Advertisement

Molecular Biology Reports

, Volume 37, Issue 8, pp 3973–3982 | Cite as

Using high competent shoot apical meristems of cockscomb as explants for studying function of ASYMMETRIC LEAVES2-LIKE11 (ASL11) gene of Arabidopsis

  • Shao-Bo Sun
  • Lai-Sheng Meng
  • Xu-Dong Sun
  • Zhen-Hua Feng
Article

Abstract

Though shoot apical meristems (SAMs) commonly exhibit low or no competence for transformation, the potent regeneration of this tissue merits further research. Especially, when shoot regeneration is recalcitrant using other tissues as explants, SAM probably is an excellent selection. In cockscomb plants, using SAMs from seedlings obtained from MS medium with 0.5 mg l−1 6-BA as explants, high frequency of transformation (approximate 20%) is obtained; whereas control SAMs performed poorly for transformation (approximate 3%). These SAMs are malformed in morphology compared to control SAMs. Further observation found that, in these SAMs, cell proliferation and/or TE formation are seen; which are not found in control SAMs. GUS assays indicated that GUS-positive blue spots at TE zones are obvious; whereas the case was contrary in control SAMs. All these data suggest that cell proliferation and/or TE formation might cause high effective transformation. This transformation system should facilitate the use of this species for studies on gene manipulation and expression. Therefore, we introduced 35S:ASL11GFP to cockscomb via Agrobacterium tumefaciens. ASYMMETRIC LEAVES2-LIKE11 (ASL11) gene of Arabidopsis is a member of the ASYMMETRIC LEAVES2 (AS2)/LATERAL ORGAN BOUNDARIES (LOB) domain gene family, and its function is largely unclear. By confocal laser scanning microscopy, we found that in most over 35S:ASL11GFP cockscomb plants, ASL11–GFP fusion protein was in discrete nuclear location. These results indicate that the T-DNA contains within the construct inserted into the host chromosomes in an integral form, and also suggest that ASL11 might be a nuclear protein and function as a potential transcription factor. Moreover, SAMs of the over 35S:ASL11GFP plants show needle-like patterns that lack organ primordial; suggesting ASL11 might be involved in sustaining indeterminate cell fate of SAMs.

Keywords

Cockscomb Shoot apical meristems (SAMs) ASYMMETRIC LEAVES2-LIKE11/LATERAL BOUNDAY DOMAIN 15 (ASL11/LBD15) 0.5 mg l−1 6-BA ASL11–GFP Transformation 

Supplementary material

11033_2010_56_MOESM1_ESM.doc (32 kb)
Supplementary material 1 (DOC 31 kb)
11033_2010_56_MOESM2_ESM.doc (78 kb)
Supplementary material 2 (DOC 78 kb)
11033_2010_56_MOESM3_ESM.doc (56 kb)
Supplementary material 3 (DOC 55 kb)
11033_2010_56_MOESM4_ESM.doc (28 kb)
Supplementary material 4 (DOC 28 kb)
11033_2010_56_MOESM5_ESM.doc (30 kb)
Supplementary material 5 (DOC 30 kb)

References

  1. 1.
    Sinha NR, Williams RE, Hake S (1993) Overexpression of the maize homeobox gene, KNOTTED-1, causes a switch from determinate to indeterminate cell fates. Genes Dev 7:787–795CrossRefPubMedGoogle Scholar
  2. 2.
    de Kathen A, Jacobsen HJ (1995) Cell competence for Agrobacterium-mediated DNA transfer in Pisum sativum L. Transgenic Res 4:184–191CrossRefGoogle Scholar
  3. 3.
    Schrammeijer B, Sijmons PC, Elzen PJM, van den Hoekema A (1990) Meristem transformation of sunflower via Agrobacterium. Plant Cell Rep 9:55–60CrossRefGoogle Scholar
  4. 4.
    Ulian EC, Smith RH, Gould JH, McKnight TD (1988) Transformation of plants via the shoot apex. In Vitro Cell Dev Biol 24:951–954CrossRefGoogle Scholar
  5. 5.
    Gould J, Banister S, Haseganwa O, Fakima M, Smith RH (1991) Regeneration of Gossypium hirsutum and G. barbadense from shoot apex tissues for transformation. Plant Cell Rep 10:12–16CrossRefGoogle Scholar
  6. 6.
    Schlappi M, Hohn B (1992) Competence of immature maize embryos for Agrobacterium-mediated gene transfer. Plant Cell 4:7–16CrossRefPubMedGoogle Scholar
  7. 7.
    Escudero J, Neuhaus G, Schlappi M, Hohn B (1996) T-DNA transfer in meristematic cells of maize provided with intracellular Agrobacterium. Plant J 10:356–360CrossRefGoogle Scholar
  8. 8.
    Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282CrossRefPubMedGoogle Scholar
  9. 9.
    Potrykus I (1990) Gene transfer to cereals: assessment and perspectives. Physiol Plant 79:125–134CrossRefGoogle Scholar
  10. 10.
    Zambryski PC (1992) Chronicles from the Agrobacterium plant cell DNA transfer story. Annu Rev Plant Physiol Plant Mol Biol 43:465–490CrossRefGoogle Scholar
  11. 11.
    Medford JI (1992) Vegetative apical meristems. Plant Cell 4:1029–1039CrossRefPubMedGoogle Scholar
  12. 12.
    Sangwan RS, Bourgeois Y, Brown S, Vasseur G, Brigitte SN (1992) Characterization of competent cells and early events of Agrobacterium-mediated genetic transformation in Arabidopsis thaliana. Planta 188:439–456CrossRefGoogle Scholar
  13. 13.
    Meng LS, Ding WQ, Hu X, Wang CY (2009) Transformation of PttKN1 gene to cockscomb. Acta Physiol Plant 31:683–691CrossRefGoogle Scholar
  14. 14.
    Shuai B, Reynaga-Peña CG, Springer PS (2002) The LATERAL ORGAN BOUNDARIES gene defines a novel, plant-specific gene family. Plant Physiol 129:747–761CrossRefPubMedGoogle Scholar
  15. 15.
    Nakazawa M, Ichikawa T, Ishikawa A, Kobayashi H, Tsuhara Y, Kawashima M, Suzuki K, Muto S, Matsui M (2003) Activation tagging, a novel tool to dissect the functions of a gene family. Plant J 34:741–750CrossRefPubMedGoogle Scholar
  16. 16.
    Chalfun-Junior A, Franken J, Mes JJ, Marsch-Martinez N, Pereira A, Angenent GC (2005) ASYMMETRIC LEAVES2-LIKE1 gene, a member of the AS2/LOB family, controls proximaldistal patterning in Arabidopsis petals. Plant Mol Biol 57:559–575CrossRefPubMedGoogle Scholar
  17. 17.
    Ori N, Eshed Y, Chuck G, Bowman JL, Hake S (2000) Mechanisms that control knox gene expression in the Arabidopsis shoot. Development 127:5523–5532PubMedGoogle Scholar
  18. 18.
    Semiarti E, Ueno Y, Tsukaya H, Iwakawa H, Machida C, Machida Y (2001) The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana regulates formation of a symmetric lamina, establishment of venation and repression of meristem-related homeobox genes in leaves. Development 128:1771–1783PubMedGoogle Scholar
  19. 19.
    Byrne ME, Simorowski J, Martienssen RA (2002) ASYMMETRIC LEAVES1 reveals knox gene redundancy in Arabidopsis. Development 129:1957–1965PubMedGoogle Scholar
  20. 20.
    Xu L, Xu Y, Dong A, Sun Y, Pi L, Xu Y, Huang H (2003) Novel as1 and as2 defects in leaf adaxial-abaxial polarity reveal the requirement for ASYMMETRIC LEAVES1 and 2 and ERECTA functions in specifying adaxial identity. Development 130:4097–4107CrossRefPubMedGoogle Scholar
  21. 21.
    Lin WC, Shuai B, Springer PS (2003) The Arabidopsis LATERAL ORGAN BOUNDARIES-domain gene ASYMMETRIC LEAVES2 functions in the repression of KNOX gene expression and in adaxial-abaxial patterning. Plant Cell 15:2241–2252CrossRefPubMedGoogle Scholar
  22. 22.
    Waites R, Selvadurai HRN, Oliver IR, Hudson A (1998) The PHANTASTICA gene encodes a MYB transcription factor involved in growth and dorsoventrality of lateral organs in Antirrhinum. Cell 93:779–789CrossRefPubMedGoogle Scholar
  23. 23.
    Schneeberger R, Tsiantis M, Freeling M, Langdale JA (1998) The ROUGH SHEATH2 gene negatively regulates homeobox gene expression during maize leaf development. Development 125:2857–2865PubMedGoogle Scholar
  24. 24.
    Timmermans MCP, Hudson A, Becraft PW, Nelson T (1999) ROUGH SHEATH2: a Myb protein that represses knox homeobox genes in maize lateral organ primordia. Science 284:151–153CrossRefPubMedGoogle Scholar
  25. 25.
    Tsiantis M, Schneeberger R, Golz JF, Freeling M, Langdale JA (1999) The maize rough sheath2 gene and leaf development programs in monocot and dicot plants. Science 284:154–156CrossRefPubMedGoogle Scholar
  26. 26.
    Bortiri E, Chuck G, Vollbrecht E, Rocheford T, Martienssen R, Hake S (2006) ramosa2 encodes a LATERAL ORGAN BOUNDARY domain protein that determines the fate of stem cells in branch meristems of maize. Plant Cell 18:574–585CrossRefPubMedGoogle Scholar
  27. 27.
    Evans MM (2007) The indeterminate gametophyte1 gene of maize encodes a LOB domain protein required for embryo sac and leaf development. Plant Cell 19:46–62CrossRefPubMedGoogle Scholar
  28. 28.
    Borghi L, Bureau M, Simon R (2007) Arabidopsis JAGGED LATERAL ORGANS is expressed in boundaries and coordinates KNOX and PIN activity. Plant Cell 19:1795–1808CrossRefPubMedGoogle Scholar
  29. 29.
    Ni RJ, Shen Z, Yang CP, Wu YD, Bi YD, Wang BC (2010) Identification of low abundance polyA-binding proteins in Arabidopsis chloroplast using polyA-affinity column. Mol Biol Rep 37:637–641CrossRefPubMedGoogle Scholar
  30. 30.
    Zhu B, Xiong AS, Peng RH, Xu J, Jin XF, Meng XR, Yao QH (2010) Over-expression of ThpI from Choristoneura fumiferana enhances tolerance to cold in Arabidopsis. Mol Biol Rep 37:961–966CrossRefPubMedGoogle Scholar
  31. 31.
    Liu C, Zhang L, Sun J, Luo Y, Wang MB, Fan YL, Wang L (2010) a simple artificial microRNA vector based on ath-miR169d precursor from Arabidopsis. Mol Biol Rep 37:903–909CrossRefPubMedGoogle Scholar
  32. 32.
    Ha CM, Jun JH, Nam HG, Fletcher JC (2007) BLADE-ON-PETIOLE1 and 2 control Arabidopsis lateral organ fate through regulation of LOB domain and adaxial–abaxial polarity genes. Plant Cell 19:1809–1825CrossRefPubMedGoogle Scholar
  33. 33.
    Meng LS, Song JP, Sun SB, Wang CY (2009) The ectopic expression of PttKN1 gene causes pleiotropic alternation of morphology in transgenic carnation (Dianthus caryophyllus L.). Acta Physiol Plant 31:1155–1164CrossRefGoogle Scholar
  34. 34.
    Guo XH, Deng KQ, Wang J, Yu DS, Zhao Q, Liu XM (2010) Mutational analysis of Arabidopsis PP2CA2 involved in abscisic acid signal transduction. Mol Biol Rep 37:763–769CrossRefPubMedGoogle Scholar
  35. 35.
    Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: B-Glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMOB J 6:3901–3907Google Scholar
  36. 36.
    Abdollahi MR, Corral-Mart′ınez P, Mousavi A, Salmanian AH, Moieni A, Segui-Simarro JM (2009) An efficient method for transformation of pre-androgenic, isolated Brassica napus microspores involving microprojectil bombardment and Agrobacterium-mediated transformation. Acta Physiol Plant 31:1313–1317CrossRefGoogle Scholar
  37. 37.
    Yasmeen A (2009) an improved protocol for the regeneration and transformation of tomato (cv Rio Grande). Acta Physiol Plant 31:1271–1277CrossRefGoogle Scholar
  38. 38.
    Meng LS, Liu HL, Cui XH, Sun XD, Zhu J (2009) ASYMMETRIC LEAVES2-LIKE38 gene, a member of AS2/LOB family of Arabidopsis, causes leaf dorsoventral alternation in transgenic Cockscomb plants. Acta Physiol Plant 31:1301–1306CrossRefGoogle Scholar
  39. 39.
    Fiuk A, Rybczynski JJ (2008) Morphogenic capability of Gentiana kurroo Royle seedling and leaf explants. Acta Physiol Plant 30:157–166CrossRefGoogle Scholar
  40. 40.
    Meng LS, Sun XD, Li F, Liu HL, Feng ZH, Zhu J (2010) Modification of flowers and leaves in Cockscomb (Celosia cristata) ectopically expressing Arabidopsis ASYMMERTIC LEAVES2-LIKE38 (ASL38/LBD41) gene. Acta Physiol Plant 32:315–324CrossRefGoogle Scholar
  41. 41.
    An G (1985) High efficiency transformation of cultured tobacco cells. Plant Physiol 79:568–570CrossRefPubMedGoogle Scholar
  42. 42.
    Aloni R (1987) Differentiation of vascular tissues. Annu Rev Plant Physiol 38:179–204CrossRefGoogle Scholar
  43. 43.
    Szekeres M, Nemeth K, Koncz-Kalman Z, Mathur J, Kauschmann A, Altmann T, Redei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85:171–182CrossRefPubMedGoogle Scholar
  44. 44.
    Yamamoto R, Demura T, Fukuda H (1997) Brassinosteroids induce entry into the final stage of tracheary element differentiation in cultured Zinnia cells. Plant Cell Physiol 38:980–983PubMedGoogle Scholar
  45. 45.
    Choe S, Dilkes BP, Gregory BD, Ross AS, Yuan H, Noguchi T, Fujioka S, Takatsuto S et al (1999) The Arabidopsis dwarf mutant is defective in the conversion of 2, 4-methylenecholesterol to campesterol in brassinosteroid biosynthesis. Plant Physiol 119:897–907CrossRefPubMedGoogle Scholar
  46. 46.
    Fukuda H (2004) Signals that control plant vascular cell differentiation. Nat Rev Mol Cell Biol 5:379–391CrossRefPubMedGoogle Scholar
  47. 47.
    Motose H, Sugiyama M, Fukuda H (2004) A proteoglycan mediates inductive interaction during plant vascular development. Nature 429:873–878CrossRefPubMedGoogle Scholar
  48. 48.
    Ito Y, Nakanomyo I, Motose H, Iwamoto K, Sawa S, Dohmae N, Fukuda H (2006) Dodeca-CLE peptides as suppressors of plant stem cell differentiation. Science 313:842–845CrossRefPubMedGoogle Scholar
  49. 49.
    Cao Z, Jia Z, Liu Y, Wang M, Zhao J, Zheng J, Wang G (2010) Constitutive expression of ZmsHSP in Arabidopsis enhances their cytokinin sensitivity. Mol Biol Rep 37:1089–1097CrossRefPubMedGoogle Scholar
  50. 50.
    Schmidt F, Marnef A, Cheung MK, Wilson I, Hancock J, Staiger D, Ladomery M (2010) A proteomic analysis of oligo(dT)-bound mRNP containing oxidative stress-induced Arabidopsis thaliana RNA-binding proteins ATGRP7 and ATGRP8. Mol Biol Rep 37:839–845CrossRefPubMedGoogle Scholar
  51. 51.
    Turner S, Gallois P, Brown D (2007) Tracheary element differentiation. Annu Rev Plant Biol 58:407–433CrossRefPubMedGoogle Scholar
  52. 52.
    Geier T, Sangwan RS (1996) Histology and chimeral segregation reveal cell-specific differences in the competence for shoot regeneration and Agrobacterium-mediated transformation in Kohleria internode explants. Plant Cell Rep 15:386–390CrossRefGoogle Scholar
  53. 53.
    Iwakawa H, Ueno Y, Semiarti E, Onouchi H, Kojima S, Tsukaya H, Hasebe M, Soma T, Ikezaki M, Machida C, Machida Y (2002) The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana, required for formation of a symmetric flat lamina, encodes a member of a novel family of proteins characterized by cysteine repeats and a leucine zipper. Plant Cell Physiol 43:467–478CrossRefPubMedGoogle Scholar
  54. 54.
    Inukai Y, Sakamoto T, Ueguchi-Tanaka M, Shibata Y, Gomi K, Umemura I, Hasegawa Y, Ashikari M, Kitano H, Matsuoka M (2005) Crown rootless1, which is essential for crown root formation in rice, is a target of an AUXIN RESPONSE FACTOR in auxin signaling. Plant Cell 17:1387–1396CrossRefPubMedGoogle Scholar
  55. 55.
    Naito T, Yamashino T, Kiba T, Koizumi N, Kojima M, Sakakibara H, Mizuno T (2007) A link between cytokinin and ASL9 (ASYMMETRIC LEAVES 2 LIKE 9) that belongs to the AS2/LOB (LATERAL ORGAN BOUNDARIES) family genes in Arabidopsis thaliana. Biosci Biotechnol Biochem 71:1269–1278CrossRefPubMedGoogle Scholar
  56. 56.
    Husbands A, Bell EM, Shuai B, Smith HMS, Springer PS (2007) LATERAL ORGAN BOUNDARIES defines a new family of DNA-binding transcription factors and can interact with specific bHLH proteins. Nucleic Acids Res 35:6663–6671CrossRefPubMedGoogle Scholar
  57. 57.
    Guo M, Thomas J, Collins G, Timmermansa MCP (2008) Direct repression of KNOX loci by the ASYMMETRIC LEAVES1 complex of Arabidopsis. Plant Cell 20:48–58CrossRefPubMedGoogle Scholar
  58. 58.
    Ueno Y, Ishikawa T, Watanabe K, Terakura S, Iwakaw H, Okada K, Machida C, Machidaa Y (2007) Histone deacetylases and ASYMMETRIC LEAVES2 are involved in the establishment of polarity in leaves of Arabidopsis. Plant Cell 19:445–457CrossRefPubMedGoogle Scholar
  59. 59.
    Theodoris G, Inada N, Freeling M (2003) Conservation and molecular dissection of ROUGH SHEATH2 and ASYMMETRIC LEAVES1 function in leaf development. Proc Natl Acad Sci USA 100:6837–6842CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Shao-Bo Sun
    • 1
  • Lai-Sheng Meng
    • 1
    • 2
  • Xu-Dong Sun
    • 2
  • Zhen-Hua Feng
    • 2
  1. 1.GanSu College of Traditional Chinese MedicineLanzhouChina
  2. 2.School of Life SciencesTongji UniversityShanghaiChina

Personalised recommendations