Teaduselt põllule geeniteadus põllumajanduses

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1 Teaduselt põllule geeniteadus põllumajanduses Evelin Loit Eesti Maaülikool 2. detsember 2011 Toila

2 Geen tunnus Geen (retsept) RNA (vahe-etapp) Rakud Ensüüm Valk (ensüüm jne) Tunnus Pigmendi molekulid Geen

3 Geenitehnoloogia kus oleme täna? 40 aastat tehnoloogia arengut esimene GM organism 1971 (Escherichea coli) esimene GM taim 1983 (tubakas) 15 aastat geenitehnoloogiat tööstuslikus kasutuses 1,7 148 miljonit ha (87x) 29 riiki

4 Umbrohumürgikindlus on eelistatuim omadus milj. Ha Umbrohumürgikindlus Putukakindlus (Bt) Herbitsiidi- ja putukakindlus 4

5 Putukakindel mais MON810 Cry geen bakterist (Bacillus thuringiensis) Toodab putukaspetsiifilist toksiini

6 Bt toksiin seob spetsiaalset putuka retseptorit 6

7 Amflora kartul paberitööstusele Tärklise molekulid Amüloos (lineaarne) Amülopektiin (hargnenud) Amflora kartul Deaktiveeritud amüloosi ensüüm

8 Pigmendi tootmine Chalcone Synthase (CHS) Petuunia toodab lillat antotsüaniini pigmenti Kalkooni süntaas on oluline ensüüm antotsüaniini tootmisel. Anthocyanins Photo credit Richard Jorgensen; Aksamit-Stachurska et al. BMC Biotechnology :25 doi: /

9 Hüpotees kui viia kaks ensüümi taime, siis saab topeltkoguse pigmenti.

10 Tegelikult, ekstra geen inhibeerib pigmendi tootmist Taimed, kus on CHS transgeen GEEN + GEEN 0 CaMV 35S pro : Photo credit Richard Jorgensen

11 Edasised suunad Herbitsiidi resistentsus Kahjurikindlus Haiguskindlus Muudetud koostis Biofarmaatsia Stressi resistentsus

12 Herbitsiidi resistentsus Täna: RoundupReady (glüfosaat) Liberty Link (glüfosinaat) Planeerimisel: Dicamba tolerantsus Ensüüm bakterist Pseudomonas

13 Glüfosaat blokeerib aminohapete sünteesi Aminohapped on valkude ehitusblokid Glüfosaat blokeerib ära ühe ensüümi (EPSP süntaas), mis vastutab aminohapete tootmise eest (trüptofaan, fenüülalaniin, türosiin) Mõned EPSP süntaasid on resistentsed glüfosaadile (üks aminohappe erinevus: Gly 96 Ala)

14 Roundup-ready taimedel on ekstra EPSP ensüüm Taimedel on 3 ensüümi koopiat: Maisi EPSPS Agrobakteri EPSPS Amp r (ampicillin resistance) Ti Plasmid Replication origin Multiple cloning site

15 Uued Bt geenid Geen/Toksiin Tootja Toote nimi Cry1Ab Monsanto Agrisure CB Yieldgard CB Cry1Ab Monsanto Yieldgard CB Cry1F Mycogen Herculex I Cry3Bb1 Monsanto Yieldgard VT Rootworm Cry34/35Ab1 Dow Herculex RW Cry1AB + Cry3BB1 Monsanto Yieldgard VT PLUS Yieldgard VT TRIPLE Cry1F + Cry34/35Ab1 Dow Herculex EXTRA

16 Valge ristik Viiruskindlus (lutserni viirus) Fragment viiruse genoomist taimes (klassikaline taime resistentsuse meetod) Saagikuse tõstmine (LXR tehnoloogia) Rohkem taime hormooni tsütokiin (vastutab taime arengu eest) Taime arengu edasilükkamine Suurem saak

17 Pikk tee regulatsioonideni Teaduse tegemine Patent 2000 Luba tegutseda ja toode luua 2001 Partneri otsimine Riisi sordi Indica valmistamine 2002 Luba, et geenikonstrukt on OK 2002 Luba, et konkreetne tehnoloogiline tulemus on OK 2003 Luba, et saadud sort on OK 2003 Luba, et saadud sort annab 1.6 g/g 2004 Luba kasvatada ja kasutada 2011?

18 Spargelkapsas Beneforté Sisaldab 2-3 korda rohkem antioksüdantide tootmist mõjutavat toitainet (glükorafaniin) Aitab hoida A, C ja E vitamiinide taset Valmis 2011

19 Vähem allergeene Karjamaa raihein Allergeenide (Lol p1 ja Lol p2) vähendamine Õietolmu allergia ja astma vähendamine Mõjutab 25% inimeste populatsioonist

20 Ligniin ja fruktaan Karjamaa raihein ja roog-aruhein Muudetud fruktaani ja ligniini biosüntees Bioetanool Parem sööda kvaliteet lihatööstuseks Vähem kasvuhoonegaasi emissiooni Keskkonnale kasu Vähendab energiakadu loomal (näiteks piimalehm toodab 90 kuni 140 kg metaani aastas, mis on võrdne 24 kuni 38 päeva kaotatud energiat)

21 Bioetanool Hirss (Panicum virgatum) Modifitseeritud ligniin Sort, mis toodab 38 % rohkem biokütust vähema eeltöötlemisega Vajab 4-5 korda vähem tsellulaasi (ensüüm, mis lagundab kiudainet)

22 Fu C et al. PNAS 2011;108: Transgeenne hirss

23 Bioetanooli mais - Enogen Lisatud bakteri geen, mis toodab ensüümi amülaas, mis lõhustab tärklise glükoosiks (esimene samm etanooli valmistamisel) Aitab tootjat pärast saagikoristust - tavaliselt tuleb ensüüm lisada väliselt 10% etanooli maisi lisamine segusse suurendab etanooli produktsiooni 10% Enogen-mais 2000 ha Kansases Syngenta (Shveits)

24 Bioetanool paplitest Modifitseeritud ligniin (ligniini tootmine alla surutud) 1 gramm kuiva puitu GM paplist annab 81% rohkem bioetanooli võrreldes tavaliste puudega Genfi ülikool

25 Õli tootmine vetikas Toormaterjal: suhkruroo biomass Kaks uut geeni: Sukroosi invertaas (sukroos glükoos ja fruktoos) Alkaani molekulide sünteesija Solazyme U.S. Patent 7,935,515 ( 515 Patent): Recombinant microalgae cells producing novel oils and relates to making triglycerides out of sugar cane.

26 Taimedes toodetud ravimid ja vaktsiinid

27 Insuliini tootmine taimedes Värvohakas (Carthamus tinctorius) Toodab seemnetes pro-insuliini valku Insuliini (SBS-1000) bioaktiivsus on võrreldav tavainsuliiniga (Humulin R) SemBioSys U.S. patent US 7,547,821 "Methods for the Production of Insulin in Plants".

28 HIV vaktsiin Tubakalehtedes 2011: I kliiniline katse (I faas) 11 tervet naist Inglismaal Pharma-Planta projekt ravimite tootmine taimedes (EL raamprojekt)

29 Riis kui vaktsiin MucoRice Ennetab koolerat Uus viis vaktsineerimiseks Soodne toota Lihtsam tarbida Seab terve keha valmis Uued mõtted: Gripivaktsiin Allergiaravim

30 Praegu väljatöötamisel

31 Põuakindlus Põuakindel HD-START valk müürloogas (Arabidopsis) Kastetud 10 päeva ilma veeta 20 päeva ilma veeta Pärast uut kastmist Kontroll Yu, H., et al. Plant Cell 2008;20:

32 Põuakindlus tuleneb suuremast juurestikust Kontroll Põuakindel Kontroll Põuakindel Yu, H., et al. Plant Cell 2008;20:

33 Transgeen: geen teisest organismist Cis-geen: geen samast organismist

34 Epigeneetiline kontroll õitsemisaja üle Pikk külm aeg Vegetatiivne areng Sügis Talv Reproduktiivne areng Kevad Mõnedel taimedel on vaja pikka külmaperioodi (vernalisatsioon), nagu talv, selleks et õitsema hakata.

35 FLC geen kontrollib õitsemist Sügis Talv Kevad FLC on õitsemise inhibiitor; kui FLC ära võtta, siis pole ka vernalisatsiooni vaja.

36 FLC takistab õitsemise aktivaatorit Tavaline taim FLC FT geen FT geeni transkriptsioon on häiritud FLC seondumisega Taime ilma flc-ta FT geen FT

37 Lisaks lugemist ne_online_preprint.pdf

38 Aitäh! Evelin Loit ;

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