**Table 1.**

*Advances in Grape and Wine Biotechnology*

parents can be determinable. The main point of the idea is based on the fact that in most of the angiosperms, the endosperm is triploid with two genome equivalents from the maternal line and one from the paternal line [5]. Our presumption was that this numeral difference in the maternal and paternal alleles causes measureable

To validate our method, pre-experiments were carried out on 12 Pinot gris seeds.

Fully matured berries were collected from Pinot gris (clone B. 10) stocks in September 2016. Seeds were removed from the berries, washed in tap water and dried in room temperature. Only fully matured, brown-coloured seeds were used in the analyses. Pinot noir, Pinot gris (clone B. 10.), Chardonnay and Cabernet sauvignon DNA (extracted formerly from leaves) were also used as controls. DNA was extracted from 12 single seeds. Prior to extraction the seeds were crushed in a mortar to powder; then, this powder was moved to a tube. Qiagen Plant Mini Kits were used for DNA extraction following the instructions of the manufacturer. The amount and quality of DNA were determined spectrophotometrically.

The SSR analysis was performed at 19 loci (see **Table 1**). PCR reaction mix was the following: 0.2 mM of each primer, 12.5 ml of Hot Start Master Mix (Quiagen) and 50 ng of template DNA, completed to the total volume of 25 ml with DNA- and

difference in the amplification of SSR alleles.

The DNA was diluted to a concentration of 10 ng/ml.

**2. Materials and methods**

*Process of double fertilisation [2].*

**Figure 1.**

**50**

*List of SSR loci [6].*

RNA-free distilled water. The following thermal profile was used: (1) 94°C for 45 min; 35 cycles of: (2) 94°C for 1 min, (3) annealing temperature (see **Table 1**) for 1 min, 73°C for 1 min; and (4) 73°C for 7 min. Each forward primer of the primer pairs was fluorescently labelled with 6FAM on the 5′ end of the DNA chain. Separation of the amplified products was carried out through capillary electrophoresis in a PE-Applied Biosystems 3100 Automated Capillary DNA Sequencer; the molecular sizes of the products were determined using Peak Scanner Software (v. 1.0; Applied Biosystems) [8]. Allele sizes and peak area were also recorded for every single allele. Data were stored in Microsoft Excel [8].
