WO2011033658A1 - PROCÉDÉ POUR CULTIVER UNE PLANTE DE LA FAMILLE DU POIS À L'AIDE DE RAYONS γ - Google Patents

PROCÉDÉ POUR CULTIVER UNE PLANTE DE LA FAMILLE DU POIS À L'AIDE DE RAYONS γ Download PDF

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Publication number
WO2011033658A1
WO2011033658A1 PCT/JP2009/066402 JP2009066402W WO2011033658A1 WO 2011033658 A1 WO2011033658 A1 WO 2011033658A1 JP 2009066402 W JP2009066402 W JP 2009066402W WO 2011033658 A1 WO2011033658 A1 WO 2011033658A1
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day
dose rate
irradiation
group
rays
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PCT/JP2009/066402
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English (en)
Japanese (ja)
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徹夫 中島
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独立行政法人 放射線医学総合研究所
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Priority to PCT/JP2009/066402 priority Critical patent/WO2011033658A1/fr
Priority to JP2010209714A priority patent/JP5483108B2/ja
Publication of WO2011033658A1 publication Critical patent/WO2011033658A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general

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  • the present invention relates to a method for cultivating legumes using ⁇ -rays, and in particular, cultivating leguminous plants including a step of irradiating a plant body with ⁇ -rays at a dose rate in a specific range after the seedling has passed. Regarding the method.
  • Legumes such as soybeans are one of the main grains of civilization and are produced about 200 million tons worldwide.
  • Leguminous plants such as soybean are widely used not only for food but also as vegetable oil and livestock feed, and in part, they are also being used as bioenergy, and their production is increasing year by year.
  • One countermeasure against such an increase in demand is to produce soybean with high productivity by genetic engineering techniques.
  • plants created by such genetic engineering techniques have become a global problem because they may adversely affect the conservation and sustainable use of biological diversity.
  • Non-patent Document 1 reports on the hormesis effect of radiation such as ⁇ rays.
  • these reports can be understood as suggesting that effects such as the promotion of growth can be obtained by irradiating a prescribed amount of radiation in the state of seeds, tuberous roots, and seedlings.
  • the dose rate and the irradiation period are not mentioned, and the effects obtained by the irradiation are various. For this reason, there is much debate about the validity of the data.
  • Non-Patent Document 2 Recently, there is a report that growth and wrinkle are reduced from about 0.4 Gy by rapid irradiation of young seedlings after germination, but there is no change in the size of wrinkles.
  • Non-Patent Document 3 There is a report that, in the contaminated area around Chernobyl, the beans grown in the presence of radionuclides in the soil are small in harvested beans.
  • the present invention is based on the discovery of conditions for increasing the production of legumes that could not be predicted at all from the technical knowledge at the time of filing of the present application. In other words, not only the total dose but also the irradiation time and dose rate were examined in detail. After the seedling time passed, the total dose was 0.5-10.0 Gy and 0.04-0.4 Gy / day. When plants were irradiated with ⁇ -rays at a dose rate in the range, the seeds were found to be larger and the average weight of the pods increased than in the case of non-irradiation, and doses in the narrower range of 0.05 to 0.08 Gy / day were found. When the plant body was irradiated with ⁇ rays at a rate, the number of pods per individual also increased, and the weight of cocoons harvested from one individual was found to increase significantly, leading to the completion of the present invention. .
  • the plant body is given a total dose of 0.5-10.0 Gy, a dose rate of 0.04-0.4 Gy / day
  • a leguminous plant cultivation method a legume plant grown by this method, and a seed obtained therefrom, characterized by irradiating with ⁇ rays preferably at a dose rate of 0.05 to 0.08 Gy / day Is.
  • seedling refers to a plant that has germinated from the seed of a seed plant and has a cotyledon remaining, and “after the seedling has passed” means that the plantlet has lost its cotyledon and is normally It refers to the season with leaves.
  • flower bud means a growth point that has undergone vegetative growth differentiated into a growth point that undergoes reproductive growth, that is, a flower or inflorescence primordium. Means that the plant has grown and is close to flowering.
  • Non-Patent Document 6 states that soybean flower bud differentiation is performed around 20 days before flowering). At the practical level, you can follow it.
  • the formation time and flowering time are different.
  • the stage before flower bud formation When viewed at an individual level, the stage before flower bud formation, the stage of flower bud formation, two or more stages of formation and flowering can coexist. Accordingly, when the terms “before flowering”, “before bud formation”, and “flower bud formation” are used herein, unless stated otherwise, if at least one flower is before flowering, “before flowering” "If at least one bud is formed, then" before bud is formed ", and if at least one flower bud is formed, it corresponds to" flower bud formation ".
  • the cocoon is enriched means a state in which the cocoon is sufficiently swollen by the seeds therein.
  • until the eyelids are enriched means until at least one eyelid is enriched, but in terms of further ensuring the effect of the method of the present invention, the irradiation of ⁇ rays is almost (80 % Or more, preferably 90% or more) is preferably carried out until the soot is enriched.
  • seed includes not only fully-ripened seeds but also immature seeds. Such immature seeds are edible in Japan and the like.
  • the dose rate and total dose are values calculated from the absorbed air dose (Gy) measured for the ability to ionize air directly or indirectly by gamma rays emitted from a radiation source (radioactive material). means.
  • the dose of gamma rays was defined by the source nuclide, radioactivity, distance from the source, etc., and was measured with a thermoluminescence dosimeter (TLD).
  • TLD thermoluminescence dosimeter
  • the cultivation method of the present invention larger seeds can be harvested than in the normal cultivation method.
  • the number of pods per individual in addition to being able to harvest large seeds, the number of pods per individual can be increased, and the yield of seeds per individual can be increased. .
  • the present invention eliminates such a barrier and enables an increase in production by a simple method.
  • FIG. 2 is a graph showing the number of pupae per individual in each group of tests conducted in 2008.
  • FIG. 4 is a graph showing the average value of the weight per cocoon for each group of tests conducted in 2009.
  • FIG. 5 is a graph showing the number of pupae per individual in each group of tests conducted in 2009.
  • FIG. 7 is a graph showing the number of pupae per individual in each group of tests conducted in both 2008 and 2009.
  • FIG. 9 is a copy of a photograph showing the migration pattern of two-dimensional electrophoresis in which the protein expression pattern in seeds harvested in 2008 was analyzed.
  • FIG. 9A shows the migration pattern of seeds harvested from the control group
  • FIG. 9B shows the migration pattern of seeds harvested from the 0.2 Gy / day dose rate group.
  • the plant body is irradiated with ⁇ rays after the seedling time has elapsed.
  • Patent Document 1 many attempts have been made to irradiate various radiations at the seed, tuberous root, and seedling stages in various plants including legumes.
  • Patent Document 1 ⁇ -rays after the seedling stage in which differentiation has progressed have been made.
  • irradiation of the plant caused an increase in legume seed production.
  • the feature of the present invention that the plant body is irradiated with ⁇ -rays at the stage after seedling is inventive with respect to the technical level at the time of filing.
  • Non-patent Documents 4 and 5 Non-patent Documents 4 and 5
  • ⁇ -irradiation after the flowering period is related to seed development. It is expected that it will have some effect on the expression of genes. Therefore, from the viewpoint of increasing the seed size, irradiation with ⁇ rays may be started after flower formation and before flowering.
  • the period of flower bud differentiation is considered to work in any of the period of flower bud differentiation, the period of wrinkle formation, or the period of wrinkle formation.
  • a large number of wrinkles are formed (eg, 50% or more of the total number of wrinkles expected per individual), more preferably until wrinkles are formed.
  • seeds will be further enlarged by ⁇ rays during the period until the pods are enriched, and irradiation of ⁇ rays should be carried out at least until the seeds in the pods begin to expand. It is more preferable to carry out until several to 10 ridges start to swell, more preferably until at least one cocoon is filled, until 80% or more of all cocoons are filled. It will be particularly preferred to do so.
  • a dose rate of 0.04 to 0.4 Gy / day preferably 0.04 to 0.4.
  • Gamma rays are irradiated at 3 Gy / day, more preferably at a dose rate of 0.05 to 0.25 Gy / day.
  • test results on the hormesis effect were reported by irradiating the seeds of various plants including legumes with gamma rays, but none of the detailed conditions other than the dose were studied, and the effects There were also various. Therefore, ⁇ -ray irradiation at a low dose rate in a specific range proposed by the present application presents for the first time a clear condition for increasing the legume seed size (1.3 times or more by dry weight). In this respect, the present invention has inventiveness with respect to the technical level at the time of filing.
  • the present invention in a particularly preferred embodiment, presents conditions for increasing the number of pods to be formed and increasing the yield of blisters per individual. That is, by irradiating the plant body with ⁇ rays at a dose rate in a narrower range of 0.05 to 0.08 Gy / day, preferably 0.05 to 0.07 Gy / day, for the predetermined period, The number of pods formed can also be increased and the yield per individual can be increased. In such an extremely narrow range, the phenomenon that seeds grow and the number of pods formed increases, which has never been known so far, and it is the first time that there is a clear condition that makes it possible to increase the production of legume seeds. Present.
  • Irradiation with ⁇ -rays may be performed continuously or intermittently, but it is preferable to perform ⁇ -ray irradiation as continuously as possible. Moreover, when irradiating at intervals, the irradiation can be performed several days apart, but it is desirable that the irradiation be performed for two days or less.
  • the total irradiation date is preferably 10 days or more, more preferably 15 days or more, still more preferably 20 days or more, and particularly preferably 25 days or more.
  • the total dose of ⁇ rays depends on the total irradiation day, but is preferably about 0.5 to 10.0 Gy, more preferably about 0.8 to 6.0 Gy. It is particularly preferably about 0.0 to 2.0 Gy.
  • the total dose is preferably 0.5 to 10.0 Gy, and the dose rate is In the case of 0.05 to 0.25 Gy / day, the total dose is preferably about 0.8 to 6.0 Gy, more preferably about 1.0 to 5.0 Gy, and the dose rate is When it is 0.05 to 0.08 Gy / day, the total dose is preferably about 1.0 to 2.0 Gy, and when the dose rate is 0.06 to 0.08 Gy / day, The total dose is preferably about 1.3 to 2.0 Gy.
  • the total dose of 10 Gy or less is used for potato germination prevention and sterilization treatment (germination suppression is about 0.05-0.15 kGy, sterilization of spoilage pathogens is 1-7 kGy, seasoning or food
  • the sterilization of the material is 10-50 kGy), which is a very low dose range, and the present invention seems to have a low barrier to its practical use.
  • the irradiation time per day is preferably determined according to the dose rate to be used, from the viewpoint of the effectiveness of irradiation and avoiding a high level of irradiation per unit time.
  • the dose rate to be used from the viewpoint of the effectiveness of irradiation and avoiding a high level of irradiation per unit time.
  • ⁇ -ray radiation source may for example 60Co, 192Ir, 169Yb, 170Tm, 75Se, 124Sb, be mentioned 153Gd or 137 Cs.
  • leguminous plants include, for example, plants such as soybean genus, genus pea, and pea genus. .
  • a plant of the genus Soybean which is considered to have a certain resistance to radiation, is preferable.
  • Japan there is an eating habit of using immature seeds such as soybeans, green beans, cowpeas, peas, peanuts, wisteria bean, jujube, winged bean, 16 cowpeas, etc. and young pods as they are. Being eaten generally.
  • soybean varieties developed to eat as “edamame” include early varieties such as Sachio Okuhara, Swan, Sayamusume, Beer Friend, Shirako, etc .; Fuki, Ryokan, Mikawajima, Nishikiaki, Mid- and early-birth varieties such as evening coolness, hot spring girls, Fukushiko, etc .; late-generation varieties such as Akiyoshi and Toyoshirome; Black bean varieties; Dadacha beans, Kurosaki tea beans, Takihime, Fukunari etc.
  • ripe seeds are used as processed foods such as tofu, miso and natto, or as raw materials for vegetable oils in Japan, and are widely used worldwide as raw materials for vegetable oils and livestock feed. Attempts have also been made to use it as bioenergy.
  • the present invention is considered to contribute to the utilization of such leguminous plants, and is considered to be effective when applied to soybean family plants, particularly early soybean species (note that genes related to early / late nature are not considered). (See Patent Documents 7 and 8).
  • the leguminous plant obtained by the method of the present invention is different from the same non-irradiated plant in terms of phenotypes such as an increase in seeds and an increase in the number of pods, as demonstrated in Examples described later.
  • phenotypes such as an increase in seeds and an increase in the number of pods, as demonstrated in Examples described later.
  • protein expression there is a difference in protein expression with respect to the same non-irradiated plant.
  • such differences are not due to differences in the genes themselves, but are thought to be because the expression of the genes was affected by gamma rays.
  • the present invention is also directed to legumes having an unprecedented character cultivated by the method described in detail above, and seeds obtained from such legumes.
  • Such legumes and seeds provide a new means of efficiently providing food or energy resources without concern about the adverse effects on biodiversity conservation and sustainable use by transgenic plants. obtain.
  • pots with 1 to 3 seedlings per pot (pot) are assigned to the control and three irradiation groups, each with about 10 seedlings, and the length distribution in each group Sorted out to be roughly aligned.
  • Installed in the field located in Hitachiomiya City, Ibaraki Prefecture).
  • control group was placed outside the gamma ray field as a pot, and was cultivated there until July 31, 2008. Another control group was placed as a pot in Chiba, away from the gamma ray field, where it was grown until July 31, 2008. Irrigation of each group was performed 2-3 times / week throughout the cultivation period. On the day following the end of the irradiation period (July 31, 2008, 79 days after seed sowing), the straw was harvested from the plant. In addition, individuals that died significantly due to insect damage or the like were excluded.
  • Test results 3-1 Number of pods and weight of seeds per pod The number of pods obtained and the weight of seeds per pod were measured for each group. The test results are as shown in FIGS.
  • FIG. 1 shows the average weight per cocoon of each group of tests conducted in 2008
  • FIG. 2 shows the number of cocoons per individual in each group of tests conducted in 2008
  • FIG. The total weight of FIG. 4 shows the average weight per cocoon of each group in the test conducted in 2009
  • FIG. 6 shows the average weight per cocoon of each group of tests conducted in both years
  • FIG. 7 shows the number of cocoons per individual in each group of tests conducted in both years.
  • FIG. 8 is a photograph showing beans harvested from each group of tests conducted in 2008.
  • the average weight per cage is 0.06Gy / day dose rate (total dose 1.5Gy) group, and 0.2Gy / day dose rate (total dose 5Gy) There was an increase in the group.
  • the dry weight of the seeds of the group was 1.3 and 1.44 times the weight, respectively.
  • the number of wrinkles per individual increased significantly compared to the control group in the 0.06 Gy / day dose rate (total dose 1.5 Gy) group.
  • the Gy / day dose rate (total dose 0.5 Gy) group and the 0.2 Gy / day dose rate (total dose 5 Gy) group were not significantly different from the control group.
  • the total weight of wrinkles per individual increased significantly compared to the control group.
  • the obtained pellet is placed in a buffer (100 mM Tris-HCl pH 8.5, 4% SDS, 2% mercaptoethanol, 20% glycerol, 2 mM PMSF) for protein collection of the granular fraction, and placed in boiling water for 3 minutes. It was. After cooling, the mixture was centrifuged (18800 g, 10 min, room temperature), the supernatant was removed, passed through a 0.22 ⁇ m PVDF membrane filter, and subjected to acetone extraction (in 20 volumes of acetone, ⁇ 20 ° C., overnight).
  • a buffer 100 mM Tris-HCl pH 8.5, 4% SDS, 2% mercaptoethanol, 20% glycerol, 2 mM PMSF
  • the sample subjected to acetone extraction is centrifuged (18800g, 10min, 4 ° C), the resulting pellet is re-stirred with 80% acetone, the lysate is placed at -20 ° C for 90min, and further centrifuged (18800g, 10min, 4 ° C).
  • the obtained pellet was dissolved in a lysis buffer (5M Urea, 2M thiourea, 2% CHAPS, 2% SB3-10, 1% DTT).
  • the lysate was subjected to ultracentrifugation (100,000 g, + 20 ° C., 1 h), and the resulting supernatant was used for electrophoresis as a granular fraction sample.
  • large bean can be harvested from existing legumes without using genetic engineering techniques, and the yield of seed per individual can be greatly increased.
  • Leguminous plants have been studied for use as energy resources and as pharmaceutical production plants in addition to their use as food, and their use in a wide range of fields is conceivable. Also, in space, radiation always exists, and it seems possible to apply the present invention by controlling it well.

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  • Life Sciences & Earth Sciences (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Botany (AREA)
  • Ecology (AREA)
  • Forests & Forestry (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Pretreatment Of Seeds And Plants (AREA)

Abstract

L'invention porte sur un procédé pour cultiver une plante de la famille du pois qui permet la récolte de graines plus grandes que celles récoltées par des procédés de culture classique. Le procédé comprend l'irradiation de la plante qui est dans un stade post-ensemencement par un rayon γ à un débit de dose de 0,04 à 0,4 Gy/jour et à la dose totale de 0,5 à 10,0 Gy.
PCT/JP2009/066402 2009-09-18 2009-09-18 PROCÉDÉ POUR CULTIVER UNE PLANTE DE LA FAMILLE DU POIS À L'AIDE DE RAYONS γ WO2011033658A1 (fr)

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PCT/JP2009/066402 WO2011033658A1 (fr) 2009-09-18 2009-09-18 PROCÉDÉ POUR CULTIVER UNE PLANTE DE LA FAMILLE DU POIS À L'AIDE DE RAYONS γ
JP2010209714A JP5483108B2 (ja) 2009-09-18 2010-09-17 γ線を利用したマメ科植物の栽培方法

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PCT/JP2009/066402 WO2011033658A1 (fr) 2009-09-18 2009-09-18 PROCÉDÉ POUR CULTIVER UNE PLANTE DE LA FAMILLE DU POIS À L'AIDE DE RAYONS γ

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103907517A (zh) * 2013-01-05 2014-07-09 赵百华 一种豌豆苗育培技术
CN105284353A (zh) * 2015-10-26 2016-02-03 和县德生农业发展有限公司 一种四季豆的春早熟栽培方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000316383A (ja) * 1999-05-07 2000-11-21 Japan Atom Energy Res Inst 低エネルギー電子線による植物の生育促進法
JP2004222551A (ja) * 2003-01-21 2004-08-12 Japan Atom Energy Res Inst 放射線処理を利用した植物への窒素固定菌着生促進法
JP2008022814A (ja) * 2006-07-25 2008-02-07 Gyoseiin Genshino Iinkai Kakuno Kenkyusho 輻射照射による海藻激成方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000316383A (ja) * 1999-05-07 2000-11-21 Japan Atom Energy Res Inst 低エネルギー電子線による植物の生育促進法
JP2004222551A (ja) * 2003-01-21 2004-08-12 Japan Atom Energy Res Inst 放射線処理を利用した植物への窒素固定菌着生促進法
JP2008022814A (ja) * 2006-07-25 2008-02-07 Gyoseiin Genshino Iinkai Kakuno Kenkyusho 輻射照射による海藻激成方法

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103907517A (zh) * 2013-01-05 2014-07-09 赵百华 一种豌豆苗育培技术
CN105284353A (zh) * 2015-10-26 2016-02-03 和县德生农业发展有限公司 一种四季豆的春早熟栽培方法

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