Please wait a minute...
Shopping Cart Email List Chinese
Advanced Search Fig/Tab
Frontiers in Biology

ISSN 1674-7984

ISSN 1674-7992(Online)

CN 11-5892/Q

Featured Articles  |  Most Downloaded  |  Most Reading
Online Submission Subscribe to Our RSS Content Feed
Front Biol    2010, Vol. 5 Issue (5) : 464-470    https://doi.org/10.1007/s11515-010-0820-0
RESEARCH ARTICLE
Pattern analysis of stem cell differentiation during in vitroArabidopsis organogenesis
Ying Hua SU1,2, Zhi Juan CHENG1,2, Yu Xiao SU1,2, Xian Sheng ZHANG1,2()
1. College of Life Sciences, Shandong Agricultural University, Tai’an 271018, China; 2. State Key Laboratory of Crop Biology, Shandong Agricultural University, Tai’an 271018, China
 Download: PDF(364 KB)   HTML
 Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract

Plant somatic cells have the capability to switch their cell fates from differentiated to undifferentiated status under proper culture conditions, which is designated as totipotency. As a result, plant cells can easily regenerate new tissues or organs from a wide variety of explants. However, the mechanism by which plant cells have such remarkable regeneration ability is still largely unknown. In this study, we used a set of meristem-specific marker genes to analyze the patterns of stem cell differentiation in the processes of somatic embryogenesis as well as shoot or root organogenesis in vitro. Our studies furnish preliminary and important information on the patterns of the de novo stem cell differentiation during various types of in vitro organogenesis.
Keywords Organ regeneration      stem cell differentiation      WUS expression      WOX5 expression     
Corresponding Author(s): ZHANG Xian Sheng,Email:zhangxs@sdau.edu.cn   
Issue Date: 01 October 2010
 Cite this article:   
Ying Hua SU,Zhi Juan CHENG,Yu Xiao SU, et al. Pattern analysis of stem cell differentiation during in vitroArabidopsis organogenesis[J]. Front Biol, 2010, 5(5): 464-470.
 URL:  
https://academic.hep.com.cn/fib/EN/10.1007/s11515-010-0820-0
https://academic.hep.com.cn/fib/EN/Y2010/V5/I5/464
Fig.1  Morphogenesis of somatic embryos and the expression of and during somatic embryogenesis. A: Morphology of embryonic callus in embryonic callus-inducing medium (ECIM) by scanning electron microscopy; B: Somatic embryos with two cotyledon primordia on the surface of embryonic callus edge in somatic embryo-inducing medium (SEIM) for 4 days; C: Somatic embryos by light microscopy in SEIM for 8 days; D and E: hybridization shows the localization of transcripts in SEIM for 24 h and 2 days; F: signals in the sections of embryonic calli in SEIM for 3 days; G: signals (, red) in SEIM for 24 h; H: (, green) and (red) signals in SEIM for 2 days; I: Both and signals localized within the shoot apical meristem (SAM) of the somatic embryos in SEIM for 4 days. Arrowheads indicate the signals of transcription in D, E, F, G, H and I. Scale bars= 80 μm (D, E, F, G, H and I), 1.3 mm (C) and 30 μm (A and B).
Fig.1  Morphogenesis of somatic embryos and the expression of and during somatic embryogenesis. A: Morphology of embryonic callus in embryonic callus-inducing medium (ECIM) by scanning electron microscopy; B: Somatic embryos with two cotyledon primordia on the surface of embryonic callus edge in somatic embryo-inducing medium (SEIM) for 4 days; C: Somatic embryos by light microscopy in SEIM for 8 days; D and E: hybridization shows the localization of transcripts in SEIM for 24 h and 2 days; F: signals in the sections of embryonic calli in SEIM for 3 days; G: signals (, red) in SEIM for 24 h; H: (, green) and (red) signals in SEIM for 2 days; I: Both and signals localized within the shoot apical meristem (SAM) of the somatic embryos in SEIM for 4 days. Arrowheads indicate the signals of transcription in D, E, F, G, H and I. Scale bars= 80 μm (D, E, F, G, H and I), 1.3 mm (C) and 30 μm (A and B).
Fig.2  Shoot regeneration and the expression of and during shoot regeneration. A: Callus induced from stage-10 pistil explants on callus induced medium (CIM) for 20 days; B: Callus transferred onto shoot induction medium (SIM) for 6 days; C: Regenerated shoots from callus on SIM for 20 days; D to F: Longitudinal section of transgenic callus on CIM for 20 days (D), and transferred to SIM for 6 days (E) and 8 days (F); G: transcription shown as red color signals () in callus on SIM for 4 days; H: transcripts shown as green signals () in a domain above the expressed region in callus on SIM for 6 days; I: The transcription status of both (green) and (red) in a pro-meristematic tissue. Arrowheads indicate the signals of expression in E, F, G, H and I. Scale bars= 2.0 mm (A, B and C) and 80 μm (D to I).
Fig.2  Shoot regeneration and the expression of and during shoot regeneration. A: Callus induced from stage-10 pistil explants on callus induced medium (CIM) for 20 days; B: Callus transferred onto shoot induction medium (SIM) for 6 days; C: Regenerated shoots from callus on SIM for 20 days; D to F: Longitudinal section of transgenic callus on CIM for 20 days (D), and transferred to SIM for 6 days (E) and 8 days (F); G: transcription shown as red color signals () in callus on SIM for 4 days; H: transcripts shown as green signals () in a domain above the expressed region in callus on SIM for 6 days; I: The transcription status of both (green) and (red) in a pro-meristematic tissue. Arrowheads indicate the signals of expression in E, F, G, H and I. Scale bars= 2.0 mm (A, B and C) and 80 μm (D to I).
Fig.3  Root regeneration and expression of the and during root regeneration. A: Callus induced from stage-10 pistil explants on callus induced medium (CIM) for 10 days; B: Callus on CIM for 20 days; C: Regenerated roots from callus on root induction medium (RIM) for 15 days; D: transcription shown as green color signals () in callus on CIM for 20 days; E and F: transcription in callus on RIM for 3 days (E) and 7 days (F); G: transcription shown as red color signals () in callus on CIM for 20 days; H and I: transcription in callus on RIM for 3 days (H) and 5 days (I). Arrowheads in Figs. E and F indicate expression, and in Fig. H and I indicate expression. Scale bars= 2.0 mm (A, B and C) and 80 μm (D to I).
Fig.3  Root regeneration and expression of the and during root regeneration. A: Callus induced from stage-10 pistil explants on callus induced medium (CIM) for 10 days; B: Callus on CIM for 20 days; C: Regenerated roots from callus on root induction medium (RIM) for 15 days; D: transcription shown as green color signals () in callus on CIM for 20 days; E and F: transcription in callus on RIM for 3 days (E) and 7 days (F); G: transcription shown as red color signals () in callus on CIM for 20 days; H and I: transcription in callus on RIM for 3 days (H) and 5 days (I). Arrowheads in Figs. E and F indicate expression, and in Fig. H and I indicate expression. Scale bars= 2.0 mm (A, B and C) and 80 μm (D to I).
Fig.4  A working model on the patterns of the stem cell formation during shoot or root organ regeneration. A: Immature zygotic embryos were used as explants to obtain embryogenic calli in embryonic callus-inducing medium (ECIM). Embryonic calli were cultured in somatic embryo-inducing medium (SEIM) to induce somatic embryos. expression (red) is induced at day 1, and gene (blue) is induced later at day 2. The expression domain is located just below the expression domain in the meristems of somatic embryos. B: Pistils were cut and transferred to callus induced medium (CIM) to induce callus. The calli cultured on CIM for 20 days were transferred onto SIM for shoot induction. expression (red) is induced at day 4, and expression (blue) is induced at around day 6. signals were visulized below the expression domain in the regenerated shoot meristems. During the root regeneration, the calli (also derived from pistil explants on CIM cultured for 20 days) were transferred onto the RIM for root induction. (pink) and (sky blue) signals distribute evenly along the edges of the whole callus on CIM, and then shows a regional pattern in callus for root induction on the root induction mediem (RIM) at about day 3. signal is localized in the quiescent center (QC), and signal is detected in the stem cells of the pro-meristematic root tissues at day 5 and at the tip of the mature roots on RIM at around day 7.
Fig.4  A working model on the patterns of the stem cell formation during shoot or root organ regeneration. A: Immature zygotic embryos were used as explants to obtain embryogenic calli in embryonic callus-inducing medium (ECIM). Embryonic calli were cultured in somatic embryo-inducing medium (SEIM) to induce somatic embryos. expression (red) is induced at day 1, and gene (blue) is induced later at day 2. The expression domain is located just below the expression domain in the meristems of somatic embryos. B: Pistils were cut and transferred to callus induced medium (CIM) to induce callus. The calli cultured on CIM for 20 days were transferred onto SIM for shoot induction. expression (red) is induced at day 4, and expression (blue) is induced at around day 6. signals were visulized below the expression domain in the regenerated shoot meristems. During the root regeneration, the calli (also derived from pistil explants on CIM cultured for 20 days) were transferred onto the RIM for root induction. (pink) and (sky blue) signals distribute evenly along the edges of the whole callus on CIM, and then shows a regional pattern in callus for root induction on the root induction mediem (RIM) at about day 3. signal is localized in the quiescent center (QC), and signal is detected in the stem cells of the pro-meristematic root tissues at day 5 and at the tip of the mature roots on RIM at around day 7.
1 Aida M, Beis D, Heidstra R, Willemsen V, Blilou I, Galinha C, Nussaume L, Noh Y S, Amasino R, Scheres B (2004). The PLETHORA genes mediate patterning of the Arabidopsis root stem cell niche. Cell , 119(1): 109–120
2 Birnbaum K D, Sánchez Alvarado A (2008). Slicing across kingdoms: regeneration in plants and animals. Cell , 132(4): 697–710
3 Brawley C, Matunis E (2004). Regeneration of male germline stem cells by spermatogonial dedifferentiation in vivo. Science , 304(5675): 1331–1334
doi: 10.1126/science.1097676
4 Cheng Z J, Zhu S S, Gao X Q, Zhang X S (2010). Cytokinin and auxin regulates WUS induction and inflorescence regeneration in vitro in Arabidopsis. Plant Cell Rep , 29(8): 927–933
doi: 10.1007/s00299-010-0879-8
5 Clark S E, Jacobsen S E, Levin J Z, Meyerowitz E M (1996). The CLAVATA and SHOOT MERISTEMLESS loci competitively regulate meristem activity in Arabidopsis. Development , 122(5): 1567–1575
6 Ding Z, Friml J (2010). Auxin regulates distal stem cell differentiation in Arabidopsis roots. Proc Natl Acad Sci U S A , 107(26): 12046–12051
doi: 10.1073/pnas.1000672107
7 Evans M J, Kaufman M H (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature , 292: 154–156
doi: 10.1038/292154a0
8 Fletcher J C, Meyerowitz E M (2000). Cell signaling within the shoot meristem. Curr Opin Plant Biol , 3(1): 23–30
doi: 10.1016/S1369-5266(99)00033-3
9 Galinha C, Hofhuis H, Luijten M, Willemsen V, Blilou I, Heidstra R, Scheres B (2007). PLETHORA proteins as dose-dependent master regulators of Arabidopsis root development. Nature , 449(7165): 1053–1057
doi: 10.1038/nature06206
10 Gallois J L, Nora F R, Mizukami Y, Sablowski R (2004). WUSCHEL induces shoot stem cell activity and developmental plasticity in the root meristem. Genes Dev , 18(4): 375–380
doi: 10.1101/gad.291204
11 Gross-Hardt R, Laux T (2003). Stem cell regulation in the shoot meristem. J Cell Sci , 116(Pt 9): 1659–1666
doi: 10.1242/jcs.00406
12 Higashi K, Shiota H, Kamada H (1998). Patterns of expression of the genes for glutamine synthetase isoforms during somatic and zygotic embryogenesis in carrot. Plant Cell Physiol , 39(4): 418–424
13 Jürgens G (2001). Apical-basal pattern formation in Arabidopsis embryogenesis. EMBO J , 20(14): 3609–3616
doi: 10.1093/emboj/20.14.3609
14 Laux T, Mayer K F X, Berger J, Jürgens G (1996). The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development , 122(1): 87–96
15 Li Q Z, Li X G, Bai S N, Lu W L, Zhang X S (2002). Isolation of HAG1 and its regulation by plant hormones during in vitro floral organogenesis in Hyacinthus orientalis L. Planta , 215(4): 533–540
doi: 10.1007/s00425-002-0796-3
16 Long J A, Moan E I, Medford J I, Barton M K (1996). A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature , 379(6560): 66–69
doi: 10.1038/379066a0
17 Lotan T, Ohto M, Yee K M, West M A L, Lo R, Kwong R W, Yamagishi K, Fischer R L, Goldberg R B, Harada J J (1998). Arabidopsis LEAFY COTYLEDON1 is sufficient to induce embryo development in vegetative cells. Cell , 93(7): 1195–1205
18 Morgan T (1901). Regeneration. New York: Macmillan
19 Müller B, Sheen J (2008). Cytokinin and auxin interaction in root stem-cell specification during early embryogenesis. Nature , 453(7198): 1094–1097
doi: 10.1038/nature06943
20 Murashige T, Skoog F (1962). A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant , 15: 473–497
doi: 10.1111/j.1399-3054.1962.tb08052.x
21 Prantl K (1874). Untersuchungen uber die Regeneration des Vegetationspunktes an Agiospermenwurzeln. Arb Bot Inst Wurzburg , 4: 546–562
22 Rieu I, Laux T (2009). Signaling pathways maintaining stem cells at the plant shoot apex. Semin Cell Dev Biol , 20(9): 1083–1088
doi: 10.1016/j.semcdb.2009.09.013
23 Sarkar A K, Luijten M, Miyashima S, Lenhard M, Hashimoto T, Nakajima K, Scheres B, Heidstra R, Laux T (2007). Conserved factors regulate signalling in Arabidopsis thaliana shoot and root stem cell organizers. Nature , 446(7137): 811–814
doi: 10.1038/nature05703
24 Scheres B (2007). Stem-cell niches: nursery rhymes across kingdoms. Nat Rev Mol Cell Biol , 8(5): 345–354
doi: 10.1038/nrm2164
25 Schoof H, Lenhard M, Haecker A, Mayer K F X, Jürgens G, Laux T (2000). The stem cell population of Arabidopsis shoot meristems in maintained by a regulatory loop between the CLAVATA and WUSCHEL genes. Cell , 100(6): 635–644
26 Shiota H, Satoh R, Watabe K, Harada H, Kamada H (1998). C-ABI3, the carrot homologue of the Arabidopsis ABI3, is expressed during both zygotic and somatic embryogenesis and functions in the regulation of embryo-specific ABA-inducible genes. Plant Cell Physiol , 39(11): 1184–1193
27 Shiota H, Tachikawa K, Watabe K, Kamada H (1999). Successful long-term preservation of abscisic acid-treated and desiccated carrot somatic embryos. Plant Cell Rep , 18: 749–753
doi: 10.1007/s002990050654
28 Sieburth L E, Meyerowitz E M (1997). Molecular dissection of the AGAMOUS control region shows that cis elements for spatial regulation are located intragenically. Plant Cell , 9(3): 355–365
29 Stone S L, Kwong L W, Yee K M, Pelletier J, Lepiniec L, Fischer R L, Goldberg R B, Harada J J (2001). LEAFY COTYLEDON2 encodes a B3 domain transcription factor that induces embryo development. Proc Natl Acad Sci U S A , 98(20): 11806–11811
doi: 10.1073/pnas.201413498
30 Su Y H, Zhao X Y, Liu Y B, Zhang C L, O’Neill S D, Zhang X S (2009). Auxin-induced WUS expression is essential for embryonic stem cell renewal during somatic embryogenesis in Arabidopsis. Plant J , 59(3): 448–460
doi: 10.1111/j.1365-313X.2009.03880.x
31 Terpstra I, Heidstra R (2009). Stem cells: The root of all cells. Semin Cell Dev Biol , 20(9): 1089–1096
doi: 10.1016/j.semcdb.2009.09.012
32 Thomson J A, Itskovitz-Eldor J, Shapiro S S, Waknitz M A, Swiergiel J J, Marshall V S, Jones J M (1998). Embryonic stem cell lines derived from human blastocysts. Science , 282(5391): 1145–1147
doi: 10.1126/science.282.5391.1145
33 Turnpenny L, Spalluto C M, Perrett R M, O’Shea M, Hanley K P, Cameron I T, Wilson D I, Hanley N A (2006). Evaluating human embryonic germ cells: concord and conflict as pluripotent stem cells. Stem Cells , 24(2): 212–220
doi: 10.1634/stemcells.2005-0255
34 van den Berg C, Willemsen V, Hendriks G, Weisbeek P, Scheres B (1997). Short-range control of cell differentiation in the Arabidopsis root meristem. Nature , 390(6657): 287–289
doi: 10.1038/36856
35 Weigel D, Jürgens G (2002). Stem cells that make stems. Nature , 415(6873): 751–754
36 Xu J, Hofhuis H, Heidstra R, Sauer M, Friml J, Scheres B (2006). A molecular framework for plant regeneration. Science , 311(5759): 385–388
doi: 10.1126/science.1121790
37 Zhao X Y, Liu M S, Li J R, Guan C M, Zhang X S (2005). The wheat TaGI1, involved in photoperiodic flowering, encodes an ArabidopsisGI ortholog. Plant Mol Biol , 58(1): 53–64
doi: 10.1007/s11103-005-4162-2
38 Zuo J, Niu Q W, Frugis G, Chua N H (2002). The WUSCHEL gene promotes vegetative-to-embryonic transition in Arabidopsis. Plant J , 30(3): 349–359
doi: 10.1046/j.1365-313X.2002.01289.x
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed