WO2019181528A1 - Plant de capsicum à haute teneur en capsinoïdes - Google Patents

Plant de capsicum à haute teneur en capsinoïdes Download PDF

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WO2019181528A1
WO2019181528A1 PCT/JP2019/009048 JP2019009048W WO2019181528A1 WO 2019181528 A1 WO2019181528 A1 WO 2019181528A1 JP 2019009048 W JP2019009048 W JP 2019009048W WO 2019181528 A1 WO2019181528 A1 WO 2019181528A1
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content
capsinoid
fruit
carotenoid
population
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PCT/JP2019/009048
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Japanese (ja)
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哲也 関
雅文 太田
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味の素株式会社
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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
    • A01H5/08—Fruits
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/82—Solanaceae, e.g. pepper, tobacco, potato, tomato or eggplant

Definitions

  • the present invention relates to a plant belonging to the genus Capsicum containing capsinoids and carotenoids at a high concentration in the mature stage and a method for producing the same.
  • Capsicum annuum L. fruits and leaves are widely used around the world as food, spices and pharmaceutical ingredients.
  • the pungent component is a group of substances consisting of 12 or more homologues such as capsaicin and dihydrocapsaicin, and is collectively called capsaicinoid.
  • capsaicin is known to have various physiological activities, for example, effects such as suppression of obesity by enhancement of energy metabolism based on promotion of adrenaline secretion (non-patent literature). 1, 2).
  • capsaicinoid has a problem that its use as a food or a food additive or a medicine is limited because its use amount is limited because of its strong pungency and invasiveness.
  • Non-patent Document 3 a spicy varieties selected and fixed in Kyoto University experimental field from the self-progeny of spicy cultivar "CH-19" (Kyoto Prefectural University Faculty of Agriculture) “CH-19 Sweet” contains almost no capsaicinoid, and instead contains a large amount of capsaicinoid-like analogs collectively called capsinoids such as capsiate and dihydrocapsiate (Non-patent Document 3).
  • This capsinoid is a compound represented by the following general formula (I) or (II).
  • the difference in the chemical structure between capsaicinoid and capsinoid is that capsaicinoid is a compound in which vanillylamine and fatty acid are amide-bonded.
  • the capsinoid is only a compound in which vanillyl alcohol and a fatty acid are ester-bonded (Non-patent Document 4).
  • n means an integer of 0 to 10, preferably an integer of 3 to 5.
  • Capsinoids are known to have the same energy metabolism promoting effect, obesity suppressing effect, immunostimulating effect and the like as capsaicinoids.
  • food compositions and pharmaceutical compositions containing capsinoids are disclosed (Patent Document 1), and obesity inhibitors, body fat accumulation inhibitors, and adrenergic secretion promoters containing capsinoids as active ingredients are disclosed. (Patent Document 2).
  • a blood circulation promoting external preparation has been reported as a cosmetic (Patent Document 3).
  • Patent Document 4 a method for selecting a plant that biosynthesizes capsinoids is disclosed, and not only Capsicum annuum containing CH-19 sweet, but also a plant belonging to the genus Capsicum widely. Baccatum (Ahi Amarillo, etc.), C.I. Chinense (Habanero, Boot Jolokia, etc.), C.I.
  • Frutescens eg, red pepper
  • C.I. It is also disclosed that pubescens (such as Lokoto) has a capsinoid biosynthesis system (Patent Document 5). Then, using the above sorting method, C.I. Zavory Hot, Aji Dulce, and Caribbean Sweet, which are varieties that produce capsinoids in chinense, are selected (Non-patent Document 5). However, in the existing selection of capsinoid products, the capsinoid content is high in immature fruits but at most 2 mg / g (dry fruit weight), it is greatly reduced in mature fruits, and at most about 0.1 mg / g (dry fruit) Weight) grade. (Non-patent document 5). Therefore, in order to ingest capsinoids, it is necessary to ingest immature fruits or ingest them after separation and purification of capsinoids from immature fruits.
  • Non-Patent Document 6 Non-Patent Document 6
  • carotenoids containing capsanthin which are reported to be contained in pepper, have many useful properties such as antioxidant activity, cancer prevention activity, glucose metabolism improvement activity, lipid metabolism improvement activity, antiallergic effect, antiobesity effect, etc. Have been shown (Non-patent Documents 6 and 7), but it is difficult to ingest unless they are from mature fruits.
  • An object of the present invention is to provide a red pepper containing a high content of capsinoid and carotenoid at the same time in the fruit.
  • capsinoid-producing varieties in which the expression or activity of an enzyme (p-AMT) that catalyzes a transamination reaction from vanillin to vanillylamine is reduced.
  • p-AMT mutants by multiplying the line containing carotenoids (for example, spicy Kinense spp.)
  • carotenoids for example, spicy Kinense spp.
  • the present invention includes the following aspects.
  • a plant belonging to the genus Capsicum having a capsinoid content of 0.2 mg or more and a carotenoid content of 0.05 mg or more per 1 g of the dry weight of the fruit.
  • [5] The plant according to any one of [1] to [4], wherein Capsicum chinense capsicum or Capsicum chinense is included in the hybrid.
  • [6] The plant according to any one of [1] to [5] obtained by the following steps: (1) A step of obtaining an F1 population by crossing a red pepper line in which p-AMT gene expression or enzyme activity is reduced with a line containing a carotenoid at the mature stage; (2) obtaining an F2 population by self-pollination of plants from the F1 population; (3) Breeding the F2 population and measuring capsinoid and carotenoid content at maturity; (4) A step of selecting a plant having a high capsinoid content and carotenoid content in the fruit at the mature stage.
  • a capsicum line in which p-AMT gene expression or enzyme activity is reduced is a line obtained by crossing at least one selected from the group consisting of Zavory Hot, Aji Dulce, Caribbean Sweet, CH19 sweet and S3212.
  • [8] The plant according to [6] or [7], wherein the line containing carotenoid in the mature stage is a line containing Capsicum chinense species or Capsicum chinense species as a hybrid.
  • the Capsicum chinense species is a line obtained by crossing at least one selected from the group consisting of habanero, infinity, and boot jorochia.
  • a method for producing a plant belonging to the genus Capsicum having a capsinoid content of 0.2 mg or more and a carotenoid content of 0.05 mg or more per 1 g of the dry weight of the fruit in the ripening stage comprising the following steps: (1) A step of obtaining an F1 population by crossing a red pepper line in which p-AMT gene expression or enzyme activity is reduced with a line containing a carotenoid at the mature stage; (2) obtaining an F2 population by self-pollination of pepper from the F1 population; (3) Breeding the F2 population and measuring capsinoid and carotenoid content at maturity; (4) A step of selecting a plant having a high capsinoid content and carotenoid content in the fruit at the mature stage.
  • capsinoids with less stimulation energy metabolism activation effect, body fat accumulation suppression effect, immune activation effect and the like, and effects such as nutritional value and antioxidant activity, natural red and yellow pigments Therefore, it is possible to provide a pepper that contains a carotenoid that is also useful as the above.
  • FIG. 1 shows the correlation between capsinoid content and carotenoid content in pepper.
  • the chili peppers reported so far do not coexist with capsinoids and carotenoids, but the chili pepper according to the present invention has been shown to be chili peppers coexisting with these useful compounds.
  • ⁇ represents an example, and ⁇ represents a comparative example and a reference example.
  • the present invention relates to a plant belonging to the genus Capsicum having a capsinoid content of 0.2 mg or more and a carotenoid content of 0.05 mg or more per 1 g of the dry weight of the fruit (sometimes abbreviated as Capsicum plant of the present invention). About.
  • fruit is preferable.
  • the “maturity period” in the present invention means “a maturity period in which the fruit exhibits red, orange, and yellow in the pericarp after the immature period when the fruit skin color is green”.
  • capsicum is classified by maturity, it is usually divided into immature fruits, intermediate fruits, and fully-ripened fruits (see: Evaluation of functional components according to ripeness and harvest time of Tabasco (Capsicum frescens var. Tabasco).
  • plants in the mature stage includes intermediate fruits and fully mature fruits.
  • immature fruits are red, orange, and yellow on the surface of the fruit with a range of less than 5% per area (fruits with green pericarp), and mature fruits with more than 5% per area (over 5% of the pericarp is red and orange) Yellow fruit).
  • the “plant in the mature stage” includes a plant that has been matured on the herb before harvesting, for example, in an immature stage, ripened and matured under arbitrary conditions.
  • the Capsicum plant of the present invention can contain capsinoids at the mature stage with excellent taste and nutrition.
  • the capsinoid content per gram dry weight of the fruit is usually 0.2 mg or more, preferably 0.5 mg or more, more preferably 1 mg or more, further preferably 2 mg or more, particularly preferably 5 mg or more, and most preferably 6 mg. That's it.
  • an upper limit is not specifically limited, It is 100 mg or less.
  • the capsinoid content per gram dry weight of the fruit at the time of ripening is usually 0.2 mg or more, preferably 0.5 mg or more, more preferably 1 mg or more, further preferably 2 mg or more, and particularly preferably 5 mg or more. Most preferably, it is 6 mg or more.
  • an upper limit is not specifically limited, It is 100 mg or less.
  • the carotenoid content per gram dry weight of the fruit is usually 0.05 mg or more, preferably 0.1 mg, more preferably 0.2 mg or more, and further preferably 0.3 mg or more. Although an upper limit is not specifically limited, It is 50 mg or less.
  • the carotenoid content per gram dry weight of the fruit at the time of ripening is usually 0.1 mg or more, preferably 0.15 mg, more preferably 0.2 mg or more, and further preferably 0.3 mg or more. Although an upper limit is not specifically limited, It is 50 mg or less.
  • the capsanthin content per gram dry weight of the fruit is usually 0.05 mg or more, preferably 0.1 mg or more, more preferably 0.15 mg, and further preferably 0.2 mg or more. Although an upper limit is not specifically limited, It is 25 mg or less.
  • the capsanthin content per gram of dry weight of the fruit at the time of ripening is usually 0.05 mg or more, preferably 0.1 mg, more preferably 0.15 mg, and further preferably 0.2 mg or more. Although an upper limit is not specifically limited, It is 25 mg or less.
  • the plant of the genus Capsicum of the present invention preferably contains a Capsicum chinense species or a Capsicum chinense species as a hybrid.
  • the plant of the genus Capsicum of the present invention is a strain that accumulates carotenoids in the mature stage, particularly a strain of Capsicum chinense species or a strain in which the strain is hybridized with a pepper strain in which p-AMT gene expression or enzyme activity is reduced. Further, it can be obtained by crossing, using a line obtained by breeding selection of a line excellent in capsinoid accumulation centering on the accumulation ability of carotenoid, or by further breeding.
  • the Capsicum plant of the present invention is obtained by the following steps: (1) A step of obtaining an F1 population by crossing a red pepper line in which p-AMT gene expression or enzyme activity is reduced with a line containing a carotenoid at the mature stage; (2) obtaining an F2 population by self-pollination of plants from the F1 population; (3) Breeding the F2 population and measuring capsinoid and carotenoid content at maturity; (4) A step of selecting a plant having a high capsinoid content and carotenoid content in the fruit at the mature stage.
  • the breeding in (3) is a concept including cultivation and breeding. That is, (3) includes a step of measuring capsaicinoid and capsinoid content in the fruit of the F2 population itself.
  • red pepper strain in which p-AMT gene expression or enzyme activity is reduced is, for example, a strain obtained by crossing at least one selected from the group consisting of Zavery Hot, Aji Dulce, Caribbean Sweet, CH19 Sweet, and S3212. Can be mentioned. Among them, Capsicum chinense species such as Aji Dulce, Zavory Hot, and Caribbean Sweet are preferable from the viewpoint of high capsinoid content.
  • p-AMT (putative aminotransferase) means an enzyme that catalyzes a transamination reaction from vanillin to vanillylamine.
  • p-AMT gene expression is decreased (function-deficient mutation) or enzyme activity is reduced. It needs to be lowered.
  • the “function-deficient mutation” in the p-AMT gene means a mutation that reduces the activity of the enzyme to such an extent that the production of capsinoid is significantly increased as compared with the corresponding wild-type strain. It is not necessary to be a mutation that eliminates. Moreover, the mutation may be such that the activity of the enzyme disappears or decreases as a result of disabling or reducing transcription from the gene or translation from the transcription product.
  • Examples of the method for detecting a mutation in the p-AMT gene include the method described in Japanese Patent No. 5428219.
  • a capsicum strain in which the expression or enzyme activity of the p-AMT gene is reduced can be selected, for example, by the method for detecting a mutation in the p-AMT gene described in the above-mentioned Japanese Patent No. 5428219.
  • strains that accumulate carotenoids at maturity include Capsicum chinense species or species that include Capsicum chinense species in hybrids.
  • At least one selected from the group consisting of habanero, boot joroquia, infinity, morgan scorpion, dorset naga, carolina reaper, brown egg, trinidad scorpion, king naga jolokia is crossed. Lines obtained in this manner are preferable, and habanero, boot jorochia, and infinity are more preferable.
  • cultivars produced by crossing lines consisting of habanero, boot jorochia, and infinity to a line selected from Aji Dulce and repeatedly selecting and breeding lines with excellent carotenoid and capsinoid accumulation and harvestability are preferred.
  • Capsinoid content is measured using high performance liquid chromatography (HPLC), UV absorption spectrum, mass spectrum, and fluorescence spectrum are used as the detection system, gas chromatography is used, and mass spectrum and flame ionization are used as the detection system. Examples include a method using a detector.
  • HPLC high performance liquid chromatography
  • UV absorption spectrum, mass spectrum, and fluorescence spectrum are used as the detection system
  • gas chromatography is used
  • mass spectrum and flame ionization are used as the detection system. Examples include a method using a detector.
  • a plant having a high capsinoid content is a plant having a capsinoid content of 0.2 mg or more per 1 g of dry weight of fruits.
  • Carotenoid content is measured using high performance liquid chromatography (HPLC), UV absorption spectrum, mass spectrum, and fluorescence spectrum are used as the detection system, gas chromatography is used, and mass spectrum and flame ionization are used as the detection system. Examples include a method using a detector.
  • a plant having a high carotenoid content is a plant having a carotenoid content of 0.05 mg or more per 1 g of dry weight of fruits.
  • breeding selection is to repeat selection of individuals and line cultivation for each selected individual, and to select and fix and make a pure line by comparing the lines in the separated generation of hybrids. It can be done by a method.
  • the cultivation method of the produced pepper is not particularly limited as long as it is a cultivation method usually used in pepper cultivation.
  • outdoor cultivation rain-free soil cultivation, rain-free hydroponics, plant factory cultivation, and multi-soil Cultivation cultivation etc. are mentioned.
  • the plant of the genus Capsicum of the present invention is a capsicum obtained by breeding selection by crossing, and does not include a capsicum by a gene recombination technique.
  • the chili pepper by the genetic recombination technique means a red pepper artificially introduced with a foreign gene by the genetic recombination technique, and does not mean a red pepper bred by crossing.
  • the present invention relates to a method for producing a plant belonging to the genus Capsicum having a capsinoid content of 0.2 mg or more and a carotenoid content of 0.05 mg or more per gram of dry weight of the fruit in the ripening stage, the method comprising the following steps: Also included: (1) A step of obtaining an F1 population by crossing a red pepper line in which p-AMT gene expression or enzyme activity is reduced with a line containing a carotenoid at the mature stage; (2) obtaining an F2 population by self-pollination of pepper from the F1 population; (3) Breeding the F2 population and measuring capsinoid and carotenoid content at maturity; (4) A step of selecting a plant having a high capsinoid content and carotenoid content in the mature stage.
  • Various definitions and contents are as described above.
  • the fruit of the plant belonging to the genus Capsicum of the present invention can be used for various processed products by processing methods such as freezing, drying, crushing, crushing, grinding, extraction, concentration, dissolution, enzyme treatment, acid-alkali treatment and the like. .
  • the above processed product is a composition containing high concentrations of capsinoids and carotenoids, and is less pungent and invasive than capsaicin, so it can be suitably used as a food or medicine or an additive thereto.
  • the above composition has the effect of lowering blood triglyceride concentration, increasing blood free fatty acid concentration, increasing blood adrenaline level, increasing blood glucose concentration and increasing oxygen consumption, thereby activating energy metabolism.
  • it is useful for improving general nutritional status and promoting health, such as suppressing obesity and improving metabolic status, such as transient hypoglycemia.
  • Examples of uses as pharmaceuticals include, for example, pharmaceuticals having various physiological activities such as energy metabolism activators, obesity inhibitors, body fat accumulation inhibitors, analgesics, adrenaline secretion promoters, endurance enhancers, etc. Can do.
  • composition when added to food, it can be used as a health food or functional food having various physiological activities as described above.
  • the form of the preparation may be, for example, a solid preparation such as a tablet, capsule, granule, powder or suppository, or a liquid preparation such as a syrup, elixir or injection.
  • HPLC measurement conditions for capsinoids are as follows. HPLC conditions: Column: YMC Trial Phenyl plus 3 ⁇ m (diameter 4.6 mm ⁇ 250 mm) Eluent: 70% methanol / 30% water (v / v) Detection: Fluorescence Ex280nm Em320nm Temperature 40 ° C
  • HPLC measurement conditions for carotenoids including capsanthin are as follows. HPLC conditions: Column: YMC Trial Phenyl plus 3 ⁇ m (diameter 4.6 mm ⁇ 250 mm) Eluent: 91.99% methanol / 8.00% water / 0.01% ammonium acetate (v / v) Detection: UV475nm Temperature 40 ° C
  • Carotenoids and capsinoids were measured in mature fruits of these habanero, infinity, and boot Jolocia p-AMT mutants. The measured values were obtained by measuring 3 samples of fruits of each strain and calculating the average value. Carotenoids were quantified by the method of non-patent literature (horticulture study 14 (4): 391-396, 2015), and the total amount of capsanthin and carotenoid was measured. Carotenoids were purchased and used as standard products (Capsanthin: cat.no. 465-42-9 manufactured by Funakoshi, beta carotene: cat. No. 7235-40-7 manufactured by Funakoshi).
  • the capsinoid was quantified by the method of Patent Document (JP 4089306B2), capsiate, dihydrocapsiate, nordihydrocapsiate was detected, and the total amount of capsinoid was calculated.
  • Capsinoids were synthesized according to the method of non-patent literature (Journal of Japanese Society for Agricultural Chemistry, Vol. 77 (2003) No. 10, p. 1008-1009). The results are shown in Table 1.
  • ND indicates a capsinoid detection limit (less than 0.1 mg / g), a carotenoid detection limit (0.05 mg / g) and a capsanthin detection limit (0.01 mg / g).
  • the capsinoid content and carotenoid content in red pepper are plotted in FIG.
  • the capsicum chinense mature fruits contained carotenoids in an amount of 0.1 mg / g dry weight or more and capsinoids in an amount of 1.0 mg / g dry weight or more.
  • Capsicum annuum p-AMT mutant mature fruit (Comparative Example 3: CH19 sweet, Comparative Example 4: TC4-2) contained carotenoids at a dry weight of 1.4 mg / g or more, but capsinoids had a detection limit (0.1 mg / G) or less.
  • capsicum annuum p-AMT mutant immature fruits (Reference Examples 1 to 3), carotenoids were less than 0.05 mg / g dry weight, and capsinoids were detected but could not coexist with carotenoids.
  • the mature fruit of Ahidurce (Comparative Example 6) contained 0.6 mg / g dry weight of carotenoids, but capsinoids were below the detection limit (0.1 mg / g).
  • the carotenoid of the Ahidulce immature fruit (Comparative Example 5) was less than 0.05 mg / g dry weight, and although capsinoid was detected, it could not coexist with the carotenoid. From Table 1 and FIG.

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  • Health & Medical Sciences (AREA)
  • Physiology (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

L'invention concerne un Capsicum annuum qui a une concentration élevée de capsinoides et caroténoïdes dans le fruit de celui-ci. L'invention concerne une plante qui appartient au genre Capsicum et a, pour 1 g de fruit sec, une teneur en capsinoïdes d'au moins 0,2 mg et une teneur en caroténoïdes d'au moins 0,05 mg.
PCT/JP2019/009048 2018-03-23 2019-03-07 Plant de capsicum à haute teneur en capsinoïdes WO2019181528A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114208659A (zh) * 2021-10-18 2022-03-22 新疆天椒红安农业科技有限责任公司 一种高色素辣椒的育种方法

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
JANG, S. ET AL.: "Substitution of a dysfunctional pAMT allele results in low-pungency but high levels of capsinoid in capsicum chinense 'Habanero'.", PLANT BREED. BIOTECH., vol. 3, no. 2, 2015, pages 119 - 128, XP055637522 *
JEONG, H. ET AL.: "Marker-assisted backcross breeding for development of pepper varieties (Capsicum annuum) containing capsinoids.", MOL. BREEDING, vol. 35, 2015, pages 226, XP035924004 *
LI, C. J. ET AL.: "Effects of virus-induced gene silencing (VIGS) of habanero putative aminotransferase (pAMT) on capsaicin and capsiate synthesis", JAPANESE JOURNAL OF PHYTOPATHOLOGY, vol. 83, no. 3, August 2017 (2017-08-01), pages 248 , 249 *
TANAKA, Y. ET AL.: "Novel loss-of-function putative aminotransferase alleles cause biosynthesis of capsinoids, nonpungent capsaicinoid analogues, in mildly pungent chili peppers (Capsicum chinense", J. AGRIC. FOOD CHEM., vol. 58, 2010, pages 11762 - 11767, XP055637524 *
WATANABE, TATSUO: "3 The history, food culture and functionality of pepper, item of ''3) Capsanthin capsorbin", FUNCTIONAL FOOD, vol. 11, no. 2, 2017, pages 101 - 105 *
YASUI, HIROYUKI ET AL.: "Capsanthin", FUNCTIONAL FOOD, vol. 5, no. 2, 2011, pages 177 - 182 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114208659A (zh) * 2021-10-18 2022-03-22 新疆天椒红安农业科技有限责任公司 一种高色素辣椒的育种方法

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