We have extended the combined use of the "pseudo-testcross" mapping strategy and RAPD markers to map quantitative trait loci (QTLs) controlling traits related to vegetative propagation in Eucalyptus. QTL analyses were performed using two different interval mapping approaches, MAPMAKER-QTL (maximum likelihood) and QTL-STAT (non-linear least squares). A total of ten QTLs were detected for micropropagation response (measured as fresh weight of shoots, FWS), six for stump sprouting ability (measured as # stump sprout cuttings, #Cutt) and four for rooting ability (measured as % rooting of cuttings, %Root). With the exception of three QTLs, both interval-mapping methods yielded similar results in terms of QTL detection. Discrepancies in the most likely QTL location were observed between the two methods. In 75% of the cases the most likely position was in the same, or in an adjacent, interval. Standardized gene substitution effects for the QTLs detected were typically between 0.46 and 2.1 phenotypic standard deviations (σp), while differences between the family mean and the favorable QTL genotype were between 0.25 and 1.07 (σp). Multipoint estimates of the total genetic variation explained by the QTLs (89.0% for FWS, 67.1 % for#Cutt, 62.7% for %Root) indicate that a large proportion of the variation in these traits is controlled by a relatively small number of major-effect QTLs. In this cross, E. grandis is responsible for most of the inherited variation in the ability to form shoots, while E. urophylla contributes most of the ability in rooting. QTL mapping in the pseudo-testcross configuration relies on withinfamily linkage disequilibrium to establish marker/trait associations. With this approach QTL analysis is possible in any available full-sib family generated from undomesticated and highly heterozygous organisms such as forest trees. QTL mapping on two-generation pedigrees opens the possibility of using already existing families in retrospective QTL analyses to gather the quantitative data necessary for marker-assisted tree breeding.

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We used allozyme markers to estimate the amount of natural hybridization between Eucalyptus grandis and E. urophylla in a 7.4-hectare commercial hybrid-seed orchard planted in Espírito Santo, Brazil. This orchard was planted in 1982 using a honeycomb design, with each hexagonal plot containing one E. grandis tree surrounded by six E. urophylla trees. There were 267 replicated hexagonal plots in the orchard. Seeds were harvested from the E. grandis clone only. The multilocus outcrossing rate estimated for the E. grandis clone averaged 70.2%, ranging from 33.0 to 99.0% among individual trees. Contaminant pollination, inferred from progeny genotypes containing alleles not present in the seven parental clones, accounted for 14.4% of the hybrid seed. Contaminant pollen was attributed to neighboring eucalyptus stands isolated from the orchard by a 400-m wide belt of native forest. Inbred and hybrid progenies were identified by their allozyme genotypes and transplanted to the field. Field growth of inbred progeny was 30% lower than that of hybrid plants at two and three years of age.
Utilizamos aloenzimas como marcadores para estimar o grau de hibridação natural entre Eucalyptus grandis e E. urophilla em um pomar de semente híbrida comercial instalado no Espírito Santo, Brasil. Esse pomar, que compreende 7,4 hectares, foi plantado em 1982 e é constituído por 267 parcelas hexagonais. Em cada parcela a matriz E. grandis, produtora de sementes, está cercada pelos seis clones de E. urophilla, doadores de pólen. A taxa de fecundação cruzada (tm), estimada simultaneamente por vários locos de aloenzimas, foi da ordem de 70,2%, variando entre 33 e 99% em árvores individuais. Genótipos encontrados na progênie contendo alelos não presentes nos sete clones parentais permitiram detectar a contaminação do pomar por pólen de outra origem. Esses genótipos constituíram 14,4% da progênie híbrida. A origem do pólen contaminante foi atribuída a povoamentos de eucaliptos vizinhos, isolados do pomar por uma faixa de floresta nativa com 400 metros de largura. Os híbridos e as plantas resultantes de autofecundação puderam ser identificados por meio de seus genótipos de aloenzimas. Todas as plantas examinadas foram transplantadas para o campo. Medições feitas no segundo e terceiro ano após o transplante mostraram que o crescimento da progênie de autofecundação foi 30% inferior ao da progênie híbrida

Quantitative trait loci (QTL) mapping of forest productivity traits was performed using an open pollinated half-sib family of Eucalyptus grandis. For volume growth, a sequential QTL mapping approach was applied using bulk segregant analysis (BSA), selective genotyping (SG) and cosegregation analysis (CSA). Despite the low heritability of this trait and the heterogeneous genetic background employed for mapping, BSA detected one putative QTL and SG two out of the three later found by CSA. The three putative QTL for volume growth were found to control 13.7% of the phenotypic variation, corresponding to an estimated 43.7% of the genetic variation. For wood specific gravity five QTL were identified controlling 24.7% of the phenotypic variation corresponding to 49% of the genetic variation. Overlapping QTL for CBH, WSG and percentage dry weight of bark were observed. A significant case of digenic epistasis was found, involving unlinked QTL for volume. Our results demonstrate the applicability of the within half-sib design for QTL mapping in forest trees and indicate the existence of major genes involved in the expression of economically important traits related to forest productivity in Eucalyptus grandis. These findings have important implications for marker-assisted tree breeding.

Self-incompatibility found in a Eucalyptus grandis clone was used to promote interspecific hybridization between E. grandis and E. urophylla clones. The interspecific hybridization achieved in an open-pollinated commercial seed orchard planted in Espírito Santo, Brazil was evaluated by the multi-locus outcrossing rate (tm) of the seed producing clone, E. grandis. The percentage of outcrossed seeds reached 95.9%. The outcrossing rate of individual trees was quite variable, but was always above 70.0%. Wright's coefficient was negative (F = -0.30) revealing an excess of heterozygous genotypes in the progeny. Genetic parameters confirmed the high degree of hybridization expected in this orchard. The 800-m wide protection belt of native tropical forest that surrounds this orchard has significantly reduced pollen contamination, in comparison to a 400-m wide belt used in another local seed orchard.
Verificamos a eficiência do uso da auto-incompatibilidade encontrada em um clone de Eucalyptus grandis para aumentar a hibridação interespecífica entre E. grandis e E. urophylla em um pomar clonal para produção de semente comercial, instalado no Espírito Santo, Brasil. O nível da hibridação interespecífica alcançada nesse pomar foi avaliado por meio da taxa de fecundação cruzada, tm, do clone de E. grandis produtor de semente. A taxa (tm) foi estimada usando-se vários locos de isoenzimas simultaneamente. A percentagem de sementes híbridas alcançou 95,9%. A taxa de fecundação cruzada das árvores individuais foi variável, mas sempre superior a 70%. O coeficiente de Wright foi negativo (F = -0,30), indicando haver um excesso de genótipos heterozygotos na progênie. Os parâmetros genéticos estimados confirmaram o alto nível de hibridação esperado nesse pomar. A barreira de proteção, constituída por uma faixa de 800 metros de floresta nativa, reduziu significativamente a contaminação do pomar por pólen de outros povoamentos de eucalipto, em comparação à faixa de 400 metros de largura utilizada em outro pomar na mesma região

-Studies on the QTL expression and the adequacy of methodologies involved in the analysis