Use of molecular markers (microsatellites) for genome dactyloscopy

salina@bionet.nsc.ru 

Description

Genome dactyloscopy (DNA finger-printing) is widely used for individuals genetic identification and applied in finding out paternity in criminalistics. Methods of genome dactyloscopy having been described and used earlier are based on laborious molecular-genetic processes with the use of radioactive-marked probes.
Genome dactyloscopy (GD) is also topical for plants. Genetic identification of various lines and cultivars allows to protect breeders authorship rights and to avoid illegal distribution of prospective breeding stocks.
Microsatellite markers have become widely spread in the analysis of wheat genome for the recent years. Our multi-year working experience in this type of markers points out their following advantages over other markers known in scientific issues: 
1. high polymorphism in common wheat genome combined with high specificity for each DNA sample, 
2. fast operation with this method, 
3. low expenses, 
4. no radioactivity. 
The suggested GD method of wheat cvs using microsatellite markers consists of a number of stages: 
1. detecting genomic DNA out of 5-10 individuals of each cv, 
2. matching primers to wheat microsatellite zones (2-4 per chromosome) based on literary data and own experience in operating the marker type, 
3. PCR for each plant individual and pair of primers towards microsatellite zones, 
4. gel electrophoresis with PCR products to determine the length of individual microsatellite locus, 
5. interpretation of results as for database on these markers obtained during research of this marker type for the last 5 years, 
6. compiling the cv passport on the data of genome dactyloscopy (see below). 
A unique combination of microsatellite loci is specific for each cv and it always allows to differentiate one cv from the other. 

Figure and Caption

Passport samples of wheat cvs based on the data of 17 microsatellite loci:

Saratovskaya 29 SKALA Tselinnaya 20 IRTYSHANKA Novosibirskaya 67 Length of marking fragmentin n.o.
      12   90-95
    12   1 96-100
12   1     101-105
  1, 12   1 12 111-115
1   5, 15     121-125
  5, 7, 8   5 5, 6, 13 126-130
15 15   13, 15 15 131-135
5, 7, 13 6, 10, 13 7, 13 10   136-140
6, 10   2, 9, 10   8, 10 141-145
9 2   2, 9 9 146-150
  9 6 6   151-155
2 14   14 14 156-160
        2 161-165
14, 17   8, 14     166-170
  17     17 171-175
3   17 17   176-180
      3   181-185
  3     3 186-190
    3     191-195
8     8   201-205
16         241-245
    16 16 16 246-250
  16       271-275
        4 311-315
  4       316-320
4     4   321-325
    4     326-330
          331-335
11   11 11   336-340
        11 341-345
  11       351-355

Wheat cvs under study are indicated below the table. Numbers of microsatellites are in figures. 

Length of marking fragment is on the right and to be determined for each certain cv after PCR with indicated markers and further finding out the length of marking fragment by electrophoresis related to length standards. 

Technical-Economic Advantages

Genome dactyloscopy based on the analysis for combination of microsatellite loci allows to differentiate the cv from thousands of others. It provides legal protection of authorship rights for produced cvs.

Fields of Application

Agriculture.

Level and Location of Practical Realisation

Full-scale molecular-genetic analysis carried out at the Institute of Cytology and Genetics, SB RAS.

Patent(s)

'Know-how'.

Commercial Proposals

Agreement on collaborative scientific-research activities.

Contacts:

Helen A. Salina, 
Candidate of Biological Sciences (PhD)
Institute of Cytology and Genetics, SB RAS
10 Lavrentiev ave.,
Novosibirsk 630090
Phone: +7(383) 333-37-19
Fax: +7(383) 333-12-78
E-mail: salina@bionet.nsc.ru