Potential for manipulating carbon metabolism in wheat

Sparks, C. A. and Castleden, C. K. and West, J. and Habash, D. Z. and Madgwick, P. J. and Paul, M. J. and Noctor, G. and Harrison, J. and Wu, R. and Wilkinson, J. and Quick, W. P. and Parry, M. A J and Foyer, C. H. and Miflin, B. J. (2001) Potential for manipulating carbon metabolism in wheat. Annals of Applied Biology, 138 (1). pp. 33-45. ISSN 0003-4746

Abstract

The transformation of wheat opens up opportunities for manipulation of key steps in plant metabolic pathways. Attempts to alter the expression of sucrose phosphate synthase (SPS) and glycine decarboxylase (GDC) in wheat leaves have been made. Using a maize SPS gene under the control of either a constitutive promoter (in p35SMSPS) or a green tissue specific promoter (in pRGMSPS), transformation of wheat (Triticum aestivum L. cv. Cadenza) led to the production of 22 independent transformed lines that contained the SPS transgene. Expression of the transgene, analysed by RT-PCR, revealed that the transcript was present in nine of the 12 p35SMSPS plants and six of the 10 pRGMSPS plants. The maize SPS protein could not, however, be detected in any of these lines. The expression of GDC, and in particular that of the P subunit, has been shown to be altered in plant species with the intermediate C3/C4 photosynthetic phenotype. Attempts have been made to make similar changes in the expression of GDC in wheat leaves by transforming them with full length cDNA sequences for the P subunit of GDC in the sense and antisense orientation. More than 70 transformed lines were produced. Of these, 11 contained the gene in the sense, and 31 in the antisense, orientation, as determined by PCR. Results with T1 generation plants suggested that the activity of GDC had been reduced in some lines transformed with antisense constructs. However, there was no evidence for the expression of the sense constructs. GDC mRNA could not be detected by RT-PCR in any of the 10 lines produced in the sense transformation experiments. Taken together, these results demonstrate the effectiveness of the technology to deliver large numbers of transgenic wheat plants. However, clearly this alone is not sufficient to guarantee the correct expression of the transgenes at sufficient levels to establish the presence of the gene product.