Page 112 - Zbornik_Konf_2019

https://doi.org/10.15414/2019.9788055220703

4 International Scientific Conference Abstracts Book
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CHARACTERIZATION OF BACTERIAL ENDOPHYTES OF AMARANTHUS CULTIVARS BY
NEW GENERATION SEQUENCING TECHNOLOGY
Matúš Kyseľ , Jana Žiarovská , Juraj Medo , Andrea Hricová 3
1
1
2
1 Slovak University of Agriculture in Nitra, Faculty of Agrobiology and Food Resources, Department of
Genetics and Plant Breeding, Nitra, Slovak Republic; E-mail.: jana.ziarovska@uniag.sk
2 Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of
Microbiology, Nitra, Slovak Republic
3 Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center Slovak Academy
of Sciences, Nitra, Slovak Republic
Endophytic bacteria are a natural part of plant tissues and are revealed to be important
for different functions in plants that lead to the production characteristics of them as well as
the plant health background. The analysis of intracellular bacterial variability has been
broadening in the last years by the methods of a high throughput sequencing approach that
provide new insights to the large ecodiversity that bacteria possess. The metagenomic
analysis is one of the very modern approaches used for microbial communities study. Here,
metagenomic sequencing was used to the preliminary characterization of two Amaranthus
cultivars – Pribina and Zobor. Endogenous metagenomic profiles of bacterial populations of
wheat cultivars and amaranth genotypes were obtained by NGS (MiSeq) technology.
Young leaves were collected and surface sterilized. DNA was extracted using the
GeneJet™ Plant Genomic Extraction Kit (Thermo Scientific) following the manufacturer
instruction. The amplification of approximately 500 bp long conserved regions of 16S rRNA
genes was performed by PCR using the universal bacterial primers that were modified for the
purposes of using them in the Illumina NextSeq sequencer. Nextera® XT Index Kit was used
to add the dual index barcodes to the amplicons. The sequencing procedure was performed on
Illumina MiSeq using MiSeq Kit V3 600 cycles kit with pair-end 300 bp sequencing
The presence of the genera Endobacter sp., Dichotomicrobium sp., Burkholderia sp. In
Pribina cultivar was obtained and cultivar Zobor was characterized by the presence of
propagated halophilic bacteria (genus Limimonas spp.), culminating thermophilic bacteria
(genera Oceanotoga spp. and Orientia spp.), and receded acidophilic bacteria (genera
Endobacter spp. and Abiotrophia spp.).
Our study revealed for the first time the endophytic bacteria analysis in amaranth
Pribina and Zobor cultivars using the Illumina sequencing protocol.

Keywords: Amaranthus, Pribina, Zobor, metagenomics.
Acknowledgement
The study was granted by VEGA 2/0109/19 – Morfo-fyziologická, genetická a biochemická odpoveď
rastlín láskavca (Amaranthus spp.) na stres vyvolaný ťažkými kovmi; VEGA 2/0025/15 – Endofytické
mikroorganizmy a ich potenciálna úloha pri zvyšovaní tolerancie drevín voči stresu and ITMS
26220220180 Budovanie Výskumného centra AgroBioTech.

|112 4 International Scientific Conference Agrobiodiversity Nutrition, Health and Quality of Human and Bees Life
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September 11–13, 2019