Towards an optimal sampling strategy for assessing genetic variation within and among white clover (Trifolium repens L.) cultivars using AFLP

Cost reduction in plant breeding and conservation programs depends largely on correctly defining the minimal sample size required for the trustworthy assessment of intra- and inter-cultivar genetic variation. White clover, an important pasture legume, was chosen for studying this aspect. In clonal plants, such as the aforementioned, an appropriate sampling scheme eliminates the redundant analysis of identical genotypes. The aim was to define an optimal sampling strategy, i.e., the minimum sample size and appropriate sampling scheme for white clover cultivars, by using AFLP data (283 loci) from three popular types. A grid-based sampling scheme, with an interplant distance of at least 40 cm, was sufficient to avoid any excess in replicates. Simulations revealed that the number of samples substantially influenced genetic diversity parameters. When using less than 15 per cultivar, the expected heterozygosity (He) and Shannon diversity index (I) were greatly underestimated, whereas with 20, more than 95% of total intra-cultivar genetic variation was covered. Based on AMOVA, a 20-cultivar sample was apparently sufficient to accurately quantify individual genetic structuring. The recommended sampling strategy facilitates the efficient characterization of diversity in white clover, for both conservation and exploitation

Análise da diversidade genética de cornichão com o uso de marcadores microssatélites Lotus corniculatus L. genetic diversity analysis through microsatellite markers

O objetivo neste trabalho foi analisar a diversidade genética de 14 materiais de Lotus corniculatus L. por meio de marcadores microssatélites. Foram analisados quatro cultivares e uma população de L. corniculatus e seus respectivos genótipos selecionados visando tolerância e sensibilidade ao alumínio. Foram utilizados 17 pares de primers que detectaram 36 alelos nos 17 locos microssatélites, com média de 2,25 alelos por loco. O resultado da análise de agrupamento com base nos índices de similaridade mostrou a formação de três grupos: um englobando germoplasmas e genótipos selecionados para sensibilidade ao alumínio e outros dois formados por genótipos selecionados visando tolerância ao alumínio tóxico. A análise molecular foi eficiente para detectar e quantificar a variabilidade entre os genótipos, acrescentando informações úteis ao programa de melhoramento. O uso de marcadores microssatélites permite a distinção de genótipos oriundos de um programa de seleção visando tolerância à toxidez por alumínio. As seleções realizadas originam genótipos polimórficos em relação às populações originais.<br>The aim of this work was to analyze the genetic diversity of 14 Lotus corniculatus L. materials through microsatellite markers. Four cultivars and one population of L. corniculatus and their respective genotypes selected were analyzed aiming aluminium tolerance and sensibility. The 17 markers used detected a total of 36 alleles, with an average of 2.25 alleles per locus. The result of the similarity analyses showed the formation of three groups: one enclosing germplasm and selected genotypes for aluminium sensibility, while the other two groups were formed by selected genotypes for aluminium tolerance. The molecular analysis was efficient in detecting and quantifying the variability among the genotypes, adding useful information to the breeding program. The utilization of microsatellite markers permitted the distinction of genotypes originated from a selection program aiming aluminium tolerance. The performed selections originate polymorphic genotypes in relation to the original populations