Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S117

[P&F.24]

Transgenic rice for food security and sustainable agriculture

Q. Zhang

Huazhong Agricultural University, ChinaKeywords: Transgenics; Rice

Rice is the most important food crop in many parts of the world.Rice improvement has achieved remarkable success in the last half-century with the yield doubled in most parts of the world andeven tripled in certain regions, which has contributed greatly tofood security globally. To secure rice production, large efforts havebeen made in the last two decades internationally in research anddevelopment of rice biotechnology, including genome sequencing,functional genomics and transgenics. From a global viewpoint, anumber of challenges need to be met for sustainable rice produc-tion: (1) increasingly severe occurrence of insects and diseases andindiscriminate pesticide applications; (2) high pressure for yieldincrease and overuse of fertilizers; (3) water shortage and increas-ingly frequent occurrence of drought, and (4) extensive cultivationin marginal lands. A new rice breeding goal, referred to as GreenSuper Rice, was recently proposed to address these challenges. Onthe premise of continued yield increase and quality improvement,Green Super Rice would possess resistances to multiple insectsand diseases, high nutrient-use efficiency, and drought resistance.Transgenic studies of rice have been conducted in large scale, andmost of the target traits are consistent with the goal of Green SuperRice. It is believed that transgenic rice will play a crucial role toensure sustainable rice production in the future. In the presen-tation, I will review the main progresses and constraints in theresearch and development of transgenic rice in a number of fronts.

doi:10.1016/j.jbiotec.2010.08.303

[P&F.25]

Moving crops biotechnology forward through new germ-linetransformation techniques

O.I. Kershanskaya ∗, S.V. Didorenko, A.S. Nurmagambetova, G.L.Esenbaeva, A.T. Tashkenova

Institute Plant Biology and Biotechnology, National Center Biotechnol-ogy, KazakhstanKeywords: Germ-line transformation; Techniques; Abiotic stressesresistance; Crop improvement

Elaboration of new germ-line biotechnology (genetic trans-formation by plant germ elements–pollen, ovary, embryo, seed)for crop transgenic plants creation enhance possibility of simpleintroduction of new valuable genes for plant disease and abioticstress resistance, genetic modification of photosynthesis, yield cropimprovement, benefice of plant biotechnology.

We have elaborated methods of genetic germ-line transfor-mation by Agrobacterium Pipetting into wheat ear stigma andsoybean pollen tubes. Agrobacterium Pipetting into wheat methodis based on unique for wheat mechanism of pollen distance trans-fer which provided by high level of flavonol glucosides which actsas inducer of the vir-zone of Ti plasmid. Biotechnological pro-tocol includes several stages: Agrobacterium culture grown andoptimization; improvement of technique for Agrobacterium pipet-ting into wheat ears stigma; determination of optimal stage ofplant development; screening of putative transgenic seeds onantibiotics resistance using selectable marker genes (nptII encodedkanamycin- and hptII encoded hygromycin resistance); biochemi-cal screening of putative transgenic plants in T1 progeny, in case of

PEPC gene introduction–assay of PEP-carboxylase activity; molecu-lar biological detection of transgenes in wheat genome in T1 and T2- generations by PCR, Real Time PCR, Northern and Southern Blot-ting, yield analyses. Totally has been produced about 5000 putativewheat transgenic seeds with efficiency 1.8–2.3% and more than10000 transgenic plants of first-fifth generations.

The method of pollen tube pathway transformation in soybeanprovides DNA transferring by cutting the stigma following pollina-tion. DNA presumably reaches the ovary by flowing down the pollentube exactly after anthesis and then integrates into the just fertil-ized but undivided zygotic cells. Approximately 3000 seeds wereproduced from the flowers treated with DNA. PPDK, desA12licBM3and FeSOD valuable genes have been used to provide in soybeanplant resistance to different abiotic stresses. Screening on antibi-otic resistance by marker (spectinomycin in FeSOD) and reporter(lichenase BM3 in PPDK and des A12lic BM3) genes and molecu-lar biological detection of transgenes by PCR analysis confirm lessthan 3% of progeny seeds tested expressed a positive reaction incomparison with wild soybean plants.

New techniques except waste time and money steps of tissueculture and regeneration are simple, economic and comparativelyeffective. New methods convert wheat and soybean genetic trans-formation into routine process, which might be successfully usedby many breeders and investigators for increasing of crops yieldand moving plant biotechnology forward.

doi:10.1016/j.jbiotec.2010.08.304

[P&F.26]

Engineering secondary metabolite production in plants usingcombinatorial genetic transformation

B. Miralpeix 1,∗, R. Hoefer 2, D. Werck 2, L. Dong 3, H.J.Boewmeester 3, P. Christou 1,4

1 Universitat de Lleida, Spain2 Université de Strasbourg, France3 Wageningen University, Netherlands4 Institució Catalana de Recerca i Estudis avancats (ICREA), SpainKeywords: Terpenoid indole alkaloid; Catharanthus rouseus;Genetic transfomation; Tobacco

Humans have used compounds derived from plants for treatingdiseases since prehistoric times. Over 25% of new drugs approvedfor human therapy in the last 30 years are based on molecules ofplant origin, and about 50% of the top selling drugs are derived fromknowledge of plant secondary metabolism.

Catharanthus roseus (Madagascar periwinkle) produces morethan 130 different secondary metabolites, including terpenoidindole alkaloids (TIAs). These compounds exhibit diverse biologicalactivities including anti-cancer, analgesic, spasmolytic, anti-inflammatory and insecticidal effects. However, such moleculesare present in minute amounts in plants such as C. roseus mak-ing their isolation from natural sources uneconomical; in addition,their total or partial chemical synthesis is impractical because oftheir complex structural features. Genetic engineering is thus aviable alternative for the production of such important moleculeson a large scale.

We had recently reported the development of a combinato-rial genetic transformation system which we used to generatea population of plants expressing diverse input transgenes.This methodology was exemplified with the engineering of thecarotenoid pathway in corn and resulted in corn germplasm accu-mulating very high levels of nutritionally important carotenoids.The approach also permited the dissection of the pathway, thusrevealing as yet unkown rate limiting steps.