The objective of this work was to analyze the effects of RAPD markers with skewed segregation on genetic linkage maps. Segregation data for 123 Citrus sinensis (L.) Osbeck cv. Pêra markers and 53 C. reticulata Blanco cv. Cravo markers in F1 progeny composed of 94 hybrids were used. Genetic linkage maps of the two varieties were constructed with non-skewed markers (p < 0.05 and p < 0.01) using the program MAPMAKER 3.0 and a pseudo-testcross strategy. The maps were compared to those constructed with all markers. Alterations in the genetic distances were observed based on the location of the skewed markers within the linkage groups. Generally, the skewed markers were located at the end of the linkage groups, sometimes forming entire linkage groups, without causing significant distance modifications. However, skewed markers located between non-skewed markers caused significant distance modifications and, in some cases, altered the order of the markers. Most of the skewed markers can be included in linkage maps, but in each case the degree of distance modification caused by each marker needs to be assessed.

O objetivo deste trabalho foi avaliar processos de seleção em uma população híbrida de citros, por meio de análise da segregação de marcadores RAPD. A segregação de 123 marcadores RAPD entre tangerina 'Cravo' (Citrus reticulata Blanco) e laranja 'Pêra' (C. sinensis (L.) Osbeck) foi analisada em uma progênie F1 composta por 94 híbridos. Composição genética, diversidade, heterozigosidade, diferenças estruturais dos cromossomos e presença de genes deletérios recessivos nos parentais foram discutidas com base nos tipos de segregação obtidos. Uma porcentagem elevada de marcadores com segregação distorcida em relação à proporção 1:1 esperada foi obtida na população F1. Muitos marcadores apresentaram segregação 3:1 em ambas as variedades e 1:3 em laranja 'Pêra' provavelmente decorrente de um processo de seleção direcional. A análise de distribuição da freqüência dos marcadores segregantes na população híbrida é um método simples que possibilita uma melhor compreensão da genética do grupo citros.

The objective of this work was to evaluate the processes of selection in a citrus hybrid population using segregation analysis of RAPD markers. The segregation of 123 RAPD markers between 'Cravo' mandarin (Citrus reticulata Blanco) and 'Pêra' sweet orange (C. sinensis (L.) Osbeck) was analysed in a F1 progeny of 94 hybrids. Genetic composition, diversity, heterozygosity, differences in chromosomal structure and the presence of deleterious recessive genes are discussed based on the segregation ratios obtained. A high percentage of markers had a skeweness of the 1:1 expected segregation ratio in the F1 population. Many markers showed a 3:1 segregation ratio in both varieties and 1:3 in 'Pêra' sweet orange, probably due to directional selection processes. The distribution analysis of the frequencies of the segregant markers in a hybrid population is a simple method which allows a better understanding of the genetics of citrus group.

Os marcadores moleculares têm sido utilizados como ferramentas em programas de melhoramento por hibridação sexual, permitindo a caracterização genética de grande número de genótipos. Os marcadores moleculares RAPD são os mais utilizados pois as técnicas empregadas são simples e de baixo custo. Avaliou-se a divergência genética entre híbridos F1 de tangerina 'Cravo' (Citrus reticulata Blanco) com laranja 'Pêra' (C. sinensis (L.) Osbeck) e estudou-se a variabilidade e a similaridade desses materiais entre si e em relação aos parentais. Reações RAPD, com 102 primers, foram realizadas em uma população de 94 híbridos e nos parentais. As análises multivariadas de divergência genética de componentes principais e agrupamento de Tocher foram realizadas, considerando-se, somente, os fragmentos polimórficos. As distâncias genéticas foram obtidas por meio do complemento aritmético do índice de Jaccard. Gráficos bidimensionais de dispersão das distâncias dos híbridos e parentais e da análise de divergência por meio de componentes principais foram construídos. Verificou-se elevada similaridade genética entre as variedades Cravo e Pêra e seus híbridos, demonstrando uma distribuição praticamente aleatória dos híbridos em relação aos parentais, porém em posições intermediárias. A análise de componentes principais apresentou aplicabilidade reduzida no estudo da divergência genética dos híbridos. O método de otimização de Tocher possibilitou a classificação dos híbridos e parentais em função de sua similaridade genética.

Molecular markers have been used as tools in breading programs of sexual hybridation, allowing the genetic characterization of a large number of genotypes. The RADP markers are the most used since the employed techniques are simple and of low cost. To evaluate the genetic divergence among F1 hybrids of 'Cravo' mandarin (Citrus reticulata Blanco) and 'Pêra' sweet orange (C. sinensis (L.) Osbeck), this study analyses the variability and similarity of the hybrids among themselves and with their parents. Random Amplified Polimorfic DNA marker analysis, with 102 primers, were applied to a population composed of 94 hybrids and their parents. Multivariate genetic divergence analysis of the principal components and Tocher grouping were carried out only considering the polymorphic fragments. Genetic distances were calculated by the arithmetic complement of the Jaccard index. Bidimensional dispersion graphs among hybrid and parent distances and of the divergence analysis by principal components were constructed. High genetic similarity among Cravo and Pêra varieties and their hybrids was verified, showing a casual distribution from the hybrids in relation to the parents, but in intermediary positions. The principal component analysis showed little applicability in the study of hybrid genetic divergence. The hybrids and parents were classified in groups based on the genetic similarity, using the Tocher optimization method.

Marcadores moleculares RAPD foram utilizados para avaliar a diversidade genética em uma população de 94 híbridos de tangerina 'Cravo' (Citrus reticulata Blanco) com laranja 'Pêra' (C. sinensis (L.) Osbeck), obtidos por polinização controlada. Nas reações de amplificação foram usados 102 "primers" decâmeros de seqüência arbitrária, que amplificaram 640 fragmentos, sendo 77,2% monomórficos entre os parentais. Utilizou-se o coeficiente de Jaccard para estimar a similaridade genética entre os híbridos, o método UPGMA para gerar o fenograma (NTSYS 1,7) e o software BOOD para determinar a consistência de cada agrupamento. Verificou-se que a laranja 'Pêra' apresenta maior heterozigosidade que a tangerina 'Cravo', sendo, respectivamente, 180 e 126 os números observados de loci em heterozigose. Houve alta similaridade genética entre os parentais, caracterizada pelo baixo polimorfismo de marcadores RAPD. Não houve a formação de agrupamentos estatisticamente significativos dos híbridos entre si e com os parentais, demonstrando que a constituição genética dos híbridos foi originada pela segregação independente das marcas RAPD, sendo quebrado o efeito de ligação em função do tamanho amostral em estudo. Híbridos com maior similaridade genética em relação à tangerina 'Cravo' e laranja 'Pêra' podem ser facilmente selecionados a partir de marcadores RAPD.

RAPD molecular markers were used to evaluate the genetic diversity in a progeny of 94 hybrids between 'Cravo' mandarin (Citrus reticulata Blanco) and 'Pêra' sweet orange (C. sinensis (L.) Osbeck), obtained by controlled pollination. One hundred two random decamer primers were screened, amplifying 640 fragments, 77.2% monomorphics between the parents. The Jaccard coefficient was used to estimate the genetic similarity among the hybrids, the UPGMA method to establish the phenogram (NTSYS 1.7) and the software BOOD to determine the significance of each cluster. The results indicated that 'Pêra' sweet orange heterozygosity degree is higher than 'Cravo' mandarin, with 180 and 126 loci in heterozygose, respectively. It was observed high genetic similarity between the parents, characterized by the low degree of RAPD polymorphism. Significant statistic groups were not established among the hybrids and the parents, showing that the hybrid genetic constitution was originated by independent RAPD markers segregation. Then, the ligation effect was broken because of the sample size. Hybrids with high genetic similarity degree in relation to 'Cravo' mandarin and 'Pêra' sweet orange could be easily selected using RAPD markers.

During some phases of of their life-cycle endophytic fungi colonize plants asymptomatically being found most frequently inside the aerial part of plant tissues. After surface disinfection of apparently healthy leaves from three varieties of mandarin orange and one tangor, and after incubation on appropriate culture medium, 407 fungal isolates were obtained, giving a total infection frequency of 81%. No fungal growth was observed from disinfected seeds, indicating that fungi are probably not transmitted via seeds. Of the fungal isolates, 27% belonged to the genus Guignardia, with 12 isolates being identified as Guignardia citricarpa Kiely, which is described as a citrus pathogen. The isolates were variable in respect to the presence of sexual structures and growth rates. Most of the isolates produces mature asci, supporting the hypothesis that they are nonpathogenic endophytes, which recently were identified as G. mangiferae. High intraspecific genetic variability (an average similarity coefficient of 0.6) was detected using random amplified polymorphic DNA (RAPD) markers generated by seven different primers. The highest similarity coefficient (0.9) was between isolates P15 and M86 and the smallest (0.22) between isolates P15 and C145. These results did not allow us to establish an association between genetic similarity of the fungal isolates and the citrus varieties from which they were obtained.