Page 140 - Zbornik_Konf_2019

https://doi.org/10.15414/2019.9788055220703

4 International Scientific Conference Abstracts Book
th
VARIABILITY OF MICROSATELLITE SEQUENCES DISTRIBUTION IN CULTURAL AND
WILD GENOTYPES OF AMARANTH (AMARANTHUS L.) GENOMES
Veronika Štefúnová, Milan Bežo, Monika Prostredná

Department of Genetics and Plant Breeding, Slovak University of Agriculture in Nitra, Slovak Republic;
E-mail.: veronika.stefunova@uniag.sk
The aim of the study was to assess variabilty and distribution of microsatellite
sequences in 15 cultural genotypes of Amaranthus caudatus L., Amaranthus cruentus L. and
Amaranthus hypochondriacus L. and 14 wild genotypes of Amaranthus hybridus L.,
Amaranthus palmeri S. Watson, Amaranthus retroflexus L., Amaranthus spinosus L. and
Amaranthus tuberculatus (Moq.) Sauer. Polymorphism between cultural and wild species of
amaranth (Amaranthus L.) was analyzed using ISSR markers.
The biological material (seeds) was obtained from the North Central Regional PI Station
(NC7), Iowa State University, Ames, USA. The DNA extraction was performed by Rogers and
Bendich optimized method. Primers (CA)6AG, (CA)6GG, (GT)6CC, (GTG)3GC, (GAG)3GC and
(GA)6CC were used for ISSR analysis. PCR was performed with 20 ng of DNA, 20 mmol.dm
-3
Tris-HCl, pH 8, 50mmol dm KCl, 1 U Taq polymerase, 3 mmol.dm MgCl2, 0.1 mmol. dm
-3
-3
-3
deoxyribonucleotides, 0.2 μmol. dm primer. The following program was used for
-3
amplification: 94 °C for 2 min, 45 cycles of 94 °C for 1 min, 50 °C for 1 min, 72 °C for 2 min and
a final extension 72 °C for 7 min. Fragments were separated on 2 % agarose gel. Based on
amplified DNA fragments, a matrix of genetic distances of the presence and position of the
DNA fragments was assembled. The interdependencies were constructed by the UPGMA
hierarchical cluster analysis based on genetically related coefficients in STATISTIKA program.
The most synthesized fragments were provided by primer (GAG)3GC. The least
synthesized fragments were provided when using (CA)6GG primer in PCR. Nei Li coefficient
values ranged from 0.36 to 0.65. The lowest Nie Li value was provided using (CA)6AG primer
in PCR.
Markers ISSR have great potential for generating a large number of informative
characters for phylogenetic analysis. Two primers (CA)6AG and (CA)6GG used in PCR revealed
clustering of four Amaranthus hypochondriacus genotypes originated in Nepal and India in a
separate cluster. Primer (GA)6CC revealed similar results, four Amaranthus hypochondriacus
genotypes in one cluster together with wild Amaranthus retroflexus genotype originated in
India. Primer (GAG)3GC revealed independent cluster with Amaranthus palmeri genotype
originated in Mexiko.
On the basis of repetitive sequences (GT)6CC and (CA)6AG distribution, genetic similarity
between Amaranthus caudatus L. genotypes and Amaranthus tuberculatus (Moq.) Sauer was
assessed. ISSR with (GTG)3GC primer revealed the genetic similarity between Amaranthus
caudatus L. and Amaranthus spinosus L. genotypes in a separate cluster.

Keywords: amaranth, Amaranthus caudatus, Amaranthu cruentus, Amaranthus hypochondriacus,
Amaranthus hybridus, Amaranthus palmeri, Amaranthus retroflexus, Amaranthus spinosus, Amaranthus
tuerculatus, ISSR, diversity.
Acknowledgement
The publication was prepared within the project VEGA 2/0109/19 'Morpho-physiological, Genetic and
Biochemical Response of Amaranth (Amaranthus spp.) to Heavy Metal Stress'.

|140 4 International Scientific Conference Agrobiodiversity Nutrition, Health and Quality of Human and Bees Life
th
September 11–13, 2019