Paulo Hideo Nakano Rangel

Analysis of genetic variability of South American wild rice populations (Oryza glumaepatula) with isozymes and RAPD markers

The knowledge of population structure and genetic diversity of wild relatives of rice is needed to investigate their evolutionary history and potential use in breeding programs. Very little is known about the wild rice species (Oryza spp.), particularly those that are native to South America. A study using isozyme and RAPD markers was conducted to estimate the level of genetic diversity of four South American wild rice populations (Oryza glumaepatula) recently collected in the Amazon forest and western Brazil rivers. F‐statistics and genetic diversity parameters calculated from isozyme and RAPD markers indicated high values for inbreeding coefficients and differentiation among the four populations. In agreement with this, a pattern of greater variation between than within populations was observed with both types of markers. These findings were corroborated by an AMOVA analysis, which indicated that a large portion of the total genetic variation was attributed to regional divergence. The partition of the AMOVA analysis among populations showed that most of the genetic diversity was due to differences among populations. This distribution pattern of genetic variation of O. glumaepatula populations is in agreement with the expectation for an autogamous species and provides important baseline data for conservation and collection strategies for this species.

Association mapping for yield and grain quality traits in rice (Oryza sativa L.)

Association analysis was applied to a panel of accessions of Embrapa Rice Core Collection (ERiCC) with 86 SSR and field data from two experiments. A clear subdivision between lowland and upland accessions was apparent, thereby indicating the presence of population structure. Thirty-two accessions with admixed ancestry were identified through structure analysis, these being discarded from association analysis, thus leaving 210 accessions subdivided into two panels. The association of yield and grain-quality traits with SSR was undertaken with a mixed linear model, with markers and subpopulation as fixed factors, and kinship matrix as a random factor. Eight markers from the two appraised panels showed significant association with four different traits, although only one (RM190) maintained the marker-trait association across years and cultivation. The significant association detected between amylose content and RM190 was in agreement with previous QTL analyses in the literature. Herein, the feasibility of undertaking association analysis in conjunction with germplasm characterization was demonstrated, even when considering low marker density. The high linkage disequilibrium expected in rice lines and cultivars facilitates the detection of marker-trait associations for implementing marker assisted selection, and the mining of alleles related to important traits in germplasm.

Determination of genetic variability of traditional varieties of Brazilian rice using microsatellite markers

The rice (Oryza sativa) breeding program of the Rice and Bean research center of the Brazilian agricultural company Empresa Brasileira de Pesquisa Agropecuaria (Embrapa) is well established and provides new cultivars every year to attend the demand for improved high yielding varieties with tolerance to biotic and abiotic stresses. However, the elite genitors used to compose new populations for selection are closely related, contributing to the yield plateau reached in the last 20 years. To overcome this limit, it is necessary to broaden the genetic basis of the cultivars using diverse germplasm such as wild relatives or traditional varieties, with the latter being more practical because they are more easily crossed with elite germplasm to accelerate the recovery of modern plant types in the breeding lines. The objective of our study was to characterize the allelic diversity of 192 traditional varieties of Brazilian rice using 12 simple sequence repeat (SSR or microsatellite) markers. The germplasm was divided into 39 groups by common name similarity. A total of 176 alleles were detected, 30 of which (from 23 accessions) were exclusive. The number of alleles per marker ranged from 6 to 22, with an average of 14.6 alleles per locus. We identified 16 accessions as a mixture of pure lines or heterozygous plants. Dendrogram analysis identified six clusters of identical accessions with different common names and just one cluster with identical accessions with the same common name, indicating that SSR markers are fundamental to determining the genetic relationship between landraces. A subset of 24 landraces, representatives of the 13 similarity groups plus the 11 accessions not grouped, was the most variable set of genotypes analyzed. These accessions can be used as genitors to increase the genetic variability available to rice breeding programs.

Microsatellite marker-mediated analysis of the EMBRAPA Rice Core Collection genetic diversity

The objectives of this study were to determine the genetic structure of 242 accessions from the EMBRAPA Rice Core Collection (ERiCC), to create a mini-core collection and to develop a multiplex panel of fluorescent labeled simple sequence repeats (SSRs). Eighty-six SSRs were used to identify 1,066 alleles, with an average number of 12.4 alleles/locus and average polymorphism information content (PIC)/locus of 0.75. A model-based clustering method recognized the structure of the accessions on two levels, according to their cultivation system and origin. The most divergent subgroup identified was the worldwide lowland accessions, with the highest values for gene diversity (0.75), average Rogers distance modified by Wright (0.80), average number of alleles/locus (11.7) and private alleles (132). A mini-core was assembled with the most divergent 24 lowland and upland accessions. This mini-core displayed an average distance of 0.86, an average number of alleles/locus of 8.4 and an average PIC/locus of 0.8. From the 86 SSRs, 24 were selected to compose six multiplex panels in order to optimize the process of rice genotyping. This set of markers distinguished all 242 accessions, and showed an average PIC of 0.80 and an average number of alleles/locus of 15.4, higher than the entire set of 86 SSRs. Since the heterogeneity found in lines and cultivars of ERiCC was higher than expected, it is necessary to analyze pooled DNA samples to get a better estimate of genetic variability. The SSR characterization of ERiCC clearly indicates that there is high genetic variability in rice accessions stored in genebanks worldwide which can be promptly explored by rice pre-breeding programs.

Genetic Structure of Wild Rice Oryza Glumaepatula Populations in Three Brazilian Biomes Using Microsatellite Markers

The existence of Oryza glumaepatula is threatened by devastation and, thus, the implementation of conservation strategies is extremely relevant. This study aimed to characterize the genetic variability and estimate population parameters of 30 O. glumaepatula populations from three Brazilian biomes using 10 microsatellite markers. The levels of allelic variability for the SSR loci presented a mean of 10.3 alleles per locus and a value of 0.10 for the average allelic frequency value. The expected total heterozygosity (He) ranged from 0.63 to 0.86. For the 30 populations tested, the mean observed (Ho) and expected heterozygosities (He) were 0.03 and 0.11within population, respectively, indicating an excess of homozygotes resulting from the preferentially self-pollinating reproduction habit. The estimated fixation index ( IS ) was 0.79 that differed significantly from zero, indicating high inbreeding within each O. glumaepatula population. The total inbreeding of the species (IT ) was 0.98 and the genetic diversity indexes among populations, ST and ST, were 0.85 and 0.90, respectively, indicating high genetic variability among them. Thus, especially for populations located in regions threatened with devastation, it is urgent that in situ preservation conditions should be created or that collections be made for ex situ preservation to prevent loss of the species genetic variability.

Ganho de produtividade pelo melhoramento genético do arroz irrigado no nordeste do Brasil

Os ganhos geneticos para produtividade obtidos pelo programa de melhoramento do arroz irrigado por inundacao na Regiao Nordeste do Brasil no periodo de 1984 a 1993 foram estimados visando avaliar a eficiencia do programa e tracar novas estrategias. Esta estimativa foi feita a partir dos dados de 59 ensaios regionais de rendimento conduzidos pelas empresas de pesquisa agropecuaria do Nordeste, em cooperacao com a Embrapa-Centro Nacional de Pesquisa de Arroz e Feijao (CNPAF), Goiânia, GO. O metodo estatistico utilizado baseia-se em medias ajustadas por modelo linear generalizado. O ganho genetico medio estimado foi de 54,9 ± 14,4 kg/ha/ano (0,8%). Nos ultimos tres anos houve uma tendencia de interrupcao dos ganhos. A pequena magnitude dos ganhos para producao nesta regiao podem ser atribuidos ao direcionamento do programa de geracao de linhagens da Embrapa para qualidade de graos e resistencia a doencas, as diferencas ambientais existentes entre Goiânia e a Regiao Nordeste e ao pequeno numero de ensaios conduzidos. A genealogia das linhagens foi tracada e verificou-se que os principais ancestrais sao os mesmos das cultivares recomendadas. A base genetica das linhagens e estreita, o que tambem pode estar contribuindo para a obtencao de pequenos ganhos geneticos para produtividade.

A new method to estimate genetic gain in annual crops

The genetic gain obtained by breeding programs to improve quantitative traits may be estimated by using data from regional trials. A new statistical method for this estimate is proposed and includes four steps: a) joint analysis of regional trial data using a generalized linear model to obtain adjusted genotype means and covariance matrix of these means for the whole studied period; b) calculation of the arithmetic mean of the adjusted genotype means, exclusively for the group of genotypes evaluated each year; c) direct year comparison of the arithmetic means calculated, and d) estimation of mean genetic gain by regression. Using the generalized least squares method, a weighted estimate of mean genetic gain during the period is calculated. This method permits a better cancellation of genotype x year and genotype x trial/year interactions, thus resulting in more precise estimates. This method can be applied to unbalanced data, allowing the estimation of genetic gain in series of multilocational trials.

Comparative linkage mapping of Oryza glumaepatula and Oryza sativa interspecific crosses based on microsatellite and expressed sequence tag markers

Molecular linkage maps representing the rice genome have been an important tool for breeding programs because they allow the elucidation of polygenic traits and are an efficient tool for monitoring wild introgressions in interspecific crosses. Common markers among rice genetic maps are important in defining the homology of chromosomes and the synteny between genomic target regions. We used 148 markers (expressed sequence tags, microsatellites and single nucleotide polymorphisms) to construct a molecular linkage map based on co-dominant markers for an interspecific backcross population using a wild rice (Oryza glumaepatula) from Brazil and performed a comparative analysis with other interspecific maps. The comparative analysis revealed a Spearman correlation index of 0.86 for marker order conservation to a previous map constructed for an interspecific cross using the same wild parent. Approximately 90% of markers common to other interspecific maps kept the same order. These results indicate that it will be possible to generate a unique genetic map using the wild donor and that it may be a helpful tool for breeding programs because plants derived from different interspecific populations can be rapidly scanned using markers associated with useful wild traits.

Hot spots for diversity of Magnaporthe oryzae physiological races in irrigated rice fields in Brazil

The objective of this work was to evaluate the Magnaporthe oryzae pathotype diversity in new commercial irrigated rice fields in the Araguaia River Valley, state of Tocantins, Brazil. The causal agent of rice blast has heavily affected rice production in the region. Despite the efforts of breeding programs, blast resistance breakdown has been recorded shortly after the release of new resistant cultivars developed for the region. Among the causes of resistance breakage is the capacity of the fungus to rapidly develop new pathotypes. A sample of 479 M. oryzae monosporic isolates was obtained and tested using the international rice blast differential set. Isolate collections were made in small areas designed as trap nurseries and in scattered sites in their vicinity. Analysis of 250 M. oryzae isolates from three trap nurseries indicated the presence of 45 international M. oryzae races belonging to seven pathotype groups (IA-IG). In the isolates tested, 61 M. oryzae pathotypes belonging to all but the IH group were detected. The new areas of irrigated rice in the Araguaia River Valley have the highest diversity of M. oryzae pathotypes reported so far in Brazil.

Adaptabilidade e estabilidade de acessos de uma coleção nuclear de arroz

The objective of this work was to determine the productive potential and the genotype by environment interaction of 550 accessions of the Embrapas Rice Core Collection. The performance, and yield adaptability and stability of each genotype was evaluated using the additive main effects and multiplicative interactions (AMMI) approach, in nine field experiments, carried out in six Brazilian states, under dry and irrigated conditions, during three years. The linear mixed model analysis was performed, and the estimates of the variance components were obtained by the residual maximum likelihood (REML), using the best linear unbiased prediction (BLUP) procedure for the prediction of the genotypic values of random effects (EBLUP) associated with each accession. The group of accessions with better performance was the one of lines and cultivars from Brazil (LCB), as expected. However, traditional varieties (VT) were identified among the best yielding genotypes, which shows the potential of this germplasm group to contribute as a new source of genetic variability to breeding programs. Superior accessions were identified for their stability, adaptability and grain yield, among which CA880078, CA840182 and CNA00091 stand out.