, 229:861 | Cite as

Heat-inducible C3HC4 type RING zinc finger protein gene from Capsicum annuum enhances growth of transgenic tobacco

  • Naheed Zeba
  • Mohammad Isbat
  • Nak-Jung Kwon
  • Mi Ok Lee
  • Seong Ryong Kim
  • Choo Bong Hong
Original Article


Capsicum annuum RING Zinc Finger Protein 1 (CaRZFP1) gene is a novel C3HC4-type RING zinc finger protein gene which was previously isolated from a cDNA library for hot pepper plants treated of heat-shock. The CaRZFP1 was inducible to diverse environmental stresses in hot pepper plants. We introduced the CaRZFP1 into the Wisconsin 38 cultivar of tobacco (Nicotiana tabacum) by Agrobacterium mediated transformation under the control of the CaMV 35S promoter. Expression of the transgene in the transformed tobacco plants was demonstrated by RNA blot analyses. There appeared no adverse effect of over-expression of the transgene on overall growth and development of transformants. The genetic analysis of tested T1 lines showed that the transgene segregated in a Mendelian fashion. Transgenic tobacco lines that expressed the CaRZFP1 gene were compared with several different empty vector lines and they exhibited enhanced growth; they have larger primary root, more lateral root, larger hypocotyls and bigger leaf size, resulting in heavier fresh weight. Enhanced growth of transgenic lines accompanied with longer vegetative growth that resulted in bigger plants with higher number of leaves. Microarray analysis revealed the up-regulation of some growth related genes in the transgenic plants which were verified by specific oligomer RNA blot analyses. These results indicate that CaRZFP1 activates and up-regulates some growth related proteins and thereby effectively promoting plant growth.


Capsicum Enhanced growth Heat inducible RING zinc finger protein Transgenic tobacco plants 



Arabinogalactan protein


Filamentous temperature-sensitive Z


Glycine rich protein


Light harvesting complex protein


Proline rich protein


The Arabidopsis Information Resource



This work was supported by grants to Choo Bong Hong from the Crop Functional Genomics Center of the twenty-first century Frontier Research Program (code# CG1434) funded by the Ministry of Science and Technology of Korea and from the Priority Research Institute supported by the Korean Research Foundation (grant no. 2005-005-J16002). Naheed Zeba and Mohammad Isbat were partially supported by BK21 Research Fellowships from the Ministry of Education and Human Resources Development, Korea. Transformation, in vitro regeneration, and cultivation of tobacco plants were provided by Kab Lim Lee and Mi Jin Lee of Planta Co., Korea.


  1. Alexandrov NN, Troukhan ME, Brover VV, Tatarinova T, Flavell RB, Feldmann KA (2006) Features of Arabidopsis gene and genome discovered using full-length cDNAs. Plant Mol Biol 60:71–87CrossRefGoogle Scholar
  2. Armengaud P, Breitling R, Amtmann A (2004) The potassium-dependent transcriptome of Arabidopsis reveals a prominent role of jasmonic acid in nutrient signaling. Plant Physiol 136:2556–2576PubMedCrossRefGoogle Scholar
  3. Arumugam TU, Davies E, Morita EH, Abe S (2007) Sequence, expression and tissue localization of a gene encoding a makorin RING zinc-finger protein in germinating rice (Oryza sativa L. ssp. Japonica) seeds. Plant Physiol Biochem 45:767–780PubMedCrossRefGoogle Scholar
  4. Bernhardt C, Tierney ML (2000) Expression of AtPRP3, a proline-rich structural cell wall protein from Arabidopsis, is regulated by Cell-Type-specific developmental pathways involved in root hair formation. Plant Physiol 122:705–714PubMedCrossRefGoogle Scholar
  5. Deng XW, Matsui M, Wei N, Wagner D, Chu AM, Feldmann KA, Quail PH (1992) COP1, and Arabidopsis regulatory gene, encodes a protein with both a zinc-binding motif and a G homologous domain. Cell 71:791–801PubMedCrossRefGoogle Scholar
  6. Ding L, Zhu JK (1997) A role for arabinogalactan-proteins in root epidermal cell expansion. Planta 203:289–294PubMedCrossRefGoogle Scholar
  7. Freemont PS, Hanson IM, Trowsdale J (1991) A novel cysteine-rich sequence motif. Cell 64:483–484PubMedCrossRefGoogle Scholar
  8. Freemont PS (2000) RING for destruction? Curr Biol 10:84–87CrossRefGoogle Scholar
  9. Haas BJ, Volfovsky N, Town CD, Troukhan M, Alexandrov N, Feldmann KA, Flavell RB, White O, Salzberg SL (2002) Full-length messenger RNA sequences greatly improve genome annotation. Genome Biol 3:1–12CrossRefGoogle Scholar
  10. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231CrossRefGoogle Scholar
  11. Isabel S, Leblanc E, Boissinot M, Boudreau DK, Grondin M, Picard FJ, Martel EA, Parham NJ, Chain PS, Bader DE, Mulvey MR, Bryden L, Roy PH, Ouellette M, Bergeron MG (2008) Divergence among genes encoding the elongation factor Tu of Yersinia species. J Bacteriol (Epub ahead of print)Google Scholar
  12. Joazeiro CA, Wing SS, Huang H, Leverson JD, Hunter T, Liu YC (1999) The tyrosin kinase negative regulator c-Cbl as a RING-type E2-dependent ubiquitin-protein ligase. Science 286:309–312PubMedCrossRefGoogle Scholar
  13. Knox JP, Linstead PJ, Peart J, Cooper C, Roberts K (1991) Developmentally regulated epitopes of cell surface arabinogalactan proteins and their relation to root tissue pattern formation. Plant J 1:317–326Google Scholar
  14. Ko JH, Seung HY, Kyung HH (2006) Upregulation of an Arabidopsis RING-H2 gene, XERICO confers drought tolerance through increased abscisic acid biosynthesis. Plant J 47:343–355PubMedCrossRefGoogle Scholar
  15. Mazumder B, Sampath P, Seshadri V, Maitra RK, DiCorleto PE, Fox PL (2003) Regulated release of L13a from the 60S ribosomal subunit as a mechanism of transcript-specific translational control. Cell 115:187–198PubMedCrossRefGoogle Scholar
  16. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Planta 157:385–391Google Scholar
  17. Park MH, Suzuki Y, Chono M, Knox JP, Yamaguchi I (2003) CsAGP1, a gibberellin-responsive gene from cucumber hypocotyls, encodes a classical arabinogalactan protein and is involved in stem elongation. Plant Physiol 131:1450–1459PubMedCrossRefGoogle Scholar
  18. Peng J, Richards DE, Moritz T, Cano-Delgado A, Harberd NP (1999) Extragenic Suppressors of the Arabidopsis gai mutation alter the dose-response relationship of diverse gibberellin responses. Plant Physiol 119:1199–1208PubMedCrossRefGoogle Scholar
  19. Pennell RI, Roberts K (1990) Sexual development in the pea is presaged by altered expression of arabinogalactan protein. Nature 344:547–549CrossRefGoogle Scholar
  20. Raab S, Hoth S (2007) A mutation in the AtPRP4 splicing factor gene suppresses seed development in Arabidopsis. Plant Biol (Stuttg) 9:447–452CrossRefGoogle Scholar
  21. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Press, New YorkGoogle Scholar
  22. Sasaki T, Matsumoto T, Yamamoto K, Sakata K, Baba T, Katayose Y, Wu J, Niimura Y, Cheng Z, Nagamura Y, Antonio BA, Kanamori H, Hosokawa S, Masukawa M, Arikawa K, Chiden Y, Hayashi M, Okamoto M, Ando T, Aoki H, Arita K, Hamada M, Harada C, Hijishita S, Honda M, Ichikawa Y, Idonuma A, Iijima M, Ikeda M, Ikeno M, Ito S, Ito T, Ito Y, Ito Y, Iwabuchi A, Kamiya K, Karasawa W, Katagiri S, Kikuta A, Kobayashi N, Kono I, Machita K, Maehara T, Mizuno H, Mizubayashi T, Mukai Y, Nagasaki H, Nakashima M, Nakama Y, Nakamichi Y, Nakamura M, Namiki N, Negishi M, Ohta I, Ono N, Saji S, Sakai K, Shibata M, Shimokawa T, Shomura A, Song J, Takazaki Y, Terasawa K, Tsuji K, Waki K, Yamagata H, Yamane H, Yoshiki S, Yoshihara R, Yukawa K, Zhong H, Iwama H, Endo T, Ito H, Hahn JH, Kim HI, Eun MY, Yano M, Jiang J, Gojobori T (2002) The genome sequence and structure of rice chromosome 1. Nature 420:312–316PubMedCrossRefGoogle Scholar
  23. Saurin AJ, Borden KLB, Boddy MN, Freemont PS (1996) Does this have a familiar RING? Trends Biochem Sci 21:208–214PubMedGoogle Scholar
  24. Schulz B, Kolukisaoglu HU (2006) Genomics of plant ABC transporters: the alphabet of photosynthetic life forms or just holes in membranes? FEBS Lett 580:1010–1016PubMedCrossRefGoogle Scholar
  25. Schumann U, Prestele J, O’Geen H, Brueggeman R, Wanner G, Giet C (2007) Requirement of the C3HC4 zinc RING finger of the Arabidopsis PEX10 for photorespiration and leaf peroxisome contact with chloroplasts. Proc Natl Acad Sci USA 104:1069–1074PubMedCrossRefGoogle Scholar
  26. Seifert GJ, Roberts K (2007) The biology of arabinogalactan proteins. Annu Rev Plant Biol 58:137–161PubMedCrossRefGoogle Scholar
  27. Seong ES, Choi D, Cho HS, Lim CK, Cho HJ, Wang MH (2007) Characterization of a stress-responsive ankyrin repeat-containing zinc finger protein of Capsicum annuum (CaKR1). J Biochem Mol Biol 40:952–958PubMedGoogle Scholar
  28. Serpe MD, Nothnagel EA (1994) Effects of Yariv phenylglycosides on Rosa cell suspension: evidence for the involvement of arabinogalactan-proteins in cell proliferation. Planta 193:542–550CrossRefGoogle Scholar
  29. Serrano M, Guzman P (2004) Isolation and gene expression analysis of Arabidopsis thaliana mutants with constitutive expression of ATL2, an early elicitor-response RING-H2 zinc-finger gene. Genetics 167:919–929PubMedCrossRefGoogle Scholar
  30. Shi H, Kim Y, Guo Y, Stevenson B, Zhu JK (2003) The Arabidopsis SOS5 locus encodes a putative cell surface adhesion protein and is required for normal cell expansion. Plant Cell 15:19–32PubMedCrossRefGoogle Scholar
  31. Spitzer C, Schellmann S, Sabovljevic A, Shahriari M, Keshavaiah C, Bechtold N, Herzog M, Müller S, Hanisch FG, Hülskamp M (2006) The Arabidopsis elch mutant reveals functions of an ESCRT component in cytokinesis. Development 133:4679–4689PubMedCrossRefGoogle Scholar
  32. Srinivasan R, Mishra M, Wu L, Yin Z, Balasubramanian MK (2008) The bacterial cell division protein FtsZ assembles into cytoplasmic rings in fission yeast. Genes Dev 22:1741–1746PubMedCrossRefGoogle Scholar
  33. Suh MC, Hong CB, Kim SS, Sim WS (1994) Transgenic tobacco plants with Bacillus thuringiensis δ-endotoxin gene resistant to Korean born tobacco budworms. Mol Cells 4:211–719Google Scholar
  34. Torii KU, McNellis TW, Deng XW (1998) Functional dissection of Arabidopsis COP1 reveals specific roles of its three structural modules in light control of seedling development. EMBO J 17:5577–5587PubMedCrossRefGoogle Scholar
  35. Vij S, Tyagi AK (2006) Genome-wide analysis of the associated protein (SAP) gene family containing A20/AN1 zinc-finger(s) in rice and their phylogenetic relationship with Arabidopsis. Mol Gen Genomics 276:171–565CrossRefGoogle Scholar
  36. Willats WGT, Knox JP (1996) A-role for arabinogalactan-proteins in plant cell expansions: evidence from studies on the interaction of β-glucosyl Yariv reagent with seedlings of Arabidopsis thaliana. Plant J 9:919–925PubMedCrossRefGoogle Scholar
  37. Workman C, Jensen LJ, Jarmer H, Berka R, Gautier L, Nielser HB, Saxild HH, Nielsen C, Brunak S, Knudsen S (2002) A new non-linear normalization method for reducing variability in DNA microarray experiments. Genome biol 3:1–16CrossRefGoogle Scholar
  38. Wu H, Wong E, Ogdahi J, Cheung AY (2000) A pollen tube growth-promoting arabinogalactan protein from Nicotiana alata is similar to the tobacco TTS protein. Plant J 22:165–176PubMedCrossRefGoogle Scholar
  39. Xu R, Li QQ (2003) A RING-H2 zinc-finger protein gene RIE1 is essential for seed development in Arabidopsis. Plant Mol Biol 53:37–50PubMedCrossRefGoogle Scholar
  40. Yang J, Sardar HS, McGovern KR, Zhang Y, Showalter AM (2007) A lysine-rich arabinogalactan protein in Arabidopsis is essential for plant growth and development, including cell division and expansion. Plant J 49:629–640PubMedCrossRefGoogle Scholar
  41. Yoo CM, Wen J, Motes CM, Sparks JA, Blancaflor EB (2008) A class one ADP-ribosylation factor GTPase-activating protein is critical for maintaining directional root hair growth in Arabidopsis thaliana. Plant Physiol 147:1659–1674PubMedCrossRefGoogle Scholar
  42. Yu HJ, Hogan P, Sundaresan V (2005) Analysis of the female gametophyte transcriptome of Arabidopsis by comparative expression profiling. Plant Physiol 139:1853–1869PubMedCrossRefGoogle Scholar
  43. Yuan J, Kight A, Goforth RL, Moore M, Peterson EC, Sakon J, Henry R (2002) ATP stimulates signal recognition particle (SRP)/FtsY-supported protein integration in chloroplasts. J Biol Chem 277:32400–32404PubMedCrossRefGoogle Scholar
  44. Zeba N, Ashrafuzzaman M, Hong CB (2006) Molecular characterization of the Capsicum annuum RING Zinc Finger Protein 1 (CaRZFP1) gene induced by abiotic stresses. J Plant Biol 49:484–490CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Naheed Zeba
    • 1
  • Mohammad Isbat
    • 2
  • Nak-Jung Kwon
    • 1
    • 3
  • Mi Ok Lee
    • 1
  • Seong Ryong Kim
    • 2
  • Choo Bong Hong
    • 1
  1. 1.School of Biological Sciences and Institute of Molecular Biology and Genetics Seoul National UniversitySeoulSouth Korea
  2. 2.Department of Life SciencesSogang UniversitySeoulSouth Korea
  3. 3.Department of BacteriologyUniversity of WisconsinMadisonUSA

Personalised recommendations