Studying polymorphism through ISSR markers of the genus Miscanthus and Salix bioenergy cultures representatives

Abstract

The study aimed to analyze the genome of miscanthus and energy willow, which are included in the collection of Institute of Bioenergy Crops and Sugar Beet NAAS of Ukraine, by the ISSR – PCR method to assess their genetic diversity and identify the level of polymorphism. The development of bioenergy in the world and in Ukraine has prompted farmers to focus on using crops as bioenergy plants that that did not used to be the object of their attention. Classical laboratory methods for PCR analysis were used in the work, namely DNA isolation, polymerase chain reaction (ISSR analysis), electrophoretic distribution of the obtained PCR products in agarose gel, statistical method. DNA isolation was performed by standard CTAB extraction method. Total DNA was isolated from vegetative organs, separately from each plant. Three ISSR primers were used to analyze the molecular genetic polymorphism of the genus Miscanthus. As a result of amplifcation, 14 loci were obtained, 13 of which were polymorphic. The locus polymorphism index ranged from 0.83 to 0.95. 100 % polymorphism was detected with the help of ISSR 2 and ISSR 4 markers, as 11 loci obtained with their participation were limorphic. The use of ISSR 1 allowed to obtain three alleles: one allele was found in M. sinensis Andersson, two alleles were found in M. sacchariflorus (Maxim.) Franch., And three alleles were found in M. giganteus JM Greef & Deuter ex Hodkinson. Two microsatellite primers ISSR2 and ISSR 4 were used to analyze the molecular genetic polymorphism of the genus Salix. in the range of 0.2–0.8. That is, there are amplicons that are present in most samples. Amplicons with a frequency of 0.2 and a length of 570 and 720 bp are found in two samples. The largest number of polymorphic loci was obtained using primer ISSR 4. A large number of alleles for M. gigantheus was detected, which confrms its hybrid origin. Also, the ISSR 4 primer can be used to differentiate members of the genus Salix, because it found amplicons with a length of 570 and 720 bp. with a frequency of 0.2, which are found in two samples – Clone of the Swedish willow and Willow wool and it is advisable to use it to differentiate members of the genus Salix. The use of the obtained data allows to estimate the genetic diversity of existing species of the genus Miscanthus and Salix for more accurate detection. Key words: genome, genus Miscanthus, genus Salix, ISSR polymorphism, loci.