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. 2019 Oct 10;9:14511. doi: 10.1038/s41598-019-50962-y

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Morphological and histological characterization of callus induction using embryos collected at different developmental stages. (a) Immature Embryo at 15 days after pollination; IE15. (c) Immature Embryo at 23 days after pollination; IE23. (e) Mature Embryo; ME. (b,d,f) Histological analysis of longitudinal sections across IE15, IE23 and ME, respectively. (g,i,k) Callus from IE15, IE23 and ME, respectively, at one month of induction. (h,j,l) Callus morphology from IE15, IE23 and ME, respectively (C15, C23 and CM), observed at one month of induction by stereoscopic microscopy. (m–o) Histological analysis of callus cross-sections from IE15, IE23 and ME, respectively, at one month of induction. (p) Line index (LI) bar plot. The dotted line represents the minimum LI value of an embryogenic batch (q) In vitro Capacity (IC) bar plot. Embryogenic categories are designated with the red color palette; grayscale palette represents the non-embryogenic categories. L1-L5: Leaf primordia in the maize embryo; Co: Coleoptile; Sc: Scutellum; SAM: Shoot Apical Meristem; RAM: Root Apical Meristem; S: suspensor; Ca: Calyptra; E: Embryo; EC: Embryogenic Callus; NEC: Non-Embryogenic callus; te: tracheary elements; pc: procambium cells; pa: parenchyma cells.