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International journal of molecular sciences
Nov 02, 2019;20 (21):

Allelic Variants for Candidate Nitrogen Fixation Genes Revealed by Sequencing in Red Clover ( L.).

Old��ich Trn��n��, David Vlk, Eli��ka Mackov��, Michaela Matou��kov��, Jana ��epkov��, Jan Ned��ln��k, Jan Hofbauer, Karel Vejra��ka, Hana Jake��ov��, Jan Jansa, Lubom��r Pi��lek, Daniela Knotov��
Author Information
  1. Old��ich Trn��n��: Agricultural Research, Ltd., Zahradn�� 1, 664 41 Troubsko, Czech Republic. trneny.oldrich@gmail.com.
  2. David Vlk: Department of Experimental Biology, Masaryk University, 625 00 Brno, Czech Republic. Vlk.DavidR@email.cz.
  3. Eli��ka Mackov��: Department of Experimental Biology, Masaryk University, 625 00 Brno, Czech Republic. mackova.e.94@gmail.com.
  4. Michaela Matou��kov��: Agricultural Research, Ltd., Zahradn�� 1, 664 41 Troubsko, Czech Republic. matouskova@vupt.cz.
  5. Jana ��epkov��: Department of Experimental Biology, Masaryk University, 625 00 Brno, Czech Republic. repkova@sci.muni.cz.
  6. Jan Ned��ln��k: Agricultural Research, Ltd., Zahradn�� 1, 664 41 Troubsko, Czech Republic. nedelnik@vupt.cz.
  7. Jan Hofbauer: Agricultural Research, Ltd., Zahradn�� 1, 664 41 Troubsko, Czech Republic. hofbauer@vupt.cz.
  8. Karel Vejra��ka: Agricultural Research, Ltd., Zahradn�� 1, 664 41 Troubsko, Czech Republic. vejrazka@vupt.cz.
  9. Hana Jake��ov��: Red Clover and Grass Breeding, 724 47 Hladk�� ��ivotice, Czech Republic. hana.jakesova@tiscali.cz.
  10. Jan Jansa: Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic. jansa@biomed.cas.cz. ORCID
  11. Lubom��r Pi��lek: Department of Zoology, Faculty of Science, University of South Bohemia, 370 05 ��esk�� Bud��jovice, Czech Republic. lpialek@yahoo.com. ORCID
  12. Daniela Knotov��: Research Institute for Fodder Crops, Ltd., 664 41 Troubsko, Czech Republic. knotova@vupt.cz.
PMID: 31684086 DOI: 10.3390/ijms20215470

Abstract

Plant-rhizobia symbiosis can activate key genes involved in regulating nodulation associated with biological nitrogen fixation (BNF). Although the general molecular basis of the BNF process is frequently studied, little is known about its intraspecific variability and the characteristics of its allelic variants. This study's main goals were to describe phenotypic and genotypic variation in the context of nitrogen fixation in red clover ( L.) and identify variants in BNF candidate genes associated with BNF efficiency. Acetylene reduction assay validation was the criterion for selecting individual plants with particular BNF rates. Sequences in 86 key candidate genes were obtained by hybridization-based sequence capture target enrichment of plants with alternative phenotypes for nitrogen fixation. Two genes associated with BNF were identified: ethylene response factor required for nodule differentiation () and molybdate transporter 1 (). In addition, whole-genome population genotyping by double-digest restriction-site-associated sequencing (ddRADseq) was performed, and BNF was evaluated by the natural N abundance method. Polymorphisms associated with BNF and reflecting phenotype variability were identified. The genetic structure of plant accessions was not linked to BNF rate of measured plants. Knowledge of the genetic variation within BNF candidate genes and the characteristics of genetic variants will be beneficial in molecular diagnostics and breeding of red clover.

Keywords

associated genes associated polymorphisms biological nitrogen fixation genome-wide association red clover

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Grants

  1. TH02010351/Technology Agency of the Czech Republic
  2. MZE-RO1718/Ministry of Agriculture of the Czech Republic
  3. Ref. 51834/2017-MZE17253/6.2.2/National programme of conservation and utilization of plant genetic resources and agro-biodiversity
  4. MUNI/A/0958/2018/Ministry of Education, Youth and Sports

MeSH Term

Alleles
Genes, Plant
Genotype
Host Microbial Interactions
Nitrogen Fixation
Phenotype
Plant Roots
Polymorphism, Genetic
Rhizobium
Sequence Analysis, DNA
Symbiosis
Trifolium
Journal Article

Word Cloud

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