Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S477

[P-P&F.16]

Plant genetic transformation for increased yield through C3 toC4 photosynthesis engineering strategy

O.I. Kershanskaya ∗, A.S. Nurmagambetova, L.A. Skvortsova, D.S.Nelidova, L.I. Rovenskaya, T.M. Mukhanov

Institute Plant Biology and Biotechnology, National Center Biotechnol-ogy, KazakhstanKeywords: transformation; C3 - C4 engineering; yield; wheat

The random and photosynthetic nature of the evolution of C4plants suggests that the genes encoding C4 enzymes evolved froma set of pre-existing genes in ancestral C3 plants that did not func-tion in photosynthesis. Modification of genes encoding maize C4enzymes in ancestral C3 wheat to allow their high-level expres-sion in green leaves must increase sustainability of crop production.Genetic improvement of wheat photosynthesis for increased yieldby application of plant transformation technologies could be con-sidered as a first step to wheat transformation for increased yield,metabolic engineering and metabolomics, thus to forward plantbiotechnology. Low efficiently results are known so far.

The ultimate objective of our research is to investigatethe advances in transformation technologies and thus establishapproaches for genetic modification of photosynthesis in wheat forincrease grain yield up to 30% through introduction of maize genesencoding the C4 photosynthesis enzymes into wheat.

Simple natural germ-line transformation technique by Pipet-ting Agrobacterium into the spikelets of wheat before anthesishas being elaborated. Using this method allowed us to produceputative transgenic wheat plants expressing two important maizegenes: phosphoenolpyruvate carboxylase (PEPC) and orthophos-phate pyruvate dikinase (PPDK). High level of expression of thesegenes in transgenic wheat plants was determined by assaying theactivity of PEPC in leaf protein extract, followed by gel electrophore-sis, Western, Southern blot and PCR analyses. High activities of theenzyme were correlated with the amounts of enzyme protein inthe leaves. Most transgenic wheat plants exhibited an enhancedphotosynthetic capacity. All transgenic lines showed superior pho-tosynthetic performance under different water regimes. Transgenicwheat lines expressing maize genes produced higher grain yield(25-30%), especially under adverse conditions.

doi:10.1016/j.jbiotec.2010.09.720

[P-P&F.17]

Guidance of EFSA in the area of GM plants and GM animals:overview and new developments

N Podevin

EFSA, ItalyKeywords: risk assessment; Europe; guidance

The European Food Safety Authority (EFSA) provides indepen-dent, objective and transparent scientific advice on Geneticallymodified organisms (GMOs) to the risk managers which are respon-sible for taking the market authorisation decisions.

In 2003, EFSA established its Panel on GMOs to provide indepen-dent scientific advice on the safety of GMOs for humans, animalsand the environment. The EFSA GMO Panel, is composed of 21 inter-nationally recognised scientists with a broad range of expertises,carries out its work either in response to requests for scientificadvice from risk managers or on its own initiative. In differentworking groups the EFSA GMO Panel is supported by around 50

external scientists with relevant expertise and the EFSA GMOunit.

To guide and assist applicants in the preparation and presen-tation of GMO market authorisation dossiers the EFSA GMO Panelhas published numerous guidance documents on the risk assess-ment of GMOs. Recently, the EFSA GMO Panel has been updatingits guidance documents on risk assessment of GM plants both inthe food/feed as the environmental area. In addition, work wasstarted in the area of GM plants for non-food and non-feed pur-poses and GM animals. The guidance documents follow specificEU legal requirements, defines data requirements and provides adetailed description of both scientific issues and principles to beconsidered when performing the risk assessment of GMOs.

doi:10.1016/j.jbiotec.2010.09.721

[P-P&F.18]

Agrobacterium tumefaciens-Mediated Transformation of Digi-talis purpurea L., and Important Pharmaceutical Crop

Naivy Pérez-Alonso 1, Borys Chong-Pérez 1,2,∗, Alina Capote 1,Anabel Pérez 1, Yovanny Izquierdo 1, Geert Angenon 2

1 Instituto de Biotecnologia de las Plantas, Universidad Central “MartaAbreu” de Las Villas, Cuba2 Laboratory of Plant Genetics, Department of Applied Biological Sci-ences Vrije Universiteit Brussel, BelgiumKeywords: Cardenolides; gus; PCR; Southern hybridization

Digitalis purpurea L., contain cardiac glycosides of major inter-est in pharmaceutical industries. Genetic transformation is a toolof special interest for these compounds, exclusively obtained fromplants. Besides, it grants the possibility to widen the knowledgeon the biosynthesis of these compounds. In this study, an effi-cient transformation protocol for D. purpurea that would permit thestable expression of transgenes was developed. A regeneration pro-tocol via somatic embryogenesis was obtained from in vitro plantleaf segments. The minimal inhibitory concentration of geneticinwas determined for each step of the protocol. Two Agrobacteriumstrains were used to test the T-DNA transfer ability, EHA 101 andC58C1pMP90, harbouring the binary vector pTJK136. The effective-ness of the method was proven by histochemical, PCR and Southernhybridization. A six days cocultivation on medium with 1.0 mg l−1

2,4-D was used to callus induction. The results showed that bothleaf explants and callus pieces were sensitive to geneticin 70 mg.l−1.Non differences were found between two A. tumefaciens strain ontransient Gus expression. Nevertheless, the strain C58C1pMP90yielded better results regarding number of transformed plants. GUShistochemical analysis of the putative transgenic tissues furtherconfirmed the transformation event. The transformation efficiencyreached 82%. PCR and Southern blot hybridization confirmed thepresence of the transgene and their stable integration in the regen-erated plants. Till date, only a few papers have been published onstudies of genetic transformation of Digitalis species. From theseresults an Agrobacterium tumefaciens-mediated genetic transfor-mation protocol of D. purpurea was by the first time developed.

doi:10.1016/j.jbiotec.2010.09.722