KR20140117825A - Specfic biomarker for selection of Gotgam Chamoe variety, Korean native Oriental Melon from cultivar Chamoe variety and uses thereof - Google Patents

Abstract

The present invention relates to a composition for selecting a species of dried persimmon oriental melon which is a Korean native species from a cultivated oriental melon species including a specific volatile substance derived from oriental melon, and to a method for selecting a species of dried persimmon oriental melon which is a Korean native species from a cultivated oriental melon species, which includes a step of isolating a volatile substance from each oriental melon of a mixed sample of a cultivated oriental melon species and a dried persimmon oriental melon species, and comparing the content of a specific volatile substance. The method for selecting species of dried persimmon oriental melon which is a Korean native species can be an extremely useful method by a volatile biomarker efficient in not only species developing and seed producing of oriental melon, but also protecting of a native species of dried persimmon oriental melon valuable as a genetic resource.

Description

[TECHNICAL FIELD] The present invention relates to a volatile-specific biomarker and a use thereof for screening Korean persimmon varieties from cultivated melon varieties,

The present invention relates to a specific biomarker for screening Korean persimmon varieties from cultivated oriental varieties and to their use. More particularly, the present invention relates to a biomarker of oriental persimmon varieties, The present invention relates to a composition for selecting persimmon melon varieties and a method for isolating volatile substances from each melon of cultivated melon varieties and persimmon melon varieties, and comparing the contents of specific volatiles, from the cultivated melon varieties to Korean persimmon varieties, The method comprising the steps of:

Melon ( Cucumis There's melo . makuwa ) is a vine plant belonging to the genus Bacillus and taxonomically melon ( Cucumis melo ) is a variant of. The domestic seed market of bacon and crops accounts for about 20% of the total vegetable seed market, with the second largest seed market followed by watermelon in the park and crops, and the annual market size and cultivation area There is a tendency to increase somewhat. Recently, attempts have been made to introduce important traits such as melon flavor and sugar content to other foliage and crops such as melon, and the importance of molecular markers is increasing for this purpose. Melon is also rich in vitamin C is good for fatigue recovery, anti-cancer effect, detoxification, is also effective in preventing food poisoning.

Although the classification of plants has been studied mainly on the morphological characteristics, the structure of plant components is gradually clarified according to the development of separation technology and instrumental analysis means, and the chemical structural features of plant components are used as an index of taxonomy by accumulation of data I have been. In this context, chemical taxonomy has developed, which refers to the use of cellular constituents or specific metabolites as a classification indicator of living organisms, and usually refers to secondary metabolites that can be used to compare chemical structures in plants or fungi . Dried persimmon varieties contain many nutrients such as calcium, carbohydrates, and vitamins and are resistant to disease. Also, it is a unique varieties of dried persimmon, and its fragrance is valuable as a breeding material that improves the quality to improve the taste of the consumers by promoting the taste of the consumers. Therefore, in order to protect the breed and lineage, Marker development is essential.

Korean Patent No. 0652501 discloses a method for determining the purity of F-1 seed in melons, and Korean Patent No. 0919753 discloses a powdery mildew resistance-associated SCAR marker useful in melons and melons and a method for selecting resistant melon cultivars using the same, However, specific biomarkers and their uses for screening Korean oriental melon cultivars from cultivated melon cultivars as in the present invention have not been disclosed.

The present invention has been made in view of the above-described needs. The present invention provides a method for separating persimmon-related substances, volatile substances, from persimmon, flesh and persimmon to separate persimmon persimmon and cultivated persimmon varieties of domestic native species, As a result of the comparison, it was found that volatile substances such as ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methylbutyl butanoate, Benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1- 3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl-2-allyl -2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanate oate, ethyl hexadecanoate, 1,3,5-heptatriene, 3-pentyl-1-cyclohexene, and The content of 4-hydroxy butanoic acid lactone was found to be significantly different between the oriental persimmon and cultivated oriental varieties.

Accordingly, the present invention has been accomplished by analyzing the content of the specific volatile matter and constructing a method for screening Korean persimmon varieties of oriental persimmon from the cultivated oriental varieties.

In order to accomplish the object of the present invention, the present invention provides a process for the production of ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methyl butyl butanoate (3 methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1-en- Oct-1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl-2-allyl-2-hydroxy-pentanedioate diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3,5- A group consisting of 1,3,5-heptatriene, 3-pentyl-1-cyclohexene and 4-hydroxy butanoic acid lactone Melons from native breeds that include one or more volatile substances selected emitter provides a composition for screening the South Korea native strains of persimmon melon varieties.

In addition, the present invention provides a method for screening Korean persimmon varieties of oriental persimmon from the cultivated oriental varieties including the step of measuring the specific volatile substance content.

Since the selection of the oriental persimmon varieties of Korean native species according to the present invention can be performed by measuring the content of the specific volatile substances developed in the present invention from each of the pericarp, Which is an effective volatile biomarker for protecting native varieties of dried persimmon.

In order to achieve the above object, the present invention provides a composition for selecting persimmon cultivars of persimmon cultivars of Korean native species from cultivated oriental cultivars including specific volatile substances derived from oriental melon.

In the present invention, specific volatile substances for selecting persimmon cultivars of Korean native species from cultivated melon cultivars include ethyl butanoate, ethyl hexanoate, 1-hexyl acetate 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate Oct-1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl- Diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethylhexadecanoate ethyl hexadecanoate, 1,3,5-heptatriene, 3-pentyl-1-cyclohexene and 4- Noik Acid lactone (4-hydroxy butanoic acid lactone), but to be at least one selected from the group consisting of, but not limited thereto.

The specific volatile substances derived from the melon of the present invention such as ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1-en-3-yl acetate oct-1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, and diethyl 2-allyl-2-hydroxy-pentanedioate. 2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3,5-heptatriene (3-pentyl-1-cyclohexene) and 4-hydroxy butanoic acid lactone (3-pentyl-1-cyclohexene) Emitter completely unknown for points that can be used to screen for Korea native strains of persimmon melon varieties.

The present inventors have proved that certain volatile substances of the present invention can be used as a volatile biomarker for screening Korean oriental melon cultivars from cultivated melon cultivars through the following proofs.

Specifically, 61 kinds of volatile substances listed in the following Tables 1 and 2 were detected by separating the volatile substances in the pericarp, flesh and the superficial region of the cultivated oriental cultivars cultivated in the packaging and the persimmon cultivars of persimmon cultivars of Korean native species, However,

Particularly, ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methylbutyl butanoate, benzyl acetate 1-en-3-yl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct- yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl-2-allyl-2-hydroxy propanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3,5-heptatriene, 1,3,5-heptatriene, ), 3-pentyl-1-cyclohexene and 4-hydroxy butanoic acid lactone were significantly different between the sweet potato and the persimmon sweet potato To check Respectively.

However, there is a problem in that the sweet potatoes such as 3-methyl-1-butanol, 3,3,6-trimethylhepta-1,5-dien-4-ol, 2-propylheptanol, N-tridecanol, 2-hexyloctanol, methyl formate, 2-methyl-1-butylacetate, ethyl-2-methylbuta Propyl butanoate, 3-methyl butylacetate, butyl octanoate, pentyl hexanoate, pentyl hexanoate, pentyl hexanoate, 2-hexenyl acetate,? -Terpinyl propanoate, isobonyl propanoate, neryl propanoate, geranyl propanoate, [(Z ) -7-methyltetradec-8-enyl] acetate, decyl benzoate, ethyl octadecanoate, ethyl isoallocholate, N-octadecane, n-docosane, 4-methyldodecanoic acid, hepta-1,2,4,6-tetraene, 5-methyl-3-octene, Decene, 1,3,7-octatriene-5-in, 1,3,5,7-cyclooctatetraene, trans-ocymene, bicyclo [4.4.0.] Deca- , 7,9-pentene, 1, (E) -8, (Z) -10-hexadecatriene and 1-phenylpropan-2-one.

Accordingly, the present invention has been made based on the above-described results, and it is therefore an object of the present invention to provide a process for the production of a specific volatile substance such as ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1-ene 3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl 2-allyl-2-hydroxyacetate, Diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3 3-pentyl-1-cyclohexene, and 4-hydroxy butanoic acid lactone), 5-heptatriene, In accordance with the method invented by the present inventors by comparison, the volatile matter content screening Korea native strains of persimmon varieties melons from a melon cultivated varieties can be used as a useful biomarker.

In the present invention, "volatile matter content analysis" refers to the determination of ethyl butanoate, ethyl hexanoate, 1- (2-ethylhexyl) Butyl acetate, 1-hexyl acetate, 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, Ethyl nonanoate, oct-1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, Ethyl hexadecanoate, 1,3,5-heptatriene, 3-pentyl-1-cyclohexene (3-pen tyl-1-cyclohexene and 4-hydroxy butanoic acid lactone in the presence or absence of volatile substances and measuring the amount of volatile substances.

The analytical methods for this purpose include gas chromatography-mass spectrometry (GC / MS), electron ionization (EI), headspace gas chromatography (HS-GC-BID) spectrometry detection, and electronic nose systems, but are not limited thereto.

In the composition according to an embodiment of the present invention, the cultivar melon variety may be, but is not limited to, melon honey.

In addition,

a) separating volatile substances from each melon of cultivated melon variety and dried persimmon melon variety; And

b) The volatile substances separated from each of the melon in step a) are subjected to a reaction with ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methyl butyl 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1- Oct-1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl- Diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 3-pentyl-1-cyclohexene, and 4-hydroxy butanoic acid lactone. )to From luer binary melon cultivar varieties, including the step of comparing the at least one volatile content is selected from the group provides a method for selecting a system in Korea native persimmon melons cultivars.

In the method according to an embodiment of the present invention, volatile substances can be separated from the pericarp, flesh or core of cultivated melon cultivars and persimmon cultivars in step a).

In the method according to an embodiment of the present invention, the benzyl acetate, linalool propanoate or 4-hydroxy butanoic acid lactone is cultivated in the cultivated melon variety and It may be separated from the pericarp, flesh,

The above oct-1-en-3-yl acetate or ethyl dodecanoate may be isolated from the perianth and flesh regions of cultivated oriental varieties and oriental persimmon cultivars However,

The ethyl nonanoate may be separated from the flesh of the cultivated melon variety and persimmon cultivar of persimmon cultivar,

The 3-pentyl-1-cyclohexene may be isolated from the perianths of cultivated melon cultivars and persimmon cultivars,

The above-mentioned 1-hexyl acetate, ethyl hexadecanoate or 1,3,5-heptatriene may be used as a mixture of cultivated melon cultivars and pulp of dried persimmon cultivars And may be separated from the site,

Ethylbutanoate, ethyl hexanoate, 3-methylbutyl butanoate, 2-buthyl-4-ethyl (ethyl) benzoate, 3-acetyloxybutan-2-yl acetate or diethyl-2-allyl-2-hydroxy- pentanedioate) may be isolated from, but not limited to, cultivated melon cultivars and persimmon cultivars of persimmon cultivars.

In the method according to an embodiment of the present invention, the cultivar melon variety may be, but is not limited to, malt honey melon.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

Experimental material

In October, 2012, three harvests of Aubergine Honey and Dried Persimmon were cultivated in Nongwoo Bio packaging. Each fruit was divided into shells (peel), pulp and seed (large) parts. After sampling, the mixture was frozen with liquid nitrogen, finely ground in a mortar and stored at -80 ° C.

Volatile matter extraction and composition analysis

The prepared melon samples were collected by solid phase microextraction (SMPE). Solid phase microextraction was carried out using polydimethylsiloxane / divinylbenzene / carboxene and 50/30 μm fibers as adsorption fibers. 2 g of the sample was placed in a 10 mL glass vial and sealed with an aluminum crimp seal. Then, head space gas was collected with polydimethylsiloxane fiber while keeping room temperature. The collected fragrance components were automatically injected and analyzed using an Agilent 6890N GC mainframe (Agilent, USA). Volatile components were analyzed using GC / MSD Polaris Q (Thermo Finnigan, USA). Gas chromatograph-mass spectrometer The column was HP-5 (30 m × 0.32 mm ID, film thickness 0.25 μm) under the analytical conditions. The temperature was maintained at 50 ° C. for 5 minutes and then increased to 250 ° C. at 2.5 ° C. / min and maintained at 250 < 0 > C for 5 minutes. The temperature of the injector was 200 ° C and the flow rate was 1.0 ml / min. The mass spectrum of each component was compared with the standard mass spectrum and the components were identified by comparing the standard foam with the GC retention time (RT).

Example  One. Cultivar  Melon variety Hawthorn honey from melon  Korean native species Dried persimmon melon  Analysis of Volatile Compounds to Select Varieties

 Volatile substances were separated from cultivated melon cultivars from pericarp, pulp and corn in order to screen Korean traditional varieties of dried persimmon. The presence and the amount of volatile substances were compared using gas chromatography mass spectrometry.

The flavor of the fruit enhances the flavor and affects consumers' preference. In this embodiment, volatile substances related to the fragrance of fruits reported in banana, peach, citrus, pineapple, apple, etc. have been extracted for each tissue of melon. The analysis of volatile substances for the selection of oriental persimmon varieties from cultivated melon varieties was conducted by using alcohol (9 kinds), esters (35 kinds), saturated hydrocarbons (5 kinds), unsaturated hydrocarbons (10 kinds) (2 kinds), and the presence or absence of 61 kinds of volatile substances were compared with each other (Table 1 and Table 2).

Table 1 and Table 2 show the results of analyzing the volatile substances for selecting the oriental persimmon varieties of Korean native species from cultivated melon cultivars. 2 - methyl - 1 - butyl acetate was the most abundant in cultivated melon cultivars and dried persimmon cultivars of Korean native species, accounting for 36 ~ 73% of total volatile matter. A total of 54 kinds of volatile substances were detected in the cultivar of Aubergine honey, mainly volatile substances, and volatile substances in persimmon were mainly detected in the persimmon cultivars of Korean native species, and 42 kinds of volatile substances It was found that the composition of volatile materials was significantly different in cultivated melon cultivars and dried persimmon cultivars of Korean native species.

It is also possible to use 3-methyl-1-butanol, 3,3,6-trimethylhepta-1,5-dien-4-ol, 2-propylheptanol, 2-butyloctanol, eugenol, Methyl-1-butyl acetate, ethyl-2-methyl butanoate, n-butyl acetate, n-butyl acetate, 2-methylpropanoate, 3-methylbutyl propanoate, pentyl acetate, propyl butanoate, 3-methyl butyl acetate, butyl octanoate, pentyl hexanoate, 2-phenylethyl formate [(Z) -7-methyltetradec-2-ene-1-yl] -propionamide, 8-ethylhexyl acetate, decyl benzoate, ethyl octadecanoate, ethyl isoallochlorate, 7-methyl heptadecane, n-nano decane, Hepta-1,2,4,6-tetraene, 5-methyl-3-octene, 2-methyl-2-decene, 1,3,7-octatri Ene-5-in, 1,3,5,7-cyclooctatetraene, trans-ocimene, bicyclo [4.4.0.] Deca-1,3,5,7,9-pentene, 1, ) -8, (Z) -10-hexadecatriene and 1-phenylpropan-2-one showed similar detection patterns in the cultivars of oriental melon cultivars and dried persimmon melons of Korean native species, Was extremely slight.

(4-hydroxybutanoic acid, 4-hydroxybutanoic acid), which is an ester, benzyl acetate, linalool propanoate, and lactones, are added to the cultivated melon and cultivar cultivars of persimmon, persimmon, lactone) were detected as specific varieties, and it was possible to select cultivated melon cultivars and dried persimmon cultivars.

The content of volatile substances of oct-1-en-3-yl acetate and ethyl dodecanoate, which are esters, in the perilla and flesh of cultivated melon cultivars and dried persimmon cultivars Were detected as specific changes. Thus, cultivated melon cultivars and dried persimmon cultivars could be selected.

The volatile matter content of ethyl nonanoate was found to be specifically changed in the flesh and the deep region of cultivated melon cultivars and dried persimmon cultivars. Therefore, cultivated melon cultivars and dried persimmon cultivars could be selected.

The content of 3-pentyl-1-cyclohexene (3-pentyl-1-cyclohexene) was found to be changed specifically in the pericarp of the cultivated melon variety and persimmon cultivar. Melon variety could be selected.

(1-hexyl acetate), ethyl hexadecanoate (ethyl hexadecanoate), and unsaturated hydrocarbons such as 1,3,5-heptatriene (1,3 , 5-heptatriene) were detected as specific varieties, and cultivated melon cultivars and dried persimmon cultivars could be selected.

Ethyl butanoate, ethyl hexanoate, 3-methylbutyl butanoate, 3-methylbutyl butanoate, and 2-butyl-4- 3-acetyloxybutan-2-yl acetate and diethyl-2-allyl-2-hydroxy-pentanedioate, (2-allyl-2-hydroxy-pentanedioate) was found to be specifically changed, and the cultivated melon variety and persimmon melon variety could be selected.

In the present invention, the presence or absence of volatile flavor substances and the amount of fruits in fruit melon in cultivated melon (cultivars) and dried persimmon melon (native varieties) were compared with each other, thereby providing a biomarker capable of discriminating native melon .

Claims (11)

Ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, oct-1-en-3-yl acetate 2-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl-2-allyl-2-hydroxy-pentanedioate Ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3,5-heptatriene, 1,3,5-heptatriene, (3-pentyl-1-cyclohexene) and 4-hydroxy butanoic acid lactone. The composition for screening the South Korea native strains of persimmon varieties melons from a melon cultivated varieties. The composition according to claim 1, wherein the cultivar melon is a sweet potato melon. (a) separating volatile substances from each melon of cultivated melon variety and dried persimmon melon variety; And
(b) volatile substances separated from each melon in step (a) are selected from the group consisting of ethyl butanoate, ethyl hexanoate, 1-hexyl acetate, 3- But are not limited to, 3-methylbutyl butanoate, benzyl acetate, 2-buthyl-4-ethyl benzoate, ethyl nonanoate, 1-en-3-yl acetate, 3-acetyloxybutan-2-yl acetate, diethyl- Diethyl-2-allyl-2-hydroxy-pentanedioate, ethyl dodecanoate, linalool propanoate, ethyl hexadecanoate, 1,3,5-heptatriene, 3-pentyl-1-cyclohexene, and 4-hydroxy butanoic acid lactone. acid lactone) Comparing the content of one or more volatiles selected from the group consisting of the cultivated oriental varieties. The method according to claim 3, wherein in step (a), volatile substances are separated from the cultivated melon cultivars and persimmon cultivars of persimmon cultivars. 4. The method according to claim 3, wherein the benzyl acetate, linalool propanoate or 4-hydroxy butanoic acid lactone is selected from the group consisting of cultivated oriental varieties, , Flesh, and water. 4. The method according to claim 3, wherein the oct-1-en-3-yl acetate or ethyl dodecanoate is selected from the group consisting of perilla and flesh of cultivated oriental varieties, Lt; RTI ID = 0.0 > site. ≪ / RTI > [4] The method according to claim 3, wherein the ethyl nonanoate is isolated from the flesh of the cultivated melon cultivar and persimmon cultivar of persimmon cultivar. 4. The method according to claim 3, wherein the 3-pentyl-1-cyclohexene is isolated from the perianths of the cultivated melon variety and persimmon melon variety. 4. The method according to claim 3, wherein the 1-hexyl acetate, ethyl hexadecanoate or 1,3,5-heptatriene is a cultivar melon variety And persimmon of persimmon cultivars. 4. The process according to claim 3, wherein the ethyl butanoate, ethyl hexanoate, 3-methylbutyl butanoate, 2-butyl-4-ethyl benzoate 3-acetyloxybutan-2-yl acetate or diethyl-2-allyl-2-ethylbenzoate, allyl-2-hydroxy-pentanedioate) is isolated from the deep-seated region of cultivated melon cultivars and dried persimmon cultivars. 4. The method according to claim 3, wherein the cultivar melon is a sweet potato melon.