CN112189628A

CN112189628A - Method for judging cold and heat property degree of plant food

Info

Publication number: CN112189628A
Application number: CN202011129815.1A
Authority: CN
Inventors: 曾绍校; 蒋菊蓉; 林少玲; 黄志伟; 郭泽镔; 郑明静; 黄艳
Original assignee: Fujian Agriculture and Forestry University
Current assignee: Fujian Agriculture and Forestry University
Priority date: 2020-10-21
Filing date: 2020-10-21
Publication date: 2021-01-08
Anticipated expiration: 2040-10-21
Also published as: CN112189628B

Abstract

The invention discloses a method for judging the cold and hot property degree of plant food, which is based on the theory of traditional Chinese medicine that hot people are cold and cold people are hot, and the theory that eating cold and cold food can clear away summer heat and improve the heat resistance of an organism is taken as an animal model, and after the plant food and the components thereof to be detected are fed to fruit flies, the expression quantity of heat stress protein genes (HSF70) of the female fruit flies after heat exposure is measured and compared with a blank control group, so that the cold and hot property degree of the plant food is quantitatively judged. The drosophila melanogaster is small in size, sensitive in biological reaction and sensitive to foods with weak cold and heat medicinal properties, so that the drosophila melanogaster is used as a measuring object. The method is simple to operate, has high sensitivity, is suitable for plant foods such as tea leaves and the like and components thereof, and can be used for researching and developing formulas of herbal tea, summer heat relieving food, sports drink and the like and verifying product quality.

Description

Method for judging cold and heat property degree of plant food

Technical Field

The invention belongs to the technical field of food biology, and relates to a method for judging the cold and heat property degree of plant food.

Background

The traditional Chinese medicine holds that the food has cold, warm and hot properties like the medicine. The four properties are summarized by the reactions of ancient people in food action on the body. Cold and cold, heat and warm have certain commonalities in action, but have clear limits. Cold is second to cold and warm is second to heat, while the peace of mind lies between cold and heat. The food with the functions of clearing heat, detoxifying, removing internal heat and the like is called cold and cool food and is suitable for people with heat syndrome or heat constitution. The food with the functions of warming channels, dispelling cold, promoting blood circulation, etc. is called warm food, and is suitable for people with cold syndrome or cold constitution. At present, more methods for judging the cold and heat properties of food are provided, but most of the methods are complex to operate and have high technical requirements.

In non-patent literature, "research on quantitative formulation based on the theory of tetralogy of traditional Chinese medicine" in the world, volume 10, first phase of traditional Chinese medicine, month 12 in 2014, pages 17 to 21, discloses a quantitative research method for attributes of traditional Chinese medicine, and the research discusses a method for quantifying food attributes based on a biological metabolic heat imaging (TMI) technology, microcalorimetry, a mathematical model building technology and the like. The method classifies Chinese medicinal materials into 11 grades or 9 grades, and also quantitatively distinguishes the Chinese medicinal materials by different colors. Although the methods have certain scientificity, the operation is more complex and the technical requirements are high.

In the non-patent literature, "multivariate analysis of correlation between cold and heat properties and six types of nutrient contents of 176 common foods", vol.19, sixth phase of Chinese medicine of precious countries, 2008, 6 months, 1467 pages 1469, a method for determining the cold and heat properties of foods by establishing a mathematical model through analyzing nutrient components in 176 foods is disclosed, and the method has certain scientificity for determining the cold and heat properties of foods, but does not make further quantitative analysis on the cold and heat properties of foods, and does not consider the relationship between the types and contents of minerals in the foods and the geographic environment, so further research is needed.

The fruit fly is a completely metamorphotic insect with short growth cycle and high sensitivity, and researches show that the preferential temperature of male fruit fly is much lower than that of female fruit fly, and the female fruit fly has stronger response to the temperature than the male fruit fly. Therefore, many researches on heat resistance are carried out by selecting female fruit flies, and in order to improve the heat resistance of the female fruit flies, the heat dissipation of a pungent and cool product, the cold dissipation of a pungent and warm product, and the cold food for a cold constitution person to eat a hot food and the cold food for a hot constitution person to eat a cold food can be carried out according to the principle of ' a hot person and a cold person to heat ' and ' treating cold with heat ' in Huangdi ' internal classic, so that the heat resistance of the female fruit flies can be improved by feeding the cold food to the female fruit flies. Conversely, if the heat resistance is to be lowered, it can be fed with a hot food. Therefore, the cold and heat properties of the plant food can be qualitatively or quantitatively analyzed. According to the research, the drosophila larvae or pupae bodies under the sun irradiation express a large amount of heat stress protein and can resist the high temperature of 40 ℃. Therefore, the expression change of the heat stress protein gene is closely related to heat resistance, and when the expression of the heat stress protein gene of female fruit flies is higher after the plant foods and the components thereof are fed, the higher the heat resistance of the female fruit flies is, the higher the cold property of the plant foods is; the lower the expression of the heat stress protein gene of female fruit flies after feeding the plant food and the components thereof, the higher the heat of the plant food is. Therefore, the degree of the cold and heat properties of the plant food can be distinguished through the expression change of the heat stress protein gene of the female fruit fly after the plant food and the components thereof are fed. At present, no relevant research for quantitatively analyzing the cold and heat properties of the vegetable food by the method is reported. Therefore, it is necessary to develop a method for determining the degree of cold and heat properties of plant food.

Disclosure of Invention

The invention aims to provide a method for judging the cold and heat property degree of plant food, which is simple to operate, low in technical difficulty and easy to implement compared with other methods for judging the cold and heat property degree of food.

In order to achieve the purpose, the invention adopts the following invention technology:

a method for judging the cold and heat property degree of plant food comprises the following steps:

(1) preparing a culture medium of a sample to be detected: adding plant food into blank culture medium (distilled water, corn starch, sucrose, agar, yeast powder, propionic acid) at a ratio of 5% of the final mass fraction (based on the blank culture medium), and mashing the sample to be tested to homogenate or extracting effective components according to actual conditions before adding. The blank medium comprises distilled water 130ml, corn starch 17g, sucrose 13g, agar 1.5g, yeast powder 1.5g, and propionic acid 0.8 ml.

(2) Female fruit flies which are not mated within 10 h are selected, are randomly divided into 2 groups after anesthesia, and 50 flies in each group are raised in a blank culture medium.

(3) After 3 days of culture in the blank culture medium, respectively transferring the female fruit flies into a culture medium of a sample to be tested and the blank culture medium for 3 days of culture, wherein the fruit flies cultured in the blank culture medium are used as a blank control group, and the raising conditions are as follows: temperature 26 ℃, humidity 50%, light: dark =12 h: 12 h.

(4) The female fruit flies cultured for 3 days were transferred to an empty bottle and subjected to uniform heat exposure for 0.5 to 4 hours in a climatic chamber at (36. + -. 1). degree.C.

(5) And (3) randomly selecting 20 female fruit flies after heat exposure, filling liquid nitrogen into the female fruit flies, freezing the female fruit flies at the temperature of minus 80 ℃ for detecting the expression of the heat stress protein regulatory gene (HSF70) in the female fruit flies, and finally judging the cold and heat property degree of the plant food relative to the expression change of the HSF70 of a blank group.

The female fruit fly has stable heat resistance change and good reproducibility.

And (4) in order to reduce the problem that the female fruit flies are heated unevenly in the heat exposure experiment process, the female fruit flies are placed on an automatic circular turntable (shown in the attached drawing) for heat exposure.

The higher the expression of the heat stress protein gene (HSF70) of the female fruit fly after the plant food and the components thereof are fed in the step (5), the higher the cold degree of the plant food and the components thereof are fed, and the lower the expression of the heat stress protein gene (HSF70) of the female fruit fly after the plant food and the components thereof are fed is, the higher the heat degree of the plant food and the components thereof are fed. The specific discrimination method is as follows:

when the expression of the heat stress protein gene (HSF70) of female fruit flies is increased by 10-50% compared with that of a blank control group after the plant food and the components thereof are fed, the plant food is cold food.

② the expression of the heat stress protein gene (HSF70) of female fruit flies after the plant food and the components thereof are fed is increased by > 50 percent compared with the blank control group, which indicates that the plant food is cold food.

And thirdly, the expression of the heat stress protein gene (HSF70) of the female fruit flies fed by the plant food and the components thereof is within-10 to 10 percent compared with that of the blank control group, which indicates that the plant food is the flat food.

And fourthly, the expression of the heat stress protein gene (HSF70) of the female fruit fly is reduced by more than 10 percent compared with that of a blank control group after the plant food and the components thereof are fed, and the plant food is warm food.

The invention has the advantages that: the existing method for judging the cold and heat degree of food is based on the biological metabolic heat imaging (TMI) technology, microcalorimetry, mathematical model building technology and the like to quantify the food attributes. The operation instrument and the process are complex and insensitive, and the experiment cost is higher. The method does not need complex instrument operation, only needs to feed the fruit flies with the plant food, judges the cold and heat property degree of the plant food by comparing blank groups and using the expression of the heat stress protein gene (HSF70) of female fruit flies, and has simple and sensitive operation process and low experiment cost.

Drawings

FIG. 1 expression levels of HSP70 in the experimental group and the blank control group in example 1. Different lower case letters in the same column represent significant differences, P < 0.05.

FIG. 2 expression levels of HSP70 in the experimental group and the blank control group in example 2. Different lower case letters in the same column represent significant differences, P < 0.05.

FIG. 3 expression levels of HSP70 in the experimental group and the blank control group in example 3. Different lower case letters in the same column represent significant differences, P < 0.05.

FIG. 4 expression levels of HSP70 in the experimental group and the blank control group in example 4. Different lower case letters in the same column represent significant differences, P < 0.05.

FIG. 5 expression levels of HSP70 in the experimental group and the blank control group in example 5. Different lower case letters in the same column represent significant differences, P < 0.05.

FIG. 6 expression levels of HSP70 in the experimental group and the blank control group in example 6. Different lower case letters in the same column represent significant differences, P < 0.05.

Fig. 7 is a side view of an automatic circular turntable.

Fig. 8 is a top view of the automated circular carousel.

Detailed Description

In order to facilitate understanding of the present invention, the technical solutions of the present invention are further explained below with reference to the specific embodiments, but it should be understood that the present invention is not limited by these contents. The method specifically comprises the following steps:

example 1

(1) Preparation of a sample (white tea) culture medium to be tested: adding water into white tea, mashing, homogenizing, leaching in boiling water (water extraction method) to obtain water soluble component, and adding into blank culture medium at a ratio of 5% (dry weight).

(4) Female fruit flies cultured for 3 days were transferred to an empty bottle and subjected to uniform heat exposure for 2h in an automatic round turntable in a climatic chamber at (36. + -. 1). degree.C.

(5) And (3) randomly selecting 20 female fruit flies after heat exposure, filling liquid nitrogen into the female fruit flies, freezing the female fruit flies at the temperature of minus 80 ℃, detecting the expression of the heat stress protein regulatory gene (HSF70) in the female fruit flies by a PCR amplification method, and finally judging the cold and heat property degree of the plant food relative to the expression change of HSF70 of a blank group.

TABLE 1

As can be seen from table 1 and fig. 1, the increase of the expression of the heat stress protein gene (HSF70) of female drosophila after white tea feeding is ≧ 50% compared with the expression of the heat stress protein gene (HSF70) of female drosophila fed with the blank medium, indicating that white tea is a cold food.

Example 2

(1) Preparation of a culture medium of a sample to be tested (high fructose corn syrup): the high fructose corn syrup is added into a blank culture medium according to the proportion that the final mass fraction is 5%.

(3) After 3 days of culture in the blank culture medium, the female fruit flies are respectively transplanted into a sample culture medium to be tested and the blank culture medium to be cultured for 3 days, wherein the fruit flies cultured in the blank culture medium are used as a blank control group (breeding conditions are that the temperature is 26 ℃, the humidity is 50 percent, and the light is dark =12 h: 12 h).

(4) Female fruit flies cultured for 3 days were transferred to an empty bottle and subjected to uniform heat exposure for 3 hours in a climatic chamber at (36. + -. 1). degree.C.

TABLE 2

As can be seen from Table 2 and FIG. 2, the expression of the heat stress protein gene (HSF70) in female fruit flies after feeding high fructose corn syrup was in the range of-10 to 10% as compared with the expression of the heat stress protein gene (HSF70) in female fruit flies after feeding the blank medium, indicating that high fructose corn syrup is a warm food.

Example 3

(1) Preparation of a sample to be tested (glucose) culture medium: adding glucose into a blank culture medium according to the proportion of 5 percent of the final mass fraction.

(4) Female fruit flies after 3 days of culture were transferred to empty bottles and subjected to a uniform heat exposure for 4h in a climatic chamber at (36. + -. 1). degree.C.

TABLE 3

As is clear from Table 3 and FIG. 3, the expression of the heat stress protein gene (HSF70) in female fruit flies fed with glucose was in the range of-10 to 10% as compared with the expression of the heat stress protein gene (HSF70) in female fruit flies fed with the blank medium, indicating that glucose is a flat food.

Example 4

(1) Preparation of a sample (rock sugar) culture medium to be detected: adding crystal sugar into a blank culture medium according to the proportion that the final mass fraction is 5%.

(4) Female fruit flies after 3 days of culture were transferred to an empty bottle and subjected to uniform heat exposure for 0.5h in a climatic chamber at (36. + -. 1). degree.C.

TABLE 4

As can be seen from table 4 and fig. 4, when the expression of the heat stress protein gene (HSF70) of female drosophila after feeding crystal sugar is increased by 10% to 50% compared with the expression of the heat stress protein gene (HSF70) of female drosophila fed with blank medium, it is indicated that crystal sugar is a cool food.

Example 5

(1) Preparation of a sample (fructose) culture medium to be tested: adding fructose into a blank culture medium according to the proportion of 5 percent of the final mass fraction.

(3) After 3 days of culture in the blank culture medium, the female fruit flies are respectively transplanted into the culture medium of the sample to be tested and the blank culture medium for 3 days of culture, wherein the fruit flies cultured in the blank culture medium are used as a blank control group (the raising conditions are that the temperature is 26 ℃, the humidity is 50 percent, and the light is dark =12 h: 12 h).

TABLE 5

As is clear from table 5 and fig. 5, when the expression of the heat stress protein gene (HSF70) in female drosophila after fructose feeding was less than 10% of the expression of the heat stress protein gene (HSF70) in female drosophila fed with the blank medium, fructose was a hot food.

Example 6

(1) Preparation of a sample (Mesona chinensis) culture medium to be tested: smashing and homogenizing the Mesona chinensis Benth, extracting water-soluble components, and adding into a blank culture medium according to the proportion (by dry weight) of 5% of the final mass fraction.

TABLE 6

As is clear from table 6 and fig. 6, the increase of ≧ 50% in the expression of the heat-stress protein gene (HSF70) in female drosophila after feeding mesona blume as compared with the expression of the heat-stress protein gene (HSF70) in female drosophila fed with the blank medium indicates that mesona blume is a cold food.

The present invention has been described in detail with reference to the above embodiments, which are only exemplary of the implementation of the present invention. It should be emphasized that the disclosed embodiments do not limit the scope of the invention. Rather, methods that are modified based on the claims are included within the scope of this invention.

Claims

1. A method for judging the cold and heat property degree of plant food is characterized in that: the specific method comprises the following steps:

(1) preparing a culture medium of a sample to be detected: adding the plant food into a blank culture medium according to the proportion of 5 percent of the final mass fraction, or before adding, mashing a sample to be tested into homogenate or extracting effective components according to actual conditions;

(2) selecting female fruit flies which are not mated within 10 h, randomly dividing the female fruit flies into 2 groups after anesthesia, and feeding the female fruit flies in a blank culture medium, wherein each group comprises 50 female fruit flies;

(3) after 3 days of culture in the blank culture medium, respectively transferring the female fruit flies into a culture medium of a sample to be tested and the blank culture medium for 3 days of culture, wherein the fruit flies cultured in the blank culture medium are used as a blank control group, and the raising conditions are as follows: temperature 26 ℃, humidity 50%, light: dark =12 h: 12 h;

(4) transferring the female fruit flies cultured for 3 days into an empty bottle, and carrying out uniform heat exposure for 0.5-4h in a climatic chamber at (36 +/-1) ° C;

(5) and (3) randomly selecting 20 female fruit flies after heat exposure, filling liquid nitrogen into the female fruit flies, freezing the female fruit flies at the temperature of minus 80 ℃ for detecting the expression of the heat stress protein regulatory gene HSF70 in the female fruit flies, and finally judging the cold and heat property degree of the plant food relative to the expression change of the HSF70 of a blank group.

2. The method of claim 1, wherein: and (4) in order to reduce the problem that the female fruit flies are heated unevenly in the heat exposure experiment process, the female fruit flies are placed on an automatic circular turntable for heat exposure.

3. The method of claim 1, wherein: the discrimination method is as follows:

when the expression level of the heat stress protein gene HSF70 of female fruit flies is increased by 10-50% compared with that of a blank control group after the plant food and the components thereof are fed, the plant food is cold food;

the expression quantity of the heat stress protein gene HSF70 of female fruit flies fed by the plant food and the components thereof is increased by more than or equal to 50 percent compared with that of a blank control group, which indicates that the plant food is cold food;

thirdly, compared with the expression level of the heat stress protein gene HSF70 of female fruit flies fed by the plant food and the components thereof and a blank control group, the expression level difference is in the range of-10 to 10 percent, which indicates that the plant food is flat food;

and fourthly, the expression quantity of the heat stress protein gene HSF70 of the female fruit fly fed by the plant food and the components thereof is reduced by more than 10 percent compared with that of a blank control group, which indicates that the plant food is warm food.