WO2015102130A1 - Novel brassica napus variety - Google Patents

Novel brassica napus variety Download PDF

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WO2015102130A1
WO2015102130A1 PCT/KR2013/012432 KR2013012432W WO2015102130A1 WO 2015102130 A1 WO2015102130 A1 WO 2015102130A1 KR 2013012432 W KR2013012432 W KR 2013012432W WO 2015102130 A1 WO2015102130 A1 WO 2015102130A1
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rapeseed
new
varieties
cabbage
present
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PCT/KR2013/012432
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French (fr)
Korean (ko)
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김신제
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주식회사 에프앤피
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Priority to PCT/KR2013/012432 priority Critical patent/WO2015102130A1/en
Publication of WO2015102130A1 publication Critical patent/WO2015102130A1/en

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    • 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
    • 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
    • A01H6/20—Brassicaceae, e.g. canola, broccoli or rucola

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  • the present invention relates to a new rapeseed variety, and more particularly to a new rapeseed variety by hybridization of red cabbage and Chinese cabbage.
  • rapeseed B. napus
  • cabbage B. rapa
  • cabbage B. oleracea
  • the origin of the rapeseed is not clear, but after natural hybridization between two closely-diploid species, B. oleracea (CC genome) and B. rapa (AA genome), the hybrid chromosomes were accidentally doubled. Can be estimated.
  • Rape seed contains various fatty acids, most of which are unsaturated fatty acids.
  • Fatty acids in oilseeds can be derived from palmitic acid (C16: 0), stearic acid (C18: 0), and oleic acid (C18: 1), linoleic acid (C18: 2), linoleninc acid (C18: 3), and erucic acid (C22: 1).
  • These various types of fatty acids are different in their physicochemical properties, so the range of application depends on the fatty acid composition of the oil extracted from plants.
  • An object of the present invention is to provide a new varieties of rapeseed with an increased content of erucic acid.
  • these problems are exemplary, and the scope of the present invention is not limited thereby.
  • a new rapeseed variety 'Daeae' is obtained by hybridization between red cabbage ( Brassica oleracea ) and Chinese cabbage ( Brassica rapa), and has the following botanical characteristics.
  • the multi-colored (a) leaf vein and leaf disease is pink; (b) the weight of the grains of 5.0g or more; (c) the content of erucic acid of 50% or more; And (d) a botanical characteristic of palmitoleic acid content of 0.1 ⁇ 1%.
  • the erucic acid content is 51% or more, 52% or more, 53% or more, 54% or more, 55% or more, 56% or more, 57% or more, 58% or more, 59% or more, At least 60%, at least 61%, at least 62% or at least 63%.
  • a cosmetic composition comprising the above-mentioned rapeseed new varieties extract as an active ingredient.
  • the new rapeseed varieties have a higher content of erucic acid than conventional rapeseed, since erusic acid is a fatty acid used as a raw material for cosmetics, the new rapeseed varieties according to an embodiment of the present invention can be used for preparing a cosmetic composition.
  • the erucic acid content 51% or more, 52% or more, 53% or more, 54% or more, 55% or more, 56% or more, 57% or more, 58% or more, 59% or more, 60 At least%, at least 61%, at least 62% or at least 63%.
  • a rapeseed plant hybridized and bred using a new rapeseed variety 'Dae-ae' according to an embodiment of the present invention as a pollen, a parent or a blotter is provided.
  • the rapeseed new varieties are as described above.
  • the advantageous property may be dry resistance, cold resistance, high probability, heat resistance, disease resistance or insect resistance.
  • 1 is a diagram showing the hybridization relationship diagram of the new varieties of rape seed according to an embodiment of the present invention.
  • FIG. 2 is a diagram illustrating seed deletions of plant 2 (A) having high self-incompatibility and plant 3 (B) having high self-pollinity among hybrid F 1 progeny.
  • Figure 3 is a view showing the growth of seedlings of the new varieties rapeseed rape according to an embodiment of the present invention.
  • Figure 4 is a diagram (right panel) to observe the growth of the rapeseed new varieties of rapeseed (left panel) and the leaf veins and leaf diseases of the multi-colored plants according to an embodiment of the present invention.
  • FIG. 5 is a diagram comparing the colors of the leaf veins and leaf disease of Daae Rapeseed (left panel) and Halla Rapeseed (right panel).
  • FIG. 6 is a diagram comparing the shape of the leaves of Daae Rapeseed and Halla Rapeseed.
  • Figure 7 is a view of the analysis of the karyotype of red cabbage (top), Chinese cabbage (bottom) and Daae rapeseed (bottom) of the present invention according to an embodiment of the present invention.
  • FIG. 8 is a diagram showing the results of molecular biology analysis using SNP (A) and RAPD (B) of Daae Rapeseed, red cabbage and Chinese cabbage according to an embodiment of the present invention:
  • M 1 kb ladder marker
  • FIG. 9 is a diagram showing RAPD analysis of various kinds of rapeseed varieties using primers for amplifying a rapeseed specific band:
  • M 1 kb ladder marker
  • Fertilized pods are incubated in immature embryos by artificially pollination with red cabbage ( B. oleracea ) as a model and artificial pollination with B. rapa ( B. rapa ). After 14-21 days from the mating day, fertilized pods were harvested, and embryos were extracted from the embryos and placed in MS medium. Incubation conditions are maintained under 25 ⁇ 1 °C, luminous intensity 40 ⁇ mol ⁇ m -2 ⁇ s -1 , photoperiod 16 hours. Plants arising from immature embryos were grown on larger culture bottles. After that, it is formalized and nurtured through the process of purification and seedling.
  • DNA fragments that are amplified as a kind of PCR reaction are distinguished from conventional PCR in that they are random.
  • RAPD is performed in a PCR reaction with a large genomic DNA as a template after generating some arbitrary short primers (8-12 nucleotides), and a semi-unique profile is obtained from the RAPD reaction by analyzing the resulting pattern. It is not known exactly where the primer is located, but the DNA sequence information is unnecessary because it binds to any of the DNA sequences of the target gene. Recently, RAPD has been used to identify and track the lineage of various plant and animal species.
  • RAPD analysis of genomic DNA was performed by the following method:
  • the PCR reaction composition was mixed with 50 ng of template DNA, 10 ⁇ PCR buffer (containing 1.5 mM MgCl 2 ), 200 ⁇ M of dNTPs, 2 units of taq DNA polymerase, 20 ⁇ M of primer for RAPD, and 20 ⁇ l of total volume with distilled water (DW). Adjust to to form a reaction solution.
  • the PCR reaction solution was placed in a PCR tube and denatured at 95 ° C. for 3 minutes, followed by 4 cycles of 1 minute at 94 ° C., 1 minute at 35 ° C., and 1 minute 30 seconds at 72 ° C., followed by 94 cycles.
  • 30 seconds at 40 °C, 20 seconds at 40 °C, 1 minute at 72 °C 1 cycle was repeated a total of 30 times and then synthesized by reacting for 7 minutes at 72 °C.
  • FIG. 1 is a diagram showing the hybridization relationship diagram of the new varieties of rape seed according to an embodiment of the present invention.
  • the present inventors used pollen by removing pollen from the buds of red cabbage ( B. oleracea ) and conducted cross-border artificial crosses using cabbage ( B. rapa ) as a copy. .
  • B. oleracea the buds of red cabbage
  • B. rapa the leaves of red cabbage
  • three plants were able to be secured.
  • Plant 1 died during wintering in order from the left of F 2 in FIG. 1, and plant number 3 was difficult due to high incompatibility. Because of its high self-pollination rate, it was possible to secure a large amount of future seed, which was named 'Dae-ae'.
  • FIG. 2 is a diagram illustrating seed deletions of plant 2 (A) having high self-incompatibility and plant 3 (B) having high self-pollinity among hybrid F 1 progeny. As shown in FIG. 1, the result of artificial breeding between red cabbage and Chinese cabbage is a photograph of seed loss of the obtained plant.
  • Figure 3 is a view showing the growth of seedlings of the new varieties rapeseed rape according to an embodiment of the present invention. As a result of observing the cultivar characteristics of the cross-hybrid multicolored hybrid according to an embodiment of the present invention, it was observed that the back of the leaf was pink in the seedling stage.
  • Figure 4 is a diagram (right panel) to observe the growth of the rapeseed new varieties Daaeae (left panel) and the color of the leaf veins and leaf disease of the Daae plant according to an embodiment of the present invention.
  • the leaf veins and folies have a pink color from the early growth to the flowering period.
  • FIG. 5 and 6 are diagrams comparing the leaf vein, leaf color and leaf shape of Daae Rapeseed and Halla Rapeseed.
  • Figure 7 is a view of the analysis of the karyotype of red cabbage (top), Chinese cabbage (hang) and rapeseed new varieties Daae (bottom) according to an embodiment of the present invention.
  • Daae Rapeseed according to an embodiment of the present invention was used as a model by removing the pollen from the buds of red cabbage ( B. oleracea ) and cabbage ( B. rapa ) was used as a copy.
  • B. oleracea red cabbage
  • cabbage B. rapa
  • FIG. 8 is a view showing the results of molecular biological analysis using SNP (top) and RAPD (bottom) of rapeseed new varieties Daaeae, red cabbage and Chinese cabbage according to an embodiment of the present invention.
  • SNP top
  • RAPD High Resolution Melting
  • FIG. 9 is a diagram illustrating RAPD analysis of various kinds of rapeseed varieties using primers for amplifying a rapeseed specific band.
  • the inventors have the interspecies artificial breeding was carried out ever cabbage (B.oleracea) and cabbage (B.rapa) to the seed size of a large development of rapeseed as ever cabbage.
  • the pollen was removed from the buds of red cabbage and used as a mother's head.
  • the new varieties Daae Rapeseed is characterized in that the back of the leaves pink color in the seedling stage (seedling stage) (Fig. 3), the leaf veins and foliar pink color from the early growth to flowering period (Fig. 4).
  • Fig. 3 seedling stage
  • Fig. 4 leaf veins
  • Fig. 4 foliar pink color from the early growth to flowering period
  • the multi-arachnoid fluid adult according to an embodiment of the present invention is thick and strong so that the plant posture is straight and the degree of deflection is small. It is confirmed that the seeds produced during the fertile breeding season are slightly larger than the general oilseed rape because the growth period is longer than 80 days.
  • Fatty acid content of the new rapeseed varieties 'Daeae' according to an embodiment of the present invention was analyzed, and the fatty acid contents of the rapeseed, mustard, cabbage and cabbage varieties described in Table 2 were analyzed for comparison.
  • Fatty acid pretreatment was performed by the same pretreatment method of 'fatty acid second method and fat analysis method' to perform gas chromatography analysis.
  • Fatty acid analysis was performed using gas chromatography (Agilent 7890 GC, Agilent Technologies), HP-INNOWAX (30m X 0.320 mm X 0.25 ⁇ m, Agilent Technologies), the detector was a flame ionization detector (flame ionization detector).
  • the inlet temperature was 225 ° C.
  • the detector temperature was 285 ° C.
  • the oven temperature was maintained at 100 ° C. for 4 minutes and then raised to 240 ° C. at 3 ° C. per minute for 15 minutes.
  • the detector temperature was 285 ° C., and nitrogen was used as a carrier gas.
  • the new rapeseed variety of the rapeseed contains 12.9% oleic acid and 61.5% erucic acid, and 0.62% of palmitoleic acid, which is small but not found in other plants. I could see that. Palmitoleic acid is known to be effective in preventing diabetes as unsaturated fatty acid, and since erucic acid is a fatty acid that is used as a raw material for cosmetics, it may be used in cosmetic production using a new rapeseed according to one embodiment of the present invention. 3).
  • Chromosome spectroscopy to determine whether the number of chromosome doubled was as follows. Take a root tip of 5 mm, place it in a sample bottle filled with 3 ml of cold distilled water, return it to an ice box containing crushed ice, and transport it indoors. Cut the root end about 2 mm and cut the Eppendorf tube. (eppendorf tube). The prepared sample was placed in 0.8% bromonaphthalene and reacted at room temperature for 40 minutes. 0.8% bromonaphthalin was used in about 20-30 root tips cut
  • HRM High Resolution Melting
  • the fluorescence signal was measured for 1 minute at 95 ° C and 1 minute at 40 ° C, then increased 0.02 ° C per second from 65 ° C to 95 ° C, and measured 25 times every 1 ° C. Melting curve analysis was performed using the manufacturer's operating software. Was performed.
  • Primer for amplification of the site including the base 99bp of FPBPN0011, a molecular marker that can distinguish red cabbage, Chinese cabbage, and rapeseed, and when amplifying the molecular marker FPBPN0011, it is interchelated between the double strands of genes to visually indicate the amount of gene amplification Labeling material (SYBR) was used.
  • RAPD of genomic DNA of red cabbage, Chinese cabbage and Daae rape was performed for molecular genetic analysis of the new rape variety of the present invention, Daae, red cabbage and cabbage.
  • the primer was used in the kit sold by Operon, and the primer information used is shown in Table 5.
  • RAPD Molecular biological analysis using RAPD was confirmed as a hybrid hybrid to amplify specific bands of red cabbage and Chinese cabbage (see FIG. 8B).
  • the bands amplified using the double B01 primer it was possible to confirm the multispecific rapeseed specific band of about 980 bp.
  • the B01 primer refers to the OPB01 primer described in Table 1 below, and A03 to B12 described in FIGS. 8 and 9 mean OPA03 to OPB-12 primers.
  • compositions of the present invention are illustrated below.
  • a cosmetic for skin whitening containing a new breed rapeseed Daae extract was prepared as an active ingredient.
  • a flexible lotion containing a new variety rapeseed Daae extract was prepared as shown in Table 6 below.
  • Nutrition cream containing a new variety of rapeseed Daae extract was prepared as shown in the following [Table 7].
  • the new rapeseed varieties 'Daeae' according to an embodiment of the present invention have a high content of erucic acid used as a cosmetic raw material and the size of cloth neutral is larger than that of conventional rapeseed varieties, which is advantageous for the cosmetic industry as well as biodiesel production. It can be used as a copy or parent for the production of varieties.
  • SEQ ID NOs: 1 and 2 are nucleic acid sequences of forward and reverse primers for amplification of FPBPN0011 markers used for high resolution fusion analysis of genomic DNA of a novel rapeseed variety 'Dae' according to one embodiment of the present invention.
  • SEQ ID NOs: 3 to 42 are nucleic acid sequences of random primers used for RAPD analysis of a novel rapeseed variety 'Daeae' according to one embodiment of the present invention.

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Abstract

The present invention provides a novel Brassica napus variety "Daae" which has a high fatty acid content, is obtained by interspecies crossing between Brassica oleracea with an increased seed size and Brassica rapa, and retains the following botanical characteristics.

Description

신규 유채 품종New Rapeseed Varieties
본 발명은 신규 유채 품종에 관한 것으로서, 더 상세하게는 적양배추와 배추의 교잡에 의한 신규 유채 품종에 관한 것이다.The present invention relates to a new rapeseed variety, and more particularly to a new rapeseed variety by hybridization of red cabbage and Chinese cabbage.
배추속에 속하는 작물들의 염색체 구조 및 진화 과정은 우장춘 박사의 ‘우의 삼각형(triangle of U)' 모델로 설명되는데 이에 따르면 유채(B. napus)는 배추(B. rapa)와 양배추(B. oleracea)의 자연교잡에 의해 발생한 것으로 알려져 있다. 유채(Brassica napus)의 기원은 명확치 않으나 가까이에서 자라고 있던 두 개의 이배체 종인 B. oleracea(CC genome)와 B. rapa(AA genome)간의 자연적인 교잡이 이루어진 후, 우연하게 잡종의 염색체가 배가되었던 것으로 추정할 수 있다. 다른 게놈을 가진 두개의 종이 짝을 맺은 후, 잡종의 염색체 수가 두 배가 됨으로써 다배체(polyploidy)의 특별한 경우인 복이배체(amphidiploids)가 생산되었고, 이렇게 두 배가 된 염색체 설정은 안정된 감수 분열(meiosis)을 하여 새로운 다배체를 재생산할 수 있게 하였으며, 염색체가 두 배로 늘어나지 않는 교잡은 생식 능력이 없는 자손을 낳음으로써 도퇴되었을 것으로 추정된다. B. napus에 대한 세포유전학적 연구 결과, 상기 종이 AA와 CC 게놈을 모두 가지고 있으며, 단일 게놈 종(monogenomic species)인 B. oleracea(CC genome)와 B. rapa(AA genome)에서 추출된 이중 이배체라는 것을 보여주었다(Nagaharu U, Japan. J. Bot., 7: 389-452, 1935). 유채는 19쌍의 염색체(2n=38)를 갖고 있으며, 전체 염기쌍은 약 1,100 Mbp로 추정된다(Arumuganathan and Earle, Mol. Biol. Rep., 9: 208-218, 1991). The chromosome structure and evolution of the crops in the cabbage are explained by Dr. Jang Chun's model of 'triangle of U', which suggests that rapeseed ( B. napus ) is the cabbage ( B. rapa ) and cabbage ( B. oleracea ) It is known to be caused by natural hybridization. The origin of the rapeseed (Brassica napus) is not clear, but after natural hybridization between two closely-diploid species, B. oleracea (CC genome) and B. rapa (AA genome), the hybrid chromosomes were accidentally doubled. Can be estimated. After pairing two species with different genomes, the number of chromosomes in the hybrid was doubled to produce amphidiploids, a special case of polyploidy, and this doubled chromosome setup resulted in stable meiosis. It is possible to reproduce new polyploids, and hybridization, in which chromosomes do not double, is presumed to have been withdrawn by producing offspring with infertility. Cytogenetic studies of B. napus show that the species has both AA and CC genomes and is a double diploid extracted from B. oleracea (CC genome) and B. rapa (AA genome), which are single genomic species. (Nagaharu U, Japan. J. Bot ., 7: 389-452, 1935). Rapeseed has 19 pairs of chromosomes (2n = 38) and total base pair is estimated to be about 1,100 Mbp (Arumuganathan and Earle, Mol. Biol. Rep ., 9: 208-218, 1991).
종자 내의 배가 발달함에 따라 세포 내의 대사가 활발해지고, 전색소체(proplastid)가 엽록체로 분화되며 광합성과 함께 녹말(starch)의 합성이 이루어진다. 엽록체에서 생산된 녹말은 세포질에서의 트리아실글리세리드(triacylglyceride; TAG)와 지방산(fatty acid)의 합성에 사용되게 된다. 이렇게 만들어진 지질의 상당 부분은 유체(oil body)를 구성하여 차후 발아시 사용될 양분 축적의 용도로 사용된다. 유채의 경우 성숙된 종자의 오일 함량은 약 40%에 달하며, 이러한 특징으로 인해 식용은 물론 바이오 디젤의 원료로 많은 각광을 받고 있다. As the embryos in the seeds develop, metabolism in the cells becomes active, the prochromids are differentiated into chloroplasts, and starch is synthesized with photosynthesis. Starch produced from chloroplasts is used to synthesize triacylglycerides (TAG) and fatty acids in the cytoplasm. A large portion of the lipids thus produced constitutes an oil body and is used for the accumulation of nutrients for later germination. In the case of rapeseed, the oil content of mature seeds reaches about 40%, which makes it a popular food and biodiesel raw material.
유채의 종자는 다양한 종류의 지방산을 포함하고 있으며, 대부분이 불포화 지방산으로 존재한다. 유채를 비롯한 유지작물에 포함된 지방산은 탄화수소의 길이와 이중결합의 수에 따라 팔미트산(palmitic acid; C16:0), 스테아르산(stearic acid; C18:0), 올레산(oleic acid; C18:1), 리놀레산(linoleic acid; C18:2), 리놀렌산(linoleninc acid; C18:3), 그리고 에루스산(erucic acid; C22:1) 등으로 구분된다. 이러한 다양한 종류의 지방산들은 각각 물리화학적 특성이 다르므로 식물에서 추출한 오일의 지방산 조성에 따라 그 적용 범위가 달라진다. Rape seed contains various fatty acids, most of which are unsaturated fatty acids. Fatty acids in oilseeds, including rapeseeds, can be derived from palmitic acid (C16: 0), stearic acid (C18: 0), and oleic acid (C18: 1), linoleic acid (C18: 2), linoleninc acid (C18: 3), and erucic acid (C22: 1). These various types of fatty acids are different in their physicochemical properties, so the range of application depends on the fatty acid composition of the oil extracted from plants.
야생종을 포함한 과거에 주로 재배되던 유채 품종은 에루스산의 함량이 높으나, 사람이나 가축에서 지질축적증(lipidosis)을 유발하는 것으로 알려져 최근에 재배되는 식용 유채는 에루스산 함량이 매우 낮은 품종들이 대부분이다. 올레인산 함량이 많은 유채 품종은 바이오디젤 연료용으로 전세계적으로 확대 재배되고 있으며 에루스산의 함량이 많은 유채 품종은 화장품 원료용 바이오 원자재로써 활용되고 있다. Rapeseed varieties cultivated mainly in the past, including wild species, have a high content of erucic acid, but recently edible rapeseeds are known to cause lipid accumulation in humans and livestock. Mostly. Rapeseed varieties rich in oleic acid are being cultivated all over the world for biodiesel fuels, and rape varieties rich in erucic acid are used as bio raw materials for cosmetic raw materials.
본 발명은 에루스산의 함량이 증가된 유채 신품종을 제공하는 것을 목적으로 한다. 그러나 이러한 과제는 예시적인 것으로, 이에 의해 본 발명의 범위가 한정되는 것은 아니다.An object of the present invention is to provide a new varieties of rapeseed with an increased content of erucic acid. However, these problems are exemplary, and the scope of the present invention is not limited thereby.
본 발명의 일 관점에 따르면, 적양배추(Brassica oleracea)와 배추(Brassica rapa) 의 종간 교잡에 의해 수득되고, 하기의 식물학적 특성을 가지는 유채 신품종 '다애'가 제공된다.According to an aspect of the present invention, a new rapeseed variety 'Daeae' is obtained by hybridization between red cabbage ( Brassica oleracea ) and Chinese cabbage ( Brassica rapa), and has the following botanical characteristics.
상기 다애는(a) 엽맥 및 엽병이 분홍색;(b) 천립중이 5.0g 이상;(c) 에루스산 함량이 50% 이상; 및(d) 팔미톨레산 함량이 0.1~1%인 식물학적 특성을 가질 수 있다.The multi-colored (a) leaf vein and leaf disease is pink; (b) the weight of the grains of 5.0g or more; (c) the content of erucic acid of 50% or more; And (d) a botanical characteristic of palmitoleic acid content of 0.1 ~ 1%.
상기 유채 신품종 다애에 있어서, 상기 에루스산 함량은, 51% 이상, 52% 이상, 53% 이상, 54% 이상, 55% 이상, 56% 이상, 57% 이상, 58% 이상, 59% 이상, 60% 이상, 61% 이상, 62% 이상 또는 63% 이상일 수 있다.In the rapeseed new varieties, the erucic acid content is 51% or more, 52% or more, 53% or more, 54% or more, 55% or more, 56% or more, 57% or more, 58% or more, 59% or more, At least 60%, at least 61%, at least 62% or at least 63%.
본 발명의 다른 일 관점에 따르면, 상술한 유채 신품종 추출물을 유효성분으로 포함하는 화장료 조성물이 제공된다.According to another aspect of the present invention, there is provided a cosmetic composition comprising the above-mentioned rapeseed new varieties extract as an active ingredient.
상기 유채 신품종은 종래의 유채에 비하여 에루스산의 함량이 우수하므로, 에루스산은 화장품등의 원료로 사용되는 지방산이므로, 본 발명의 일 실시예에 따른 유채 신품종을 화장품 조성물 제조에 이용할 수 있다.Since the rapeseed new varieties have a higher content of erucic acid than conventional rapeseed, since erusic acid is a fatty acid used as a raw material for cosmetics, the new rapeseed varieties according to an embodiment of the present invention can be used for preparing a cosmetic composition.
본 발명의 다른 일 관점에 따르면, 적양배추(Brassica oleracea) 와 배추(Brassica rapa)의 종간 교잡에 의하여 수득되고, 하기의 식물학적 특성 가운데 하나 이상의 특성을 갖는 유채를 선별하는 단계를 포함하는, 유채 신품종의 육종방법이 제공된다: According to another aspect of the present invention, the rapeseed obtained by species hybridization of the red cabbage ( Brassica oleracea ) and Chinese cabbage ( Brassica rapa ), comprising the step of selecting a rapeseed having one or more of the following botanical characteristics New breeding methods are provided:
(a) 엽맥 및 엽병이 분홍색;(a) the veins and lobes are pink;
(b) 천립중이 5.0g 이상; (b) at least 5.0 g of natural grains;
(c) 에루스산 함량이 50% 이상; 및(c) an erucic acid content of at least 50%; And
(d) 팔미톨레산 함량이 0.1~1%.(d) Palmitoleic acid content 0.1-1%.
상기 육종방법에 있어서, 상기 에루스산 함량은, 51% 이상, 52% 이상, 53% 이상, 54% 이상, 55% 이상, 56% 이상, 57% 이상, 58% 이상, 59% 이상, 60% 이상, 61% 이상, 62% 이상 또는 63% 이상일 수 있다.In the breeding method, the erucic acid content, 51% or more, 52% or more, 53% or more, 54% or more, 55% or more, 56% or more, 57% or more, 58% or more, 59% or more, 60 At least%, at least 61%, at least 62% or at least 63%.
본 발명의 다른 일 관점에 따르면, 본 발명의 일 실시예에 따른 유채 신품종 '다애'를 꽃가루, 모본 또는 부본으로 사용하여 교잡육종한 유채 식물체가 제공된다.According to another aspect of the present invention, a rapeseed plant hybridized and bred using a new rapeseed variety 'Dae-ae' according to an embodiment of the present invention as a pollen, a parent or a blotter is provided.
상기 유채 신품종 다애는 상술한 바와 같다.The rapeseed new varieties are as described above.
본 발명의 다른 일 관점에 따르면, 상기 유채 신품종 '다애'를 꽃가루, 모본 또는 부본으로 하여 다른 유채 품종과 교잡하는 교잡단계; 및According to another aspect of the present invention, a hybridization step of hybridizing with other rapeseed varieties by using the new rapeseed varieties 'Daeae' as pollen, mother or book; And
상기 교잡에 의해 생성된 F1 중 천립중이 5.0 g 이상이고, 에루스 함량이 50% 이상이며, 상기 다른 유채 품종의 유리한 특성을 함께 가지는 F1 개체를 선별하는 단계를 포함하는 제1항 또는 제2항의 유채 신품종 '다애'를 이용한 유채 신품종의 제조방법이 제공된다.And the cheonripjung of the F 1 generated by the hybridization 5.0 g or more, the loose and the content is more than 50%, of claim 1 comprising the step of selecting the F 1 individual having with the advantageous properties of the other of rapeseed cultivars or A method for producing a new rapeseed variety using the new rapeseed varieties 'Dae' is provided.
상기 제조방법에 있어서, 상기 유리한 특성은 내건성, 내냉성, 다수확성, 내열성, 내병성 또는 내충성일 수 있다.In the above production method, the advantageous property may be dry resistance, cold resistance, high probability, heat resistance, disease resistance or insect resistance.
상기한 바와 같이 이루어진 본 발명의 일 실시예에 따르면, 종자의 크기 및 지방산의 함량이 증대된 유채 신품종을 구현할 수 있다. 물론 이러한 효과에 의해 본 발명의 범위가 한정되는 것은 아니다.According to one embodiment of the present invention made as described above, it is possible to implement a new type of rapeseed with increased seed size and fatty acid content. Of course, the scope of the present invention is not limited by these effects.
도 1은 본 발명의 일 실시예에 따른 유채 신품종 다애의 교잡관계도를 나타낸 도이다. 1 is a diagram showing the hybridization relationship diagram of the new varieties of rape seed according to an embodiment of the present invention.
도 2는 교잡종 F1 후대 중 자가불화합성이 높은 2번 식물체(A)와 자가수분율이 높은 3번 식물체(B)의 종자 결실을 관찰한 도이다. FIG. 2 is a diagram illustrating seed deletions of plant 2 (A) having high self-incompatibility and plant 3 (B) having high self-pollinity among hybrid F 1 progeny.
도 3은 본 발명의 일 실시예에 따른 유채 신품종 다애의 어린묘 육성 모습을 나타낸 도이다. Figure 3 is a view showing the growth of seedlings of the new varieties rapeseed rape according to an embodiment of the present invention.
도 4는 본 발명의 일 실시예에 따른 유채 신품종 다애의 육성모습(좌측 판넬)과 다애 식물체의 엽맥 및 엽병의 색을 관찰한 도(우측 판넬)이다.Figure 4 is a diagram (right panel) to observe the growth of the rapeseed new varieties of rapeseed (left panel) and the leaf veins and leaf diseases of the multi-colored plants according to an embodiment of the present invention.
도 5는 다애 유채(좌측 판넬)와 한라 유채(우측 판넬)의 엽맥 및 엽병의 색을 비교한 도이다.5 is a diagram comparing the colors of the leaf veins and leaf disease of Daae Rapeseed (left panel) and Halla Rapeseed (right panel).
도 6은 다애 유채와 한라 유채의 잎의 모양을 비교한 도이다.6 is a diagram comparing the shape of the leaves of Daae Rapeseed and Halla Rapeseed.
도 7은 본 발명의 일 실시예에 따른 적양배추(상단), 배추(하단) 및 본 발명의 다애 유채(하단)의 핵형을 분석한 도이다.Figure 7 is a view of the analysis of the karyotype of red cabbage (top), Chinese cabbage (bottom) and Daae rapeseed (bottom) of the present invention according to an embodiment of the present invention.
도 8은 본 발명의 일 실시예에 따른 다애 유채, 및 적양배추와 배추의 SNP(A) 및 RAPD(B)를 이용한 분자생물학적 분석 결과를 나타내는 도이다: 8 is a diagram showing the results of molecular biology analysis using SNP (A) and RAPD (B) of Daae Rapeseed, red cabbage and Chinese cabbage according to an embodiment of the present invention:
M: 1 kb ladder 마커;M: 1 kb ladder marker;
1: 적양배추(B. olarecea red);1: red cabbage ( B. olarecea red);
2: 배추(B. rapa);2: Chinese cabbage ( B. rapa );
3: 유채 다애(B. napus 'Dae').3: Rapeseed Tea ( B. napus 'Dae').
도 9는 다애 유채 특이 밴드를 증폭하는 프라이머를 이용하여 여러 종류의 유채품종을 RAPD 분석한 도이다:FIG. 9 is a diagram showing RAPD analysis of various kinds of rapeseed varieties using primers for amplifying a rapeseed specific band:
M: 1 kb ladder 마커;M: 1 kb ladder marker;
1: 적양배추(B. olarecea red);1: red cabbage ( B. olarecea red);
2: 배추(B. rapa); 2: Chinese cabbage ( B. rapa );
3: 유채 다애(B. napus 'Dae');3: rapeseed daae ( B. napus 'Dae');
4: 내한;4: cold winter;
5: 영산;5: youngsan;
6: 탐미;6: taste;
7: 한라;7: Halla;
8: 강유;8: oil;
9: 선망;9: envy;
10: 리산드라; 및10: risandra; And
11: 제주한라.11: Jeju Halla.
일반적인 방법Common way
이하 본 발명에서 사용된 일반적인 방법을 상세히 설명한다.Hereinafter, the general method used in the present invention will be described in detail.
1. 이종간 교잡 및 미숙배배양1. Cross-border hybridization and immature culture
적양배추(B. oleracea)를 모본으로 배추(B. rapa)를 부본으로 인공수분하여 수정이 된 꼬투리를 미숙배상태에서 기내배양을 유도한다. 교배일로부터 14~21일 후 수정된 꼬투리를 채취하여 그로부터 배를 적출하여 MS 배지에 치상하였다. 기내배양 조건은 25±1℃, 광도 40 μmol· m-2· s-1, 광주기 16시간 하에서 관리한다. 미숙배로부터 발생한 식물체는 더 큰 배양병에 치상하여 배양하였고. 이후 순화과정 및 육묘과정을 거쳐 정식 및 육성한다.Fertilized pods are incubated in immature embryos by artificially pollination with red cabbage ( B. oleracea ) as a model and artificial pollination with B. rapa ( B. rapa ). After 14-21 days from the mating day, fertilized pods were harvested, and embryos were extracted from the embryos and placed in MS medium. Incubation conditions are maintained under 25 ± 1 ℃, luminous intensity 40 μmol · m -2 · s -1 , photoperiod 16 hours. Plants arising from immature embryos were grown on larger culture bottles. After that, it is formalized and nurtured through the process of purification and seedling.
2. RAPD(Random Amplified Polymorphic DNA)를 이용한 분자유전학적 분석2. Molecular Genetic Analysis Using Random Amplified Polymorphic DNA (RAPD)
PCR 반응의 일종으로서 증폭되는 DNA 단편이 무작위적이라는 점에서 통상적인 PCR과 구분된다. RAPD는 몇몇 임의의 짧은 프라이머(8-12 뉴클레오티드)를 생성한 후, 큰 게놈 DNA를 주형으로 한 PCR 반응으로 수행되는데, 반응 결과 생성된 패턴을 분석함으로써 RAPD 반응으로부터 반-독특한 프로파일이 수득된다. 프라이머가 정확히 어느 곳인지는 모르지만, 표적 유전자의 DNA 서열 중 어느 곳인가에 결합하기 때문에, 상기 DNA 서열 정보는 불필요하다. 최근에, RAPD는 다양한 식물 및 동물 종의 계통도를 확인하고 추적하는데 사용되어 왔다.DNA fragments that are amplified as a kind of PCR reaction are distinguished from conventional PCR in that they are random. RAPD is performed in a PCR reaction with a large genomic DNA as a template after generating some arbitrary short primers (8-12 nucleotides), and a semi-unique profile is obtained from the RAPD reaction by analyzing the resulting pattern. It is not known exactly where the primer is located, but the DNA sequence information is unnecessary because it binds to any of the DNA sequences of the target gene. Recently, RAPD has been used to identify and track the lineage of various plant and animal species.
본 발명에서는 하기의 방법을 통해 게놈 DNA에 대한 RAPD 분석을 수행하였다:In the present invention, RAPD analysis of genomic DNA was performed by the following method:
1) PCR 반응액 조성1) PCR reaction solution composition
PCR 반응 조성물은 주형 DNA 50 ng, 10×PCR 완충액(1.5 mM MgCl2 포함), dNTPs 200 μM, taq DNA 중합효소 2 Unit, RAPD용 프라이머 20 μM을 혼합하고 증류수(D.W.)로 총 부피를 20 ㎕로 조정하여 반응액을 조성한다. The PCR reaction composition was mixed with 50 ng of template DNA, 10 × PCR buffer (containing 1.5 mM MgCl 2 ), 200 μM of dNTPs, 2 units of taq DNA polymerase, 20 μM of primer for RAPD, and 20 μl of total volume with distilled water (DW). Adjust to to form a reaction solution.
2) PCR 반응의 수행2) conducting PCR reactions
상기 PCR 반응액을 PCR 튜브에 넣고 95℃에서 3분간 변성시킨 후, 94℃에서 1분, 35℃에서 1분, 72℃에서 1분 30초를 한 사이클로 하여 총 4회 반복실시한 후, 다시 94℃에서 30초, 40℃에서 20초, 72℃에서 1분을 한 사이클로 하여 총 30회 반복실시한 후 72℃에서 7분간 반응시켜 합성한다. The PCR reaction solution was placed in a PCR tube and denatured at 95 ° C. for 3 minutes, followed by 4 cycles of 1 minute at 94 ° C., 1 minute at 35 ° C., and 1 minute 30 seconds at 72 ° C., followed by 94 cycles. 30 seconds at 40 ℃, 20 seconds at 40 ℃, 1 minute at 72 ℃ 1 cycle was repeated a total of 30 times and then synthesized by reacting for 7 minutes at 72 ℃.
3) 전기영동3) Electrophoresis
2.0%(w/v) 아가로스 겔에 상기 PCR 반응물 20 ㎕를 적재한 후 0.5 X TBE 버퍼하에서 200 V의 전압으로 1시간 30분 동안 전기영동을 수행한다. 이 때 마커로는 1Kb ladder를 6 ㎕ 적재하여 사용한다. 전기영동이 끝난 겔은 EtBr 용액으로 염색한 후, 형광위에 올려놓고 imager를 이용하여 이미지를 수득한다.20 μl of the PCR reaction was loaded on a 2.0% (w / v) agarose gel, followed by electrophoresis for 1 hour and 30 minutes at a voltage of 200 V under 0.5 X TBE buffer. At this time, 6 µl of 1Kb ladder is used as a marker. After the electrophoresis gel is stained with EtBr solution, the gel is placed on the fluorescence to obtain an image using an imager.
이하, 첨부된 도면들을 참조하여 본 발명의 실시예를 설명하면 다음과 같다. 그러나 본 발명은 이하에서 개시되는 도면에 도시된 실시예에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 수 있는 것으로, 이하의 도면에 도시된 실시예는 본 발명의 개시가 완전하도록 하며, 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이다. 또한 설명의 편의를 위하여 도면에서는 구성 요소들이 그 크기가 과장 또는 축소될 수 있다.Hereinafter, an embodiment of the present invention will be described with reference to the accompanying drawings. However, the present invention is not limited to the embodiments shown in the drawings disclosed below, but may be implemented in various forms. The embodiments shown in the following drawings make the disclosure of the present invention complete, and It is provided to fully inform the knowledge of the scope of the invention. In addition, the components may be exaggerated or reduced in size in the drawings for convenience of description.
도 1은 본 발명의 일 실시예에 따른 유채 신품종 다애의 교잡관계도를 나타낸 도이다. 본 발명자는 적양배추처럼 종자 크기가 큰 유채를 육성하기 위하여, 적양배추(B. oleracea)의 꽃봉오리로부터 화분을 제거하여 모본으로 사용하였으며 배추(B. rapa)를 부본으로 종간 인공교배를 실시하였다. 그 결과, 3개의 식물체를 확보할 수 있었는데, 도 1의 F2의 왼쪽으로부터 순서대로 1번 식물체는 월동하는 중 고사하였으며, 2번은 자가불화합성이 높아 자식종자의 확보가 어려웠으며, 3번 식물체는 자가수분률이 높아 다량의 후대종자 확보가 가능하였으며, 이를 '다애'라 명명하였다.1 is a diagram showing the hybridization relationship diagram of the new varieties of rape seed according to an embodiment of the present invention. In order to cultivate rapeseeds with a large seed size, such as red cabbage, the present inventors used pollen by removing pollen from the buds of red cabbage ( B. oleracea ) and conducted cross-border artificial crosses using cabbage ( B. rapa ) as a copy. . As a result, three plants were able to be secured. Plant 1 died during wintering in order from the left of F 2 in FIG. 1, and plant number 3 was difficult due to high incompatibility. Because of its high self-pollination rate, it was possible to secure a large amount of future seed, which was named 'Dae-ae'.
도 2는 교잡종 F1 후대 중 자가불화합성이 높은 2번 식물체(A)와 자가수분율이 높은 3번 식물체(B)의 종자 결실을 관찰한 도이다. 상기 도 1에 개시된 바와 같이 적양배추와 배추의 종간 인공교배 결과, 수득한 식물체의 종자결실을 관찰한 사진이다. FIG. 2 is a diagram illustrating seed deletions of plant 2 (A) having high self-incompatibility and plant 3 (B) having high self-pollinity among hybrid F 1 progeny. As shown in FIG. 1, the result of artificial breeding between red cabbage and Chinese cabbage is a photograph of seed loss of the obtained plant.
도 3은 본 발명의 일 실시예에 따른 유채 신품종 다애의 어린묘 육성 모습을 나타낸 도이다. 본 발명의 일 실시예에 따른 종간 교잡종인 다애의 품종적 특성을 관찰한 결과, 유묘기(seedling stage)에 잎 뒷면이 핑크빛을 띠는 것을 관찰할 수 있었다. Figure 3 is a view showing the growth of seedlings of the new varieties rapeseed rape according to an embodiment of the present invention. As a result of observing the cultivar characteristics of the cross-hybrid multicolored hybrid according to an embodiment of the present invention, it was observed that the back of the leaf was pink in the seedling stage.
도 4는 본 발명의 일 실시예에 따른 유채 신품종 다애의 육성모습(좌측 판넬) 및 다애 식물체의 엽맥 및 엽병의 색을 관찰한 도(우측 판넬)이다. 본 발명의 일 실시예에 따른 종간 교잡종인 다애의 품종적 특성을 관찰한 결과, 생육 초기부터 개화기 생육까지 엽맥 및 엽병이 핑크빛을 띄는 것을 확인할 수 있었다. Figure 4 is a diagram (right panel) to observe the growth of the rapeseed new varieties Daaeae (left panel) and the color of the leaf veins and leaf disease of the Daae plant according to an embodiment of the present invention. As a result of observing the varietal characteristics of the cross-hybrid multi-a hybrid species according to an embodiment of the present invention, it was confirmed that the leaf veins and folies have a pink color from the early growth to the flowering period.
도 5 및 도 6은 다애 유채와 한라 유채의 엽맥, 엽병의 색 및 잎의 모양을 비교한 도이다. 본 발명의 일 실시예에 따른 종간 교잡종인 다애의 품종적 특성을 한라 유채와 비교하였다. 자엽폭, 꽃색 및 종피색은 차이가 없었고 잎은 익상전렬로 차이가 없었으나 다애 유채의 엽맥 및 엽병이 분홍빛을 띠고, 잎테두리의 톱니꼴 모양이 다애 유채가 더 밋밋하고 잎의 윗부분이 원형에 가까웠다. 5 and 6 are diagrams comparing the leaf vein, leaf color and leaf shape of Daae Rapeseed and Halla Rapeseed. The cultivar characteristics of the cross-hybrid Daae, according to an embodiment of the present invention, were compared with Halla rape. Cotyledon width, flower color, and endothelial color did not differ, and the leaves did not differ in pterygium, but the leaf veins and leaf diseases of Daae rapeseed were pinkish, and the serrated shape of the leaf rim was smoother and the upper part of the leaf was circular Close to
도 7은 적양배추(상단), 배추(중단) 및 본 발명의 일 실시예에 따른 유채 신품종 다애(하단)의 핵형을 분석한 도이다. 본 발명의 일 실시예에 따른 다애 유채는 적양배추(B. oleracea)의 꽃봉오리로부터 화분을 제거하여 모본으로 사용하였으며 배추(B. rapa)를 부본으로 이용하였다. 이에, 다애의 염색체 수가 적양배추 및 배추의 배수체여부를 확인한 결과, 양친으로 사용된 적양배추(2n=18)와 배추(2n=20)의 배수체인 2n=38개로 확인되었다.Figure 7 is a view of the analysis of the karyotype of red cabbage (top), Chinese cabbage (hang) and rapeseed new varieties Daae (bottom) according to an embodiment of the present invention. Daae Rapeseed according to an embodiment of the present invention was used as a model by removing the pollen from the buds of red cabbage ( B. oleracea ) and cabbage ( B. rapa ) was used as a copy. Thus, as a result of confirming whether the number of chromosomes of Daae is reddish cabbage and Chinese cabbage, it was confirmed that 2n = 38 which is a drainage of red cabbage (2n = 18) and cabbage (2n = 20) used as parent.
도 8은 본 발명의 일 실시예에 따른 유채 신품종 다애, 적양배추 및 배추의 SNP(상단) 및 RAPD(하단)를 이용한 분자생물학적 분석 결과를 나타내는 도이다. 고해상도 융해(HRM, High Resolution Melting) 방법을 이용하여 모본 적양배추, 부본 배추 및 다애 유채를 분석한 결과, 본 발명의 일 실시예에 따른 다애 유채가 상술한 모본 및 부본의 이형접합체임을 확인하였다. 또한, RAPD 방법을 이용하여 다애 유채의 게놈 DNA를 분석하였으며, 그 결과 다애는 적양배추와 배추의 특이밴드를 증폭하는 것을 확인할 수 있었다. 또한, 상기 도 8에서 유채다애 특이적인 밴드를 증폭시킨 프라이머 B01을 이용하여 내한, 영산, 탐미, 한라, 강유, 선망, 리산드라, 제주한라 품종과 다애 유채를 비교 분석한 결과 다애 유채만이 다애 유채 특이밴드가 증폭됨이 확인되었다. 8 is a view showing the results of molecular biological analysis using SNP (top) and RAPD (bottom) of rapeseed new varieties Daaeae, red cabbage and Chinese cabbage according to an embodiment of the present invention. As a result of analyzing the mother red cabbage, the scallop cabbage and the pear rapeseed using the High Resolution Melting (HRM) method, it was confirmed that the pear rapeseed according to an embodiment of the present invention is a heterozygote of the above-described base and the copy. In addition, the genomic DNA of Daae Rapeseed was analyzed using RAPD method. As a result, Daae amplified specific bands of red cabbage and cabbage. In addition, using the primer B01 amplified rapeseed-a specific band in FIG. 8 as a result of comparing and analyzing the cold, Youngsan, Tammy, Halla, Kangyu, envy, Rissandra, Jeju Halla varieties and Daae rapeseed It was confirmed that the rapeseed singular band was amplified.
도 9는 다애 유채 특이 밴드를 증폭하는 프라이머를 이용하여 여러 종류의 유채품종을 RAPD 분석한 도이다. FIG. 9 is a diagram illustrating RAPD analysis of various kinds of rapeseed varieties using primers for amplifying a rapeseed specific band.
이하, 실시예 및 실험예를 통하여 본 발명을 더 상세히 설명한다. 그러나 본 발명은 이하에서 개시되는 실시예 및 실험예에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 수 있는 것으로, 이하의 실시예 및 실험예는 본 발명의 개시가 완전하도록 하며, 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the present invention is not limited to the examples and experimental examples disclosed below, but may be embodied in various different forms. The following examples and experimental examples are provided to make the disclosure of the present invention complete and the general knowledge. It is provided to fully inform those who have the scope of the invention.
실시예 1: 양배추와 배추의 교잡과 신규 유채 식물체의 생산Example 1 Hybridization of Cabbage and Chinese Cabbage and Production of New Rapeseed Plants
본 발명자는 적양배추처럼 종자 크기가 큰 유채를 육성하기 위하여 적양배추(B.oleracea)와 배추(B.rapa)를 종간 인공교배를 실시하였다. 적양배추의 꽃봉오리로부터 화분을 제거하여 모본으로 사용하였으며 배추를 부본으로 하여 인공수분을 시킨 후 유산지봉지를 씌워 오염을 방지하였다. The inventors have the interspecies artificial breeding was carried out ever cabbage (B.oleracea) and cabbage (B.rapa) to the seed size of a large development of rapeseed as ever cabbage. The pollen was removed from the buds of red cabbage and used as a mother's head.
인공교배일로부터 14~21일 후 형성된 꼬투리를 퇴화하기 전에 수득하여 기내에서 미숙배배양을 수행하였으며, 이로부터 3개의 식물체를 확보할 수 있었다(도 1 참조). 그 중 1번은 월동하는 중 고사하였고, 2번은 자가불화합성이 높아 자식종자의 확보가 어려웠으며(도 2의 A 참조), 3번 식물체는 자가수분률이 높아 다량의 후대종자 확보가 가능하였으며, 이를 '다애'라 명명하였다(도 2의 B 참조). Obtained before degeneration of the pods formed 14 to 21 days after the artificial breeding day, immature culture was performed in-flight, and three plants were secured therefrom (see FIG. 1). One of them was dead during the wintering, the second was difficult to secure the child seed due to its incompatibility (see A of Fig. 2), the third plant was able to secure a large number of later seeds because of the high self-pollination rate, This was named 'Dae Ae' (see B in Fig. 2).
실시예 2: 다애의 품종적 특성Example 2: Variety Characteristics of Darae
본 발명의 일 실시예에 따른 종간 교잡종인 '다애'는 후대 식물체 육성 결과 식물체간의 균일성이 확인되었다. 신품종 다애 유채는 유묘기(seedling stage)에 잎 뒷면이 핑크빛을 띄고(도 3), 생육 초기부터 개화기 생육까지 엽맥 및 엽병이 핑크빛을 띄는 것이 특징이다(도 4). 다애 유채를 한라 유채와 비교한 결과 자엽폭, 꽃색 및 종피색은 차이가 없었고 잎은 익상전렬로 차이가 없었으나 다애 유채의 엽맥 및 엽병이 분홍빛을 띠고, 잎테두리의 톱니꼴 모양이 다애 유채가 더 밋밋하고 잎의 윗부분이 원형에 가까웠다(도 5 및 도 6). 'Daeae,' a hybrid between species according to one embodiment of the present invention, was confirmed to have uniformity among plants as a result of plant growth. The new varieties Daae Rapeseed is characterized in that the back of the leaves pink color in the seedling stage (seedling stage) (Fig. 3), the leaf veins and foliar pink color from the early growth to flowering period (Fig. 4). As a result of comparing Daae rapeseed with Halla rapeseed, there was no difference in cotyledon width, flower color and longitudinal color, and the leaves did not differ in pterygium. Was flatter and the upper part of the leaf was closer to the circle (FIGS. 5 and 6).
또한 본 발명의 일 실시예에 따른 다애 유체 성체는 주지가 굵고 강하여 식물체 자세가 곧추서고 처짐정도가 적은 편이다. 만추대성으로 개화기까지 생육기간이 80일 이상으로 길기 때문에 왕성한 영양번식을 하며 생산된 종자는 일반유채보다 다소 큰 것으로 확인되었다. In addition, the multi-arachnoid fluid adult according to an embodiment of the present invention is thick and strong so that the plant posture is straight and the degree of deflection is small. It is confirmed that the seeds produced during the fertile breeding season are slightly larger than the general oilseed rape because the growth period is longer than 80 days.
표 1 다애 유채와 한라 유채의 식물학적 특성 비교
형질 표 현 형 태 다애 유채 한라 유채
1 2 3 4 5 6 7 8 9 표현 형태 표현 형태
1 자엽의 폭 좁음 중간 넓음 5 5
2 잎테두리의 톱니꼴 모양 없음 약간있음 중간 많다 2 3
3 잎의 열편 없음,원형 없음,장란형 약간있음 중간 아주많음 6 6
4 잎 색 담록 녹 농록 8 7
5 개화기 조숙 중숙 만숙 5
6 꽃색 담황 황 적황 5 5
7 성숙기 조숙 중숙 만숙 7 4
8 종피색깔 갈색 적색 흑색 노란색 4 4
9 춘파성 매우약함 약함 중간 강함 2 5
Table 1 Comparison of Botanical Characteristics of Daae Rapeseed and Halla Rapeseeds
characteristics Expression form Daae Rapeseed Halla Rapeseed
One 2 3 4 5 6 7 8 9 Expression Expression
One Cotyledon narrowness middle breadth 5 5
2 Serrated serpentine none Some middle many 2 3
3 Lobe of leaves None, round None, oblong Some middle Very many 6 6
4 Leaf color Damnok rust Dark green 8 7
5 bloom prematurity Mature Full 5
6 Flower color Pale yellow sulfur Red yellow 5 5
7 Maturity prematurity Mature Full 7 4
8 Species Color Brown Red black yellow 4 4
9 Spring Very weak weakness middle Strong 2 5
실시예 3: 천립중 비교Example 3: Comparison of Grain Weight
다애 품종의 종자크기를 알아보기 위하여 천립중을 비교한 결과, 배추(3.0 g) 및 양배추(4.95 g) 그리고 일반적인 유채품종(강유 5.1 g, 선망 4.9 g, 한라 4.1 g)보다 천립중이 큰 5.45 g으로 확인되었다(표 2). In order to determine the seed size of Daae varieties, the results were compared with Chinese grains. It was confirmed (Table 2).
표 2
종류 천립중(g) 출처
유채 강유 5.14 국립종자원
내한 2.90 농진청
탐미 4.00 농진청
한라 2.90 농진청
대화 4.10 국립종자원
선망 4.20 국립종자원
다애 5.45 (주)에프앤피
갓 청갓 2.17 경기도농업기술원
미소 2.20 에프앤피
배추 일반적인 배추 3.00 -
봄맛배추 3.65 경기도농업기술원
양배추 적양배추 4.95 경기도농업기술원
TABLE 2
Kinds (G) source
Rapeseed Oil 5.14 National Species Resources
Cold 2.90 Rural Development Administration
Taste 4.00 Rural Development Administration
Halla 2.90 Rural Development Administration
Conversation 4.10 National Species Resources
Envy 4.20 National Species Resources
Darae 5.45 F & P
lampshade Cheonggat 2.17 Gyeonggi-do Agricultural Research and Extension Services
smile 2.20 F & P
cabbage Common cabbage 3.00 -
Spring cabbage 3.65 Gyeonggi-do Agricultural Research and Extension Services
cabbage Red Cabbage 4.95 Gyeonggi-do Agricultural Research and Extension Services
실시예 4: 지방산 함량 분석Example 4: Fatty Acid Content Analysis
본 발명의 일 실시예에 따른 유채 신품종인 '다애'의 지방산 함량을 분석하였으며, 비교를 위해 하기 표 2에 기재된 유채, 갓, 배추 및 양배추 품종의 지방산 함량을 분석하였다.Fatty acid content of the new rapeseed varieties 'Daeae' according to an embodiment of the present invention was analyzed, and the fatty acid contents of the rapeseed, mustard, cabbage and cabbage varieties described in Table 2 were analyzed for comparison.
각 식물의 종자에 종자량의 8~10배의 황산나트륨을 넣은 후, 막자사발에서 곱게 파쇄한 후, 속시렛 추출법(Soxhlet extraction method) 또는 헥산(hexane)을 이용하여 지방을 추출하였다. Into the seeds of each plant was put 8-10 times the amount of sodium sulfate, and then finely crushed in a mortar, fat was extracted using Soxhlet extraction method (hexane) or hexane (hexane).
지방산 전처리는 ‘지방산 제2법 및 지방분석법’의 동일한 전처리 방법으로 수행하여 가스 크로마토그래피(gas chromatography) 분석을 수행하였다. 지방산 분석은 가스 크로마토그래피(Agilent 7890 GC, Agilent Technologies)를 사용하였으며, HP-INNOWAX(30m X 0.320 mm X 0.25 μm, Agilent Technologies), 검출기는 수소불꽃이온화검출기(flame ionization detector)를 사용하였다. 주입구 온도는 225℃, 검출기 온도는 285℃로 하였으며, 오븐 온도는 100℃에서 4분간 유지한 후 분당 3℃씩 240℃까지 올려 15분 유지하였다. 검출기 온도는 285℃로 하였으며, 캐리어 가스(carrier gas)로 질소를 이용하였다. Fatty acid pretreatment was performed by the same pretreatment method of 'fatty acid second method and fat analysis method' to perform gas chromatography analysis. Fatty acid analysis was performed using gas chromatography (Agilent 7890 GC, Agilent Technologies), HP-INNOWAX (30m X 0.320 mm X 0.25 μm, Agilent Technologies), the detector was a flame ionization detector (flame ionization detector). The inlet temperature was 225 ° C., the detector temperature was 285 ° C., and the oven temperature was maintained at 100 ° C. for 4 minutes and then raised to 240 ° C. at 3 ° C. per minute for 15 minutes. The detector temperature was 285 ° C., and nitrogen was used as a carrier gas.
그 결과, 본 발명의 일 실시예에 따른 유채 신품종인 다애가 12.9% 올레인산 및 61.5%의 에루스산을 함유하는 것을 확인할 수 있었고, 소량이기는 하나 다른 식물에서는 발견되지 않은 팔미톨레산이 0.62% 함유되어 있음을 알 수 있었다. 팔미톨레산은 불포화지방산으로서 당뇨병 예방효과가 있는 것으로 알려져 있고, 에루스산은 화장품등의 원료로 사용되는 지방산이므로, 본 발명의 일 실시예에 따른 유채 신품종을 이용하여 화장품 제조에 이용할 수 있을 것이다(표 3 참조). As a result, it was confirmed that the new rapeseed variety of the rapeseed, according to one embodiment of the present invention, contains 12.9% oleic acid and 61.5% erucic acid, and 0.62% of palmitoleic acid, which is small but not found in other plants. I could see that. Palmitoleic acid is known to be effective in preventing diabetes as unsaturated fatty acid, and since erucic acid is a fatty acid that is used as a raw material for cosmetics, it may be used in cosmetic production using a new rapeseed according to one embodiment of the present invention. 3).
표 3 지방산 함량 분석결과
지방산 함량(%) 내한 영산 한라 S16 S19 S20 S21 미소 (갓) 다애
C16:0 팔미트산(palmitic acid) 4.1 4.0 6.4 7.7 4.6 6.2 6.4 2.3 6.32
C16:1 팔미톨레산(palmitoleic acid) - - - - - - - - 0.62
C18:0 스테아린산(stearic acid) 2.2 1.9 1.9 1.6 1.8 2.01 1.57 0.9 1.63
C18:1 올레산(oleic acid) 58.8 63.4 61.3 59.1 63.1 55.2 55.3 8.0 12.9
C18:2 리놀레산(linoleic acid) 17.4 19.1 23.0 23.2 20.2 20.6 19.2 15.8 4.4
C18:3 리놀렌산(linolenic acid) 8.1 7.3 - - 6.7 - 1.5 9.4 5.4
C20:0 아라키드산(arachidic acid) 0.8 0.7 5.3 7.7 0.6 8.36 11.9 0.8 1.53
C20:1 Cis-11-Eicosenoic 3.8 1.3 - - 1.1 - - 5.2 1.06
C22:0 비헨산(behenic acid) 0.4 - - - 0.3 - - 1.2 1.47
C22:1 에루스산 (eruic acid) 3.0 0.6 - - - - - 49.2 61.5
C24:0 리그노세린산(lignoceric acid) - - - - - - - 0.5 -
C24:1 네르본산(nervonic acid) - - - - - - - 2.2 -
TABLE 3 Fatty Acid Content Analysis Results
Fatty acid content (%) Cold Youngsan Halla S16 S19 S20 S21 Smile (freshly) Darae
C16: 0 palmitic acid 4.1 4.0 6.4 7.7 4.6 6.2 6.4 2.3 6.32
C16: 1 palmitoleic acid - - - - - - - - 0.62
C18: 0 stearic acid 2.2 1.9 1.9 1.6 1.8 2.01 1.57 0.9 1.63
C18: 1 oleic acid 58.8 63.4 61.3 59.1 63.1 55.2 55.3 8.0 12.9
C18: 2 linoleic acid 17.4 19.1 23.0 23.2 20.2 20.6 19.2 15.8 4.4
C18: 3 linolenic acid 8.1 7.3 - - 6.7 - 1.5 9.4 5.4
C20: 0 arachidic acid 0.8 0.7 5.3 7.7 0.6 8.36 11.9 0.8 1.53
C20: 1 Cis-11-Eicosenoic 3.8 1.3 - - 1.1 - - 5.2 1.06
C22: 0 behenic acid 0.4 - - - 0.3 - - 1.2 1.47
C22: 1 eruic acid 3.0 0.6 - - - - - 49.2 61.5
C24: 0 lignoceric acid - - - - - - - 0.5 -
C24: 1 nervonic acid - - - - - - - 2.2 -
실시예 5: 염색체수 분석Example 5: Chromosome Number Analysis
염색체수 배가 여부를 확인하기 위한 염색체 검경 순서는 다음과 같았다. 근단(root tip)을 5 mm 정도 채취하여 차가운 증류수를 3 ㎖ 정도 채운 샘플병에 넣고, 다시 잘게 부순 얼음이 들어 있는 아이스 박스에 넣어 실내로 운반한 다음 근단을 약 2 mm 정도 절단하여 에펜도르프 튜브(eppendorf tube)에 넣는다. 준비된 샘플을 0.8% 브로모나프탈린(bromonaphthalene)에 넣고 상온에서 40분 정도 반응시켰다. 0.8% 브로모나프탈린은 1 ㎖ 용액에 상기 2 mm로 절단한 근단 20-30개 정도를 사용하였다. 반응이 끝나면 0.8% 브로모나프탈린 용액을 제거하고, 멸균수에 3회 정도 침지시켰다. 멸균수를 제거하고, 고정액(ethanol-glacial acetic acid)을 첨가하여 상온에서 하루 고정시켰다. 다음날 멸균수로 3회 세척해 준 다음, 효소액(2% 셀룰라아제(cellulase)/1% 펙티나아제(pectinase)/0.5% 펙토리아제(pectolyase))을 넣고, 37℃에서 50분 동안 해리시켰다. 파스퇴르 피펫(Pasteur pipette)을 이용하여 근단을 빨아들이고, 깨끗한 슬라이드에 2개만 떨어뜨린다. 펙터 종이(filter paper)로 근단에 한 층 정도의 막이 있을 정도만 물기를 제거한 후, 아세토카민(acetocarmine) 염색 시약을 떨어트렸다. 근단세포가 액체와 함께 고루 퍼져나가도록 핀셋으로 고루 펴주었다. 준비된 슬라이드를 위상차 현미경(Carl Zeiss)을 사용하여 400 배율에서 염색체수를 조사하고, 1,000 배율에서 유액을 투입하여 사진을 촬영하였다.Chromosome spectroscopy to determine whether the number of chromosome doubled was as follows. Take a root tip of 5 mm, place it in a sample bottle filled with 3 ml of cold distilled water, return it to an ice box containing crushed ice, and transport it indoors. Cut the root end about 2 mm and cut the Eppendorf tube. (eppendorf tube). The prepared sample was placed in 0.8% bromonaphthalene and reacted at room temperature for 40 minutes. 0.8% bromonaphthalin was used in about 20-30 root tips cut | disconnected by the said 2 mm in 1 ml solution. After the reaction, the 0.8% bromonaphthalin solution was removed and soaked three times in sterile water. Sterile water was removed and fixed at room temperature for one day by adding ethanol-glacial acetic acid. After washing three times with sterile water the next day, the enzyme solution (2% cellulase / 1% pectinase / 0.5% pectolyase) was added and dissociated at 37 ℃ for 50 minutes. A Pasteur pipette is used to soak the proximal end and drop only 2 onto a clean slide. The filter paper was drained only to the extent that there was a layer of membrane near the end, and then acetocarmine staining reagent was dropped. Myxocytes were spread evenly with tweezers to spread evenly with the liquid. The prepared slides were examined by chromosome number at 400 magnification using a phase difference microscope (Carl Zeiss), and photographed by adding an emulsion at 1,000 magnification.
그 결과, 종간 교잡종 '다애'의 염색체수를 분석한 결과 양친으로 사용된 적양배추(2n=18)와 배추(2n=20)의 배수체인 2n=38개로 확인되었다(도 7).As a result, as a result of analyzing the number of chromosomes of the hybrid species 'Daeae', it was confirmed that 2n = 38, which is a drainage of red cabbage (2n = 18) and cabbage (2n = 20) used as parents (FIG. 7).
실시예 6: SNP 및 RAPD 분석Example 6: SNP and RAPD Analysis
6-1: SNP 분석 6-1: SNP Analysis
고해상도 융해(HRM, High Resolution Melting) 방법은 품종을 구분할 수 있는 유전자좌와 연관된 분자마커를 개발하고 LC480(Roche)을 이용한 각 식물 개체의 유전자형을 분석하기 위해 사용되었다. PCR은 5 ㎕의 주형 DNA, 10 pmole의 각 프라이머(표 4), 10 ㎕의 SYBR mixture, 2 ㎕의 MgCl2 20 ㎕을 총 부피로 하고, 수행되었다. PCR은 94℃에서 3분 동안의 반응과 94℃에서 15초 53~58℃에서 25초 및 72℃에서 45초로 구성되는 45주기의 반응, 그리고 72℃에서 5분 동안의 반응으로 구성되었다. 형광신호는 95℃에서 1분, 40℃에서 1분간 처리한 다음 65℃~95℃까지 초당 0.02℃씩 증가시키고 1℃ 올라갈 때 마다 25번 측정되었고, 융해곡선 분석은 제조사의 작동 소프트웨어를 이용하여 수행되었다.The High Resolution Melting (HRM) method was used to develop molecular markers associated with genetically distinct loci and to analyze the genotype of each plant individual using LC480 (Roche). PCR was performed with 5 μl template DNA, 10 pmole of each primer (Table 4), 10 μl SYBR mixture, 2 μl MgCl 2 20 μl in total volume. PCR consisted of a reaction for 3 minutes at 94 ° C., a 45 cycle reaction consisting of 15 seconds at 94 ° C., 25 seconds at 53-58 ° C., and 45 seconds at 72 ° C., and a reaction at 72 ° C. for 5 minutes. The fluorescence signal was measured for 1 minute at 95 ° C and 1 minute at 40 ° C, then increased 0.02 ° C per second from 65 ° C to 95 ° C, and measured 25 times every 1 ° C. Melting curve analysis was performed using the manufacturer's operating software. Was performed.
표 4 분석에 사용한 프라이머
프라이머 명칭 서열번호 핵산서열 증폭산물(bp)
FPBPN0011-F 1 5'-GCATGGTGTGGATGAAGTTG-3' 99bp
FPBPN0011-R 2 5'-TTTACCTAAGCATCAGAGGATGG-3'
Table 4 Primer used for analysis
Primer Name SEQ ID NO: Nucleic acid sequence Amplification Product (bp)
FPBPN0011-F One 5'-GCATGGTGTGGATGAAGTTG-3 ' 99bp
FPBPN0011-R 2 5'-TTTACCTAAGCATCAGAGGATGG-3 '
적양배추, 배추, 유채를 구분할 수 있는 분자마커인 FPBPN0011의 염기 99bp를 포함하는 부위의 증폭을 위한 프라이머 및 상기 분자마커 FPBPN0011 증폭시 유전자의 이중가닥 사이에 결합(interchelating)되어 유전자 증폭량을 시각적으로 나타내는 표지물질(SYBR)을 이용하였다. 우선, FPBPN0011 분자마커의 PCR 증폭하였으며, 그 후 상기 PCR 증폭 산물에 대해 온도를 가하여 FPBPN0011의 이중 가닥을 단일 가닥으로 풀어줌으로써 FPBPN0011의 이중 가닥 사이에 존재하는 표지물질의 양이 줄어들게 하였다. 그리고 온도의 상승에 따라 상기 PCR 증폭 산물의 FPBPN0011의 이중 가닥 사이에 존재하는 표지물질의 양이 감소하는 경향을 이용하여 적양배추, 배추, 유채의 FPBPN0011에 대한 ‘온도 대 표지물질의 양’에 관한 융해곡선을 결정하였다. 그 결과, 도 8의 A에 나타난 바와 같이 모본 적양배추 및 부본 배추와 다애 유채를 분석한 결과 다애 유채가 적양배추와 배추의 이형접합체로 확인되었다(도 8의 A 참조).Primer for amplification of the site including the base 99bp of FPBPN0011, a molecular marker that can distinguish red cabbage, Chinese cabbage, and rapeseed, and when amplifying the molecular marker FPBPN0011, it is interchelated between the double strands of genes to visually indicate the amount of gene amplification Labeling material (SYBR) was used. First, PCR amplification of the FPBPN0011 molecular marker, and then the temperature was applied to the PCR amplification product to release the double strand of FPBPN0011 into a single strand to reduce the amount of labeling material present between the double strands of FPBPN0011. In addition, 'temperature versus amount of labeling substance' for FPBPN0011 of red cabbage, Chinese cabbage and rapeseed was decreased by increasing the amount of the labeling substance present between the double strands of FPBPN0011 of the PCR amplification product as the temperature increased. The melting curve was determined. As a result, as shown in FIG. 8A, as a result of analyzing the mother red cabbage, the scallop cabbage, and the pear rapeseed, the pear rapeseed was identified as a heterozygote of the red cabbage and the Chinese cabbage (see FIG. 8A).
6-2: RAPD 분석6-2: RAPD Analysis
본 발명의 신규 유채 품종인 다애와 적양배추 및 배추와의 분자유전학적 분석을 위하여 적양배추 및 배추, 다애 유채의 게놈 DNA에 대한 RAPD를 수행하였다. 이때 프라이머는 Operon사에서 판매하는 키트를 사용하였으며, 사용한 프라이머 정보는 표 5에 기재된 바와 같다. RAPD of genomic DNA of red cabbage, Chinese cabbage and Daae rape was performed for molecular genetic analysis of the new rape variety of the present invention, Daae, red cabbage and cabbage. At this time, the primer was used in the kit sold by Operon, and the primer information used is shown in Table 5.
RAPD를 이용한 분자생물학적 분석결과 다애는 적양배추와 배추의 특이밴드를 증폭하는 교잡종으로 확인되었다(도 8의 B 참조). 이중 B01 프라이머를 이용하여 증폭된 밴드 중 약 980 bp의 다애 유채 특이밴드를 확인할 수 있었다. B01 프라이머를 이용하여 내한, 영산, 탐미, 한라, 강유, 선망, 리산드라, 제주한라 품종과 다애 유채를 비교 분석한 결과 다애 유채만이 다애 유채 특이밴드가 증폭됨이 확인되었다(도 9). 상기 B01 프라이머는 하기 표 1에 기재한 OPB01 프라이머를 의미하며, 도 8 및 도 9에 기재되어 있는 A03~B12는 OPA03~OPB-12 프라이머를 의미한다.Molecular biological analysis using RAPD was confirmed as a hybrid hybrid to amplify specific bands of red cabbage and Chinese cabbage (see FIG. 8B). Among the bands amplified using the double B01 primer, it was possible to confirm the multispecific rapeseed specific band of about 980 bp. Using the B01 primer to compare the analysis of cold, Youngsan, Tammy, Halla, Kangyu, envy, Rissandra, Jeju Halla varieties and Daae rapeseed was confirmed that only the Daae rapeseed amplification band of Daae rapeseed (Fig. 9). The B01 primer refers to the OPB01 primer described in Table 1 below, and A03 to B12 described in FIGS. 8 and 9 mean OPA03 to OPB-12 primers.
표 5 분석에 사용한 프라이머
프라이머 명칭 서열번호 핵산서열 프라이머 명칭 서열번호 핵산서열
OPA-01 3 5'-CAGGCCCTTC-3' OPB-01 23 5'-GTTTCGCTCC-3'
OPA-02 4 5'-TGCCGAGCTG-3' OPB-02 24 5'-TGATCCCTGG-3'
OPA-03 5 5'-AGTCAGCCAC-3' OPB-03 25 5'-CATCCCCCTG-3'
OPA-04 6 5'-AATCGGGCTG-3' OPB-04 26 5'-GGACTGGAGT-3'
OPA-05 7 5'-AGGGGTCTTG-3' OPB-05 27 5'-TGCGCCCTTC-3'
OPA-06 8 5'-GGTCCCTGAC-3' OPB-06 28 5'-TGCTCTGCCC-3'
OPA-07 9 5'-GAAACGGGTG-3' OPB-07 29 5'-GGTGACGCAG-3'
OPA-08 10 5'-GTGACGTAGG-3' OPB-08 30 5'-GTCCACACGG-3'
OPA-09 11 5'-GGGTAACGCC-3' OPB-09 31 5'-TGGGGGACTC-3'
OPA-10 12 5'-GTGATCGCAG-3' OPB-10 32 5'-CTGCTGGGAC-3'
OPA-11 13 5'-CAATCGCCGT-3' OPB-11 33 5'-GTAGACCCGT-3'
OPA-12 14 5'-TCGGCGATAG-3' OPB-12 34 5'-CCTTGACGCA-3'
OPA-13 15 5'-CAGCACCCAC-3' OPB-13 35 5'-TTCCCCCGCT-3'
OPA-14 16 5'-TCTGTGCTGG-3' OPB-14 36 5'-TCCGCTCTGG-3'
OPA-15 17 5'-TTCCGAACCC-3' OPB-15 37 5'-GGAGGGTGTT-3'
OPA-16 18 5'-AGCCAGCGAA-3' OPB-16 38 5'-TTTGCCCGGA-3'
OPA-17 19 5'-GACCGCTTGT-3' OPB-17 39 5'-AGGGAACGAG-3'
OPA-18 20 5'-AGGTGACCGT-3' OPB-18 40 5'-CCACAGCAGT-3'
OPA-19 21 5'-CAAACGTCGG-3' OPB-19 41 5'-ACCCCCGAAG-3'
OPA-20 22 5'-GTTGCGATCC-3' OPB-20 42 5'-GGACCCTTAC-3'
Table 5 Primer used for analysis
Primer Name SEQ ID NO: Nucleic acid sequence Primer Name SEQ ID NO: Nucleic acid sequence
OPA-01 3 5'-CAGGCCCTTC-3 ' OPB-01 23 5'-GTTTCGCTCC-3 '
OPA-02 4 5'-TGCCGAGCTG-3 ' OPB-02 24 5'-TGATCCCTGG-3 '
OPA-03 5 5'-AGTCAGCCAC-3 ' OPB-03 25 5'-CATCCCCCTG-3 '
OPA-04 6 5'-AATCGGGCTG-3 ' OPB-04 26 5'-GGACTGGAGT-3 '
OPA-05 7 5'-AGGGGTCTTG-3 ' OPB-05 27 5'-TGCGCCCTTC-3 '
OPA-06 8 5'-GGTCCCTGAC-3 ' OPB-06 28 5'-TGCTCTGCCC-3 '
OPA-07 9 5'-GAAACGGGTG-3 ' OPB-07 29 5'-GGTGACGCAG-3 '
OPA-08 10 5'-GTGACGTAGG-3 ' OPB-08 30 5'-GTCCACACGG-3 '
OPA-09 11 5'-GGGTAACGCC-3 ' OPB-09 31 5'-TGGGGGACTC-3 '
OPA-10 12 5'-GTGATCGCAG-3 ' OPB-10 32 5'-CTGCTGGGAC-3 '
OPA-11 13 5'-CAATCGCCGT-3 ' OPB-11 33 5'-GTAGACCCGT-3 '
OPA-12 14 5'-TCGGCGATAG-3 ' OPB-12 34 5'-CCTTGACGCA-3 '
OPA-13 15 5'-CAGCACCCAC-3 ' OPB-13 35 5'-TTCCCCCGCT-3 '
OPA-14 16 5'-TCTGTGCTGG-3 ' OPB-14 36 5'-TCCGCTCTGG-3 '
OPA-15 17 5'-TTCCGAACCC-3 ' OPB-15 37 5'-GGAGGGTGTT-3 '
OPA-16 18 5'-AGCCAGCGAA-3 ' OPB-16 38 5'-TTTGCCCGGA-3 '
OPA-17 19 5'-GACCGCTTGT-3 ' OPB-17 39 5'-AGGGAACGAG-3 '
OPA-18 20 5'-AGGTGACCGT-3 ' OPB-18 40 5'-CCACAGCAGT-3 '
OPA-19 21 5'-CAAACGTCGG-3 ' OPB-19 41 5'-ACCCCCGAAG-3 '
OPA-20 22 5'-GTTGCGATCC-3 ' OPB-20 42 5'-GGACCCTTAC-3 '
하기에 본 발명의 조성물을 위한 제조예를 예시한다.The preparation examples for the compositions of the present invention are illustrated below.
<제조예 1> 화장품의 제조Preparation Example 1 Preparation of Cosmetics
본 발명의 일 실시예에 따른 신품종 유채 다애 추출물을 유효성분으로 함유하는 피부 미백용 화장품을 제조하였다. According to an embodiment of the present invention, a cosmetic for skin whitening containing a new breed rapeseed Daae extract was prepared as an active ingredient.
<1-1> 유연 화장수<1-1> flexible lotion
신품종 유채 다애 추출물을 함유한 유연 화장수는 하기 [표 6]의 조성과 같이 제조하였다.A flexible lotion containing a new variety rapeseed Daae extract was prepared as shown in Table 6 below.
표 6
원료 함량(중량%)
신품종 유채 다애 추출물 10.00
1,3-부틸렌글리콜 1.00
디소듐이디티에이 0.05
알란토인 0.10
디포타슘글리시리제이트 0.05
시트릭애씨드 0.01
소듐시트레이트 0.02
글리세레스-26 1.00
알부틴 2.00
하이드로제네이티드캐스터오일 1.00
에탄올 30.00
보존제 미량
착색제 미량
착향제 미량
정제수 잔량
Table 6
Raw material Content (% by weight)
New Breed Rapeseed Extract 10.00
1,3-butylene glycol 1.00
Disodium ID 0.05
Allantoin 0.10
Dipotassium glycylizate 0.05
Citrix Acid 0.01
Sodium citrate 0.02
Glyceres-26 1.00
Arbutin 2.00
Hydrogenated Castor Oil 1.00
ethanol 30.00
Preservative a very small amount
coloring agent a very small amount
Flavor a very small amount
Purified water Remaining amount
<2-2> 영양 크림<2-2> nutrition cream
신품종 유채 다애 추출물을 함유한 영양크림은 하기 [표 7]의 조성과 같이 제조하였다. Nutrition cream containing a new variety of rapeseed Daae extract was prepared as shown in the following [Table 7].
표 7
원료 함량(중량%)
신품종 유채 다애 추출물 10.0
1,3-부틸렌 글리콜 7.
글리세린 1.0
D-판테놀 0.1
식물 추출물 3.2
마그네슘알루미늄실리케이트 0.3
PEG-40 스테아레이트 1.2
스테아릭애씨드 2.0
폴리소르베이트 60 1.5
친유형글리세릴스테아레이트 2.0
소르비탄세스퀴올리에이트 1.5
세테아릴알코올 3.0
미네랄오일 4.0
스쿠알란 3.8
카르릴릭/카프릭트리글리세라이드 2.8
식물성 오일 1.8
디메치콘 0.4
디포타슘글리시리제이트 미량
알란토인 미량
소듐 히아루로네이트 미량
토코페릴아세테이트 적량
트리에탄올아민 적량
보존제 적량
착향제 적량
정제수 잔량
TABLE 7
Raw material Content (% by weight)
New Breed Rapeseed Extract 10.0
1,3-butylene glycol 7.
glycerin 1.0
D-panthenol 0.1
Plant extracts 3.2
Magnesium Aluminum Silicate 0.3
PEG-40 Stearate 1.2
Stearic acid 2.0
Polysorbate 60 1.5
Lipophilic glyceryl stearate 2.0
Sorbitan sesquioleate 1.5
Cetearyl Alcohol 3.0
Mineral oil 4.0
Squalane 3.8
Carlylic / Capric Triglycerides 2.8
vegetable oil 1.8
Dimethicone 0.4
Dipotassium glycylizate a very small amount
Allantoin a very small amount
Sodium hyaluronate a very small amount
Tocopheryl Acetate Quantity
Triethanolamine Quantity
Preservative Quantity
Flavor Quantity
Purified water Remaining amount
본 발명은 도면에 도시된 실시예를 참고로 설명되었으나 이는 예시적인 것에 불과하며, 당해 기술분야에서 통상의 지식을 가진 자라면 이로부터 다양한 변형 및 균등한 다른 실시예가 가능하다는 점을 이해할 것이다. 따라서 본 발명의 진정한 기술적 보호 범위는 첨부된 특허청구범위의 기술적 사상에 의하여 정해져야 할 것이다.Although the present invention has been described with reference to the embodiments shown in the drawings, this is merely exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.
본 발명의 일 실시예에 따른 신규 유채 품종 '다애'는 화장품 원료로 사용되는 에루스산의 함량이 높고 천중립의 크기가 종래 유채 품종에 비해 크기 때문에, 화장품 산업은 물론 바이오디젤 생산 등에 유리한 유채 품종의 생산을 위한 부본 또는 모본으로 활용될 수 있다.The new rapeseed varieties 'Daeae' according to an embodiment of the present invention have a high content of erucic acid used as a cosmetic raw material and the size of cloth neutral is larger than that of conventional rapeseed varieties, which is advantageous for the cosmetic industry as well as biodiesel production. It can be used as a copy or parent for the production of varieties.
서열번호 1 및 2는 본 발명의 일 실시예에 따른 신규 유채 품종 '다애'의 게놈 DNA를 대상으로 고해상도 융해 분석을 위해 사용된 FPBPN0011 마커의 증폭을 위한 포워드 및 리버스 프라이머의 핵산서열이다.SEQ ID NOs: 1 and 2 are nucleic acid sequences of forward and reverse primers for amplification of FPBPN0011 markers used for high resolution fusion analysis of genomic DNA of a novel rapeseed variety 'Dae' according to one embodiment of the present invention.
서열번호 3 내지 42는 본 발명의 일 실시예에 따른 신규 유채 품종 '다애'의 RAPD 분석을 위해 사용된 무작위 프라이머의 핵산서열들이다.SEQ ID NOs: 3 to 42 are nucleic acid sequences of random primers used for RAPD analysis of a novel rapeseed variety 'Daeae' according to one embodiment of the present invention.

Claims (7)

  1. 적양배추(Brassica oleracea) 와 배추(Brassica rapa) 의 종간 교잡에 의해 수득되고, 하기의 식물학적 특성을 가지는 유채 신품종 '다애':A new rapeseed variety 'Daeae', obtained by cross-breeding of red cabbage ( Brassica oleracea ) and Chinese cabbage ( Brassica rapa ) and having the following botanical characteristics:
    (a) 엽맥 및 엽병이 분홍색;(a) the veins and lobes are pink;
    (b) 천립중이 5.0g 이상; (b) at least 5.0 g of natural grains;
    (c) 에루스산 함량이 50% 이상; 및(c) an erucic acid content of at least 50%; And
    (d) 팔미톨레산 함량이 0.1~1%.(d) Palmitoleic acid content 0.1-1%.
  2. 제1항에 있어서,The method of claim 1,
    상기 에루스산 함량이 60% 이상인, The erucic acid content is more than 60%,
    유채 신품종 '다애'.Rapeseed new varieties 'Daeae'.
  3. 제1항 또는 제2항의 유채 신품종 '다애' 추출물을 유효성분으로 포함하는 화장료 조성물.Claim 1 or Claim 2 cosmetic composition comprising a new varieties of 'rapae' extract as an active ingredient.
  4. 적양배추(Brassica oleracea) 와 배추(Brassica rapa) 의 종간 교잡에 의하여 수득되고, 하기의 식물학적 특성 가운데 하나 이상의 특성을 갖는 유채를 선별하는 단계를 포함하는, 유채 신품종의 육종방법:A method for breeding a new breed of rapeseed, obtained by breeding a species of red cabbage ( Brassica oleracea ) and a cabbage ( Brassica rapa ) and comprising selecting a rapeseed having one or more of the following botanical characteristics:
    (a) 엽맥 및 엽병이 분홍색;(a) the veins and lobes are pink;
    (b) 천립중이 5.0g 이상; (b) at least 5.0 g of natural grains;
    (c) 에루스산 함량이 50% 이상; 및(c) an erucic acid content of at least 50%; And
    (d) 팔미톨레산 함량이 0.1~1%.(d) Palmitoleic acid content 0.1-1%.
  5. 제1항 또는 제2항의 유채 신품종 '다애'를 꽃가루, 모본 또는 부본으로 사용하여 교잡육종한 유채 식물체.Rapeseed plants hybridized and bred using the new rapeseed varieties 'Dae-ae' according to claim 1 or 2 as pollen, a parent or a book.
  6. 제1항 또는 제2항의 유채 신품종 '다애'를 꽃가루, 모본 또는 부본으로 하여 다른 유채 품종과 교잡하는 교잡단계;A hybridization step of hybridizing with other rapeseed varieties using the new rapeseed varieties 'Daeae' according to claim 1 or 2 as pollen, a mother tree or a stalk;
    상기 교잡에 의해 생성된 F1 중 천립중이 5.0 g 이상이고, 에루스산 함량이 50% 이상이며, 상기 다른 유채 품종의 유리한 특성을 함께 가지는 F1 개체를 선별하는 단계를 포함하는 제1항 또는 제2항의 유채 신품종 '다애'를 이용한 유채 신품종의 제조방법.And the cheonripjung of the F 1 generated by the hybridization 5.0 g or more, erucic acid, and the content is more than 50%, of claim 1 comprising the step of selecting the F 1 individual having with the advantageous properties of the other of rapeseed cultivars or Method for producing a new rapeseed varieties using the new rapeseed varieties 'Daeae' of claim 2.
  7. 제6항에 있어서, The method of claim 6,
    상기 유리한 특성은 내건성, 내냉성, 다수확성, 내열성, 내병성 또는 내충성인, 제조방법.The advantageous property is a dry resistance, cold resistance, high probability, heat resistance, disease resistance or insect resistance.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104839009A (en) * 2015-05-19 2015-08-19 镇江鑫源达园艺科技有限公司 Method for improving breeding quality of white brassica rapa
CN113388694A (en) * 2021-07-13 2021-09-14 延边大学 RAPD reaction system and kit of euonymus persiciformis

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20000061752A (en) * 1999-03-30 2000-10-25 신종암 New Cultivar by Interspecific Crossing between Porphyra tenera Kjellman var. tamatsuensis Miura and Porphyra yezoensis Ueda form. narawaensis Miura and Method thereof
JP2006211945A (en) * 2005-02-03 2006-08-17 Ts Shokubutsu Kenkyusho:Kk New cruciferous plant and method for rearing the same
KR20070073652A (en) * 2007-02-02 2007-07-10 바이오스펙트럼 주식회사 A composition for improving wrinkle on skin comprising plant extracts
US20100222605A1 (en) * 2007-06-13 2010-09-02 Syngenta Participations Ag New hybrid system for brassica napus

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20000061752A (en) * 1999-03-30 2000-10-25 신종암 New Cultivar by Interspecific Crossing between Porphyra tenera Kjellman var. tamatsuensis Miura and Porphyra yezoensis Ueda form. narawaensis Miura and Method thereof
JP2006211945A (en) * 2005-02-03 2006-08-17 Ts Shokubutsu Kenkyusho:Kk New cruciferous plant and method for rearing the same
KR20070073652A (en) * 2007-02-02 2007-07-10 바이오스펙트럼 주식회사 A composition for improving wrinkle on skin comprising plant extracts
US20100222605A1 (en) * 2007-06-13 2010-09-02 Syngenta Participations Ag New hybrid system for brassica napus

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HEATH, D. W. ET AL.: "Synthesis of high erucic acid rapeseed (Brassica napus L.) somatic hybrids with improved agronomic characters", THEORETICAL AND APPLIED GENETICS, vol. 91, 1995, pages 1129 - 1136 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104839009A (en) * 2015-05-19 2015-08-19 镇江鑫源达园艺科技有限公司 Method for improving breeding quality of white brassica rapa
CN113388694A (en) * 2021-07-13 2021-09-14 延边大学 RAPD reaction system and kit of euonymus persiciformis

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