DESIGN OF SPECIFIC IDENTIFICATION METHODOF FRUIT RAW MATERIAL BASED ON MOLECULAR GENETIC ANALYSIS

• Main Author: Golubtsova Yu.V.
• Format: Article
• Language: Russian
• Published: Kemerovo State University 2017-03-01
• Series: Техника и технология пищевых производств
• Subjects: fruit raw material; species identification; plant material; molecular genetic analysis; sequencing of fruit raw material
• Online Access: http://fptt.ru/stories/archive/44/17.pdf

Species identification of fresh fruit raw material plays an important role in the quality control of finished products. The article presents the results of research aimed at the design of species identification method of fruit raw material: Ribes uva-crispa (gooseberry of «The Cooperator» variety), Rosa majalis Herrm (Rosa majalis), Prunus fruticose (ground cherry of «The Altaiskaya Lastochca» variety), Actinidia deliciosa (kiwi delicacy) applying molecular genetic analysis. Using a variety of software packages, as well as GenBank NCBI database for each of the objects of fruit raw material at the level of generic differentiation we managed to find a suitable DNA segment for further development, on its basis, of universal primers - section 18S rDNA. A unified alignment matrix for each of the objects of fruit raw material with the help of GeneDoc program has been developed; recheck of each matrix for the presence of read errors or other controversial single nucleotide substitutions has been carried out. The authors have conducted an algorithmic sequence analysis and the search for optimum positions of primer setting using PrimerQuest program with the indication of the maximum size of the amplicon, readable by a primer pair not exceeding 300 bp. The optimum parameters of the amplification process are the following: primers’ volume of 0.5 l; amplification conditions: 95 °C, 60 seconds; 62 °C, 45 seconds; 72 °C, 30 seconds; 30 cycles have been determined. The selected amplification mode is confirmed by the results of PCR with all samples of fruit raw material with visualization in the form of electrophoregram in 1% gel. Additional investigation of primers’ specificity using the BLAST algorithm has been done. It has been found that all sequences of fragments readable with each of the pairs of primers designed coincide with deposited in Genbank sequences of investigated raw material.