ISHS Acta Horticulturae 473: International Symposium on the Importance of Varieties and Clones in the production of Quality Wine
PCR/RAPD TO DETERMINE THE GENOTYPE OF WOODEN ROOTSTOCK CUTTINGS |
| Authors: | T. Wolf, K. Eimert, E. Bleser, R. Ries |
| Abstract:
Traditionally, grapevine varieties are identified by morphological characteristics, e.g. leaf shape and size, type and density of hairs on shoot tips and leaves. This method depends both on green shoots and in many cases on specific stages of plant development. Furthermore also environmental factors affect morphological characteristics of plants. To become independent of environmental influence and plant development, molecular biological techniques e.g. RFLP and RAPD/PCR were soon adapted to grapevines. Most previous studies on grapevine identification dealt with scion varieties, whereas publications on rootstocks are quite rare. Rootstocks, treated as disbudded material, do not have enough morphological characteristics for variety identification. Therefore the aim of this study was to develop a procedure to identify rootstock varieties from dormant cuttings. During these investigations a method for DNA-extraction from one-year old grapevine cuttings was developed by adapting LODHI's DNA-extraction procedure. This modified method allows a fast reliable and inexpensive DNA-extraction. It yields good DNA quality and purity. DNA, extracted by this procedure, was amplified by RAPD/PCR reaction and fractionated on an agarose-gel. Overall 48 RAPD (random amplified polymorphic DNA) -primers were tested to find polymorphisms useful for the identification of the seven most common rootstock varieties in Germany (Teleki 8B, Kober 125AA, Kober 5BB, 5C Geisenheim, SO4, 3309C and Boerner). It proved to be possible to distinguish all seven rootstocks using two primers only. Therefore the method is suitable for routine variety determination of dormant cuttings. | |
Download Adobe Acrobat Reader (free software to read PDF files) | |
