EP2811822A1 - PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI - Google Patents

PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI

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Publication number
EP2811822A1
EP2811822A1 EP13702652.2A EP13702652A EP2811822A1 EP 2811822 A1 EP2811822 A1 EP 2811822A1 EP 13702652 A EP13702652 A EP 13702652A EP 2811822 A1 EP2811822 A1 EP 2811822A1
Authority
EP
European Patent Office
Prior art keywords
plant
tomato
plants
allele
mutant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP13702652.2A
Other languages
German (de)
English (en)
Inventor
Rudolf Verhoef
Dörthe Bettina DRÄGER
Arie Vogelaar
Zeger Otto Van Herwijnen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Rijk Zwaan Zaadteelt en Zaadhandel BV
Original Assignee
Rijk Zwaan Zaadteelt en Zaadhandel BV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rijk Zwaan Zaadteelt en Zaadhandel BV filed Critical Rijk Zwaan Zaadteelt en Zaadhandel BV
Priority to EP13702652.2A priority Critical patent/EP2811822A1/fr
Publication of EP2811822A1 publication Critical patent/EP2811822A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/82—Solanaceae, e.g. pepper, tobacco, potato, tomato or eggplant
    • A01H6/825—Solanum lycopersicum [tomato]
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/08—Fruits
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae

Definitions

  • the present invention relates to a tomato plant
  • invention further relates to a marker and the use of the marker for identifying tomato plants with this phenotype.
  • the invention also relates to the seeds and progeny of such plants and to propagation material for obtaining such plants. Furthermore the invention relates to the use of the plants, seeds and propagation material that show the
  • Tomato is a vegetable crop grown worldwide in all conditions and climates, both in protected cultivation or in open field. Growing tomato plants requires a lot of labour and attention, during which the plants are handled
  • Handling the plants includes activities such as grafting, planting, pruning, winding, and of course harvesting.
  • Another aspect of a tomato plant is the typical tomato smell that it emits. This can become rather
  • Non-glandular trichomes are usually regarded as , hairs' and do not produce, store, or secrete specific biochemical compounds.
  • a glandular trichome typically consists of a stalk, made up of one or more cells, and one or more glandular cells at the tip of the stalk that form the glandular head.
  • glandular trichomes are identified in tomato and related Solanum species, namely types I, IV, VI, and VII. These types differ in size and length of the stalks, and in number of secretory cells that form the glandular head (McDowell et al . , Plant Physiology Vol. 155, 524-539 (2011)) .
  • glandular head are classified as secreting glands, while multicellular glands are classified as storage glands.
  • Type VI trichomes are composed of four disc cells, or glandular cells, at the end of a one- or two-celled stalk.
  • the four disc cells form the multicellular glandular head (Fig. 1) .
  • Biochemical compounds that are produced by the various glandular trichomes in tomato comprise terpenoids, flavonoids, fatty acids, alkaloids, and acyl sugars such as acyl glucoses and acyl sucroses.
  • chlorophyll however is not among the compounds known to be produced or secreted by glandular hairs in tomato.
  • the produced compounds are known to play important roles in attracting and repelling various insects and in determining susceptibility to certain diseases.
  • many aspects of the roles of these metabolites are still unclear, and extensive research is ongoing to determine more precisely the functionality of glandular trichomes and the substances they excrete.
  • the new tomato plant was analysed for phenotypical characteristics.
  • the glandular trichomes in cultivated tomato (Solanum lycopersicum) that are most abundant are type I and type VI.
  • the typical glandular heads of type VI trichomes, which are made up of four glandular cells, were not present among the trichomes (Fig. 2B) , only the stalks could still be
  • the areas were 100 - 10,000 x reduced in plants of the invention as compared to other plants, including wild type and isogenic plants ( Figures 3, 8, and 11) .
  • the sesquiterpenes showed a similar range of 100 - 10,000 x area reduction in plants of the invention ( Figures 4, 9, and 12).
  • the compound cis-3-hexanal shows a significant 3-fold reduction for plants of the invention when the year is taken into account, which was calculated using an unbalanced ANOVA approach, and hexanal is about 40-fold reduced ( Figures 5, 10, 13, and 14). Plants belonging to the invention therefore showed a significant reduction in terpenes, sesquiterpenes and aldehydes, in addition to absence of type VI glands, absence of green staining, and absence or strong reduction of typical tomato smell.
  • Chlorophyll typically shows two absorbance peaks, one at around between 420 and 460 nm and one between around 620 and 700 nm, both of which were clearly absent in all extracts. Therefore it had to be concluded that the absence of green staining of tomato plants of the invention is not due to a change in
  • Carotenoids which account for or contribute to many coloured plant tissues, were not found among the substances that were analysed. Carotenoids have an
  • plants of the invention do not contain flavonoids that are present in the green vegetative parts of prior art tomato plants.
  • the plant does not comprise or has at least a strongly reduced number of the glandular cells of type VI trichomes ,
  • the plant has a strongly reduced level of volatiles, in particular terpenes, sesquiterpenes and aldehydes ,
  • the plant does not express or is at least strongly reduced in the typical tomato smell
  • the plant does not comprise or has at least a strongly reduced level of flavonoids.
  • the mutant allele was named tghVI (tomato
  • chromosome 9 of the tomato genome between the physical positions 1750800 bp and 4517648 bp, preferably between positions 1939546 bp and 2970346 bp, most preferably at about position 2454946 bp on the physical SL2.30 map of the Solanum lycopersicum genome.
  • the mutant tghVI allele can be identified by the presence of a molecular SNP marker that is linked to the tghVI allele.
  • a SNP marker linked to the tghVI allele is located at 1750800 bp on chromosome 9 of the physical Solanum lycopersicum map SL2.30 and is a nucleotide change from A to G.
  • the presence of the mutant tghVI allele in a plant of the invention is preferably also associated with a molecular SNP marker located on chromosome 9 at 1750800 bp on the public SL2.30 map of the Solanum lycopersicum genome, wherein the SNP is a nucleotide change from A to G.
  • the absence of the SNP marker does not mean that the mutant allele is not present.
  • a plant without the SNP marker but with the phenotype of the invention caused by the mutant allele can thus still be a plant of the invention.
  • the present invention thus provides a tomato plant (Solanum lycopersicum) comprising a mutant allele which allele is linked to the following phenotypic features:
  • the plant does not comprise or has at least a strongly reduced number of the glandular cells of type VI trichomes , - the plant does not cause green staining upon contact,
  • the plant has a strongly reduced level of volatiles, in particular terpenes, sesquiterpenes and aldehydes ,
  • the plant does not express or is at least strongly reduced in the typical tomato smell
  • the plant does not comprise or has at least a strongly reduced level of flavonoids.
  • Plants of the invention have modified type VI trichomes which modification leads to absent or nonfunctional type VI glandular cells or type VI glandular cells that do not function properly.
  • the four glandular cells i.e. the complete glandular head, are preferably completely absent.
  • the trait of having modified type VI trichomes that lack all of the four glandular cells that make up the glandular head can for example be obtained from plants, representative seeds of which were deposited with the NCIMB under number NCIMB 41845, but the trait of lacking four glandular cells or having non-functional or mal ⁇ functioning glandular cells could also be obtained from other, yet unknown sources.
  • glandular cells of type VI trichomes and at the same time would generate a complete absence of staining and a strong reduction or lack of a typical tomato smell, and the
  • tomato plants of the invention leads to major advantages during tomato growing.
  • the necessity to treat the skin with a substance in advance of handling tomato plants to prevent staining is removed.
  • the use of protective clothing or gloves when working in a tomato crop is largely reduced.
  • no rigorous methods to remove the staining after handling tomato plants are
  • tomato plants of the invention will substantially facilitate tomato growing.
  • the mutant tghVI allele is obtainable from a tomato plant comprising said mutant tghVI allele, representative seed of which was deposited with the NCIMB under deposit number NCIMB 41845.
  • the tomato plant of the invention carrying the mutant tghVI allele is obtainable by crossing a first tomato plant with a second tomato plant, wherein one of the said plants is grown from seeds of which a
  • one of the said plants used for crossing to obtain a tomato plant of the invention is carrying the tghVI allele.
  • Selection for plants comprising the tghVI allele can be done on phenotype, i.e. determining the presence of one or more of the phenotypic aspects of the trait, which are the absence of green staining, the absence or near absence of flavonoids, the reduction of volatiles, and the absence of a typical tomato smell.
  • selection can be done on the presence of modified type VI trichomes that lack glandular heads.
  • Selection can alternatively be done genetically on the presence of the relevant mutant tghVI allele, for example by using one or more molecular markers.
  • molecular markers results in a reliable outcome, and can be done in a very early plant stage.
  • the presence of the mutant tghVI allele also leads to the absence of the four glandular cells that form the glandular heads of type VI trichomes.
  • the leaves of tomato plants of the invention are thus absent in fully developed type VI trichomes, and only possess type VI trichomes that lack glandular heads.
  • Other glandular as well as non- glandular trichomes besides type VI can however be detected on a tomato plant of the invention.
  • the absence of glands of type VI trichomes is linked with the absence of green staining from the green vegetative parts of the plant upon contact.
  • the green vegetative parts comprise the leaves and the stems of a plant.
  • the presence of the tghVI allele preferably further leads to the absence of a typical tomato smell in a tomato plant of the
  • the absence of green staining and the absence of a typical tomato smell are the result of a modification in the composition of at least flavonoids and volatiles, due to the absence of glands of type VI trichomes.
  • the mutant tghVI allele of the invention is present in homozygous form.
  • the trait of the invention is monogenic, i.e. caused by a single gene, and is inherited in a recessive way.
  • plants that carry the genetic trait, i.e. the mutant tghVI allele can suitably be identified among descendants from a cross between a plant not carrying the trait, and a plant that does carry the said trait, by growing F2 plants from seeds that are the result of the initial cross and a selfing step, and selecting plants showing the desired trait.
  • Selecting the plants can be done phenotypically by determining the absence of green staining, and/or the absence of a typical tomato smell, and/or the absence of flavonoids and/or the absence or reduction of volatiles and/or the absence of the glandular cells of type VI trichomes or can be done through identification of the mutant allele, for example by means of the marker defined herein .
  • Determining absence of green staining is done in comparison to a control.
  • the control to determine the absence of green staining is any existing tomato plant, or an isogenic tomato plant, i.e. a plant that is
  • selecting the plants in the F2 can be done phenotypically by determining the absence of the four glandular cells that form the glandular heads of type VI trichomes.
  • the absence of glandular heads of type VI trichomes as mentioned throughout this application is determined on the green vegetative parts of the plant.
  • the absence of glands of type VI trichomes should not be
  • the invention furthermore relates to a cell of a tomato plant as claimed.
  • a cell of a tomato plant as claimed.
  • Such cell may be either in isolated form or may be part of the complete tomato plant or parts thereof and then still constitutes a cell of the invention because such a cell harbours in its genetic constitution the genetic information that leads to the characteristics that define the tomato plant of the invention.
  • Each cell of tomato plants of the invention carries the genetic
  • Each cell thus carries the mutant tghVI allele in its genome .
  • the invention also relates to tissue of a plant as claimed.
  • the tissue can be undifferentiated tissue or already differentiated tissue.
  • Undifferentiated tissues are for example stem tips, anthers, petals, pollen and can be used in micropropagation to obtain new plantlets that are grown into new plants of the invention that have the above identified characteristics.
  • the invention according to a further aspect thereof relates to seeds of a plant as claimed.
  • the seeds do not show the genetic trait of the tomato plant of the invention, they harbour the genetic information that when a plant is grown from the seeds makes this plant a plant of the invention that has the above identified characteristics.
  • the invention also relates to progeny of the plants, cells, tissues and seeds of the invention.
  • progeny can in itself be plants, cells, tissues or seeds.
  • progeny is intended to mean the first and all further descendants from a cross with a plant of the invention that comprises the mutant tghVI allele that leads to the absence of the glandular heads of type VI trichomes.
  • Progeny of the invention are descendants of any cross with a plant of the invention that carries the trait that leads to the absence of the glandular heads of type VI trichomes or to the other characteristics identified above .
  • Progeny also encompasses plants that carry the trait of the invention and are obtained from other plants or progeny of plants of the invention by vegetative propagation or multiplication.
  • the invention thus further relates to seed of the claimed plant and to parts of the plant that are suitable for sexual reproduction.
  • Such parts are for example selected from the group consisting of microspores, pollen, ovaries, ovules, embryo sacs and egg cells.
  • the parts are for example selected from the group consisting of microspores, pollen, ovaries, ovules, embryo sacs and egg cells.
  • invention relates to parts of the plant that are suitable for vegetative reproduction, in particular cuttings, roots, stems, cells, and protoplasts.
  • tissue culture of the claimed plant comprises regenerable cells.
  • tissue culture can be derived from leaves, pollen, embryos, cotyledons, hypocotyls, meristematic cells, roots, root tips, anthers, flowers, seeds and stems.
  • the invention furthermore relates to hybrid seed and to a method of producing hybrid seed comprising crossing a first parent plant with a second parent plant and harvesting the resultant hybrid seed, wherein said first parent plant and/or said second parent plant is the plant as claimed.
  • the tghVI allele is present in homozygous form and the plant expresses the phenotype described above.
  • the invention also relates to inbreds and doubled haploids of tomato plants of the invention.
  • the invention relates to tomato plants of the invention that carry the mutant tghVI allele, and having acquired said allele by introduction from a suitable source, either by conventional breeding, or genetic modification, in particular by cisgenesis or transgenesis .
  • Cisgenesis is genetic modification of plants with a natural gene, coding for an (agricultural) trait, from the crop plant itself or from a sexually compatible donor plant.
  • Transgenesis is genetic modification of a plant with a gene from a non-crossable species or a synthetic gene.
  • the invention also relates to the germplasm of plants of the invention.
  • the germplasm is constituted by all inherited characteristics of an organism and according to the invention encompasses at least the tghVI allele of the invention.
  • the germplasm can be used in a breeding programme for the development of tomato plants that are lacking in or have a reduction in green staining from the green vegetative parts of the plant upon contact, and/or are lacking a typical tomato plant aroma, and/or are absent in flavonoids from the green vegetative parts of the plant, and/or are reduced in volatile compounds, and/or are lacking glands of tomato type VI trichomes.
  • the invention also relates to a tomato fruit that is produced by a plant of the invention.
  • the invention further relates to a food product, comprising the fruit of a tomato plant as claimed, or parts thereof.
  • the invention also relates to a food product in processed form.
  • the invention relates to a method for production of a tomato plant comprising the tghVI allele, comprising
  • the parent that provides the trait of the invention is not necessarily a plant grown directly from the deposited seeds.
  • the parent can also be a progeny plant from the seed or a progeny plant from seeds that are identified to have the trait of the invention by other means .
  • the invention relates to a method for production of a tomato plant comprising the tghVI allele, comprising
  • the invention additionally provides a method of introducing an additional desired trait into a tomato plant comprising the tghVI allele, comprising:
  • the invention includes a tomato plant produced by this method.
  • the selection step is suitably done by means of marker analysis with the SNP marker
  • selection for plants comprising the tghVI allele is done in the Fl .
  • selection for the trait of the invention is started in the F2 of a cross or alternatively of a backcross.
  • Selection in the Fl is suitably done by means of the marker.
  • Selection in the F2 can also be done on the basis of the phenotype.
  • selection for plants comprising the tghVI allele is started in the F3 or a later generation.
  • the plant comprising the mutant tghVI allele is a plant of an inbred line, a hybrid, a doubled haploid, or of a segregating population.
  • the invention further provides a method for the production of a tomato plant comprising the tghVI allele using a doubled haploid generation technique to generate a doubled haploid line comprising the said mutant tghVI allele that leads to the absence of the glandular heads of type VI trichomes, a modification in the composition of flavonoids and volatiles, absence of green staining, and/or absence of a typical tomato plant smell.
  • the invention furthermore relates to hybrid seed and to a method for producing hybrid seed comprising
  • the invention relates to a method for producing a hybrid tomato plant comprising crossing a first parent tomato plant with a second parent tomato plant and harvesting the resultant hybrid tomato seed, in which the first parent tomato plant and/or the second parent tomato plant comprises the mutant tghVI allele.
  • the invention also relates to a method for the production of a tomato plant comprising modification in the composition of flavonoids and volatiles, absence of the glandular heads of type VI trichomes, absence of green staining, and/or absence of a typical tomato plant smell, by using a seed that comprises the mutant tghVI allele in its genome for growing the said tomato plant.
  • the seeds are suitably seeds of which a representative sample was
  • the invention also relates to a method for seed production comprising growing tomato plants from seeds of which a representative sample was deposited with the NCIMB under deposit number NCIMB 41845, allowing the plants to produce seeds, and harvesting those seeds. Production of the seeds is suitably done by crossing or selfing.
  • the invention relates to a method for the production of a tomato plant comprising the mutant tghVI allele by using tissue culture.
  • the invention furthermore relates to a method for the production of a tomato plant comprising the mutant tghVI allele by using vegetative reproduction.
  • the invention relates to a method for the production of a tomato plant comprising the mutant tghVI allele by using a method for genetic modification to introgress the mutant tghVI allele into the tomato plant.
  • Genetic modification comprises transgenic modification or transgenesis , using a gene from a non-crossable species or a synthetic gene, and cisgenic modification or cisgenesis, using a natural gene, coding for an (agricultural) trait, from the crop plant itself or from a sexually compatible donor plant.
  • the invention also relates to a breeding method for the development of tomato plants that comprise modification in the composition of flavonoids and volatiles, absence of the glandular heads of type VI trichomes, absence of green staining, and/or absence or strong reduction of a typical tomato smell, wherein germplasm comprising the mutant tghVI allele is used.
  • Representative seed of said plant comprising the mutant tghVI allele and being representative for the germplasm was deposited with the NCIMB under deposit number NCIMB 41845.
  • the invention relates to a method for the production of a tomato plant comprising the mutant tghVI allele wherein progeny or propagation material of a plant comprising the mutant tghVI allele is used as a source to introgress the mutant tghVI allele into another tomato plant.
  • Representative seed of said plant comprising the mutant tghVI allele was deposited with the NCIMB under deposit number NCIMB 41845.
  • the invention provides preferably a tomato plant showing modification in the composition of flavonoids and volatiles, absence of the glandular heads of type VI trichomes, absence of green staining, and/or absence or strong reduction of a typical tomato plant smell, which plant is obtainable by any of the methods herein described.
  • a mutant allele can be identified by the use of a molecular marker.
  • a mutant allele can alternatively be identified by the position on a genetic map, or by
  • the , genetic trait' is the trait or characteristic that is conferred by the mutant tghVI allele.
  • the genetic trait can be identified phenotypically, for example by performing visual observations or biochemical analyses.
  • Genetic maps can vary according to the method by which they are assembled. A person skilled in the art knows how to compare and combine genetic maps, whereby differences between genetic maps can be eliminated or minimized.
  • positions as used herein are physical positions based on the public physical map of the tomato genome SL2.30 (version of August 2010, http://solgenomics.net/organism/
  • solanum_lycopersicum/genome solanum_lycopersicum/genome
  • accession number NCIMB 41845 accession number NCIMB 41845.
  • Figure 1 Type VI trichome Solanum lycopersicum
  • Figure 2 A: Wildtype TR306-1 type VI trichomes with glandular head. B: mutant 3432-1 type VI trichomes with glandular heads absent.
  • Figure 6 Absorbance of tomato staining particles. The order of the accessions in the legenda is the order of the graph lines from top to bottom.
  • Figure 7 Acyl sugar measurements. The results are given for glucose in nmol/cm 2 leaf disc.
  • Figure 12 Combined data of Fig. 4 and Fig. 9
  • Figure 13 Combined data of Fig. 5 and Fig. 10
  • the treated seeds were germinated and the resulting plants were grown in a greenhouse to produce M2 seeds.
  • M2 seeds were harvested and bulked in one pool.
  • the resulting pool of M2 seeds was used as starting material to identify individual M2 plants that were absent in green staining upon contact with the green
  • the efficacy of the genetic modification procedure was assessed by determining the occurrence of bleached plants, which is indicative for chlorophyll loss due to modifications in genes directly or indirectly involved in the formation or accumulation of chlorophyll.
  • M2 tomato seeds were germinated in soil and grown to small plantlets. Subsequently, approximately 7000 randomly chosen plants were transferred for a seedling screen.
  • the green plant parts of the mutant plants were visually observed by use of a binocular, and compared with normal wild type (WT) tomato plants.
  • WT normal wild type
  • the microscopic observation showed that the glandular heads of type VI trichomes, which are made up of four glandular cells, were not present among the glands of the mutant tomato plants. It was therefore concluded that there was linkage between the absence of type VI glands and the absence of green staining.
  • the F2 segregated in a manner that corresponds with a monogenic recessive inheritance of the trait of the invention. Both the absence of green staining and the absence of four glandular cells of type VI trichomes
  • the trait of the invention could be brought into a wild type tomato plant by crossing the wild type plant with a plant of the invention and selecting for the desired phenotype in the F2 of that cross, either by selection on absence of green staining, or by selection on absence of four glandular cells of type VI trichomes.
  • the SNP marker that correlated most closely to the mutant allele is located on chromosome 9 at 1750800 bp on the public SL2.30 map of the Solanum lycopersicum genome, wherein the SNP is a nucleotide change from A to G.
  • This molecular marker can be used to identify the presence of the mutant allele in plants grown from seeds as deposited under NCIMB number 41845.
  • the new mutant allele was provisionally named tghVI (tomato glandular hairless VI).
  • Other plants that comprise the same mutant tghVI allele of the invention might also be linked to the A/G SNP on position 1750800 bp of the public SL2.30 map, but can optionally also be linked to another molecular SNP marker or any other molecular marker that is polymorphic in a certain population.
  • Plants of the invention were analysed for a large number of volatiles, and compared with a tomato plant that did not contain the mutant tghVI allele of the invention.
  • a tomato plant to be measured was placed in a plastic bag for 10 minutes. Next, the plants were touched to allow the release of the relevant compounds to be analysed.
  • a number of tomato plants were analysed for the green staining. Included were plants of the invention, isogenic plants lacking the mutant allele of the invention, wild type (normal) tomato plants, and plants from the art that contain the hi (hairless) mutant allele (Fig. 6) .
  • Plants were analysed at about 7-8 weeks after sowing.
  • a metal spatula was used to wipe along the stem of each plant in the experiment, thus covering a surface area of approximately 75cm 2 .
  • the resulting substance present on the spatula was dissolved in 1.0 ml ethanol. After drying the spatula, this procedure was repeated several times for each plant on different positions covering the whole stem surface .
  • the extracts from the individual plants were centrifuged for 5 minutes at 13000 rpm to remove debris, after which each extract was diluted with an equal volume of ethanol.
  • a UV-VIS spectrophotometer was used to measure the absorbance of the solutions.
  • the first absorbance peak is around between 210-290 nm, which is not in the visible part of the spectrum.
  • the second absorbance peak can range
  • Flavonoids are a class of polyphenols, the presence of which can be
  • the phenolics are measured against a
  • Acylated sugars are sugar compounds such as glucose or sucrose that are covalently bound to a fatty acid via an ester bond.
  • Example 6 Twelve leaf discs were taken with a cork borer of
  • Discs were taken from the 2 nd , 3 rd and 4 th leaf counted from the shoot apex of a plant, avoiding main veins, from plants of about 10-12 weeks old. Discs were placed in vials with 2 ml dichloromethane and rotated softly for 30 sec to dissolve surface compounds.
  • 0.1 ml of leaf surface extract was evaporated under nitrogen gas and redissolved in 0.5 ml 0.1M NaOH solution in methanol. Samples were heated to 60°C in a water bath for 15 min to allow saponification, i.e. breaking of the ester bond with NaOH and heat. Samples were then cooled and filtered.
  • Tomato plant (Solanum lycopersicum L.) comprising a mutant allele that leads to modification in the composition of flavonoids and volatiles as a result of the absence of the glandular heads of type VI trichomes, as compared to a plant not carrying the said genetic trait, which mutant allele is as comprised in a tomato plant representative seed of which was deposited with the NCIMB under deposit number NCIMB 41845.
  • Tomato plant as described in clause 1 or 2 obtainable by crossing a first tomato plant with a second tomato plant, wherein one of the said plants is grown from seeds as deposited with the NCIMB under deposit accession number NCIMB 41845, or a progeny plant thereof, and
  • Tomato plant as described in clause 1 or 2 obtainable by crossing a first tomato plant with a second tomato plant, wherein one of the said plants is carrying the said genetic trait that results in modification in the composition of flavonoids and volatiles as a result of the absence of the glandular heads of type VI trichomes, and selecting, preferably in the F2 generation, for plants that are modified in the composition of flavonoids and volatiles as a result of the absence of the glandular heads of type VI trichomes, as compared to a plant not carrying the said genetic trait.
  • Tomato plant as described in any of the clauses 1 to 4, wherein the mutant allele leads to the absence of green staining from the green vegetative parts of the plant upon contact and/or the absence or reduction of a typical tomato aroma.
  • a plant produced from the propagation material comprises the mutant allele as defined in clause 1 or 2.
  • Tomato plant as described in any of the clauses 1-5 and 7, seed as described in clause 6, or propagation material as described in clause 8, wherein the said mutant allele in the genome of seeds of which a representative sample was deposited as NCIMB 41845 is linked to a molecular SNP marker on position 1750800 of the public physical map SL2.30, wherein the SNP is a nucleotide change from A to G.
  • Food product comprising the tomato fruit as described in clause 10, or parts thereof, optionally in processed form.

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Abstract

La présente invention porte sur une plante de tomate (Solanum lycopersicum L.) dans laquelle les têtes glandulaires des trichomes de type VI sont absentes et qui possède une composition modifiée de flavonoïdes et de volatils. En outre, l'odeur typique de la tomate est absente ou presque. La plante comporte un allèle mutant, qui peut être obtenu à partir d'une plante de tomate, dont la graine représentative a été déposée sous le numéro d'enregistrement NCIMB 41845. Ledit allèle mutant se trouve sur le chromosome 9 entre les positions 1750800 bp et 4517648 bp, de préférence entre les positions 1939546 bp et 2970346 bp, surtout approximativement sur la position 2454946 bp sur une carte physique du génome de Solanum lycopersicum.
EP13702652.2A 2012-02-07 2013-02-07 PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI Withdrawn EP2811822A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP13702652.2A EP2811822A1 (fr) 2012-02-07 2013-02-07 PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP12154257.5A EP2625952A1 (fr) 2012-02-07 2012-02-07 Plant de tomate absent dans la coloration verte
PCT/EP2013/052391 WO2013117623A1 (fr) 2012-02-07 2013-02-07 Plante de tomate comportant un allèle mutant de tghvi
EP13702652.2A EP2811822A1 (fr) 2012-02-07 2013-02-07 PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI

Publications (1)

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EP2811822A1 true EP2811822A1 (fr) 2014-12-17

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EP12154257.5A Withdrawn EP2625952A1 (fr) 2012-02-07 2012-02-07 Plant de tomate absent dans la coloration verte
EP13702652.2A Withdrawn EP2811822A1 (fr) 2012-02-07 2013-02-07 PLANTE DE TOMATE COMPORTANT UN ALLÈLE MUTANT DE tghVI

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EP12154257.5A Withdrawn EP2625952A1 (fr) 2012-02-07 2012-02-07 Plant de tomate absent dans la coloration verte

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EP (2) EP2625952A1 (fr)
JP (1) JP6371224B2 (fr)
KR (1) KR102076019B1 (fr)
CN (1) CN104334010B (fr)
AU (1) AU2013218080B2 (fr)
CA (1) CA2862098C (fr)
IL (1) IL232919B (fr)
WO (1) WO2013117623A1 (fr)

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US20220389442A1 (en) * 2017-12-08 2022-12-08 Rijk Zwaan Zaadteelt En Zaadhandel B.V. Tbrfv resistant tomato plant

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Publication number Priority date Publication date Assignee Title
US20060134786A1 (en) * 2004-09-30 2006-06-22 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
EP2534942A1 (fr) * 2011-06-16 2012-12-19 Rijk Zwaan Zaadteelt en Zaadhandel B.V. Plant de tomate absent dans la coloration verte

Non-Patent Citations (1)

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Title
See references of WO2013117623A1 *

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CN104334010A (zh) 2015-02-04
AU2013218080B2 (en) 2018-08-09
CN104334010B (zh) 2017-06-20
JP2015509359A (ja) 2015-03-30
CA2862098A1 (fr) 2013-08-15
EP2625952A1 (fr) 2013-08-14
AU2013218080A1 (en) 2014-06-19
WO2013117623A1 (fr) 2013-08-15
IL232919A0 (en) 2014-07-31
KR102076019B1 (ko) 2020-02-11
JP6371224B2 (ja) 2018-08-08
KR20140119703A (ko) 2014-10-10
CA2862098C (fr) 2022-08-16
IL232919B (en) 2020-02-27

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