WO2007061146A1 - Procede de production de choux chinois transgenique utilisant un tissu de tige de fleur, et produit resultant a resistance accrue a la pourriture mole - Google Patents

Procede de production de choux chinois transgenique utilisant un tissu de tige de fleur, et produit resultant a resistance accrue a la pourriture mole Download PDF

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WO2007061146A1
WO2007061146A1 PCT/KR2005/003960 KR2005003960W WO2007061146A1 WO 2007061146 A1 WO2007061146 A1 WO 2007061146A1 KR 2005003960 W KR2005003960 W KR 2005003960W WO 2007061146 A1 WO2007061146 A1 WO 2007061146A1
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chinese cabbage
plant
lane
transgenic
soft rot
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PCT/KR2005/003960
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English (en)
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Jong Kee Kim
Byung Ho Hwang
Soo Seong Lee
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Chung-Ang University Industry-Academy Cooperation Foundation
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Priority to JP2008542213A priority Critical patent/JP5020253B2/ja
Publication of WO2007061146A1 publication Critical patent/WO2007061146A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
    • C12N15/8205—Agrobacterium mediated transformation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8281—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for bacterial resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance

Definitions

  • the present invention relates to a method for producing a transgenic Chinese cabbage plant from a flower stalk tissue of Chinese cabbage and a transgenic Chinese cabbage produced thereby, having high resistance to soft rot More particularly, the present invention relates to a method in which a foreign gene is introduced into a flower stalk tissue of Chinese cabbage and then the transformed tissue is cultured to a plantlet, and a transgenic Chinese cabbage plant which harbors therein a foreign gene associated with soft rot resistance so as to show superior resistance to soft rot.
  • Chinese cabbage is a leafy vegetable which consists mainly of water, with edible leaves rich in calcium and vitamin C. Optimally growing at 18 ⁇ 22°C Chinese cabbage has affinity for low temperatures and shows poor heat tolerance. It grows well under weak light while its roots are widely arranged. Morphologically, Chinese cabbage has 2/5 spiral phyllotaxis and leaves in a basal rosette. Showing self incompatibility, Chinese cabbage needs cross fertilization and features inbred depression and heterosis. Conventionally, the varietals improvement of Chinese cabbage has been achieved by hybridization, mainly using Fl heterosis. However, this method for varietals improvement has come to the end due to the limitation of available genetic resources. Recent great advances in tissue culturing and genetic engineering technology have allowed the construction of a new road for the varietals improvement of celery cabbage.
  • tissue culture technology is useful to maintain and proliferate these individuals.
  • Various plant transduction techniques that have recently been developed, have lead to the development of new plant varieties, but they suffer from low regeneration efficiency.
  • tissue culture techniques that assure newly developed plants of high regeneration efficiency so as to effectively utilize plant transduction techniques.
  • plant transformation Since the first success with tobacco, plant transformation has been established as a potent method of finding functions of genes of interest as well as of introducing useful foreign genes into plant cells to create new varieties.
  • Examples of the transformation techniques of introducing genes into plant cells include electroporation, direct gene transfer, micro-injection, tissue injection, use of PEG, Ca +2 , or Agrobaderium. New plant varieties are acquired through the selection of transformant cells and the regeneration of transgenic plants.
  • plant transformation mediated by Agrobacterium tumefiiciens takes advantage of the mechanism in which Agwbacterium transfers its own DNA into plant genomes, and has become the most common method for the introduction of foreign genes into plant cells and the subsequent regeneration of transgenic plants, finding various applications in general research fields including genetics, biochemistry, embryology, and physiology as well as botany.
  • A. tumefiiciens and AgrObacteriutn rhizogenes both Gram- negative soil bacteria, are known to naturally infect the wound sites in dicotyledonous plants, causing the formation of crown gall tumors and hairy root tumors, respectively, by transformation with their own genes.
  • rhizogenes is attributed to the specific plasmids that they carry, for example, the Ti (tumor inducing) plasmid in A. tumefiiciens) and the Ri (root inducing) plasmid in A. rhizogenes.
  • the tumor induction by these microorganisms results from the transfer of a particular DNA segment (T-DNA) of the Ti or Ri plasmid into the nucleus of infected cells where it is then stably integrated into the host genome and transcribed. It is known that the transfer of T-DNA (transfer DNA) is controlled by the expression of virulent genes harbored in the Ti plasmid.
  • campestris var. para plant transformation has been successfully achieved in many cases.
  • leaf cytoplasms of B. campestris var. rapa were successfully transformed directly with cauliflower mosaic virus genes by Tanaka et al. (1985) and Paszkowski et al., (1986) to afford transgenic plants.
  • Ohlsson and Eriksson (1988) reported that a cytoplasm derived from the hypocotyls of B. campest ⁇ s var. oleijera was infected with Agrobacterium to give a kanamycin-resistant callus.
  • a transgenic plant was not obtained. There are successes in obtaining transgenic plants from Chinese cabbage(B. campestris).
  • a method comprising: introducing a transformation vector carrying a gene associated with soft rot resistance into an Agrobacterium sp. strain; inoculating a flower stalk tissue of Chinese cabbage with the microorganism; culturing the inoculated flower stalk tissue to a planflet; quantitatively and qualitatively analyzing the plantiet for transformation with the gene through PCR, Southern blotting and RT-PCR; and assay the transgenic Chinese cabbage plant for soft rot resistance.
  • the method according to the present invention exhibits a transformation efficiency of 2.8%, which is two or more times improved over 0.4 ⁇ 0.8% obtained by conventional methods using cotyledon or hypocotyl. It is expected that the method of the present invention can be applied to plants other than celery cabbage, thereby constructing a plant transformation system more efficient than that established by conventional methods. Also, the transgenic Chinese cabbage plant produced by the method of the present invention is far superior in soft rot resistance to wild type.
  • FIG. 1 is a schematic gene map showing a T-DNA portion of the dual vector pCAMBIA2300-BcPGJP2.
  • FIG. 2 depicts regeneration from adventitious shoots formed from the flower stalk tissue segments of Chinese cabbage to transgenic plants.
  • FIG.3 is a photograph of electrophoresed PCR products showing the presence of an npt ⁇ gene in a transgenic Chinese cabbage plant.
  • FIG. 4 is a photograph of Southern blots showing the hybridization of the genomic DNA of the To transgenic plant with an nptll gene probe.
  • FIG. 5 is a photograph of electrophoresed RT-PCR products showing the expression of the nptll gene in the transgenic plant
  • FIG. 6 is a photograph of electrophoresed RT-PCR products showing the expression of the BcPGIPl gene in the transgenic plants.
  • FIG. 7 shows comparison of soft rot resistance between a wild-type Chinese cabbage plant and a transgenic plant
  • a transgenic Chinese cabbage plant highly resistant to soft rot is provided.
  • the production of the transgenic Chinese cabbage plant starts with the construction of a vector carrying a gene conferring resistance to the plant
  • the vector is introduced into a strain of Agrobacterium sp. Then, the strain is allowed to infect a flower stalk tissue of celery cabbage, followed by tissue culturing the infected flower stalk tissue.
  • Genomic DNA is prepared from a leaf tissue regenerated from the flower stalk tissue and analyzed through PCR, DNA gel blotting, and RT-PCR to determine whether the Chinese cabbage is transgenic or not
  • the celery cabbage, if transgenic, is analyzed for resistance to soft rot.
  • a method for producing a transgenic Chinese cabbage plant characterized in that a flower stalk tissue of Chinese cabbage is infected with a microorganism which harbors a vector carrying a foreign gene of interest, and is then tissue-cultured.
  • the foreign gene useful in the present invention may be BcPGIFl (polygalacturonase-inhibiting protein 2) that shows potent resistance to soft rot, or may be determined depending on necessity and purpose.
  • BcPGIFl polygalacturonase-inhibiting protein 2
  • the microorganism suitable for harboring the foreign DNA-introduced vector may be a strain of Agrobacterium spp. or may belong to a different genus according to transformation efficiency.
  • a transgenic Chinese cabbage plant having high soft rot resistance is produced by the method in which Chinese cabbage is infected with Agrobacterium sp. containing a vector carrying a heterogeneous gene directed to potent soft rot resistance.
  • a better understanding of the present invention may be obtained through the following examples which are set forth to illustrate, but are not to be construed as the limit of the present invention.
  • a PGlFl gene obtained from a J3-11M-54 lineage of celery cabbage, was inserted into pCAMBIA2300, a plant transformation vector, at a position between the restriction enzyme sites BamH land Kpn J followed by introducing the vector into Agrobacterium tumefijciens LBA4404 for the transformation of celery cabbage.
  • a BcPGlFl gene of J3-11M-54 lineage was inserted into pCAMBIA2300, a plant transformation vector, at a position between the restriction enzyme sites BamH land Kpn I followed by introducing the vector into Agrobacterium tumefadens LBA4404, which was then used for the transformation of tobacco and Chinese cabbage(FIG.1).
  • FIG. 1 is a schematic gene map showing a T-DNA portion of the dual vector pCAMBIA2300-BcPG/P2, where NFm stands for Neomycin phosphotransferase II, 35S Pro for CaMV 35S promoter, and NOS for Nopaline synthase gene terminator. Transformation of Celery cabbage
  • BcPGIPl was expressed in J3- 11M-3, J3-11M-44 and J3-11M-54 lineages of celery cabbage, which had been bred in the Horticultural Crops Quality Lab. of Applied Plant Science Department, Industrial Science College, ChungAng University, Korea.
  • a flower stalk tissue of Chinese cabbage was utilized. 4-week-old seeding of J3-11M-3, J3-11M-44 and J3-11M-54 lineages of Chinese cabbage were treated at 4 ° Gfor 4 to 6 weeks in a green house and bolted.
  • the parenchyma of the bolted flower stalk was cult into cylindrical segments, each 5 mm long, which were than sterilized in 70% EOH for 30 sec, washed once with sterile water, immersed in 1% sodium hypochloride for 10 min, and washed three times with sterile water.
  • the sterilized flower stalk tissue segments were immersed for 10 min in an Agrobacterium culture which had been cultured for 36 hours with agitation, followed by filtering off the culture solution through filter papers.
  • the segmental tissues were cultured in a selection medium (MS medium 4.4g/L, sucrose 30g/L, phyto-agar 8g/L, AgNC%4.0mg/L, NAA 1.0mg/L, BAP 4.0mg/L, KM 25mg/L, Cefotaxime 200mg/L, pH 5.7) at 23 "(for 16 hours under light and then for 8 hours in the absence of light At intervals of 15 days, sub-culturing was conducted 4 ⁇ 6 times with callus tissues cut from J3-11M-44.
  • MS medium 4.4g/L, sucrose 30g/L, phyto-agar 8g/L, AgNC%4.0mg/L, NAA 1.0mg/L, BAP 4.0mg/L, KM 25mg/L, Cefotaxime 200mg/L, pH 5.7
  • the callus tissues were sub-cultured 4 ⁇ 6 times in a selection medium having a low kanamycin concentration (MS medium 4.4g/L, sucrose 30g/L, phyto-agar 8g/L, AgNO 3 4.0mg/L, NAA 1.0mg/L, BAP 4.0mg/L, KM lOmg/L, Cefotaxime 200mg/L, pH 5.7).
  • a selection medium having a low kanamycin concentration MS medium 4.4g/L, sucrose 30g/L, phyto-agar 8g/L, AgNO 3 4.0mg/L, NAA 1.0mg/L, BAP 4.0mg/L, KM lOmg/L, Cefotaxime 200mg/L, pH 5.7.
  • the completely differentiated plantiets which were obtained by inducing the tissues to root for 2-3 weeks, were transplanted in a flower pot, 7 cm in diameter, containing sterile bed soil a diameter of 7cm. After being covered with vinyl wrap to maintain moist conditions, the plantiets were allowed to grow at 23 "C for 5 ⁇ 7 days with a cycle of a light condition for 16 hours and a dark condition for 8 hours. When the aerial portion normally grew and the roots were well developed, the plantiets were transplanted into a container 25 cm in diameter and cultivated in a greenhouse.
  • FIG. 2 depicts a regeneration process from adventitious shoots formed from the flower stalk tissue segments of Chinese cabbage to transgenic plants, showing shoots formed after the growth of the segments for 5 ⁇ 6 weeks in panel A, shoots transferred into a rooting medium in panel B, transgenic explants in panel C and transgenic plants with flower buds in panel D.
  • EXAMPLE 2 To Determine Whether a Putative Transgenic Chinese cabbage Carries a Foreign Gene
  • Example 2 To determine whether the transgenic Chinese cabbage plant prepared in Example 1 carries nptH and BcPGIP2, PCR and DNA gel blotting were performed.
  • PCR was performed. While 100 ng of the genomic DNA isolated from the leaves serves as a template, it, along with a set of npt H primers (forward) 5'-ATGGGGATTGAACAAGATGGATTGGe 1 and (reverse) 5'-
  • TCAGAAGAACTCGTCAAGAAGGCGATAG-3' underwent a PCR, which started with 94 "C pre-denaturation for 5 min and carried out with 30 cycles of denaturing temperature at 94 °C for 1 min, annealing temperature at 60 °C for 1 min and extending temperature at 72 ° C for 1 min in the presence of AccuPower PCR premix (Bioneer), finally followed by 72°C extension for an additional 10 min. Qn 1.2% agarose gel, the PCR reaction was electrophoresed to identify a DNA fragment 798 bp in size, and the results are shown in FIG. 3.
  • FIG. 3 is a photograph of PCR analysis showing a lkb plus DNA ladder (M), a PCR product of a control plant (lane 1), PCR products of transgenic plants (lanes 2 to 15) wherein lane 2/T 0 -I, lane 3;To-2, lane 4;To-3, lane 5;To4, lane 6;To-6, lane 7;To-8, lane 8;To-9, lane 9/T 0 -IO, lane 10/T 0 -Il, lane ll;To-12, lane 12/T0-15, lane 13;To-16, lane 14;To-17, and lane 15;T ⁇ -19, and a positive control (vector pCAMBIA 2300-BcPGIP2) (lane P).
  • M lkb plus DNA ladder
  • EXPERIMENTAL EXAMPLE 2 DNA Gel Blotting for Determining the Introduction of nptU Transgenic plants regenerated from J3-11M-44 lineage were analyzed using a
  • DNA gel blotting method DNA gel blotting method.
  • 200 mg of leaves were taken from each of the transgenic plants and ground using liquid nitrogen and a mortar.
  • 700 ⁇ « of an extraction buffer (1.25% SDS, 0.1M Tris-HQ, 0.5M NaQ, 0.05M EDTA, 0.38% sodium bisulfite, pH 8.0) that was preheated to 65 ° C.
  • the DNA thus obtained was transferred into a new 1.5mL vial, washed twice with 70% EOH, dried and suspended in a TE buffer (1OmM Tris-Q pH 8.0, ImM EDTA).
  • FIG. 4 along with lamda ONA/ HincM (lane M) and a wild type control J3-11M-44 (lane Q, genomic DNAs from the transgenic plants are run on lanes 1 to 19 wherein lane l;To-l, lane 2;To-2, lane 3;To-3, lane 4;To-6, lane 5;To-8, lane 6;To-9, lane 7/T 0 -IO, lane 8/T 0 -Il, lane 9;To-12, lane lO/To-15, lane ll;To-16, lane 12/To-17,lane 13;To-194ane 14;T(r20, lane 15;To-21, lane 16;T(r34, lane 17;To-36, lane 18;To-38, and lane 19
  • T 0 -I, Tff-2, TQ-3 and To-20 and another identical band pattern found in To-6, To-8, To-9, T 0 -IO, T 0 -H, TQ-12, To-15, T 0 -Io, To-17, To-21, To-34, TO-36, T ⁇ -38 and To-39.
  • EXPERIMENTAL EXAMPLE 3 RT-PCR for Determining the Expression of npt U and BcPGlPl The transgenic celery cabbages To-6, T 0 -IO, To-12, To-15, To-16, To-17, To-20, and T 0 -
  • RNA samples 21 were analyzed for the expression of npt H and BcPGlPl through RT-PCR.
  • total RNA was prepared from leaves of each transgenic plant using an RNeasy Plant mini kit (Ojazen) and quantitatively analyzed using a spectrophotometer. After being synthesized from 5 ⁇ g of each total RNA with the aid of a Superscript l ⁇ Strand Synthesis Kit (Invitrogen U.S.A.), a 1 st strand cDNA was used as a template for RT-PCR which was carried out for determining the expression of npt ⁇ . in a manner similar to that of Experimental Example 1.
  • BcPGIPl a set of the following primers was used:
  • the RT-PCR started with 94 ° C pre-denaturation for 5 min and was performed with 30 cycles of denaturing temperature at 94 °C for 1 min, annealing temperature at 58 ° C for 1 min and extending temperature at 72 ° C for 1 min, finally followed by 72 "C extensions for an additional 10 min.
  • an actin gene of Chinese cabbage was amplified using a set of the following BcActin primers:
  • FIG.5 is an electrophoresis photograph of RT-PCR products from the wild-type plant J3-11M-44 (lane 1) and the transgenic plants (lanes 2 to 9), wherein lane 2;To-6, lane 3;To-lO, lane 4;To-12, lane 5;T ⁇ -15, lane 6;T(rl6, lane 7;T(rl7,lane 8;To-20, and lane 9;To-21, along with a lkb plus DNA ladder (lane M), showing the expression of npt ⁇ gene in the transgenic plants.
  • HG. 6 is a photograph of electrophoresed RT-PCR products from wild-type plants (lanes 1 to 3) and transgenic plants (lanes 4 to 11) wherein lane 4;To-6, lane 5;To-lO, lane 6;To-12,lane 7;T ⁇ rl5, lane 8;To-16, lane 9;To-17, lane 10;T(r20 and lane 11/1 ⁇ 21, along with a lkb plus DNA ladder (lane M), showing the expression of BcPGIPl in the transgenic plants.
  • both npt E and BcPGIPl are normally expressed in the transgenic plants.
  • Example 1 The Chinese cabbage transformed in Example 1 was analyzed for resistance to soft rot 60 days after transplantation, the transgenic Chinese cabbage plants of J3-11M-44 lineage, which were found to carry and express the BcPGIP2 gene as measured by PCR, Southern blotting and RT-PCR analyses, and a wild type Chinese cabbage plant were exposed to pathogens causing soft rot
  • leaves taken from the transgenic and wild type Chinese cabbage plants were inoculated with the pathogens at a density of 1.2XlO 5 cfu/ ⁇ Jt, using a point inoculation method (Park et al., 2004), so as to quantitatively measure the susceptibility thereof to soft rot
  • a point inoculation method Park et al., 2004
  • the transgenic plant To-21 was found to develop disease lesion 12 hours after inoculation, and be improved in soft rot resistance by 54.6% 12 hours after inoculation, by 52.5% 18 hours after inoculation, by 29.1% 24 hours after inoculation, by 19.7% 30 hours after inoculation, and by 20.3% 36 hours after inoculation, compared to the wild type.
  • transgenic Chinese cabbage having potent resistance to soft rot can be prepared by a method utilizing a flower stalk tissue in accordance with the present invention.
  • the method is also applicable to plants other than celery cabbage, thereby constructing a plant transformation system more efficient than that established by conventional methods. Therefore, the present invention finds useful applications in the crop industry.

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Abstract

L'invention porte sur un procédé de production de choux chinois transgénique utilisant un tissu de tige de fleur, ce qui donne un choux chinois transgénique présentant une résistance accrue à la pourriture mole. Le procédé présente un rendement de transformation de 2,8 % d'au moins 2 fois supérieur au rendement de 0,4-0,8 % obtenu par le procédé classique utilisant un cotylédon ou un hypocotyle. Ledit procédé devrait pouvoir s'appliquer à d'autres plantes que le choux chinois et permettre de réaliser un système de transformation de plantes plus efficace que celui obtenu par les procédés classiques. Le choux transgénique ainsi produit est très supérieur au type sauvage pour ce qui est de la résistance à la pourriture mole.
PCT/KR2005/003960 2005-11-22 2005-11-23 Procede de production de choux chinois transgenique utilisant un tissu de tige de fleur, et produit resultant a resistance accrue a la pourriture mole WO2007061146A1 (fr)

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JP2008542213A JP5020253B2 (ja) 2005-11-22 2005-11-23 花茎組織を用いたハクサイの形質転換体の製造方法及びそれから生産される軟腐病抵抗性の向上した形質転換体

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JP2016140318A (ja) * 2015-02-03 2016-08-08 住友ゴム工業株式会社 ゴムノキの再生方法、ゴムノキの増殖方法、シュートの誘導方法、シュートの伸長方法、シュートの発根方法、及び幼植物の馴化方法

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