eggplant

ParthenocarpyProduction of fruit in absence of fertilizationFruits are SEEDLESSSeedlessness is advantageous for:consumersgrowers

Genetic engineering of parthenocarpy ineggplantUntransformed control GMDefH9-iaaM eggplant

ParthenocarpicfruitsEffect of the DefH9-iaaM gene in a genetic background with aparthenocarpic tendency: Control = Tal1/1 untransformedTal1/1 iaaM=GM with the DefH9-iaaM geneDefH9-iaaM eggplant fruits from either emasculatedor from self pollinated flowers

Conclusion‣DefH9-iaaM gene confers parthenocarpic fruit development(facultative parthenocarpy)‣Transgenic marketable parthenocarpic fruits are obtainedunder enviromental condition prohibitive for untransformedcontrol plants (proof of concept)Rotino G.L., Perri E., Zottini M., Sommer H., and Spena A. (1997) Genetic engineeringof parthenocarpic plants. Nature Biotech. 15, 1398-1401.

Greenhouse winter production oftransgenic parthenocarpic hybridsTrial carried out at Vittoria (RG), SicilyMaterialsTransgenic hybrids: : P1, P3 and P4Untrasformed hybrids: : C1, C2 and cv Talina F1C1 ctr P1C2 ctr P3 and P4Plastic (PE)-woodgreenhouse

35Min. Max. Aver.30Temperature during the winter trial2520°C15105014-nov28-nov12-dic26-dic9-gen23-gen6-feb20-feb6-mar20-mar3-apr17-apr1-mag15-mag29-magGM fruit growth is enhancedL= untransformed not sprayedc=untransformed sprayedr= GM not sprayedl c r

Winter production140012001000sprayednot sprayedg/plant8006004002000P1 C1 P3 P4 C2 Tal F1GenotypesC1=untransformed control of P1C2= untransformed control of P3 and P4P1, P3, P4: DefH9-iaaM eggplantsTalF1= genetic background with parthenocarpic tendency

Total Production(February-may)70006000sprayednot sprayed5000g/plant40003000200010000P1 C1 P3 P4 C2 Tal F1Genotypes

Number of fruits/plantWinter production9876sprayednot sprayedN°543210P1 C1 P3 P4 C2 Tal F1Genotypes

Number of fruits/plantTotal production40353025sprayednot sprayedN°20151050P1 C1 P3 P4 C2 Tal F1Genotypes

HARVESTGM not sprayedControl not sprayedGM sprayedControl sprayed

Conclusion‣Transgenic hybrids give a significantly higher productionthan controls sprayed with phytohormones‣No need of phytohormones spray‣Reduction of 9% of the estimated total cultivation costsGM seedless eggplant fruitDonzella. G., Spena A. and Rotino, G.L. (2000) Transgenic parthenocarpic eggplants:superior germplasm for increased winter production. Mol. Breed. 6, 79-86.

Greenhouse “early” spring and openfield production of transgenicparthenocarpic eggplant hybrids

Greenhouse locations: Monsampolo del Tronto(AP)Pontecagnano (SA)Open field location : Monsampolo del Tronto(AP)MaterialsGreenhouse: Transgenic parthenocarpic hybrids(P1; P3; P5) were compared with correspondingcontrols (C1; C2) and cultivar Talina (SG-Sandoz)Open field: Transgenic hybrids P1 (long shape)and P10 (oval shape) were compared withcorresponding controls C1 and C10

Greenhouse : early production per plant,average of two locations700600500g4003002001000P1 C1 P2 P5 C2 TalinaHybrids

Greenhouse: total productionper planta2500 aa ag200015001000500bb0P1 C1 P2 P5 C2 TalinaHybrids

Greenhouse n° fruits/plant early production2.091.551.401.261.120.570.430.280.140.00P1 C1 P2 P5 C2 TalinaHybrids

Greenhouse n° fruits/plant total production109876543210P1 C1 P2 P5 C2 TalinaHybrids

ConclusionsGreenhouse cultivation for early spring production‣DefH9-iaaM gene causes parthenocarpic fruit development‣Marketable parthenocarpic fruits are obtained undercultivation conditions prohibitive for control plants‣GM Hybrids have given a significantly higher productionthan controls‣Fruits are seedlessAcciarri N, Restaino F., Vitelli G., Perrone D., Zottini M., Pandolfini T., Spena A., Rotino G.L. (2002)Genetically modified parthenocarpic eggplants: improved fruit productivity under both greenhouse and openfield cultivation. BMC Biotechnology 2(4). http://www.biomedcentral.com/1472-6750/2/4/

Open field: early production140012001000g /p lan t8006004002000P1 C1 P10 C10Hybrids

Open field: total production350030002500g/plant2000150010005000P1 C1 P10 C10Hybrids

Open field: early production3,02,52,0n° fruits/plant1,51,00,50,0P1 C1 P10 C10Hybrids

Open field: total production10,09,08,07,0n° fruits/plant6,05,04,03,02,01,00,0P1 C1 P10 C10Hybrids

ConclusionsOpen field trial‣ Higher “early” production‣ Seedless fruits‣ Better planning of the harvests‣ High fruit quality for both fresh market and industryAcciarri N, Restaino F., Vitelli G., Perrone D., ZottiniM., Pandolfini T., Spena A., Rotino G.L. (2002)Genetically modified parthenocarpic eggplants:improved fruit productivity under both greenhouseand open field cultivation. BMC Biotechnology 2(4).http://www.biomedcentral.com/1472-6750/2/4/

101418384246Increment of Eq. diameteremasc././selfed120% of selfed fruit100806040Tal 1-1 Ctr EmTal F1 Ctr EmTal 1-1 #1-1 Em202222630346Days after fruit setGrowth of GM pollinated and emasculated fruits is identicalGrowth of Talina (TalF1) emasculated fruits is curtailed in comparisonto pollinated fruits

Increment of Pol. diameteremasc././selfed% of selfed fruit200180160140120100806040Tal 1-1 CtrTal F1 CtrTal 1-1 #1-1202 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38Days after fruit setGrowth of GM pollinated and emasculated fruits is identicalGrowth of Talina (TalF1) emasculated fruits is curtailed in comparisonto pollinated fruits

DefH9-iaaM gene is expressed during both flower and fruit developmentThis finding is consistent with the faster growth of parthenocarpic fruits

How do we obtain F1 transgenic parthenocarpicseeds?All the trials were performed with the DefH9-iaaM gene at thehemizygous stateDefH9-iaaM was expressed and gave satisfactory results when inserted(by cross pollination) in different genetic backgroundSeeds have been obtained from aged DefH9-iaaM transgenic plantThe delayed female fertility allows to produce homozygous lines byselfingHomozygous lines for DefH9-iaaM are employed as male parents formass multiplication