Inference of genetic diversity in popcorn S3 progenies.

Molecular markers are a useful tool for identification of complementary heterotic groups in breeding programs aimed at the production of superior hybrids, particularly for crops such as popcorn in which heterotic groups are not well-defined. The objective of the present study was to analyze the genetic diversity of 47 genotypes of tropical popcorn to identify possible heterotic groups for the development of superior hybrids. Four genotypes of high genetic value were studied: hybrid IAC 125, strain P2, and varieties UENF 14 and BRS Angela. In addition, 43 endogamous S3 progenies obtained from variety UENF 14 were used. Twenty-five polymorphic SSR-EST markers were analyzed. A genetic distance matrix was obtained and the following molecular diversity parameters were estimated: number of alleles, number of effective alleles, polymorphism information content (PIC), observed and expected heterozygosities, Shannon diversity index, and coefficient of inbreeding. We found a moderate PIC and high diversity index, indicating that the studied population presents both good discriminatory ability and high informativeness for the utilized markers. The dendrogram built based on the dissimilarity matrix indicated six distinct groups. Our findings demonstrate the genetic diversity among the evaluated genotypes and provide evidence for heterotic groups in popcorn. Furthermore, the functional genetic diversity indicates that there are informative genetic markers for popcorn.

Genetic diversity of popcorn genotypes using molecular analysis.

In this study, we analyzed dominant molecular markers to estimate the genetic divergence of 26 popcorn genotypes and evaluate whether using various dissimilarity coefficients with these dominant markers influences the results of cluster analysis. Fifteen random amplification of polymorphic DNA primers produced 157 amplified fragments, of which 65 were monomorphic and 92 were polymorphic. To calculate the genetic distances among the 26 genotypes, the complements of the Jaccard, Dice, and Rogers and Tanimoto similarity coefficients were used. A matrix of Dij values (dissimilarity matrix) was constructed, from which the genetic distances among genotypes were represented in a more simplified manner as a dendrogram generated using the unweighted pair-group method with arithmetic average. Clusters determined by molecular analysis generally did not group material from the same parental origin together. The largest genetic distance was between varieties 17 (UNB-2) and 18 (PA-091). In the identification of genotypes with the smallest genetic distance, the 3 coefficients showed no agreement. The 3 dissimilarity coefficients showed no major differences among their grouping patterns because agreement in determining the genotypes with large, medium, and small genetic distances was high. The largest genetic distances were observed for the Rogers and Tanimoto dissimilarity coefficient (0.74), followed by the Jaccard coefficient (0.65) and the Dice coefficient (0.48). The 3 coefficients showed similar estimations for the cophenetic correlation coefficient. Correlations among the matrices generated using the 3 coefficients were positive and had high magnitudes, reflecting strong agreement among the results obtained using the 3 evaluated dissimilarity coefficients.

Construction of an integrated genetic map for Capsicum baccatum L.

Capsicum baccatum L. is one of the five Capsicum domesticated species and has multiple uses in the food, pharmaceutical and cosmetic industries. This species is also a valuable source of genes for chili pepper breeding, especially genes for disease resistance and fruit quality. However, knowledge of the genetic structure of C. baccatum is limited. A reference map for C. baccatum (2n = 2x = 24) based on 42 microsatellite, 85 inter-simple sequence repeat, and 56 random amplified polymorphic DNA markers was constructed using an F2 population consisting of 203 individuals. The map was generated using the JoinMap software (version 4.0) and the linkage groups were formed and ordered using a LOD score of 3.0 and maximum of 40% recombination. The genetic map consisted of 12 major and four minor linkage groups covering a total genome distance of 2547.5 cM with an average distance of 14.25 cM between markers. Of the 152 pairs of microsatellite markers available for Capsicum annuum, 62 were successfully transferred to C. baccatum, generating polymorphism. Forty-two of these markers were mapped, allowing the introduction of C. baccatum in synteny studies with other species of the genus Capsicum.

Functional molecular markers (EST-SSR) in the full-sib reciprocal recurrent selection program of maize (Zea mays L.).

This study aimed to improve grain yield in the full-sib reciprocal recurrent selection program of maize from the North Fluminense State University. In the current phase of the program, the goal is to maintain, or even increase, the genetic variability within and among populations, in order to increase heterosis of the 13th cycle of reciprocal recurrent selection. Microsatellite expressed sequence tags (EST-SSRs) were used as a tool to assist the maximization step of genetic variability, targeting the functional genome. Eighty S1 progenies of the 13th recur-rent selection cycle, 40 from each population (CIMMYT and Piranão), were analyzed using 20 EST-SSR loci. Genetic diversity, observed heterozygosity, information content of polymorphism, and inbreeding co-efficient were estimated. Subsequently, analysis of genetic dissimilarity, molecular variance, and a graphical dispersion of genotypes were conducted. The number of alleles in the CIMMYT population ranged from 1 to 6, while in the Piranão population the range was from 2 to 8, with a mean of 3.65 and 4.35, respectively. As evidenced by the number of alleles, the Shannon index showed greater diversity for the Piranão population (1.04) in relation to the CIMMYT population (0.89). The genic SSR markers were effective in clustering genotypes into their respective populations before selection and an increase in the variation between populations after selection was observed. The results indicate that the study populations have expressive genetic diversity, which cor-responds to the functional genome, indicating that this strategy may contribute to genetic gain, especially in association with the grain yield of future hybrids.

Use of elephant grass for energy production in Campos dos Goytacazes-RJ, Brazil.

Elephant grass is a tropical forage plant widely distributed throughout Brazil. It was first exclusively used in the livestock sector as cattle feed. The grass is characterized by its high productivity and photosynthetic capacity and is considered as an alternative source of renewable energy. Here, we estimated the general combining ability of the parents and specific combining ability of the hybrids based on morpho-agronomic biomass-quality traits. The experiment was conducted in a randomized block design with 3 replicates. The diallel was composed of 16 hybrids and 2 groups of genitors. In the diallel analysis of variance, we observed a significant difference among treatments. A significant difference was observed among genitors for dry matter production (DMP). For the general combining ability of group 1, the traits leaf blade width, DMP, height, percentage of neutral detergent fiber, percentage of hemicellulose, percentage of lignin, percentage of acid detergent fiber, and percentage of cellulose were significant. For the estimates of general combining ability of DMP, parents Porto Rico 534-B, Vruckwona, Taiwan A-146, and Mercker S. E. A. were 0.4748, 3.2819, 1.1659, and 0.4317. The parents of Mercker S. E. A. and Porto Rico 534-B produced the highest percentage of detergent fiber and percentage of lignin with values of 0.1482 and 0.0856. Thus, parents Vruckwona, Porto Rico 534-B, and Taiwan A-146 are promising for integration into breeding programs. The best hybrid combinations for DMP were 1 x 5, 1 x 8, 2 x 6, 3 x 7, and 4 x 5.

Genetic-molecular characterization of backcross generations for sexual conversion in papaya (Carica papaya L.).

The low number of improved cultivars limits the expansion of the papaya crop, particularly because of the time required for the development of new varieties using classical procedures. Molecular techniques associated with conventional procedures accelerate this process and allow targeted improvements. Thus, we used microsatellite markers to perform genetic-molecular characterization of papaya genotypes obtained from 3 backcross generations to monitor the inbreeding level and parental genome proportion in the evaluated genotypes. Based on the analysis of 20 microsatellite loci, 77 genotypes were evaluated, 25 of each generation of the backcross program as well as the parental genotypes. The markers analyzed were identified in 11 of the 12 linkage groups established for papaya, ranging from 1 to 4 per linkage group. The average values for the inbreeding coefficient were 0.88 (BC1S4), 0.47 (BC2S3), and 0.63 (BC3S2). Genomic analysis revealed average values of the recurrent parent genome of 82.7% in BC3S2, 64.4% in BC1S4, and 63.9% in BC2S3. Neither the inbreeding level nor the genomic proportions completely followed the expected average values. This demonstrates the significance of molecular analysis when examining different genotype values, given the importance of such information for selection processes in breeding programs.

Use of microsatellite markers in molecular analysis of segregating populations of papaya (Carica papaya L.) derived from backcrossing.

Brazil is the world leader in papaya production. However, only a small number of cultivars are registered for commercial planting, mainly owing to delays in obtaining cultivars and the high costs of the field phase of breeding programs. These costs can be reduced when molecular tools are combined with conventional breeding methods. In the present study, we conducted a molecular analysis of a self-fertilized population of a first backcrossing generation of BC1S1 papaya plants via microsatellite markers both to monitor the level of homozygosity and the gene/allele transfer that confers the Golden trait (fruit color) and to assess the parental genomic proportion in the genotypes studied. Based on the analysis of 20 polymorphic microsatellite loci, 19 genotypes with the Golden trait belonging to BC1S1 were evaluated in addition to the parental genotypes. Genetic distance was estimated through weighted index. The genotypes were then grouped using the hierarchical nearest neighbor method, and the analysis of principal coordinates was used to measure the proportion of parental genomes in the segregating genotypes. The mean value of the inbreeding coefficient was 0.36. The analysis of the principal coordinates revealed that on average, 64% of the recurrent parent genome was present in the population. Together, the analyses allowed the selection of 3 individuals for the next backcross cycle (33BC1S1-18, 34BC1S1-16, and 37BC1S1-10). These individuals had a higher proportion of the recurrent parent and were grouped close to the recurrent parent in the cluster analysis.

Full-sib reciprocal recurrent selection in the maize populations Cimmyt and Piranão.

We estimated the genetic gains of the 12th cycle of reciprocal recurrent selection for maize traits of agronomic interest. We used 23 ISSR molecular markers in an attempt to maximize genetic variability among and within populations based on selection of S(1) progenies. To this end, 138 full-sib families were evaluated in a randomized block design in two environments (the municipalities of Campos dos Goytacazes and Itaocara, in the State of Rio de Janeiro, Brazil), with replications within sets. Direct selection for grain yield was used for the selection of the families. To assess genetic diversity among and within populations, we examined plants produced from part of the S(1s) seeds from the parents that originated the 42 full-sib families that were selected from the agronomic traits. Direct selection for grain yield provided good gains for the traits evaluated, with estimated improvement of -0.87 days for days to flowering, 0.35 plants, 1.79 ears per plot, 0.58 g per 100-grain weight, 308.21 g ear weight per plot, and 261.83 kg/ha grain yield. Application of molecular markers at the stage of superior progeny selection led to increased genetic distance among populations, which is a very important factor for utilization of heterosis and providing greater longevity to the reciprocal recurrent selection program.

Multivariate analysis to determine the genetic distance among backcross papaya (Carica papaya) progenies.

Morpho-agronomic and molecular (RAPD and ISSR markers) data were used to evaluate genetic distances between papaya backcross progenies in order to help identify agronomically superior genotypes. Thirty-two papaya progenies were evaluated based on 15 morpho-agronomic characteristics, 20 ISSR and 19 RAPD primers. Manhattan, Jaccard and Gower distances were used to estimate differences based on continuous and binary data and combined analyses, respectively. Except for production, there were significant differences in the continuous variables among the genotypes. The molecular analysis revealed 193 dominant markers (ISSR and RAPD), being 53 polymorphic loci. Among the various clusters that were generated, the one based on a combined analysis of morpho-agronomic and molecular data gave the highest cophenetic correlation (0.72) compared to individual analysis, consistently allocating the progenies into six groups. We found that the Gower algorithm was more coherent in the discrimination of the genotypes, demonstrating that a combination of molecular and agronomic data is valuable for studies of genetic dissimilarity in papaya.

Use of molecular markers in reciprocal recurrent selection of maize increases heterosis effects.

We examined the effect of incorporation of molecular markers on variability between and within populations in order to maximize heterotic effects and longevity of a maize reciprocal recurrent selection program. Molecular variability was quantified by inter-simple sequence repeat (ISSR) markers between and within the maize populations Cimmyt and Piranão in the 10th cycle of a reciprocal recurrent selection program. Forty-two S(1) progenies of each population were analyzed, these being families of full-sibs selected according to their agronomic traits. Thirteen primers were selected, which produced 140 bands; 114 of them were polymorphic and 26 monomorphic. Based on UPGMA grouping analysis and by genetic distances, it was possible to identify "contaminant" progenies. These progenies belong to the Piranão or Cimmyt groups, but cluster in the opposite heterotic group. Identification of "contaminant" progenies is relevant for selection, because, besides identifying genotypes that should be eliminated at the recombination stage, it allows increased heterosis expression in crosses between more genetically distinct individuals. After the elimination of the "contaminant" progenies and those that were allocated between the heterotic groups, a new statistical analysis was carried out, which demonstrated increased genetic distances between the populations. It was concluded that the application of molecular markers in reciprocal recurrent selection programs allows the optimization of the monitoring of genetic variability within and between populations, favoring recombination between more distant progenies, besides ensuring increased longevity of the reciprocal recurrent selection program.

Genetic characterization of papaya plants (Carica papaya L.) derived from the first backcross generation.

The limited number of papaya varieties available reflects the narrow genetic base of this species. The use of backcrossing as a breeding strategy can promote increases in variability, besides allowing targeted improvements. Procedures that combine the use of molecular markers and backcrossing permit a reduction of the time required for introgression of genes of interest and appropriate recovery of the recurrent genome. We used microsatellite markers to characterize the effect of first-generation backcrosses of three papaya progeny, by monitoring the level of homozygosity and the parental genomic ratio. The homozygosity level in the population ranged from 74 to 94%, with a mean of 85% for the three progenies (52-08, 52-29 and 52-34). The high level of inbreeding found among these genotypes increases the expectation of finding more than 95% fixed loci in the next generation of self-fertilization of superior genotypes. The mean proportion of the recurrent parent genome found in first-generation backcross progeny was 50.1%; 52-34 had a larger genomic region in common with the recurrent genitor and the lowest level of homozygosity. The progeny 52-08 was genetically closest to the donor genitor, and it also had the highest level of homozygosity. We found that linking conventional procedures and molecular markers contributed to an increase in the efficiency of the breeding program.