Volume 7, Issue 8, Pages 1388-1392 (August 2014) Short-Root1 Plays a Role in the Development of Vascular Tissue and Kranz Anatomy in Maize Leaves  Thomas L. Slewinski, Alyssa A. Anderson, Simara Price, Jacob R. Withee, Kimberly Gallagher, Robert Turgeon  Molecular Plant  Volume 7, Issue 8, Pages 1388-1392 (August 2014) DOI: 10.1093/mp/ssu036 Copyright © 2014 The Authors. All rights reserved. Terms and Conditions

Figure 1 Whole-Plant and Leaf Vascular Phenotypes Associated with the zmshr1 Mutant. Wild-type (WT) (B, D, F, I, J, K) and zmshr1 mutant (C, D, G, H, L–R) maize. (A) Photograph of 3-week-old maize plants. zmshr1 mutants plants are smaller than WT siblings, have thinner leaves, and reduced root growth. (B–E) Light micrographs of cleared leaf tissue (IKI-stained to reveal starch) showing that the regular parallel vein pattern (B, D) is disrupted in mutant leaves (C, D). (F–H) Free-hand cross-sections of fresh leaf tissue illuminated with UV light. (H) Stained with berberine-aniline blue staining to show suberization of BS and lignified xylem. Mutants display altered M cell development and plastid density (G, H) when compared to WT (F). Yellow arrows show an additional mutant parenchyma-like, mesophyll cell layer. Minor veins in mutants leaves frequently have an incomplete BS/endodermal cell layer (L–O) which normally fully encircles minor veins in WT leaves (I–K). (I) and (J) show WT BS layers in cleared IKI-stained leaves (I) and in cross-section illuminated by UV light (J). (K) Transmission electron micrograph (TEM) of WT minor vein showing normal Kranz anatomy and plastid structures. (L–O) zmshr1 mutant leaf showing incomplete Kranz anatomy and direct contact between xylem elements and M cells. (M) Cleared, IKI-stained leaves also shown in transverse cross section illuminated by UV light without staining (N) and with berberine-aniline blue staining (O) to illuminate lignin within the xylem elements and suberin in the BS cell walls. Black solid triangle shows incomplete Kranz anatomy along a minor vein. (L) TEM of a minor vein in a mutant leaf. Open triangles show absence of BS cells where M cells are in direct contact with xylem elements. Asterisk shows empty BS cell. (P–U) BS cells without associated veins. IKI starch stained mutant leaves in paradermal view in (P) and (T) and in transverse cross section in (Q) and (S). (R) Transverse cross-section illuminated by UV light with berberine-aniline blue staining to show suberization of BS with no associated veins. (R) TEM of mutant leaf. Red arrows show isolated BS cells not associated with vascular cores. M, mesophyll; BS, bundle sheath; X, xylem; XP, xylem parenchyma. Scale bars in (B, C, P) = 400 μm; (D, E, I, M) = 225 μm; (F, G, T) = 60 μm; (Q, S) = 50 μm; (H, J, N, O, U) = 40 μm; (K, L, R) = 15 μm. Molecular Plant 2014 7, 1388-1392DOI: (10.1093/mp/ssu036) Copyright © 2014 The Authors. All rights reserved. Terms and Conditions