Colletotrichum lindemuthianum, causal agent of anthracnose in the common bean, has wide genetic variability. Differential bean cultivars and morphological and physiological characteristics were used to analyze 74 isolates of C. lindemuthianum collected in two counties in the state of Minas Gerais, Brazil. Six different races were found, with a predominance of race 65 at both locations. Isolates were classified according to their sensitivities to the fungicide thiophanate-methyl, normally used in the control of common bean anthracnose. In all, ≈10% of isolates were resistant to the fungicide in vitro. Characteristics such as indexes of mycelia growth rate, colony diameter, sporulation capacity, and percentage of germination demonstrated the high genetic variability of C. lindemuthianum. We also observed variation in conidial cytology. The conidia of most isolates showed septa formation after germination, in contrast to septa absence, previously reported in the literature. Sexual and asexual reproduction were evaluated for mechanisms that may contribute in the generation of variability in C. lindemuthianum. Conidial anastomosis tubes were commonly found, indicating that asexual reproduction can help increase variability in this species. Information from this study confirmed high variability in C. lindemuthianum and will guide future studies in basic knowledge and applied technologies.

Anthracnose caused by Colletotrichum lindemuthianum is an important disease of common bean, resulting in major economic losses worldwide. Genetic diversity of the C. lindemuthianum population contributes to its ability to adapt rapidly to new sources of host resistance. The origin of this diversity is unknown, but sexual recombination, via the Glomerella teleomorph, is one possibility. This study tested the hypothesis that Glomerella strains that are frequently recovered from bean anthracnose lesions represent the teleomorph of C. lindemuthianum. A large collection of Glomerella isolates could be separated into two groups based on phylogenetic analysis, morphology, and pathogenicity to beans. Both groups were unrelated to C. lindemuthianum. One group clustered with the C. gloeosporioides species complex and produced mild symptoms on bean tissues. The other group, which belonged to a clade that included the cucurbit anthracnose pathogen C. magna, caused no symptoms. Individual ascospores recovered from Glomerella perithecia gave rise to either fertile (perithecial) or infertile (conidial) colonies. Some pairings of perithecial and conidial strains resulted in induced homothallism in the conidial partner, while others led to apparent heterothallic matings. Pairings involving two perithecial, or two conidial, colonies produced neither outcome. Conidia efficiently formed conidial anastomosis tubes (CATs), but ascospores never formed CATs. The Glomerella strains formed appressoria and hyphae on the plant surface, but did not penetrate or form infection structures within the tissues. Their behavior was similar whether the beans were susceptible or resistant to anthracnose. These same Glomerella strains produced thick intracellular hyphae, and eventually acervuli, if host cell death was induced. When Glomerella was co-inoculated with C. lindemuthianum, it readily invaded anthracnose lesions. Thus, the hypothesis was not supported: Glomerella strains from anthracnose lesions do not represent the teleomorphic phase of C. lindemuthianum, and instead appear to be bean epiphytes that opportunistically invade and sporulate in the lesions.

Native and exotic Brazilian plants collected in the State of Minas Gerais were evaluated for their anticancer potential. Methanol extracts from leaves of 51 plant species were tested for cytotoxicity against four tumor cell lines: B16 (murine skin), HL-60 (human leukemia), MCF-7 (human breast), and HCT-8 (human colon). Plant extracts that exhibited IC(50) values less than 30 microg/ml against any tumor cell line were tested on sea urchin egg development and mouse erythrocytes. In addition, all extracts were evaluated for their general toxicity using the brine shrimp lethality assay. The most active extracts against the tumor cells were those obtained from Lantana fucata, Copaifera langsdorffii, and Momordica charantia. These three extracts inhibited sea urchin development from the first cleavage, but those from C. langsdorffii and M. charantia were very active against mouse erythrocytes. Only the L. fucata extract presented no hemolytic activity. Consequently, although the extracts of L. fucata, M. charantia, and C. langsdorffii could be useful in the development of new anticancer products, the first of these extracts is the most promising since it did not present unspecific toxicity, as suggested by negative results obtained with brine shrimp lethality and mouse erythrocytes assays.

ABSTRACT. Glomerella cingulata f. sp phaseoli is the sexual phase of the fungus Colletotrichum lindemuthianum, the causal agent of common bean anthracnose. This fungus is of great concern, because it causes large economic losses in common bean crops. RAPD markers of five populations of G. cingulata f. sp phaseoli from two Brazilian states were analyzed to determine if this population possesses the sexual reproductive potential to generate the genetic variation that is observed in this phytopathogen. We identified 128 polymorphic bands, amplified by 28 random primers. The estimates of genetic similarity in this analysis ranged from 0.43 to 1.00, and the dendrogram generated from analysis of all genotypes displayed five principal groups, coinciding with the five populations. Genetic differentiation was observed between the populations (G ST = 0.6455); 69% of the overall observed genetic variation was between individual populations and 31% of the variance was within the sub-populations. We identified significant levels of linkage disequilibrium in all populations. However, the values of the disequilibrium ranged from low to moderate, indicating that this pathogen maintains a genetic structure consistent with sexual reproduction. The mean contribution of sexual reproduction was determined by comparison of the amplitudes of genetic similarity of isolates from sexual and asexual phases. These results support the hypothesis that recombination plays an important role in determining the amplitude of variability in this pathogen population and that this determination occurs on a fine scale.

©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 18 (4): gmr18357 A.V. Tiago et al. 2 (SSRY47), with a mean of 0.680. The expected and observed heterozygosity showed an average of between 0.717 and 0.688, for SSRY126 and SSRY47, respectively. The heterozygosity values observed were higher than those expected in five of the six populations, generating negative values of the fixation index (-0.070). Among the six populations, Alta Floresta and Cuiabá had the highest percentage of polymorphic loci (100%). The groupings obtained by UPGMA, Structure and PCoA among the six populations were concordant in allocating the individuals into two genetic groups. We found considerable genetic diversity among the samples, evidenced by the high values in the diversity indices. These high values are possibly related to the management of the fields and the exchange of propagative material among the farmers. Therefore, it is proposed that both populations be conserved since they have potential that could be used for genetic improvement of this essential crop.

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Cassava (Manihot esculenta) is a traditional crop in tropical and subtropical regions that is used for human consumption and in animal feed. This studied cassava landraces cultivated in northern Mato Grosso State, Brazil, to detect the variability in family farms, aiming at the preservation and use potential of these genetic resources. A total of 120 M. esculenta individuals were evaluated at the rate of 40 per location (population), where each landrace was represented by 10 plants. The 14 microsatellite markers examined showed genetic diversity. The average polymorphic information content (PIC) was 0.621 and the average number of alleles per locus was seven. Genetic diversity parameters indicated that H o was higher than H e for the three populations, showing negative fixation indices and a lack of inbreeding. AMOVA revealed greater molecular variation within the populations (92%). Bayesian analysis and the ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 18 (3): gmr18315 E.C.M. Pedri et al. 2UPGMA clustering method resulted in two main groups formed with the individuals distributed randomly; i.e., regardless of collection site (location). The gene flow found in this study is a consequence of the introduction and exchange of genetic material (landraces) performed by the farmers, who act on their farms as maintainers of local diversity. Manihot esculenta cultivation in northern Mato Grosso State ensure the on-farm conservation of genetic variability of the species and constitutes a source of genetic resources such as genes of resistance and adaptation, which can be exploited and utilized in breeding programs.

income mainly from cassava cultivation and flour production for the market, direct their variety choices to those that are most productive. Brava variety was the most frequent (found in eight family units) and was considered the most profitable for the production of flour Network analysis showed that propagule circulation and information occurs between the residents and also with other communities of the region, which are important sources of new varieties. Two farmers were identified as the most active in this network, showing potential as key elements for the circulation of propagating material. According to the cluster analysis using the genetic data, the most recently introduced varieties (Baixinha, Liberatona, Broto roxo, Mansa, Ramo branco, Carneiro and Cuiabana) are separated from those introduced a long time ago. The varieties pointed out as bitter by the farmers were also grouped together. The results showed the importance of traditional farmers in maintaining a high genetic diversity of manioc varieties, despite the directing of the choice of varieties to meet market needs.