**3. Results**

DNA extraction and amplification in 18 loci (out of the 19) were successful in all of the 12 seeds. In VVMD5 the amplification was weak, so the results were unevaluated. In all of the remaining 18 loci, the maternal and parental alleles were determined (**Figure 2**); according to our presumption, the allele with the higher area value was supposed to be maternal. Ratio of the quantity of maternal and paternal alleles was computed based on area values. In some loci (where 'Pinot gris' has a homozygote genotype and the majority of the seeds showed also homozygote genotype— VMC4G6 and VMCNG1E1), this ration was excluded from the further analyses.

Based on the remaining 16 loci, the average ratio of maternal and paternal alleles ranged from 1.89 (VMC5E9) to 2.58 (VrZag25), which confirms our presumption.

Maternal and paternal haplotypes were determined by the separation of maternal and paternal alleles based on the method described previously (**Table 2**).

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

#### **Figure 2.**

*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).*


#### **Table 2.**

*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].*

**53**

**Author details**

provided the original work is properly cited.

Gyöngyi Knolmajerné Szigeti and János Májer

*Reconstruction of Parental SSR Haplotypes from a Single Grape Seed*

from the pollinator, which is surely different from 'Pinot gris'.

PCR reaction optimised for other plant parts can be applied.

'Pinot gris' (blue coloured, italic letters in **Table 2**). These alleles were inherited

for SSR analyses. The amplification of DNA is successful in most of the loci; the

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

The method could be suitable for the analyses of archaeological grapevine seeds,

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.

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

The file from **Figure 1** is created under the Creative Commons Attribution

Such amount of DNA can be extracted from a single grape seed, which is suitable

*DOI: http://dx.doi.org/10.5772/intechopen.85685*

**4. Discussion**

the use of databases.

([11, 12]).

**Licencing**

licence [13].

with the following limitations:

carbonisation processes [10].

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Gizella Győrffyné Jahnke\*, Zóra Annamária Nagy, Csaba Németh,

NARIC Research Institute for Viticulture and Enology, Hungary

\*Address all correspondence to: jahnke.gizella@szbki.naik.hu

'Pinot gris' (blue coloured, italic letters in **Table 2**). These alleles were inherited from the pollinator, which is surely different from 'Pinot gris'.
