plants Article Investigation of Root Morphological Traits Using 2D-Imaging among Diverse Soybeans (Glycine max L.) Pooja Tripathi 1,† , Jamila S. Abdullah 1,† , Jaeyoung Kim 2 , Yong-Suk Chung 2 , Seong-Hoon Kim 3 , Muhammad Hamayun 4 and Yoonha Kim 1, *   Citation: Tripathi, P.; Abdullah, J.S.; Kim, J.; Chung, Y.-S.; Kim, S.-H.; Hamayun, M.; Kim, Y. Investigation of Root Morphological Traits Using 2D-Imaging among Diverse Soybeans (Glycine max L.). Plants 2021, 10, 2535. https://doi.org/10.3390/ plants10112535 Academic Editor: Daniel K. Gladish Received: 3 September 2021 Accepted: 19 November 2021 Published: 21 November 2021 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). 1 Department of Applied Biosciences, Kyungpook National University, Daegu 41566, Korea; tripathipooza21@gmail.com (P.T.); abdullajamila@gmail.com (J.S.A.) 2 Department of Plant Resources and Environment, Jeju National University, Jeju 63243, Korea; baron7798@jejunu.ac.kr (J.K.); yschung@jejunu.ac.kr (Y.-S.C.) 3 National Agrobiodiversity Center, National Institute of Agricultural Sciences, RDA, Jeonju 54874, Korea; shkim0819@korea.kr 4 Department of Botany, Abdul Wali Khan University, Mardan 23200, Pakistan; hamayun@awkum.edu.pk * Correspondence: kyh1229@knu.ac.kr; Tel.: +82-53-950-5710 † These authors also contributed equally to this work. Abstract: Roots are the most important plant organ for absorbing essential elements, such as water and nutrients for living. To develop new climate-resilient soybean cultivars, it is essential to know the variation in root morphological traits (RMT) among diverse soybean for selecting superior root attribute genotypes. However, information on root morphological characteristics is poorly understood due to difficulty in root data collection and visualization. Thus, to overcome this problem in root research, we used a 2-dimensional (2D) root image in identifying RMT among diverse soybeans in this research. We assessed RMT in the vegetative growth stage (V2) of 372 soybean cultivars propagated in polyvinyl chloride pipes. The phenotypic investigation revealed significant variability among the 372 soybean cultivars for RMT. In particular, RMT such as the average diameter (AD), surface area (SA), link average length (LAL), and link average diameter (LAD) showed significant variability. On the contrary RMT, as with total length (TL) and link average branching angle (LABA), did not show differences. Furthermore, in the distribution analysis, normal distribution was observed for all RMT; at the same time, difference was observed in the distribution curve depending on individual RMT. Thus, based on overall RMT analysis values, the top 5% and bottom 5% ranked genotypes were selected. Furthermore, genotypes that showed most consistent for overall RMT have ranked accordingly. This ultimately helps to identify four genotypes (IT 16538, IT 199127, IT 165432, IT 165282) ranked in the highest 5%, whereas nine genotypes (IT 23305, IT 208266, IT 165208, IT 156289, IT 165405, IT 165019, IT 165839, IT 203565, IT 181034) ranked in the lowest 5% for RMT. Moreover, principal component analysis clustered cultivar 2, cultivar 160, and cultivar 274 into one group with high RMT values, and cultivar 335, cultivar 40, and cultivar 249 with low RMT values. The RMT correlation results revealed significantly positive TL and AD correlations with SA (r = 0.96) and LAD (r = 0.85), respectively. However, negative correlations (r = −0.43) were observed between TL and AD. Similarly, AD showed a negative correlation (r = −0.22) with SA. Thus, this result suggests that TL is a more vital factor than AD for determining SA compositions. Keywords: root morphological traits; root surface area; root length; root diameter; WinRHIZO 1. Introduction The soybean is an affordable protein-rich source of food that also has high economic and environmental values. Therefore, soybean is one of three major crops worldwide [1]. Climate is rapidly changing, and as a result, temperature increase has led to a substantial yield decline in soybean. Thus, huge economic losses occur yearly [2]. Drought is among Plants 2021, 10, 2535. https://doi.org/10.3390/plants10112535 https://www.mdpi.com/journal/plants