Planta

, Volume 232, Issue 4, pp 925–936

The oil palm metallothionein promoter contains a novel AGTTAGG motif conferring its fruit-specific expression and is inducible by abiotic factors

  • Vahid Omidvar
  • Siti Nor Akmar Abdullah
  • Amir Izadfard
  • Chai Ling Ho
  • Maziah Mahmood
Original Article

Abstract

The 1,053-bp promoter of the oil palm metallothionein gene (so-called MSP1) and its 5′ deletions were fused to the GUS reporter gene, and analysed in transiently transformed oil palm tissues. The full length promoter showed sevenfold higher activity in the mesocarp than in leaves and 1.5-fold more activity than the CaMV35S promoter in the mesocarp. The 1,053-bp region containing the 5′ untranslated region (UTR) gave the highest activity in the mesocarp, while the 148-bp region was required for minimal promoter activity. Two positive regulatory regions were identified at nucleotides (nt) −953 to −619 and −420 to −256 regions. Fine-tune deletion of the −619 to −420 nt region led to the identification of a 21-bp negative regulatory sequence in the −598 to −577 nt region, which is involved in mesocarp-specific expression. Gel mobility shift assay revealed a strong interaction of the leaf nuclear extract with the 21-bp region. An AGTTAGG core-sequence within this region was identified as a novel negative regulatory element controlling fruit-specificity of the MSP1 promoter. Abscisic acid (ABA) and copper (Cu2+) induced the activity of the promoter and its 5′ deletions more effectively than methyl jasmonate (MeJa) and ethylene. In the mesocarp, the full length promoter showed stronger inducibility in response to ABA and Cu2+ than its 5′ deletions, while in leaves, the −420 nt fragment was the most inducible by ABA and Cu2+. These results suggest that the MSP1 promoter and its regulatory regions are potentially useful for engineering fruit-specific and inducible gene expression in oil palm.

Keywords

Biolistic transformation Fruit-specific promoter Gel mobility shift assay Inducible expression Oil palm metallothionein Transient expression assay 

Abbreviations

UTR

Untranslated region

GUS

β-Glucuronidase

GFP

Green fluorescent protein

EMSA

Electrophoretic mobility shift assay

MT

Metallothionein

ERE

Ethylene-responsive element

ABRE

Abscisic acid-responsive element

MeJa-RE

Methyl jasmonate-responsive element

WUN-RE

Wounding-responsive element

Cu2+-RE

Copper-responsive element

Supplementary material

425_2010_1220_MOESM1_ESM.doc (33 kb)
Supplementary material 1 (DOC 33 kb)
425_2010_1220_MOESM2_ESM.doc (136 kb)
Supplementary material 2 (DOC 135 kb)
425_2010_1220_MOESM3_ESM.doc (34 kb)
Supplementary material 3 (DOC 34 kb)
425_2010_1220_MOESM4_ESM.doc (38 kb)
Supplementary material 4 (DOC 37 kb)

References

  1. Acevedo-Hernandez GJ, Leon P, Herrera-Estrella LR (2005) Sugar and ABA responsiveness of a minimal RBCS light-responsive unit is mediated by direct binding of ABI4. Plant J 43:506–519CrossRefPubMedGoogle Scholar
  2. Agius F, Amaya I, Botella MA, Valpuesta V (2005) Functional analysis of homologous and heterologous promoters in strawberry fruits using transient expression. J Exp Bot 56:37–46Google Scholar
  3. Atkinson RG, Bolitho KM, Wright MA, Iturriagagoitia-Bueno T, Reid SJ, Ross GS (1998) Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato. Plant Mol Biol 38:449–460CrossRefPubMedGoogle Scholar
  4. Bratic AM, Majiv DB, Samardzic JT, Maksimovic VR (2009) Functional analysis of the buckwheat metallothionein promoter: tissue specificity pattern and up-regulation under complex stress stimuli. Plant Physiol 166:996–1000CrossRefGoogle Scholar
  5. Chatthai M, Osusky M, Osuska L, Yevtushenko D, Misra S (2004) Functional analysis of a Douglas-fir metallothionein-like gene promoter: transient assays in zygotic and somatic embryos and stable transformation in transgenic tobacco. Planta 220:118–128CrossRefPubMedGoogle Scholar
  6. Clendennen SK, May GD (1997) Differential gene expression in ripening banana fruit. Plant Physiol 115:463–469CrossRefPubMedGoogle Scholar
  7. Cobbett C, Goldsbrough P (2002) Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasis. Annu Rev Plant Biol 53:159–182CrossRefPubMedGoogle Scholar
  8. Dong JZ, Dunstan DI (1996) Expression of abundant mRNA during somatic embryogenesis of white spruce [Picea glauca (Moench) Voss]. Planta 199:459–466CrossRefPubMedGoogle Scholar
  9. Endo T, Shimada T, Fujii H, Moriguchi T, Omura M (2007) Promoter analysis of a type 3 metallothionein-like gene abundant in Satsuma mandarin (Citrus unshiu Marc.) fruit. Sci Hortic 12:207–214CrossRefGoogle Scholar
  10. Foley RC, Liang ZM, Singh KB (1997) Analysis of type 1 metallothionein cDNAs in Vicia faba. Plant Mol Biol 33:583–591CrossRefPubMedGoogle Scholar
  11. Fordham-Skelton AP, Lilley C, Urwin PE, Robinson NJ (1997) GUS expression in Arabidopsis directed by 5′-regions of the pea metallothionein-like gene PsMTA. Plant Mol Biol 34:659–668CrossRefPubMedGoogle Scholar
  12. Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucl Acid Res 27:297–300CrossRefGoogle Scholar
  13. Hobo T, Asada M, Kowyama Y, Hattori T (1999) ACGT-containing abscisic acid response element (ABRE) and coupling element 3 (CE3) are functionally equivalent. Plant J 19:679–689CrossRefPubMedGoogle Scholar
  14. Hsieh HM, Huang PC (1998) Promoter structure and activity of type 1 rice metallothionein-like gene. DNA Seq 9:9–17PubMedGoogle Scholar
  15. Hsieh HM, Liu WK, Chang A, Huang PC (1996) RNA expression patterns of a type 2 metallothionein-like gene from rice. Plant Mol Biol 32:525–529CrossRefPubMedGoogle Scholar
  16. Hu CY, Bol G, Xia LH, Biao YZ (2006) Isolation and characterization of El I gene promoter from tomato. Agric Sci China 5:661–669Google Scholar
  17. Hwang SH, Lee IA, Yie SW, Hwang DJ (2008) Identification of an OsPR10a promoter region responsive to salicylic acid. Planta 227:1141–1150CrossRefPubMedGoogle Scholar
  18. Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405CrossRefGoogle Scholar
  19. Kawashima I, Kennedy TD, Chino M, Lane BG (1992) Wheat Ec metallothionein genes like mammalian Zn2+ metallothionein genes are conspicuously expressed during embryogenesis. Eur J Biochem 209:971–976CrossRefPubMedGoogle Scholar
  20. Lang BG, Kajioka R, Kennedy TD (1987) The wheat germ Ec protein is a zinc-containing metallothionein. Biochem Cell Biol 65:1001–1005CrossRefGoogle Scholar
  21. Ledger SE, Gardner RC (1994) Cloning and characterization of five cDNAs for genes differentially expressed during fruit development of kiwifruit (Actinidia deliciosa var. deliciosa). Plant Mol Biol 25:877–886CrossRefPubMedGoogle Scholar
  22. Lescot M, Dehais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouze P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acid Res 30:325–327CrossRefPubMedGoogle Scholar
  23. Lü S, Gu H, Yuan X, Wang X, Wu AM, Qu L, Liu JY (2007) The GUS reporter-aided analysis of the promoter activities of a rice metallothionein gene reveals different regulatory regions responsible for tissue-specific and inducible expression in transgenic Arabidopsis. Trans Res 16:177–191CrossRefGoogle Scholar
  24. Masani AMY, Ho CL, Parveez GKA (2008) Construction of PHB and PHBV transformation vectors for bioplastics production in oil palm. J Oil Palm Res 2:37–55Google Scholar
  25. Murashige T, Skoog F (1962) A revised method for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  26. Naik SN, Goud VV, Rout PK, Dalai AK (2010) Production of first and second generation biofuels: a comprehensive review. Renew Sustain Energ Rev 14:578–597CrossRefGoogle Scholar
  27. Nash J, Luehrsen KR, Walbot V (1990) Bronze-2 gene of maize: reconstruction of a wild-type allele and analysis of transcription and splicing. Plant Cell 2:1039–1049CrossRefPubMedGoogle Scholar
  28. Omidvar V, Siti Nor Akmar A, Marziah M, Maheran AA (2008) A transient assay to evaluate the expression of polyhydroxybutyrate genes regulated by oil palm mesocarp-specific promoter. Plant Cell Rep 27:1451–1459CrossRefPubMedGoogle Scholar
  29. Pastuglia M, Roby D, Dumas C, Cock JM (1997) Rapid induction by wounding and bacterial infection of S gene family receptor-like kinase in Brassica oleracea. Plant Cell 9:1–13Google Scholar
  30. Quinn JM, Merchant S (1995) Two copper-responsive elements associated with the Chlamydomonas Cyc6 gene function as targets for transcriptional activators. Plant Cell 7:623–628CrossRefPubMedGoogle Scholar
  31. Rasmussen TB, Donaldson LA (2006) Investigation of the endosperm-specific sucrose synthase promoter from rice using transient expression of reporter genes in guar seed tissue. Plant Cell Rep 25:1035–1042CrossRefPubMedGoogle Scholar
  32. Rauser WE (1999) Structure and function of metal chelators produced by plants: the case for organic acids, amino acids, phytin, and metallothioneins. Cell Biochem Biophysiol 31:19–48CrossRefGoogle Scholar
  33. Reid SJ, Ross GS (1997) Up-regulation of two cDNA clones encoding metallothionein-like proteins in apple fruit during cool storage. Plant Physiol 100:183–189CrossRefGoogle Scholar
  34. Ren Y, Zhao J (2009) Functional analysis of the rice metallothionein gene OsMT2b promoter in transgenic Arabidopsis plants and rice germinated embryos. Plant Sci 176:528–538CrossRefGoogle Scholar
  35. Robinson NJ, Tommey AM, Kuske C, Jackson PJ (1993) Plant metallothioneins. Biochem J 295:1–10PubMedGoogle Scholar
  36. Rouster J, Leah R, Mundy J, Cameron-Mills V (1997) Identification of a methyl jasmonate-responsive region in the promoter of a lipoxygenase-1 gene expressed in barley grain. Plant J 11:513–523CrossRefPubMedGoogle Scholar
  37. Siti Nor Akmar A, Zubaidah R (2007) Regulatory sequences for regulation of gene expression in plants and other organisms, and compositions, products and methods related thereto. United State Patent US 7,173,120 B2Google Scholar
  38. Siti Nor Akmar A, Cheah SC, Murphy DJ (2002) Isolation and characterization of two divergent type 3 metallothioneins from oil palm (Elaeis guineensis). Plant Physiol Biochem 40:255–263CrossRefGoogle Scholar
  39. Terzaghi WB, Cashmore AR (1995) Light-regulated transcription. Annu Rev Plant Physiol Plant Mol Biol 46:445–474CrossRefGoogle Scholar
  40. Whitelaw CA, Huquet JA, Thurman DA (1995) The isolation and characterization of type II metallothionein-like genes from tomato (Lycopersicon esculentum L.). Plant Mol Biol 29:685–687CrossRefGoogle Scholar
  41. Wong HL, Sakamoto T, Kawasaki T, Umemura K, Shimamoto K (2004) Down-regulation of metallothionein, a reactive oxygen scavenger, by the small GTPase OsRac1 in rice. Plant Physiol 135:1447–1456CrossRefPubMedGoogle Scholar
  42. Yamagata H, Yonesu K, Hirata A, Aizono Y (2002) TGTCACA motif is a novel cis-regulatory enhancer element involved in fruit-specific expression of the cucumisin gene. J Biol Chem 277:11582–11590CrossRefPubMedGoogle Scholar
  43. Yi SY, Sun AQ, Sun Y, Yang JY, Zhao CM, Liu J (2006) Differential regulation of Lehsp23.8 in tomato plants: analysis of a multiple stress-inducible promoter. Plant Sci 171:398–407CrossRefGoogle Scholar
  44. Yin T, Wu H, Zhang S, Liu J, Lu H, Zhang L, Xu Y, Chen D (2009) Two negative cis-regulatory regions involved in fruit-specific promoter activity from watermelon (Citrullus vulgaris S.). J Exp Bot 60:169–185CrossRefPubMedGoogle Scholar
  45. Zhang H, Huang Z, Xie B, Chen Q, Tian X, Zhang X, Zhang H, Lu X, Huang D, Huang R (2004) The ethylene-, jasmonate-, abscisic acid- and NaCl-responsive tomato transcription factor JERF1 modulates expression of GCC box-containing genes and salt tolerance in tobacco. Planta 220:262–270CrossRefPubMedGoogle Scholar
  46. Zhou J, Goldsbrough PB (1995) Structure, organization and expression of the metallothionein gene family in Arabidopsis. Mol Gen Genet 248:318–328CrossRefPubMedGoogle Scholar
  47. Zhou GK, Xu YF, Liu JY (2005) Characterization of a rice class II metallothionein gene: tissue expression patterns and induction in response to abiotic factors. J Plant Physiol 162:686–696CrossRefPubMedGoogle Scholar
  48. Zubaidah R, Siti Nor Akmar A (2003) Development of a transient promoter assay system for oil palm. J Oil Palm Res 15:62–69Google Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Vahid Omidvar
    • 1
    • 2
  • Siti Nor Akmar Abdullah
    • 1
    • 2
  • Amir Izadfard
    • 1
  • Chai Ling Ho
    • 3
  • Maziah Mahmood
    • 3
  1. 1.Department of Agriculture Technology, Faculty of AgricultureUniversity Putra Malaysia (UPM)SerdangMalaysia
  2. 2.Laboratory of Plantation CropsInstitute of Tropical Agriculture, UPMSerdangMalaysia
  3. 3.Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular SciencesUniversity Putra Malaysia (UPM)SerdangMalaysia

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