Using of Molecular Markers in Plants Breeding

Currently, depending on the technique of execution and the characterized part of the genome, there are a large number of different DNA markers: RFLP, AFLP, RAPD, СAPS, SSR, SCAR, SNP, et al. Repeating SINE and LINE sequences can be such a genetic marker. They are retrotransposons or mobile genetic elements that are widely represented in the genomes of the entire eukaryotic kingdom and make up a large portion of genomic DNA (up to half of the plant genome). One of the earlier methods of DNAmarkers getting is Restriction Fragment Length Polymorphism, also named as RFLP. One of the earlier methods of DNA-markers getting is Restriction Fragment Length Polymorphism, also named as RFLP. The principle of this method founded on detection of specific nucleotide sequences in genomic DNA, with help of “blothybridization”. This method of analysis is in demand in our days, but in identification and differentiation of plants it has changed by methods, which basing on PCR-amplification of DNA with using of random selected primers, or primers, that compliment to known plots of genomic [1-3].

RAPD-markers. One of the most popular types of markers in mapping of plants genomics is RAPD (Random Amplified Polymorphic DNA). The method is in amplification of DNA of object during PCR with random primer, which has length of 10– 11 nucleotides. Method of detection of RAPD-markers is easy by technic execute. If the protocol has optimized, RAPD is convenient, easy and suitable for analyze of plants genomics. The method was used successfully for genome mapping of much agricultures, varieties marking and polymorphism of species. Using of RAPDmethod is especially perspective for genes mapping of quantitative attributes of plants [4,5].

AFLP (Amplified Restriction Fragment Length Polymorphism) – technology of getting random molecular markers with specific primers. According to this DNA are processing with combination of 2 restriction enzymes. Specific adapters are lysing with “sticky” ends of restrictive fragments. After this fragment are amplify with using primers that complimentary to sequence of adapter and to site of restriction, and additionally bearing one or more randomly selected bases at their 3’-ends. The set of fragments obtained depends on the restriction enzymes and randomly selected nucleotides at the 3’-ends of the primers [6].

SCAR markers. SCAR (Sequence Characterized Amplified Region) or an amplified region characterized by sequencing. Theseare PCR-based molecular markers derived from RAPD or ISSR fragments, which are devoid of most of the disadvantages of RAPD markers and are applicable for a variety of studies

According to numerous observations, the data obtained using RAPD analysis using random primers are quite relative due to the sensitivity of this method to reaction conditions. Therefore, to study polymorphic RAPD fragments and create specific DNA markers based on them, it became necessary to switch to the classical PCR method using a pair of long primers complementary to a specific sequence [5, 6].

CAPS. The CAPS (Cleaved Amplified Polymorphic Sequences) methodology, as well as SCAR, belongs to the STS (Sequence Tagged Site) group, since it is based on the amplification of strictly defined fragments of the genome with a known sequence. The principle of the method is as follows: genomic DNA is amplified using a pair of highly specific primers, then the resulting fragment is processed with a restriction endonuclease; differences between genomes manifest themselves in the form of different numbers and lengths of restriction fragments during agarose gel electrophoresis.

SSR-markers. Microsatellites, also called Simple Sequence Repeats (SSRs), are short tandem repeats of simple nucleotide motifs that contain from one to ten nucleotides in a repeating unit. They are found in all eukaryotic genomes, and in plants the most common repeats are (A) n, (AT) n and (GA) n motifs, where n varies between 10 and 80. Microsatellites, first obtained using PCR, are highly polymorphic markers for individual loci which are classified as dispersed tandem repeating sequences [6, 7].

The future is impossible without conducting biotechnological and molecular researches. One of the priority areas of research In Sumy National Agrarian University – Polymerase Chain Reaction (Pcr): An Alternative of Immunological Studies with the Determination of Disease Resistance Gene in Winter Bread Wheat Genotypes. The Purpose of the Research – breeding lines differentiation of bread winter wheat with resistance against diseases with Lr, Sr, Pm, Ug, Sr1AR genes for new varieties origination using molecular diagnostic methods. Materials of this Research – seeds of Triticum aestivum (cultivars-carriers of wheatrye translocations) and hybrid material newly created of bread winter wheat. The Tasks of the Work: to identify by means of the polymerase chain reaction of 1BL/1RS and 1AL/1RS wheat-rye translocations in bread winter wheat genotypes; to investigate the genetic effects of 1BL/1RS combinations and 1AL/1RS translocations in bread winter wheat genotypes; to carry out the genotype selection of bread winter wheat with a mix of signs from carriers of various wheat-rye translocations.

The perspectives for molecular research of bread wheat

• The identification of bread winter wheat lines with resistance to fungal diseases (Erysiphe graminis, Tilletia caries, Ustilago tritici, Puccinia recondita, Puccinia graminis in particular the TTKSK strain Ug99 strain, etc.) with an introgressive rye component (1RS);

• Combination in one genotype of the 1RS chromosome shoulder on genomes of different orders to get new-generation wheat lines that will provide the index increasing of the genetic potential realization of the grain crop productivity;

• Establishing the reasons for the significant presence of the pathogens on winter wheat;

• Recommendations for breeding for resistance and production of resistant varieties;

• Comparison of classical methods with molecular diagnostics

None.

No Conflicts of Interest.

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