eucalyptus saligna DNA, digested with Eco RI, electrophoresed and blotted on to nylon membranes (Hybond-N, Amersham, UK), using the Southern blotting procedure, and UV irradiated for one minute for DNA fixation. The hybridizations were carried out overnight with digoxigenin labeled random inserts of E. saligna DNA by using the Genius Kit (Boehringer Mannheim). Detection of the DNA-DNA hybrids was performed in the presence of 0.5% blocking agent and the substrates NBT/BCIP were replaced by 0.26 mM AMPPD in the final alkaline assay buffer (50 mul/cm2). After membrane incubation for five minutes at room temperature in a sealed plastic bag, the AMPPD solution was retrieved and stored at 4-degrees-C for reuse. A Kodak X-BRAF QA-S film was pressed firmly onto the bag containing the wet membrane, exposed for two to six hours and then developed. After use, the probes were stripped off and the blots reutilized, three times so far, with the same results.

Mantispa minuta Fabricius (Neuroptera : Mantispidae) on egg sacs of the orb-weaver spider Parawixia bistriata (Araneidae) in forests of Eucalyptus grandis in Brazil

This is the first report of the interaction between the two entomophagous agents Mantispa minuta Fabricius (Mantispidae) and Parawixia bistriata Rengger, an orb-weaver spider (Araneidae). From egg sacs kept in the laboratory we observed the emergence of adult neuropterans followed by the emergence of a high number of spiderlings. This association illustrates the interactions among the invertebrate predators coexisting in Eucalyptus plantations managed without chemical treatment, in the State of São Paulo, Brazil.

Morphological and anatomical studies of the seedes and seedlings of Eucalyptus citriodora and E. maculata

Morphological and anatomical aspccts of secds and seedlings of Eucalyptus citriodora Hook and E. maculara Hook were studied in detail and faund to be e.xtremely similar. However. the external characteristics of the secd. the secd coat anatomy and sorne features of the scedlings appeared to be very useful in the identificatian of these two economica11y important species.

Bird diversity and abundance in forest fragments and Eucalyptus plantations around an Atlantic forest reserve, Brazil

Little of Brazil's remaining Atlantic forest is protected, so it is important to assess how well the region's wildlife can persist in areas/habitats outside reserves. We studied bird diversity and abundance during 546 point counts in the Sooretama/Linhares reserve, 200 point counts in 31 forest fragments (10-150 h), and 50 point counts in < 30-year-old Eucalyptus plantations, within 7 km of the reserve. Only eight bird species were recorded in Eucalyptus, and this impoverishment, as compared to some Eucalyptus plantations elsewhere in Brazil may be a result of intensive clearance of understory vegetation. Species diversity in forest fragments was significantly lower than in the reserve. Twelve, mostly non-forest or edge species, were significantly commoner in the fragments, but nineteen species were frequent in the reserve but rare or absent in forest fragments. These included two Pyrrhura parakeets, a Brotogeris parakeet, a trogon Trogon, a jacamar Galbula, woodpeckers Piculus and Campephilus, Myrmotherula antwrens, and Hemithraupus and Tachyphonus tanagers. Bird species richness at points in forest fragments did not decline with fragment size, distance from the reserve, or forest quality. However, forest in fragments was more heavily degraded than forest within the reserve and poor forest quality may be the cause of declines in some species. Whilst protection of forest within reserves is a priority, management of forest fragments may aid conservation of some threatened species.

Morphological and anatomical studies of the seeds and seedlings of Eucalyptus pilularis and E. umbra

Morphological and anatomical aspects of seeds and seedlings of Eucalyptus pilularis Sm. and E. umbra R.T. Baker were studied in detail and fovnd to be similar, wlth the exception of sorne anatomical features of the testa and the average length of the fertile seeds, which is significantly different for the two species.

Eucalyptus ESTs related to genes for oxidative stress

Oxidative stress generating active oxygen species has been proved to be one of the underlying agents causing tissue injury after the exposure of Eucalyptus (Eucalyptus spp.) plants to a wide variety of stress conditions. The objective of this study was to perform data mining to identify favorable genes and alleles associated with the enzyme systems superoxide dismutase, catalase, peroxidases, and glutathione S-transferase that are related to tolerance for environmental stresses and damage caused by pests, diseases, herbicides, and by weeds themselves. This was undertaken by using the eucalyptus expressed-sequence database (https//forests.esalq.usp.br). The alignment results between amino acid and nucleotide sequences indicated that the studied enzymes were adequately represented in the ESTs database of the FORESTs project.

Density and spatial distribution of Atta sexdens rubropilosa and Atta laevigata colonies (Hym., Formicidae) in Eucalyptus spp. forests

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Acclimation to short-term low temperatures in two Eucalyptus globulus clones with contrasting drought resistance

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Sensitivity of mycobacteria to different species of eucalyptus l'herit

Sensitivity profile of 10 mycobacteria of clinical interest to eight different species of Eucalyptus L'Herit was studied. A Proportion Method Indirect Test was performed to detect resistant bacilli utilizing 5 and 10 mg/ml of essential oils incorporated to Lowestein-Jensen medium. Mycobaterium tuberculosis and other slow growing mycobacteria were more sensitive to essential oils than faster growing ones. The most effective essential oils, in decreasing order, were Eucalyptus citriodora, Eucalyptus maculata and Eucalyptus tereticornis.

Cytogenetic analysis of seven species of Eucalyptus L'Hér. (Myrtaceae)

Seven species of the genus Eucalyptus were studied cytogenetically (E. deanei, E. dunni, E. grandis, E. maculata E. propinqua, E. saligna and E. tereticornis). The species showed a symmetrical karyotype with 2n=22 chromosomes, with chromosome length ranging from 0.58 μm to 1.39 μm. Karyotypic analysis indicated homogeneity of morphology and of chromosome number for most of the species of this genus studied here, although casual disploid species with 2n=24 have been found in previous studies. According to these data, a basic number of x=11 was established for this genus. The evolutionary tendency probably occurred by structural alterations (deletions, duplications, additions and translocations) and in some cases by aneuploid chromosome alterations.

Anatomical and biochemical characterization of the calcium effect on eucalyptus urophylla callus morphogenesis in vitro

Calcium chloride concentrations from 0.0 to 12.12 mM were added to the culture medium and calcium content in calluses were determined directly by X-ray fluorescence spectrometry, a non-destructive method, allowing the processing of the same tissue for histological analysis. A multivariate statistical analysis (PCA - Principal Components Analysis) grouped the treatments into 5 blocks and indicated the most responsive group. Lack of calcium supply caused a complete absence of a morphogenic process and tissue collapse. An increase in calcium concentration gave higher total protein and sugar contents, an increase in peroxidase specific activity and changes in the histological characteristics. It was possible to verify that calcium stimulated globular somatic embryo formation at concentration of 6.62 mM.