Evaluation of root traits in F2-progeny of interspecific hybrid between Lotus corniculatus “Super-Root” and tetraploid Lotus japonicus

  • Rinda Puspasari
  • Masatsugu Hashiguchi
  • Ryoma Ushio
  • Genki Ishigaki
  • Hidenori Tanaka
  • Ryo AkashiEmail author
Regular Article



“Super-Root” (SR) is a fast-growing root culture established in the legume forage species Lotus corniculatus that displays vigorous growth and high embryogenic capacity, uniquely allowing for continuous root cloning, direct somatic embryogenesis, and mass regeneration of plants under hormone-free culture conditions. This study’s primary aim was to verify the possibility of introducing the SR trait to another legume species as a means of enhancing root traits. Our previous research produced a F1-generation hybrid between the colchicine-doubled form of the diploid Lotus japonicus ‘Miyakojima’ MG-20 and SR-derived Lotus corniculatus. The current study’s focus was to evaluate the root phenotype of the F2-generation to confirm the introduction of SR characteristics to Lotus japonicus.


We assessed 242 rooted F2-hybrid progeny using cluster analysis, concentrating on four morphological root traits: root fresh weight, total root length, maximum root length, and root diameter. Principal Component Analysis (PCA) was used to determine the correlations among eight traits of the F2-progeny of the WinRhizo root morphological measurement data and to identify what traits contributed most to their root variation.


The 242-hybrid progeny and both parents were grouped into eight different clusters using a similarity phenogram. One cluster consisted of lines 36, 82, 86 and 184 that considerably exceeded the RFW, TRL, and MRL values of the L. japonicus and SR parents.


The results confirmed the introduction of SR traits to the F2-hybrid progeny and identified potential candidates for breeding and gene isolation that will be conducted to expand the scope of this research.


Interspecific hybrids Lotus corniculatus Root traits Super-Root 



Maximum root length


Root diameter


Root fresh weight




Total root length



This work was partially supported by the Genesis Research Institute, Incorporated and the National BioResource Project (NBRP) of the Japan Agency for Medical Research and Development (AMED). We would like to sincerely thank Professor Franz Hoffmann, from the University of California Irvine, for his kind pre-reviewing.


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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Interdisciplinary Graduate School of Agriculture and EngineeringUniversity of MiyazakiMiyazakiJapan
  2. 2.Faculty of AgricultureUniversity of MiyazakiMiyazakiJapan
  3. 3.Center for the Promotion of Institutional ResearchUniversity of MiyazakiMiyazakiJapan

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