**4. Discussion**

*Advances in Grape and Wine Biotechnology*

Maternal and paternal haplotypes were determined by the separation of mater-

Based on our results, it can be established that most of the seeds are originating from the selfing of 'Pinot gris', but cross-fertilisation is appearing in some cases, such as in the case of seed no. 4, where in six loci the paternal allele is absent from

**Locus Maternal haplotype Paternal haplotype 'Pinot gris' genotype** VMC8A7 **158 158** 158:158 VVMD28 **236 236** 218:236 VrZag79 **242 248** 242:248 VVMD7 **240** *238* 240:244 VMC1C10 **157 157** 157:157 VVS2 **150 134** 134:150 VMC3D12 **236** *234* 200:236 VMC5G8 **315 311** 311:315 Scu06vv **164 172** 164:172 VMC5E9 **220** *218* 216:220 VMC7G3 **116** *114* 116:116 VrZag21 **205** *195* 199:205 VMC4G6 **122 122** 122:122 VMC1F10 **190 208** 190:208 VrZag25 **237 225** 225:237 VMC2H4 **206** *204* 206:224 VMCNG1E1 **124 124** 124:124 VVIM10 **335 335** 335:339

*Example for the determination of maternal and paternal allele in the case of seed no. 11 and VMC4A1 locus:* 

*maternal allele, 270; paternal allele, 282 (maternal genotype, 270:278).*

*Determination of parental haplotypes in the case of seed no. 4. (Alleles of 'Pinot gris' are red coloured, bold* 

*letters; alleles not in 'Pinot gris' genotype are blue coloured, italic letters) [9].*

nal and paternal alleles based on the method described previously (**Table 2**).

**52**

**Table 2.**

**Figure 2.**

Such amount of DNA can be extracted from a single grape seed, which is suitable for SSR analyses. The amplification of DNA is successful in most of the loci; the PCR reaction optimised for other plant parts can be applied.

Our method is safely applicable for the determination of parental haplotypes. Based on the determined haplotypes, the parental identity could be determined by the use of databases.

The method could be suitable for the analyses of archaeological grapevine seeds, with the following limitations:

The quality and quantity of the extracted DNA could be poor because of the degradation; it depends on the age of the seeds and the environmental effects, e.g. carbonisation processes [10].

Mutations can occur in the SSR loci during the time, which can be manifested in the occurrence of the so-called null alleles. Null alleles result in amplification failures, which can be rid by the use of shortened primers in the PCR reactions ([11, 12]).
