ISSR Primer Selection for Genetic Variability Analyses with Jaborandi (Pilocarpus microphyllus Stapf ex Wardlew., Rutaceae)

Abstract

Pilocarpus microphyllus Stapf ex Wardlew. (jaborandi) is native to the North and Northeast regions of Brazil. Intensive exploitation for the extraction of pilocarpine, a valuable alkaloid for the pharmaceutical industry, has killed off or brought about a loss of vigor of naturally occurring plant populations. As a result, the species is officially listed as an endangered species of the Brazilian flora. Genetic diversity is required for the adaptation of populations to environmental changes, and maintaining it is a central objective for biological conservation. ISSRs are dominant markers widely used in genetic diversity studies of endangered species, allowing for the identification of genotypes and cultivars, as well as helping in phylogenetic studies based on DNA fingerprinting. This study presents an ISSR primer selection for genetic structure analyses of natural populations and cultivated collections of P. microphyllus.

Genetic diversity of ten Moroccan populations of Tetraclinis articulata as revealed by Inter Simple Sequence Repeat (ISSR) markers

“…An essential step before the analysis of the genetic diversity of a population is the development of appropriate markers to avoid those that either fail to amplify or generate only a few fragments and low polymorphism levels (Rocha et al, 2014). The selected primers have proved to be a reliable molecular tools for inter-populations genetic diversity studies of T. articulata and other conifers (Liu et al, 2013;Sánchez-Gómez et al, 2013;Tam and Hoa, 2006).…”

Seasonal change in main alkaloids of jaborandi (Pilocarpus microphyllus Stapf ex Wardleworth), an economically important species from the Brazilian flora

“…ISSR markers are known to be highly polymorphic in P . microphyllus [ 12 ] and will be informative for future studies on the genetic diversity of wild populations.…”

“…Eleven polymorphic ISSR primers used were from previous studies [ 12 ] ( Table 1 ). The samples were amplified by the polymerase chain reaction (PCR) using a modified version of the protocol [ 22 ].…”

“…However, since 2008, P . microphyllus has been listed as an endangered species in the Brazilian flora [ 11 , 12 ].Propagation of cultivars for research and industrial applications is therefore of fundamental importance for the species’ biological conservation and for reducing pressure on wild populations. Many low-income communities depend on harvesting P .…”

Caracterização e seleção de marcadores moleculares em Croton linearifolius Mull. Arg. como subsídio para estudos genéticos

“…As variações no percentual de iniciadores RGA e ISSR identificados com padrões de amplificação polimórficos é também observado em outros estudos disponíveis na literatura, a exemplo dos resultados de Chagas et al (2015) que selecionaram seis de 29 iniciadores ISSR testados em Elaeis guineensis, bem como dos resultados de Rocha et al (2014) que selecionaram 34 de 48 iniciadores testados em Pilocarpus microphyllus. A variação observada no percentual de iniciadores selecionados, no número de marcas e no nível de polimorfismo decorre, entre outros fatores, da particularidade das espécies investigadas, bem como do número de indivíduos testados.…”