Biologia reprodutiva em acessos de Lippia alba (Mill.) N.E. Brown (Verbenaceae)

Abstract

The reproductive biology of a species is closely related to the genetic basis of a population and is one of the main factors related to the dispersion and success of angiosperms. Lippia alba (Verbenaceae) is known by several common names as erva cidreira, falsa melissa, chá de tabuleiro, being widely used in folk medicine mainly due to its essential oil components. The species can be easily found from tropical to temperate regions, having broad phenotypic plasticity. Recent studies indicate a possible autopoliploidy origin for the species having diploids (2n = 30), aneuploids (2n = 38), triploids (2n = 45), tetraploids (2n = 60) and hexaploid (2n = 90) individuals. Despite the importance of Lippia alba, little information about the species' reproductive system is available. The goal of the present work was to study the reproduction strategy of the species to better understand the origin of the five cytotypes as well as to help the elucidation of the genetic basis of the species. Inflorescences of forty-one individuals were isolated from March 2015 to December 2016. The number of produced seeds was compared between isolated and nonisolated inflorescences. Fresh seeds from 20 accessions were collected and the measures of length and width were taken. Then the seeds were placed to germinate in petri dishes with moistened filter paper under temperatures ranging from 18°C-25°C-40°C (in 24 hour cycles). To estimate the DNA content, a single seed at a time was chopped in Marie nuclear extraction buffer at 4°C. The solution was filtered through a 30 μm filter and stained with propidium iodide. For genotyping, DNA was extracted from seeds collected in isolated and non-isolated inflorescences. Leaves were used for DNA extraction of the mother plant of eight accessions that were compared using microsatellite primers. For a cytohistological analysis of the buds’ development, samples were collected before the anthesis and were fixed in glutaraldehyde solution for 24h. Comparative morphological analyzes between diploid, triploid and tetraploid accessions were also performed, using five floral measures. Seeds originating from diploids had a higher germination index when compared to those from the polyploids and the ones originated from isolated inflorescences generally did not germinate. The set of results allowed to conclude that Lippia alba has preferentially an allogamic reproductive behavior, being different between diploids and polyploids. The cytohistological and cytometric analyzes did not allow to exclude the occurrence of apomixis reproduction. The results suggest that Lippia alba has alternative ways of reproduction.