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Cloning, functional characterization and heterologous expression of TaLsi1, a wheat silicon transporter gene

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Abstract

Silicon (Si) is known to be beneficial to plants, namely in alleviating biotic and abiotic stresses. The magnitude of such positive effects is associated with a plant’s natural ability to absorb Si. Many grasses can accumulate as much as 10% on a dry weight basis while most dicots, including Arabidopsis, will accumulate less than 0.1%. In this report, we describe the cloning and functional characterization of TaLsi1, a wheat Si transporter gene. In addition, we developed a heterologous system for the study of Si uptake in plants by introducing TaLsi1 and OsLsi1, its ortholog in rice, into Arabidopsis, a species with a very low innate Si uptake capacity. When expressed constitutively under the control of the CaMV 35S promoter, both TaLsi1 and OsLsi1 were expressed in cells of roots and shoots. Such constitutive expression of TaLsi1 or OsLsi1 resulted in a fourfold to fivefold increase in Si accumulation in transformed plants compared to WT. However, this Si absorption caused deleterious symptoms. When the wheat transporter was expressed under the control of a root-specific promoter (a boron transporter gene (AtNIP5;1) promoter), a similar increase in Si absorption was noted but the plants did not exhibit symptoms and grew normally. These results demonstrate that TaLsi1 is indeed a functional Si transporter as its expression in Arabidopsis leads to increased Si uptake, but that this expression must be confined to root cells for healthy plant development. The availability of this heterologous expression system will facilitate further studies into the mechanisms and benefits of Si uptake.

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References

  • Arsenault-Labrecque G, Menzies JG, Bélanger RR (2011) Effect of silicon absorption on soybean 1 resistance to Phakopsora pachyrhizi in different cultivars. Plant Dis 96(1):37–42

  • Bélanger RR, Benhamou N, Menzies JG (2003) Cytological evidence of an active role of silicon in wheat resistance to powdery mildew (Blumeria graminis f. sp. tritici). Phytopathology 93:402–412

    Article  PubMed  Google Scholar 

  • Bragg JN, Jackson AO (2004) The C-terminal region of the Barley stripe mosaic virus protein participates in homologous interactions and is required for suppression of RNA silencing. Mol Plant Pathol 5:465–481

    Article  PubMed  CAS  Google Scholar 

  • Caron L, Rousseau F, Gagnon É, Isenring P (2000) Cloning and functional characterization of a cation-Cl cotransporter-interacting protein. J Biol Chem 275:32027–32036

    Article  PubMed  CAS  Google Scholar 

  • Chérif M, Asselin A, Bélanger RR (1994) Defense responses induced by soluble silicon in cucumber roots infected by Phytium ssp. Phytopathology 84:236–242

    Article  Google Scholar 

  • Chiba Y, Mitani N, Yamaji N, Ma JF (2009) HvLsi1 is a silicon influx transporter in barley. Plant J 57:810–818

    Article  PubMed  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • Cooke J, Leishman MR (2011) Silicon concentration and leaf longevity: is silicon a player in the leaf dry mass spectrum? Funct Ecol 25:1181–1188

    Article  Google Scholar 

  • Corrales I, Poschenrieder C, Barceló J (1997) Influence of silicon pretreatment on aluminium toxicity in maize roots. Plant Soil 190:203–209

    Article  CAS  Google Scholar 

  • Côté-Beaulieu C, Chain F, Menzies JG, Kinrade SD, Bélanger RR (2009) Absorption of aqueous inorganic and organic silicon compounds by wheat and their effect on growth and powdery mildew control. Environ Exp Bot 65:155–161

    Article  Google Scholar 

  • Diogo RVC, Wydra K (2007) Silicon-induced basal resistance in tomato against Ralstonia solanacearum is related to modification of pectic cell wall polysaccharide structure. Physiol Mol Plant Pathol 70:120–129

    Article  CAS  Google Scholar 

  • Epstein E (1999) Silicon. Annu Rev Plant Physiol Plant Mol Biol 50:641–664

    Article  PubMed  CAS  Google Scholar 

  • Fauteux F, Rémus-Borel W, Menzies JG, Bélanger RR (2005) Silicon and plant disease resistance against pathogenic fungi. FEMS Microbiol Lett 249:1–6

    Article  PubMed  CAS  Google Scholar 

  • Ghanmi D, McNally DJ, Benhamou N, Menzies JG, Bélanger RR (2004) Powdery mildew of Arabidopsis thaliana: a pathosystem for exploring the role of silicon in plant-microbe interactions. Physiol Mol Plant Pathol 64:189–199

    Article  CAS  Google Scholar 

  • Guével MH, Menzies J, Bélanger RR (2007) Effect of root and foliar applications of soluble silicon on powdery mildew control and growth of wheat plants. Eur J Plant Pathol 119:429–436

    Article  Google Scholar 

  • Hodson MJ, White PJ, Mead A, Broadley MR (2005) Phylogenetic variation in the silicon composition of plants. Ann Bot 96:1027–1046

    Article  PubMed  CAS  Google Scholar 

  • Johansen LK, Carrington JC (2001) Silencing on the spot. Induction and suppression of RNA silencing in the agrobacterium-mediated transient expression system. Plant Physiol 126:930–938

    Article  PubMed  CAS  Google Scholar 

  • Liang Y (1999) Effects of silicon on enzyme activity and sodium, potassium and calcium concentration in barley under salt stress. Plant Soil 209:217–224

    Article  CAS  Google Scholar 

  • Liang Y, Zhu J, Li Z, Chu G, Ding Y, Zhang J, Sun W (2008) Role of silicon in enhancing resistance to freezing stress in two contrasting winter wheat cultivars. Environ Exp Bot 64:286–294

    Article  CAS  Google Scholar 

  • Ma JF (2004) Role of silicon in enhancing the resistance of plants to biotic and abiotic stresses. Soil Sci Plant Nutr 50:11–18

    Article  CAS  Google Scholar 

  • Ma JF (2010) Si transporters in higher plant. In: Jhon PT, Bienert PG (eds) MIPs and their role in the exchange of materials. Landes Bioscience, Texas, pp 99–109

    Chapter  Google Scholar 

  • Ma JF, Takahashi E (2002) Soil fertilizer and plant silicon research in Japan. Elsevier, Amsterdam

    Google Scholar 

  • Ma JF, Yamaji N (2006) Silicon uptake and accumulation in higher plants. Trends Plant Sci 11:392–397

    Article  PubMed  CAS  Google Scholar 

  • Ma JF, Yamaji N (2008) Functions and transport of silicon in plants. Cell Mol Life Sci 65:3049–3057

    Article  PubMed  CAS  Google Scholar 

  • Ma JF, Higashitani A, Sato K, Takeda K (2003) Genotypic variation in silicon concentration of barley grain. Plant Soil 249:383–387

    Article  CAS  Google Scholar 

  • Ma JF, Tamai K, Yamaji N, Mitani N, Konishi S, Katsuhara M, Ishiguro K, Murata Y, Yano M (2006) A silicon transporter in rice. Nature 440:688–691

    Article  PubMed  CAS  Google Scholar 

  • Ma JF, Yamaji N, Mitani N, Kazunori T, Konishi S, Fujiwara T, Katsuhara M, Yano M (2007) An efflux transporter of silicon in rice. Nature 448:209–212

    Article  PubMed  CAS  Google Scholar 

  • Mitani N, Ma JF (2005) Uptake system of silicon in different plant species. J Exp Bot 56:1255–1261

    Article  PubMed  CAS  Google Scholar 

  • Mitani N, Chiba Y, Yamaji N, Ma JF (2009a) Identification and characterization of maize and barley Lsi2-like silicon efflux transporters reveals a distinct silicon uptake system from that in rice. Plant Cell 21:2133–2142

    Article  PubMed  CAS  Google Scholar 

  • Mitani N, Yamaji N, Ma JF (2009b) Identification of maize silicon influx transporters. Plant Cell Physiol 50:5–12

    Article  PubMed  CAS  Google Scholar 

  • Mitani N, Yamaji N, Ago Y, Iwasaki K, Ma JF (2011) Isolation and functional characterization of an influx silicon transporter in two pumpkin cultivars contrasting in silicon accumulation. Plant J 66:231–240

    Article  PubMed  CAS  Google Scholar 

  • Mitani-Ueno N, Yamaji N, Ma JF (2011) Silicon efflux transporters isolated from two pumpkin cultivars contrasting in Si uptake. Plant Signal Behav 6:991–994

    Article  PubMed  CAS  Google Scholar 

  • Richmond KE, Sussman M (2003) Got silicon? The non-essential beneficial plant nutrient. Curr Opin Plant Biol 6:268–272

    Article  PubMed  CAS  Google Scholar 

  • Rodrigues FÁ, Datnoff LE, Korndörfer GH, Seebold KW, Rush MC (2001) Effect of silicon and host resistance on sheath blight development in rice. Plant Dis 85:827–832

    Article  CAS  Google Scholar 

  • Sangster AG, Hodson MJ, Ling LEC (2009) Biomineralisation/environment interactions in conifers: Illustrated by hemlock, Tsuga canadensis (L.) Carr. Quat Int 193:3–10

    Article  Google Scholar 

  • Takahashi E, Ma JF, Miyake Y (1990) The possibility of silicon as an essential element of higher plants. Comments Agric Food Chem 2:99–122

    CAS  Google Scholar 

  • Takano J, Wada M, Ludewig U, Schaaf G, vonWiren N, Fujiwara T (2006) The Arabidopsis major intrinsic protein NIP5; 1 is essential for efficient boron uptake and plant development under boron limitation. Plant Cell 18:1498–1509

    Article  PubMed  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  • Tocquin P, Corbesier L, Havelange A, Pieltain A, Kurtem E, Bernier G, Perilleux C (2003) A novel high efficiency, low maintenance, hydroponic system for synchronous growth and flowering of Arabidopsis thaliana. BMC Plant Biol 3:2

    Article  PubMed  Google Scholar 

  • Yamaji N, Mitani N, Ma JF (2008) A transporter regulating silicon distribution in rice shoots. Plant Cell 20:1381–1389

    Article  PubMed  CAS  Google Scholar 

  • Zhang C, Wang L, Nie Q, Zhang W, Zhang F (2008) Long-term effects of exogenous silicon on cadmium translocation and toxicity in rice (Oryza sativa L.). Environ Exp Bot 62:300–307

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (NSERC), and the Canada Research Chairs Program to R.R. Bélanger and by Grant Genomics for Agricultural Innovation IPG-0006 from the Ministry of Agriculture, Forestry and Fisheries of Japan (to J.F.M.) and Grant-in-Aid for Scientific Research 21248009 and 22119002 on Innovative Areas from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to J.F.M.).

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Correspondence to Richard R. Bélanger.

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Jonatan Montpetit, Julien Vivancos and Namiki Mitani-Ueno contributed equally to this work.

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11103_2012_9892_MOESM1_ESM.doc

Supplementary Fig. 1: Graphical representation of a Kyte and Doolittle hydrophobicity profile generated from the predicted amino acid sequences of the rice and wheat silicon transporter genes. Transmembrane segments (TM) and NPA boxes are positioned on the graph. X-axis represents the numbering in amino acids (DOC 44 kb)

Supplementary Table 1: Primer sequences (EPS 686 kb)

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Montpetit, J., Vivancos, J., Mitani-Ueno, N. et al. Cloning, functional characterization and heterologous expression of TaLsi1, a wheat silicon transporter gene. Plant Mol Biol 79, 35–46 (2012). https://doi.org/10.1007/s11103-012-9892-3

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  • DOI: https://doi.org/10.1007/s11103-012-9892-3

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