Seleção de primers issr para Alcantarea nahoumii (Leme) J. R. Grant (Bromeliaceae), desenhos de primers trap e rt-qpcr para Abacaxi (Ananas comosus (L.) Merrill) visando a transferibilidade para Alcantarea nahoumii (Bromeliaceae)

Abstract

The Brazilian Atlantic Forest comprises a remarkably heterogeneous vegetation, including several related ecosystems, such as those that occur in rocky outcrops. The Bromeliaceae family has stood out as one of the most diverse families, frequent in these rocky environments. This family includes several species: some with great ornamental potential and threatened with extinction, such as Alcantarea nahoumii and others of great economic importance, such as the popular pineapple. Despite being highly vulnerable to the risk of extinction, there are no studies on genetic diversity with molecular markers for A. nahoumii, and no studies on gene expression. Given the above and aiming at molecular studies, this work aimed to amplify the genomic DNA of A. nahoumii with ISSR (Inter Simple Sequence Repeat) primer, design TRAP (Target Region Amplification Polymorphism) and RT-qPCR (Reverse Transcription Quantitative PCR) using the pineapple reference genome aiming at transferability to A. nahoumii, whose genome has not yet been sequenced. For the selection of ISSR primers, 10 primers were used for amplification of A. nahoumii DNA. For the design of TRAP and RT-qPCR primers, the selected EST sequences were those that encode enzymes involved in the process of embryogenesis and abiotic stress. These EST sequences correspond to highly conserved genomic regions among plants, which increases the probability of transferability between species of the same family. Of the 10 ISSR primers tested, 4 showed a good amplification pattern and could be used for future genotyping and genetic variability studies of A. nahoumii. 144 TRAP fixed primers were developed, of which 43 were indicated as the most promising. In addition, 255 pairs of RT-qPCR primers were designed, of which 130 met the ideal parameters. Obtaining primers is of paramount importance for molecular research. Through simple techniques, making use of bioinformatics programs, it becomes possible to acquire satisfactory results regarding the amplification of the desired DNA sequences.