MUTAGENIZED TOBACCO PLANT AS SEED CULTURE FOR THE PRODUCTION OF OIL FOR ENERGETIC, INDUSTRIAL AND ALIMENTARY USES

• US 20100058655A1
• Filed: 08/27/2007
• Published: 03/11/2010
• Est. Priority Date: 03/14/2007
• Status: Active Grant

• Current Assignee: Aep-Advanced Ecopower Patents SA
• Original Assignee: Aep-Advanced Ecopower Patents SA
• Inventors: Fogher, Corrado
• Granted Patent: US 8,563,827 B2
• Time in Patent Office: Days
• US Class Current: 44/589
• CPC Class Codes: A01H 1/06 Processes for producing mutations, e.g. treatment with chemicals or with radiation specific mutations prepared by genetic engineering on plant cell or plant tissues C12N15/00 ; process for producing transgenic plants C12N15/82
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02E 50/30 Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

The present invention relates to the development of tobacco plants, modified through mutagenesis techniques, interspecific hybridisation followed by poliploidisation and recombinant DNA technologies, characterised by the fact of being capable of producing a very high amount of seeds and their use for the production of oil for energetic and industrial scopes, such as combustion oil, biodiesel and lubricating oil, and for animal and human alimentation.

Claims

1. A mutagenised tobacco plant characterised in that it produces a seed quantity of at least 20 quintals per hectare.

2. The plant according to claim 1, wherein said seed quantity is of about 50 quintals per hectare, or of about 70 quintals per hectare, or of about 90 quintals per hectare.

3. The plant according to claim 1, characterised in that it is further modified in the fatty acid metabolism by genetic transformation and by the fact that it produces a seed containing an oil having a iodine title lower or equal to 120.

4. The plant according to claim 1, characterised in that it is further modified in the fatty acid metabolism by genetic transformation and by the fact that it produces a seed containing an oil percentage comprised between about 40% to about 60% of the seed.

5. The plant according to claim 3, modified by transformation with one or more expression vectors comprising a seed specific expression cassette comprising, ordered from 5′

to 3′

a DNA sequence coding for a promoter of a plant gene specific for the expression in seed storage organs;

a DNA sequence coding for the signal sequence of a plant protein capable of directing said protein to the endoplasmic reticulum (ER);

a DNA sequence coding for a signal sequence capable of directing the protein to the plastid or a DNA sequence coding for a signal sequence able to anchor the protein to the ER;

a DNA sequence coding for a protein selected from the group comprising the genes acetyl-CoA carboxylase (ACCase), diacyl-glycerol acyltransferase (DGAT), lysophosphatidate acyl transferase (LPAT), phosphatidate phosphohydrolase (PAPase) acyl protein carrier (ACP), malonyl-CoA;

ACP transacylase, ketoacyl-ACP synthase (KAS), ketoacyl-ACP reductase, 3-hydroxyacyl-ACP dehydrase, enoyl-ACP reductase, stearoyl-ACP desaturase, acyl-ACP thioesterase, glycerol-3-phosphateacyltransferase, 1-acyl-sn-glycerol-3-phosphate acyltransferase, cytidine-5-diphosphate-diacylglycerol synthase, phosphatidyl glycerophosphate synthase, phosphatidyl glycerol-3-phosphate phosphatase, FAD1-8 desaturase, phosphatidic acid phosphatase, monogalactosyl diacyl glycerol synthase, digalactosyl diacyl glycerol synthase, sulpholipid biosynthesis protein, long chain acyl-coA synthase, glycerol-3-phosphate acyltransferase (GPAT), diacylglycerol cholinephospho transferase, phosphatidylinositol synthase, acil-CoA diacylglycerol acyltransferase, acyl-ACP desaturase, lineoyl desaturase, sphingolipid desaturase, oleate 12-desaturase, fatty acid acetylenase, fatty acid epoxygenase, diacylglycerol kinase, cholinephosphate cytidyl transferase, choline kinase, phospholipase, phosphatidylserine decarboxylase, phosphatidylinositol kinase, ketoacyl-CoA syntase, CER transcription factor, oleosin, 3-ketoacyl-CoA thiolase, acyl-CoA dehydrogenase, enoyl-CoA hydratase, acyl-CoA oxidase, and a polyadenilation signal.

6. The plant according to claim 1, characterised in that it is further modified by genetic transformation for insects, herbicides and fungi resistance.

7. The use of the plant according to claim 1, for the production of seeds for the manufacturing of tobacco oil, fuel oils, biodiesel, animal dietary supplements, solid fuels, human dietary supplements, lubricants.

8. The use of the plant according to claim 1, for the production of biomass for biochemical conversion or for thermochemical conversion by direct combustion, carbonisation, pyrolisis, gasification, anaerobic digestion, aerobic digestion, alcoholic fermentation or steam explosion procedures.

9. A method for the production of the plants according to claim 1, comprising

a) subjecting to mutagenesis seeds produced by initial crossings between individuals of the same species belonging to wild type or to selected varieties;

b) allowing said seeds to germinate and selecting the plants of the M2-M4 generations according to the following parameters;

i) presence of one or more phenotypic manifest characteristics selected from;

plant height 80-120 cm, leaves with a thin lamina and upright bearing, compact inflorescence, flowers number higher than 100, capsules number higher than 100, number of seeds per capsule higher than 5,000, lignified and robust stalk, deep roots;

ii) stability of the selected character in the generations following M2;

iii) verification of the hereditariness of the selected character;

c) allowing to germinate the seeds selected at b) and regenerating the plants starting from the leaf mesophyll callus induced in vitro at the presence of phytohormones and selecting the plants maintaining the characters selected at step-b) in the R0-R2 generations;

d) seeding in open field the plants selected at c) and further selecting plants producing at least 20 quintals of seed per hectare.

10. The method according to claim 9 wherein a) is substituted by

a′

) producing mutagenised seeds by carrying out interspecific crossings in the Nicotiana genus, followed by backcrossing of the F1 individuals and by amphidiploids induction by treating the vegetative apex of the plants produced by said crossings with colchicine.

11. Method according to claim 9 further comprising:

e) transforming the plants obtained at step-d) with vectors comprising expression cassettes expressing in plant genes for insects, herbicides and/or fungal diseases resistance and selecting the thus transformed plants in the T0-T4 generations for insects, herbicides and/or fungal diseases resistance and/or;

f) transforming the plants obtained at d) with one or more vectors comprising expression cassettes expressing in seed genes of the fatty acids metabolism followed by selection of the transformed plants in the T0-T4 generations for characteristics such as total oil content of the seeds and fatty acid composition of the same;

g) crossing the plants obtained at e) and/or f) and selecting the resulting progeny for one or more of the characteristics selected in the group;

high seed production ability in a quantity higher than 40 quintals per hectare, seed oil content higher than the 42%, iodine title in the oil obtained from the seeds of said progeny lower tan 120, fatty acids composition of the oil contained in said progeny seeds variable with respect to the wild type plant, insect resistance, herbicides resistance, fungi resistance.

12. The seed of the plants according to claim 1.

13. The use of the seed according to claim 12 for the manufacturing of tobacco oil, fuel oils, biodiesel, animal dietary food supplements, solid fuels, dietary food supplements for humans.

19. Biodiesel comprising oil of tobacco seeds according to claim 12, transesterified with methanol.

14. A method for the extraction of oil from tobacco seeds wherein the oil yield is between the 70 and the 95% of the oil contained in said seeds comprising:

a) mechanically extracting said oil by pressing with helical press and obtaining oil and a residual oilcake;

b) filtering said oil extracted in a) with paper or cloth filters.

15. The method according to claim 14 further comprising:

c) chemically extracting by the use of solvents the residual oil contained in the oilcake obtained in a).

16. The method according to claim 14 wherein said oil is extracted form seeds.

17. The method according to claim 15 further comprising:

d) refining the oil obtained in a) or b), and c).

18. Tobacco'"'"'s seed oil obtainable with the extraction method according to claim 15 characterised in that it has an iodine value lower than or equal to 120.

20. Food supplement for human use characterised in that it comprises tobacco'"'"'s seeds oil obtainable with the process according to claim 17.

21. Feed supplement for animal use comprising the oilcake obtainable in a) of claim 14.

22. Solid fuel comprising the oilcake obtainable in a) of claim 14.

23. Industrial lubricant comprising the tobacco'"'"'s seed oil obtainable from claim 14.