WO1996012402A1 - Riz resistant aux herbicides et procede d'amelioration genetique afferent - Google Patents

Riz resistant aux herbicides et procede d'amelioration genetique afferent Download PDF

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Publication number
WO1996012402A1
WO1996012402A1 PCT/JP1994/001757 JP9401757W WO9612402A1 WO 1996012402 A1 WO1996012402 A1 WO 1996012402A1 JP 9401757 W JP9401757 W JP 9401757W WO 9612402 A1 WO9612402 A1 WO 9612402A1
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WO
WIPO (PCT)
Prior art keywords
rice
resistant
herbicide
seeds
breeding
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.)
Ceased
Application number
PCT/JP1994/001757
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English (en)
Japanese (ja)
Inventor
Yoshio Shigematsu
Masami Ogawa
Noriko Nakata
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.)
Sankyo Co Ltd
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Sankyo Co Ltd
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 Sankyo Co Ltd filed Critical Sankyo Co Ltd
Priority to PCT/JP1994/001757 priority Critical patent/WO1996012402A1/fr
Priority to AU79488/94A priority patent/AU7948894A/en
Publication of WO1996012402A1 publication Critical patent/WO1996012402A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/93—Ligases (6)
    • 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
    • A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4636—Oryza sp. [rice]
    • 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
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • 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
    • A01H5/10—Seeds

Definitions

  • the present invention relates to a rice and its seed resistant to a herbicide having a fatty acid synthesis inhibitory action, a method of breeding the same, and a selective control of grass weeds by applying the herbicide to a field where the rice is cultivated. On how to enable.
  • JP-A-64-85024 discloses that rice callus is cultured in a medium containing a herbicide such as bensulfuron-methyl or glyphosate, and the rice is resistant to the herbicide.
  • a herbicide such as bensulfuron-methyl or glyphosate
  • benzulfuron-methyl which is the subject of the above-mentioned patent gazette, is originally a safe herbicide for rice, and has a drawback that its efficacy is also weak for barnyardgrass.
  • glyphosate is an effective herbicide for foliage treatment, and therefore cannot be used as a soil treatment herbicide.
  • Herbicides that exhibit a fatty acid synthesis inhibitory action are known to have a strong herbicidal activity against grass weeds in both soil treatment and foliage treatment. Is also known to be effective. At present, their use is limited due to their severe chemical damage to grasses in general. Attempts to confer corn resistance to these herbicides are disclosed in PCT International Patent Publication No. WO 9216101. However, there have been no successful reports on rice.
  • the present invention provides rice that exhibits resistance to a herbicide having a fatty acid synthesis inhibitory action, and enables selective control of grass weeds by applying the herbicide to a field where the rice is cultivated.
  • the aim is to provide a method.
  • the present inventors cultured rice cells in a medium containing a herbicide exhibiting a fatty acid synthesis inhibitory action, selected rice culture cells that were resistant to the herbicide, and re-differentiated them to resist the herbicide. Sex rice and its seeds were created.
  • the present invention relates to a rice and its seed resistant to a herbicide having a fatty acid synthesis inhibitory action; a rice and its seed resistant to a herbicide having an acetyl-CoA carboxylase inhibitory action; aryloxy phenoxy fatty acid , Derivatives of aryloxyphenoxy fatty acids, such as salts, esters, and amides, N-benzoyl-N-phenylamino acids, and derivatives of N-benzylamino acids, salts, esters, and amides of N-phenylamino acids And seeds thereof which are resistant to herbicides selected from cyclohexanediones and derivatives thereof such as aryloxyphenoxy fatty acids, salts, esters and amides of aryloxyphenoxy fatty acids.
  • Rice and seeds resistant to the selected herbicide selected from aryloxyphenoxy fatty acids and their ester derivatives Rice and seeds resistant to herbicides; rice and seeds resistant to phenolic sub-mouth, rice and seeds resistant to fluazifop butyl and seeds: rice and seeds resistant to quizalofopethyl
  • the seed a rice broth is cultured in a medium containing a herbicide having an action of inhibiting fatty acid synthesis, and a rice culture cell resistant to the herbicide is selected and redifferentiated.
  • Herbicide-resistant rice and its seed breeding method and cultivating the rice seed and crossing it with other varieties of rice, and breeding a herbicide-resistant herbicide-resistant rice seed
  • a rice cell in a medium containing a herbicide having an inhibitory effect on a cetyl-CoA carboxylase Culturing, selecting rice culture cells resistant to the herbicide, and redifferentiating the rice culture cells.
  • Breeding method of rice and seeds resistant to the herbicide, and cultivating the rice seeds A method of breeding rice seeds resistant to a herbicide exhibiting acetyl-CoA carboxylase inhibitory activity by crossing rice varieties with other varieties of rice; aryloxy phenoxy fatty acids, aryloxy fenoki Derivatives of salts of fatty acids, such as esters, amides, etc., N-benzoyl-N-phenylamino acids, salts of N-benzoyl-N-phenylamino acids, derivatives such as esters, amides, and cyclohexanediones Culturing rice cells in a medium containing a herbicide selected from the group consisting of: selecting rice culture cells resistant to the herbicide; and redifferentiating the cultured rice cells.
  • aryloxyphenoxy fatty acids such as N-benzoyl-N-phenylamino acids, N-benzoyl-N-phenylamino acid salts, esters, amides, and other herbicides selected from cyclohexanedione.
  • a method for breeding seeds culturing rice cells in a medium containing a herbicide selected from derivatives of aryloxyphenoxy fatty acids, salts, esters and amides of aryloxyphenoxy fatty acids; A rice cultivation cell which is resistant to the herbicide, and a method for breeding the rice and the seed thereof, wherein Cultivate rice seeds and cross them with other varieties of rice. Resistant to herbicides selected from derivatives such as aryloxy carboxylic acids, salts of aryloxy carboxylic acids, esters and amides.
  • Rice seed breeding method rice cells are cultured in a medium containing a herbicide selected from aryloxyfuninoxy fatty acids and ester derivatives thereof, and rice culture cells resistant to the herbicide are selected.
  • a method of breeding rice culturing rice cells in a medium containing fluazifop butyl, selecting rice culture cells resistant to the herbicide, and redifferentiating the cells; Breeding method for cultivated rice and its seeds, cultivating the rice seeds and crossing them with other varieties of rice, breeding rice seeds resistant to fluazifopbutyl; Breeding of rice and seeds resistant to quizalofopethyl, characterized by culturing rice cells, selecting rice culture cells resistant to the herbicide, and redifferentiating them.
  • Fig. 1 shows the growth of calli selected from rice (Nipponbare) on a medium supplemented with fluazihob butyl or cetoxydim.
  • FIG. 2 shows the inhibitory effect of the herbicide on the acetyl-CoA carboxylase of Nipponbare-selective strain ( ⁇ ) and the selected No. 17 strain (Oka) yellow-flowered sprout.
  • R / S indicates a drug concentration ratio that inhibits both enzyme activities by 50%.
  • the herbicides having an inhibitory action on fatty acid synthesis used in the present invention include phenoxaprop (Ifenoxap rop), dicloofop (diclofop), clofop lclofop), and triphop ( trifop and fenthiaprop, uazifop, haloxyfop, haloxyfop, quizalofop and chlorazifop, and allyloxyphenoxy fatty acids: fenoxaprop-ethyl ) (Ethyl (RS) -2- [4- (6-chloro-l, 3-benzoxazol-2-yloxy) phenoxy] ropionate), diclofop-methyl (Methyl (RS) -2- [ 4- (2, 4- dichlorophenoxy) phenoxy] propionate), clofop-isobutyl (Iso
  • Cultured rice cells can be induced as callus by culturing ripe seeds, immature seeds, roots, anthers or pollen in a suitable medium after sterilization.
  • the basal medium any known medium can be used. In particular, it is possible to grow well using the MS or N6 basal medium. Vitamins can be obtained by using those contained in the basal medium.
  • the addition of amino acids is effective in maintaining the regeneration ability, and the addition of 1 to 50 mM proline is particularly effective.
  • As a carbon source 1 to 5% of sucrose may be added, and up to 5% of sorbitol-mannitol may be added as an osmotic pressure regulator.
  • a medium containing 2,4-dichlorophenoxyacetic acid as a hormone at a concentration of 0.1 to lO rn gZ, particularly at a concentration of l to 4 mgZ £, is advantageous.
  • other auxins such as naphthalene acetic acid, indole acetic acid, and indole acetic acid, and a small amount of cytokinin such as force-ricein and benzyladenine may be added.
  • the force is usually induced on a solid medium, or a liquid medium may be used.
  • the callus can be maintained and proliferated using a medium having the same composition in either a bright place or a dark place, but usually in a dark place. [When using a control medium, add 0.1 to 0.6% gel light or 0.3 to 2% agar to the medium.
  • callus cultured in solid culture or cells cultured in liquid culture can be used.
  • treatment with ultraviolet rays, X-rays, gamma rays, or treatment with a chemical agent such as ethyl methanesulfonate is suddenly performed. Mutations may be induced.
  • resistant cells For the selection of resistant cells, a method of culturing callus nodules and small pieces in a medium containing a herbicide can be used.
  • this method requires a great deal of effort to produce callus nodules and small pieces, and different types of resistant cells coexist in the same nodule.
  • the obtained resistant calli tends to become a chimera, which is a mixture of different cells.
  • use liquid media By repeating the above procedure, resistant cells can be obtained, but they are the same in that they tend to be chimeras.
  • sensitive cells may be contaminated. This can be eliminated by gradually increasing the drug concentration and selecting.
  • a method of relatively easily obtaining a mutant strain close to a clone is a method of selecting or shredding a cell clump finer than proliferated cells, and spreading this on a solid medium containing a drug, followed by culturing. Specifically, callus cultured in a solid medium or a liquid medium is passed through a sieve having an opening of about 0.1 to 2 mm, preferably about 0.5 mm, to obtain small clumps of dozens of cells.
  • the drug concentration is determined from the concentration that inhibits the growth of unselected rice.
  • Herbicides that inhibit fatty acid synthesis generally inhibit the growth of rice cultured cells at 0.05 to lmgZ, and completely halt the growth at lmg / or more.Therefore, when selecting drug concentration at a single point, Even if selection is performed by gradually increasing the concentration, eventually add 1 mg Z £ or more of the drug to obtain a resistant cell line.
  • the differentiation medium may be a medium containing no auxin, which is basically the same as the callus culture medium, or a solid medium containing 2,4-dichlorophenoxyacetic acid of 0.1 mg or less or naphthalene acetic acid of 1 mgZ or less.
  • a medium the selected cells are transplanted into the medium and cultured in a light place for 10 days to 2 months to differentiate the plant. At this time, it is more effective to add 1 to OmgZfi cytokinins.
  • Differentiated rice plants are grown in another fresh medium until the plant height is 5 to 20 cm and the roots are sufficiently elongated. This is transplanted to a pot filled with soil, and the humidity is gradually lowered to acclimate it before cultivation outdoors or in a greenhouse. All of the rice thus obtained is resistant to herbicides that inhibit fatty acid synthesis. However, in the next generation individuals that germinated from the seeds obtained from this individual, usually resistant and susceptible individuals are separated at a ratio of 3: 1. Therefore, it is necessary to select resistant individuals again.
  • the resistant rice individuals show normal growth, but Since the individuals do not grow at all, resistant rice individuals can be easily selected.
  • a resistant rice body can be easily selected by a method in which the herbicide is applied to the rice body one to three weeks after germination of the seed, and the susceptible rice individual dies.
  • the fixed rice resistance trait can be introduced into other varieties of rice by a method known to those skilled in the art of backcrossing after crossing.
  • the above-mentioned resistance trait is dominantly inherited, it is also possible to cultivate rice seeds in which the resistance traits are fixed and to breed rice of another variety to obtain drug-resistant F1 rice seeds.
  • Rice seeds having fixed resistance traits can be cultivated by a normal method of direct sowing and transplanting.
  • the herbicide By applying the herbicide to the field where the rice was cultivated, the rice seeds are not harmful to rice. Weeds can be controlled selectively.
  • the treatment can be carried out over a wide period from pre-emergence to heading, and the drug is formulated according to the purpose by mixing the soil before sowing or transplanting, or treating the soil or foliage after emergence or transplantation. Can be administered. If desired, it is possible to simultaneously control broadleaf and weeds belonging to the family Calyptaceae by mixing with chemicals such as vensulfuron methyl benzozone.
  • the rice and seeds resistant to the herbicide having the fatty acid synthesis inhibitory action of the present invention were obtained from Sankyo Agricultural Research Laboratory (located in Yasu-cho, Yasu-gun, Shiga, Japan). Those who are breeding, preserving, and managing in 041), and who intend to conduct them for testing or research, can receive a sale.
  • the rice plant created by the present invention is resistant to herbicides having a fatty acid synthesis inhibitory action, the rice plant is not harmed by the treatment with this herbicide and can selectively control grass weeds. Can be. That is, by cultivating the rice according to the present invention, it is possible to effectively control gramineous weeds in fields, such as the rice breeder, which have been difficult to control conventionally.
  • Rice (Nipponbare) Brown rice, from which rice husks have been removed from ripe seeds, is sterilized by soaking it in 70% ethanol for 1 minute, then in a 2% aqueous solution of hypochlorous acid for 15 minutes, and washing it with sterile water several times. Then, the embryos were placed on a callus-derived solid medium 30 ml with the embryos facing upward.
  • the callus induction medium is based on N6 medium and contains 12 mM proline, 100 mg / £ casamino acid, 2 mg Z £ 2,4-dichloromouth phenoxyacetic acid, 3% sucrose, 0.25% gel light. It was added and the one prepared to ⁇ 5.7 was used.
  • the callus grown in the liquid culture was passed through a sieve with a mesh of 0.5 mm while being spooned to obtain small aggregated cells. These cells are added per 100m of sterile water 12.
  • Cultured rice cells were inhibited by about 85% growth in a medium with a phenoxapropethyl concentration of 0.1 mgZ_e, and could hardly grow in a 0.3 mgZ medium. After culturing for 1 to 3 months in the medium described above, 21 resistant cell lines were obtained from 80 g of fresh weight cells. This was transplanted to a solid medium of the same composition and grown.
  • Figure 1 shows the growth on the other two herbicide-supplemented media. Many strains grew well in the medium containing fluazifop butyl and were resistant. Although growth was inferior in the medium containing setoxydim, resistance was observed in some of them.
  • No. 17 which had strong resistance to phenoxysubupet and fluazihobbutyl in the above assay and was also resistant to sethoxydim was selected and transplanted to a differentiation medium. And cultured under 27, 3000 lux light.
  • the differentiation medium is based on N6 medium, 0.2% casamino acid, 2% sucrose, 3% A solid medium supplemented with Toll, 1 ⁇ naphthalene acetic acid, and 10 ⁇ rice was used.
  • the cells began to germinate at about day 10, and then stem and leaf differentiation and rooting occurred.
  • the rice was transplanted to a fresh differentiation medium and further cultured, so that healthy rice seedlings with a plant height of about 15 cm were obtained.
  • the redifferentiated seedlings were transplanted to a sterilized soil filled in a plastic container of 6 cm on a side. After cultivation for two weeks while gradually lowering the humidity, the seeds were transplanted to a 100-liter Pellet bottle and cultivated by a conventional method to obtain a large number of fertile seeds.
  • the seeds obtained as described above were germinated, and 10 seeds were sown per 30 m of a 0.5% agar medium containing the above herbicide. This was cultivated for one week in a light place at 27 ° C. and 300 lux. Table 1 shows the results. Approximately 3-4 of the plants germinated from the next-generation seeds showed normal growth in the presence of phenoxapropethyl.
  • Example 2 Selection was performed in the same manner as in Example 1, and 17 2 cell lines capable of growing in a medium supplemented with 2 mg / £ phenolic submouth were selected from 89 g of freshly cultured cells.
  • the selected strain was transplanted to the same differentiation medium as in Example 1 to regenerate the plant.
  • the regenerated plant was cultivated in the usual manner to obtain rice (Koshihikari) seeds resistant to the herbicide.
  • rice Koshihikari
  • Table 2 shows the results. Resistance of these rices was fixed in the same manner as in Example 1. When this resistant fixed line was cultivated in the field, it showed the same growth and morphology as unselected Koshihikari, and the same yield was obtained.
  • fenoxaprobuethyl was added in an amount of 1 kg / ha. After soil treatment or when the plant height has reached about 5 cm, 500 mg of fenoxaprophetyl solution was sprayed onto the foliage to 0.5 kgZha. Cloths and subsequent growth were investigated. As a result, as shown in Table 3, weeds could be effectively controlled by any of the treatments.
  • Herbicidal effect is normal growth (0>, about 20% dying (1), about 40% dying (2), about 60% dying (3), about 80% dying (4), complete dying Evaluation was made on a scale of (5).

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Abstract

L'invention porte sur une variété de riz et ses graines, rendue résistante à un herbicide présentant une action inhibitrice de la synthèse d'acides gras ainsi que sur un procédé d'amélioration génétique du riz et de ses graines visant à lui conférer cette résistance. Le procédé consiste à cultiver des cellules de riz dans un milieu contenant l'herbicide, à sélectionner des cellules de riz résistantes à l'herbicide et à les redifférentier. Le procédé d'amélioration génétique visant à rendre le riz résistant à un herbicide présentant une action inhibitrice de la synthèse d'acides gras consiste à cultiver sa graine et à la croiser avec un riz d'une autre variété. L'invention porte sur une méthode de lutte contre les mauvaises herbes des graminées consistant dans l'épandage d'un herbicide ayant une activité inhibitrice de la synthèse d'acides gras sur le champ dans lequel est cultivé une variété de riz résistante à un herbicide. Dans la mesure où le riz produit en application de la présente invention est résistant à cette sorte d'herbicide, il ne pâtira pas de troubles de nature chimique occasionnés par cet herbicide et il sera possible de juguler les mauvaises herbes des graminées de façon sélective.
PCT/JP1994/001757 1994-10-19 1994-10-19 Riz resistant aux herbicides et procede d'amelioration genetique afferent Ceased WO1996012402A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/JP1994/001757 WO1996012402A1 (fr) 1994-10-19 1994-10-19 Riz resistant aux herbicides et procede d'amelioration genetique afferent
AU79488/94A AU7948894A (en) 1994-10-19 1994-10-19 Herbicide-resistant rice and method of breeding the same

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP1994/001757 WO1996012402A1 (fr) 1994-10-19 1994-10-19 Riz resistant aux herbicides et procede d'amelioration genetique afferent

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60210929A (ja) * 1984-03-06 1985-10-23 モレキユラー・ジネテイクス・インコーポレーテツド 耐除草剤性植物

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS60210929A (ja) * 1984-03-06 1985-10-23 モレキユラー・ジネテイクス・インコーポレーテツド 耐除草剤性植物

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KODANSHA, 1 April 1992, BY YASUYUKI YAMADA, "Plant Particle, Cell Engineering Manual", 1st, p. 181-189. *
PHYSIOLOGIA PLANTARUM, Vol. 91, (25.07.94), FRANCOIS J. TARDIF and STEPHEN B. POWLES, p. 488-494. *
THEORETICAL AND APPLIED GENETICS, Vol. 63, (1982), B.R. THOMAS and D. PRATT, p. 169-176. *

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