Methodology for kariological study of Brazilian Palms

Abstract

In the laboratory of cytogenetics of the DBAA-UNESP we are studying the karyotipe of some Brazilian Palms. To determine the best protocol, methods of seed germination, inhibition of mitosis, time to pick up the roots and staining were analyzed. The results shown that the seed germination in sphagnum is effective to achieve good roots. The best time to collect the root tips is between 11 to 12 AM., when there are more cell metaphases. The inhibition of mitosis cycle at metaphases may be effective both with 8-hydroxiquinoleine (0,03% -5 hours) or with cold water (0°C - 18-20 hours). The staining with Giemsa 2% showed the best chromosome figures in the metaphases. Now, to get good metaphases slides the method in use in the lab is: 1) seed germination in sphagnum at room temperature and high humidity; 2) The major roots are cut maintaining at least 5 cm, because this technic allows new emergence of roots, increasing the number of roots collected per germinated seed, that is very important in some species with poor germination rates; 3) To get the mitosis inhibition we are using cold water (0°C) treatment for 18-20 hours, following the standard protocols for conservation and hydrolysis; or enzyme digestion with pectinasecellulase 4) the staining procedures are made using Giemsa 2%. The Brazilian palms species studied and their respective chromosome number were: Aiphanes acanthophylla (2n=30), A. caryotaefolia (2n=30), Syagrus quinquifaria (2n=32), S. coronata (2n=32), S. romanzoffiana (2n=32), Euterpe edulis (2n=36), E. oleracea (2n=36), Copernicia prunifera (2n=36), Scheelea lauromuelleriana (2n=32) and Bactris gasipaes (2n=30).