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P-52<br />

Establishment of virus-resistance in grapevine rootstocks<br />

G. Buchholz, M. Krieger, T. Manthey, C. Dubois, M. Sinn, J. Kramm, G.M. Reustle*<br />

RLP AgroScience GmbH, AlPlanta, - Institute for Plant Research, Neustadt / W, Germany<br />

*Corresponding author: goetz.reustle@agroscience.rlp.de<br />

Grapevine is affected by a multitude of viruses, leading to a loss of yield <strong>and</strong> quality of the<br />

grapes. The most destructive virus diseases worldwide are grapevine fanleaf <strong>and</strong> grapevine<br />

leafroll. An effective <strong>and</strong> sustainable protection against harmful viruses is to breed resistant elite<br />

<strong>and</strong> rootstock varieties. However there is no natural source of resistance available to be<br />

introduced by classic crossbreeding <strong>program</strong>mes. For this reason, RLP AgroScience is applying<br />

transgenic strategies (induction of gene silencing, expression of scFv-antibodies) to establish<br />

virus resistance both in elite <strong>and</strong> in rootstock varieties. In a project funded by the EU (FP5) <strong>and</strong><br />

in cooperation <strong>with</strong> the Federal Association of Vine Plant Producers <strong>and</strong> a grapevine nursery,<br />

various gene constructs were developed, expressing i) the core region of the movement protein,<br />

ii) inverted repeats <strong>and</strong> iii) scFv-antibodies, in order to establish virus resistance against<br />

nepoviruses (GFLV, ArMV, RpRSV). Almost 100 transgenic lines of the rootstocks (SO4,<br />

Binova, 125AA, Richter 110) containing the different gene constructs have since been<br />

developed. The complete integration of the constructs has been confirmed in most of the<br />

transgenic lines by molecular characterisation. Between one <strong>and</strong> five copies of the transgenes<br />

were detected. In Northern analysis, transcripts of the relevant sequences were found in some but<br />

not in all transgenic lines. Detection of siRNA resulting from RNA-silencing of the virus-derived<br />

sequences is in process. Virus resistance of the transgenic lines is evaluated by laboratory<br />

screening methods, developed at AgroScience (dual culture of nematodes <strong>and</strong> c<strong>and</strong>idate lines,<br />

detection of silencing by the GFP-sensor). Furthermore, transgenic lines are cultivated in a<br />

Saran-house equipped <strong>with</strong> soil containers, which were inoculated <strong>with</strong> Grapevine Fanleaf Virus<br />

(GFLV)-infected nematodes Xiphinema index. An overview about molecular analysis <strong>and</strong> the<br />

evaluation experiments for virus resistance of the transgenic lines will be presented.<br />

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