KR101791563B1 - Method for delaying the ripening of fruits using sound treatment - Google Patents

Abstract

The present invention relates to a method for delaying the maturation of fruit by a sound wave of a specific band, and more specifically, to a method of delaying maturation of fruit by suppressing the expression of an ethylene biosynthesis gene, And it can be used for long-term storage and long-term storage.

Description

[0001] The present invention relates to a method for delaying the ripening of fruits using sound waves,

The present invention relates to a method for delaying maturity of a fruit by sonic wave processing in a specific range.

Fruits are classified into two types according to respiration pattern during ripening. Tomato, apple, peach and others are belong to Climacteric type, and respiration increases rapidly during aging, and thus the amount of ethylene is rapidly increased. On the other hand, non-climacteric type fruits such as grapes and citrus have little change in respiration, and thus the generation of ethylene is minimal.

Ethylene is a plant hormone involved in various processes such as fruit ripening, seed germination, fruit and flower, aging of leaves and desertification of leaves and flowers. Ethylene is an essential hormone for fruit ripening and prevents the production of ethylene. If the ethylene is not recognized, the fruit is not matured properly. If ethylene production is suppressed, the aging is delayed and the storage period is prolonged. Fruits grow rapidly in the presence of ethylene and reach a state of maturation suitable for eating. After this stage, the quality degradation process called aging proceeds. Techniques for inhibiting the synthesis and action of ethylene for the maintenance and long-term storage of fruits and crops have been studied for a long time and have been widely used commercially (Postharvest Biology and Technology, vol. 43, no. 1, 27 (2007); Journal of experimental botany, vol.57, no.12, pp.3313-3325 (2006)).

Japanese Patent Laid-Open Publication No. 2005-117919 relates to a method for inhibiting ethylene production in plants, wherein one or two or more metal ions selected from the group consisting of Ni, Cu and Co are absorbed from the outside of the plant, And inhibiting the formation of ethylene in the plant. However, the technology to inhibit plant ethylene production through sonication is unknown.

Accordingly, the inventors of the present invention have completed the present invention by confirming that the maturation of the fruit is delayed by treating plants with a sound wave of a certain band to inhibit ethylene synthesis.

Japanese Patent Application Laid-Open No. 2005-117919

Postharvest Biology and Technology, vol.43, no.1, pp.23-27 (2007) Journal of experimental botany, vol.57, no.12, pp.3313-3325 (2006)

It is an object of the present invention to provide a method for delaying the maturation of fruits using sound waves.

The present invention provides a method of delaying the maturation of fruits using sound waves.

Specifically, the present invention provides a method for delaying the maturation of fruit using a sound wave comprising treating the plant with a sound wave of 900 Hz to 1200 Hz, preferably 900 Hz to 1100 Hz.

The sound wave process preferably processes sound waves of 900 Hz to 1100 Hz with a sound volume of 50 to 100 dB for 4 to 8 hours and a sound volume of 80 dB (decibel) for 6 hours at 1000 Hz But is not limited thereto.

In one embodiment of the present invention, the inventors of the present invention conducted experiments to determine the optimum range and processing time for the maturation delayed tomatoes in a range of 0 Hz, 250 Hz, 500 Hz, 800 Hz, 1 kHz and 1.5 kHz A single sonic wave was processed for 1, 3, 6, 18 hours and the color change of tomato was observed. As a result, the incubation delay effect was the best when treated at 1 kHz singe wave condition for 6 hours, and the delayed incubation effect at other single sound condition and treatment time was minimal (Fig. 1). In addition, when treated with a 1 kHz single sound wave condition for 6 hours, the delayed effect was about 20 ~ 30% as compared with the control (Fig. 2).

The term "sound wave" used in the present invention refers to a phenomenon in which a compressed portion having a relatively high air density formed by the vibration of an object and a lean portion having a low air density are propagated to the outside. Depending on the frequency, the sound waves are divided into audible waves having frequencies of about 20 to 20,000 Hz, infrasonic waves having a frequency of 20 Hz or less, and ultrasonic waves having a frequency of 20000 Hz or more. Generally, music is composed of composite sound waves. In the present invention, sound waves of a specific range other than the complex sound waves are used. Specific sonic processing was performed on the plant through the speakers installed in the noiseless growth phase using the Adoba Audition 3.0 software (Adobe System Company) program.

The term " ripening of fruit "as used in the present invention refers to the fact that fruit in the initial solid state undergoes a process such as reduction of organic acid, increase of pigment and increase of respiration to produce sweetness, aroma, And hemicellulose is decomposed to indicate the process of softening the fruit. In this specification, maturity of fruit is used in the same sense as aging of fruit. Changes in skin color are important indicators of the degree of ripening. In the case of tomatoes, the skin changes from green to orange to red.

The method of delaying the maturity of the fruit by using the sound wave of the present invention may be to delay the maturation of the fruit by suppressing the expression of the ethylene biosynthesis-related gene by treating the sound wave.

The ethylene biosynthesis-related gene may be any one or more genes selected from the group consisting of LeACS2, LeACS4, and LeACO1.

The plant may be an fruit of a plant. In one embodiment of the present invention, tomato fruit in the mature green stage was harvested and sonicated to tomato fruit.

The fruit is preferably any one selected from the group consisting of tomatoes, apples, pears, bananas, peaches, plums, blueberries, mangoes, apricots, kiwi, pineapples, kiwi, persimmons, melons, avocados and figs, Most preferred, but not limited thereto.

As used herein, the term "ethylene" refers to a hormone that promotes aging of fruit, flowers, and leaves, and is referred to as a ripening hormone or an aging hormone. The inhibition of ethylene production slows down the aging process and thus the plant's shelf life, which is very useful for the maintenance of long-term storage and long-term storage of fruits. For this reason, techniques for inhibiting the synthesis and action of ethylene have been widely used commercially (Postharvest Biology and Technology, vol.43, no.1, pp.23-27 (2007); J. Experimental Botany, vol.57, no. 12, pp. 333-3325 (2006)). Since the maturation and maturation of agricultural products after harvest are closely related to the production of ethylene, the present invention analyzes the expression pattern of key genes involved in the synthesis of ethylene after the sonication to determine whether or not the sonication can inhibit ethylene biosynthesis Respectively.

In the first step of ethylene biosynthesis pathway, the precursor S-adenosyl methionine (SAM) is converted to ACC (1-aminocycloprapane-1-carboxylic acid) by ACC synthase (ACS) And ACC, which is converted in the second step, synthesizes ethylene as the final product by ACC oxidase (ACO). Two enzymes, ACS and ACO, play a key role in ethylene biosynthesis. Ethylene biosynthesis in tomato fruit is controlled by two systems, System 1 and System 2. System 1 is auto-inhibitory in mature growth and is present at basal levels in all tissues, including non-ripening fruit. LeACS1 (Lycopersicon esculentum ACS1) and LeACS6 (Lycopersicon esculentum ACS6) two genes are associated with one system. System 2 and functions while the fruits and petals of aging, here are LeACS2 (Lycopersicon esculentum ACS2) and LeACS4 (Lycopersicon esculentum ACS4). Two genes are involved. The ripening of respiratory spikelets and senescence of petals are promoted by System 2, and the expression of ACS and ACO genes is essential. The inventors of the present invention observed that ACS and ACO gene expression was suppressed by treating various wave ranges of Arabidopsis thaliana in the prior art. LeACO2 and LeACS4 genes are also closely related to the ripening of tomato fruit (Leuc01 gene). (The New phytologist, vol.203, no.1, pp.206-218 (2014); Science, vol.306, no.5701, pp.1513-1515 (2004); Journal of experimental botany, vol.60, no.12, pp.3311-3336 (2009)). Among ACO genes, ACO1 and ACO4 genes are known to be closely related to ripening. In particular, ACO1 gene is a major gene among ACO genes expressed in aged tomato fruit.

In one embodiment of the present invention, in order to investigate the effect of sound waves by molecular biologically identifying the cause of the delay of incubation, tomatoes that have been subjected to the sonication treatment are periodically sampled while preserving the fruits, and the ethylene synthesis related genes Were analyzed by Q-PCR (Quantitative PCR, Real-time PCR). In particular, we analyzed the expression patterns of five genes, LeACS1, LeACS6, LeACS2, LeACS4 and ACO, which play a key role in ethylene biosynthesis associated with aging. As a result, the LeACS1 and LeACS6 genes, which are involved in System 1, which is present at a basal level in all tissues including non-ripnening fruit, are not significantly affected by sonication (Fig. 3). However, the expression levels of LeACS2, LeACS4, and LeACO1 genes, which are representative genes involved in System 2, were significantly reduced by sonication (Figs. 4 and 5).

Therefore, it was concluded that the treatment of plants with the sound wave according to the present invention resulted in a decrease in the expression of LeACS2, LeACS4 and LeACO1 genes, resulting in a decrease in ethylene synthesis and a delay in the overall aging process.

Using the method of delaying maturation of fruit using the sound wave of the present invention can delay the maturing of short fruit after the harvest and shorten the shelf life of the fruit so as to reduce the economic loss, keeping freshness and long-term storage.

FIG. 1 is a graph comparing the degree of ripening of tomato fruit after 14 hours of storage at room temperature for 6 hours in a variety of single sonic waves (0, 250 Hz, 500 Hz, 800 Hz, 1 kHz, 1.5 kHz).
Fig. 2 is a photograph showing a comparison of the color change of tomato fruits by sonic treatment: Fig.
No processing, no processing of sound waves; And
Sonication, 1 kHz single sound wave for 6 hours.
3 is a diagram showing the expression patterns of LeACS1A and LeACS6 genes by sonication:
Control, do not process sound waves; And
Sound treatment, 1 kHz single sound wave for 6 hours.
FIG. 4 is a diagram showing the expression patterns of LeACS2 and LeACS4 genes by sonication:
Control, do not process sound waves; And
Sound treatment, 1 kHz single sound wave for 6 hours.
5 is a diagram showing the expression pattern of LeACO1 gene by sonic treatment:
Control, do not process sound waves; And
Sound treatment, 1 kHz single sound wave for 6 hours.

Hereinafter, the present invention will be described in more detail in the following examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

< Example  1> sonicated Tomato fruit  Ready

<1-1> Preparation of tomato

In the greenhouse of the Yongin farmhouse in Gyeonggi-do, around mid-April 2014, about 50 days after blooming, the harvested tomatoes ( Solanum lycopersicum ) were used for the experiment.

<1-2> Sound wave processing

Tomatoes selected in Example <1-1> were appropriately placed in a silent growth chamber (manufactured by Kagaku Kagaku Kogyo Co., Ltd.) equipped with a loudspeaker and treated with sound waves. For the sound processing, five kinds of single sound waves of 250 Hz, 500 Hz, 800 Hz, 1 kHz and 1.5 kHz were processed for 1, 3, 6 and 18 hours, , dB) was fixed at 80 dB.

< Example  2> Tomato fruit  Mature delay effect test

In the mature green stage tomatoes selected to select the optimum range and processing time for delayed maturation, a single sound wave of various frequencies such as 250 Hz, 500 Hz, 800 Hz, 1 kHz, , 6, and 18 hours, and then the color change of tomato was observed periodically while keeping at room temperature. Tomatoes with sonic waves were observed at regular temperature and stored at 24 ℃. The photographs were photographed using a digital camera (EOS550D, Canon, Japan) at the same place. Changes in skin color are important indicators of the degree of ripening. In the case of tomatoes, the skin changes from green to orange to red.

As a result of observing the color change of tomato according to the time of single sonication, the treatment effect of single sonic wave for 1, 3, and 18 hours was not good or the result was not better than the control group. Therefore, in the following experiment, the processing time of a single sound wave is set to 6 hours.

FIG. 1 graphically shows the ratios of the tomato skin color after 14 days from room temperature storage after treating each single sound wave for 6 hours. Though the degree of color change of tomatoes was different according to the treated sonic waves, the delayed effect was the best when treated with 1 kHz single sound wave for 6 hours (Fig. 1).

FIG. 2 is a photograph showing changes in the skin color after 2 days, 5 days, 7 days, 10 days and 14 days while keeping the tomato fruit treated for 6 hours at 1 kHz singular sound condition at room temperature. As a result of statistical analysis of the pericarp color of tomato fruit, about 20 ~ 30% of delayed maturation was observed at 1 kHz compared with no treatment (Fig. 2).

< Example  3> Analysis of Expression Pattern of Ethylene Synthesis Related Gene by Sonication

In order to investigate the effect of sonic wave by investigating the cause of delayed maturation, we analyzed the mutation pattern of ethylene-related gene closely related to maturation by Q-PCR Quantitative PCR, Real-time PCR).

<3-1> Selection of gene to analyze expression pattern

Two enzymes, 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and ACC oxidase (ACO), play a key role in ethylene biosynthesis.

Ethylene biosynthesis in tomato fruit is controlled by two systems, System 1 and System 2. System 1 is located at a basal level in all tissues, including non-ripening fruit, which operates on mature growth and irrespective of maturation. Two genes, LeACS1 and LeACS6, are associated with System 1. System 2 functions during aging of fruit and petals, in which two genes, LeACS2 and LeACS4, are involved and the expression of ACS and ACO genes is essential.

There have been many reports that the LeACO1 gene is closely related to the maturation of tomato fruit with LeACS2 and LeACS4 genes (Journal of Experimental Botany, vol. 53, No. 377, pp. 2057-2063 (2002); Science , Journal of Experimental Botany, vol. 60, no. 12, pp. 3311-3336 (2009); The New Phytologist, vol. 203, vol. 306, no. 26, pp. 1513-1515 no. 1, pp. 206-218 (2014)). The ACO1 gene is a major gene among the ACO genes expressed in aged tomato fruit.

Therefore, we analyzed the expression patterns of 5 genes, LeACS1, LeACS6, LeACS2, LeACS4 and ACO, which play a key role in ethylene biosynthesis associated with maturation.

<3-2> Tomato fruit  Sound wave processing condition

The tomato fruit was sonicated under the condition of sonic wave treatment which showed the best effect in <Example 2>. Specifically, tomatoes were treated with a 1 kHz single sound wave for 6 hours after harvesting according to the experimental method described in Example 1. The sonicated tomato fruits were sampled periodically while being stored, and the expression patterns of the five genes of Example <3-1> were analyzed by Q-PCR.

More specifically, tomato fruit of the Mature Green stage was treated at 1 kH for 6 hours and then stored for 14 days. The sample was periodically sampled and immediately frozen in liquid nitrogen and stored at -121 ° C. until used in the experiment. Tomato RNA was extracted with Plant RNeasy extraction kit (Qiagen) using 100 mg of the crushed tomato powder. CDNA was synthesized with ampfiRivert Platinum cDNA Synthesis Master Mix (GenDEPOT, South Korea) using 2 ㎍ of total RNA extracted. Real-time quantitative PCR (qPCR; StepOnePlus ™ Real-Time PCR Systems, Applied Biosystems) was performed with 30 ng of synthesized cDNA using SYBR Premix Ex TaqTM kit (TaKaRa). At this time, 40 cycles were carried out under the conditions of holding stage (95 ° C for 1 minute), cycling stage (95 ° C for 5 seconds, 60 ° C for 34 seconds), and melting cycle curve (95 ° C for 15 seconds , 60 ° C for 1 minute, and 95 ° C for 15 seconds.

<3-3> Analysis of Expression Pattern of Ethylene Biosynthesis Related Gene

First, the expression of LeACS1 and LeACS6 genes related to the system 1 is maintained at a certain level in the mature green period, and the expression level is remarkably decreased when the aging starts. As shown in FIG. 3, the LeACS1 gene was increased up to day 1 after the sonication, and the LeACS6 gene was increased up to 2 days after the sonication, and then decreased thereafter. However, when treated with sonic waves, expression of LeACS1 and LeACS6 genes was repeatedly increased and decreased without a certain pattern. As a result, the influence of the sound waves on the expression of the LeACS1 and LeACS6 genes was estimated to be very weak (Fig. 3).

On the other hand, the expression pattern of LeACS2 and LeACS4 gene related to system 2 regulating fruit ripening and petal senescence differed by sonication. Specifically, the expression level of the LeACS2 gene was increased up to 3,000 times and the expression level of LeACS4 gene was increased to about 1,400 times at 12 to 14 days, Respectively. On the other hand, the expression of LeACS2 and LeACS4 genes decreased until 7 days after sonication, but the expression level of LeACS2 and LeACS4 increased slightly after that, but the increase of expression level was very small compared with no treatment 4). Therefore, we can estimate that the expression of LeACS2 and LeACS4 genes decreased and the amount of ethylene produced decreased accordingly.

In addition, as shown in FIG. 5, the ACO1 gene expression increases with the increase of ethylene synthesis as the ripening of the tomato fruit proceeds at the time of the sonic treatment, and the ACO1 gene expression is increased ) Treatment. The decrease in the expression of LeACS2 and LeACS4 genes and the decrease in the expression of ACO1 gene were thought to be responsible for inhibition of ethylene synthesis by sonication.

These results suggest that the LeACS1 and LeACS6 genes involved in System 1, which is present at a basal level in all tissues, including non-ripening fruits, regardless of maturation, Were not significantly affected. However, the expression levels of LeACS2, LeACS4 and LeACO1 genes, which are representative genes involved in System 2, were significantly reduced by sonication.

Therefore, it is considered that the synthesis of ethylene is decreased due to the remarkable decrease of expression of LeACS2, LeACS4 and LeACO1 genes in the sonication, and thus the overall aging process is delayed.

Claims (6)

A method for delaying the maturation of tomato fruit using sonic waves comprising treating the plant with a sonic wave of 900 Hz to 1200 Hz.
The method according to claim 1, wherein the sonic wave processing is a sonic wave of 900 Hz to 1100 Hz for 4 to 8 hours with a sonic size of 50 to 100 dB.
The method according to claim 1, wherein the method comprises delaying the maturation of the tomato fruit by inhibiting the expression of the ethylene biosynthesis-related gene by treating the sonic wave.
4. The method according to claim 3, wherein the ethylene biosynthesis-related gene is any one or more genes selected from the group consisting of LeACS2, LeACS4, and LeACO1.
The method according to claim 1, wherein the plant is tomato fruit of a plant. delete

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