Design and study of SCAR-markers for populations Mykizha Oncorhynchus (Parasalmo) mykiss from Kamchatka land.

 

M.N. Melnikova, S.D. Pavlov and N.V. Antipova

 

Department of Ichthyology, LomonosovMoscow State University, Moscow, 119899 Russia.

E-mail: melnik-06@mail.ru

 

Abstract: The species Oncorhynchus (Parasalmo) mykiss has a complex population structure, wide distribution, and high temp of microevolution. Also, this species is the one of the main object of aquaculture and fish farming. Thatís the one of the reason, why many scientists looking for different studies of mykiss genome.

As have been showed in our previous studies, the kamchatka mykiss has a low level of genome variability. The sequencing and restriction analysis of different genes (cytb; ND3; ND4; ATP6; ATP8; ITS1; ITS2; GH1; GH2; and others) of nuclear and mtDNA has obtain the lack of prospects of widespread salmonic gene markers forkamchatka mykiss populations.

All of this turns us to searching of new genetic markers for Oncorhynchus (Parasalmo) mykiss. One of methods of search of polymorphic sequences is the RAPD-method (Randomly Amplified Polymorphic DNA). However the RAPD-analysis reveals anonymous sites of genome and has low reproducibility of results. But, the SCAR-markers (Sequence Characterized Amplified Region) has originated from polymorphic RAPD-markers, and there are deprived the majority of this lacks and some of them can be used as unique markers of mykiss genome.

In our study, the searching of polymorphic RAPD-fragments for different mykiss populations has been done (including the resident and anadromous forms from western and east coast of Kamchatka, Shantarian and harvest samples). After, with these fragments, the SCAR-primers we designed.

We received 10 heterogeneous products in the size 100-800 bp, having population character of distinctions. These fragments were cut out, cloned and sequenced. The new SCAR-primers has a specific character and good repeatability. They give a single amplify product and identify the certain sites of DNA. In total 7 polymorphic SCAR-markers have been received.

 

key words: amplification, markers, polymorphism, genome, species, resident and anadromous forms.

 

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