JPS63160509A - Method for measuring activity of agricultural seed - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a method for measuring the activity of agricultural and forestry seeds.

Prior Art When preserving harvested agricultural and forestry seeds, a critical issue is whether the activity of the seeds, that is, the ability to germinate and grow, is preserved. Conventionally known methods for measuring the activity of seeds include a method of absorbing water at a constant temperature and examining the germination rate, a method of staining with a dye, and a method of measuring substances such as sugar eluted from the seeds at a constant G. .

All of these conventionally used methods require time and effort to measure, and for example, methods for determining germination rates vary depending on the type of seed. It usually takes 3-7 days. Also, the method of staining and the method of measuring eluted substances are
Requires 2-24 hours pre-treatment.

On the other hand, according to previous research, when seeds are allowed to absorb water and placed in an environment where they can germinate, adenosine triphosphate (hereinafter referred to as AT
It has been revealed that the amount of ATP (denoted as P) increases rapidly in a short period of time, and it has been suggested that this increase in ATP is closely related to the initiation of germination. In the present invention, as a result of research focusing on the phenomenon that ATPI increases after water absorption, we found that ATPfi at the initial stage of water absorption
It was found that there was a correlation between the increase in the number of seeds and the germination rate of the seeds. Next, a concrete explanation will be given using an experimental example.

[Experimental example 1. ] After harvesting in 1984, sun-dried Akihikari brown rice was heated to 80°C, and samples with different germination rates were created by varying the heating time. This sample was allowed to absorb water at 30°C, taken out every 2 hours, frozen, and crushed. 10d of distilled water was added to 10 grains of rice, and extracted by heating at 100°C for 10 minutes. The amount of ATP was measured. The experimental results are shown in FIGS. 2 and 3 of the attached drawings.

[Experimental Example 26] Koshihikari harvested in 1983 and 1984 and stored as brown rice at low temperature and room temperature was used in Experimental Example 1. ATPffi was measured under the same extraction conditions as above, but with different water absorption times. The experimental results are shown in Tables 1 and 2 of the attached drawings.

Table 1 shows the germination rate of brown rice (Koshihikari) with different storage periods and storage methods, and the amount of ATP at each 30-minute water absorption time, and Table 2 shows the ATP amount at each water absorption time for the data in Table 1.
The results of linear regression analysis of TPffi and germination rate values are shown.

From Table 1, Table 2, and Figure 2, it can be seen that the germination rate is large and small.
There is a clear difference in the pattern of increasing TP,
It was found that there is a correlation between the germination rate and the rate of increase in ATP, especially at the early stage of water imbibition (Figure 3). Therefore, for brown rice with different germination rates every 30 minutes after water absorption,
When TPffi was measured, as shown in Table 1, the greater the germination rate, the greater the rate of increase in ATP itself.

When we investigated the correlation between ATP& and germination rate at each water absorption time (Table 2), we found that ATPf between 1.0 and 1.5 hours of water absorption.
It was found that the correlation coefficient between fi and germination rate is large.

Based on these findings, it has been found that the activity of seeds can be measured by measuring the amount of ATP at the initial stage of water absorption.

Therefore, an object of the present invention is to provide a method that can quickly measure the activity of seeds by measuring the amount of increase in ATP immediately after water absorption.

Means for Solving Problem 1. As stated above, the object of the present invention is to shorten as much as possible the labor and time required for the conventional pretreatment and measurement of seed activity measurements. - In order to achieve this objective, the method for measuring the activity of seeds according to the present invention involves making the seeds absorb water at a constant temperature, freezing the seeds immediately after the water absorption to stop the biological enzyme reaction, and then A low-temperature grinding means is used to grind the seeds at a low temperature, the seeds are ground under conditions that suppress enzyme reactions, ATP is extracted from the ground seeds, the extracted ATP is reacted with an enzyme, and the ATP concentration is determined from the amount of luminescence. ,
The method is characterized in that the activity level of the seeds is measured by calculating and comparing the rate of increase or pattern of increase in ATP based on the obtained ATP degree.

In the method for measuring the activity of seeds of the present invention configured as described above, the seeds whose activity is to be measured are sequentially transferred to and processed in each treatment step, and each treatment step has a method for measuring the activity of seeds. Optimal treatment conditions are set depending on the type of seed to be treated, and therefore, the treatment operation can be performed relatively easily, and the activity can be measured quickly and accurately regardless of the type of seed.

Embodiments Hereinafter, embodiments of the present invention will be described with reference to FIG. 1 of the accompanying drawings.

FIG. 1 schematically shows an embodiment of a seed activity measuring device implementing the method according to the present invention.

The names and functions of each block are shown below.

Block 1 - water absorption means. constant temperature, e.g. 20-30
It is configured to allow the seeds to absorb water at ℃.

A timer can be provided to set the water absorption means for the seeds, so that the seeds that have absorbed water can be taken out at regular intervals.

Block 2 - Freezing means. The seeds absorbed by the water absorption means 1 are immediately frozen. It consists of a freezer and a device for spraying refrigerants such as liquid nitrogen. This freezing means 2 stops the bioenzymatic reaction of the seeds being carried out during water absorption, and facilitates the subsequent crushing operation.

Block 3 - Grinding means. The frozen seeds are crushed to improve the ATP extraction efficiency. In order to suppress physiological reactions, the ambient temperature of this crushing means 3 is set to a low temperature of 0° C. or lower.

Block 4 - Water injection device. Pre-operation to extract ATP. In order to suppress physiological reactions, the water injection temperature is maintained at approximately O'C.

Block 5 - heating device. ATP is extracted from the ground sample. It is preferable to perform the ATP extraction operation at a constant temperature, for example, by heating in boiling water, and by this heating, the ATP is extracted.
At the same time as extracting P, the enzyme reaction is stopped and deactivated. In this case, it is desirable to cool rapidly after heating, since substances that inhibit ATP-dependent fluorescent enzyme reactions will also be extracted if heated too much. Further, in order to obtain optimal conditions for ATP extraction depending on the seed, the water injection date, heating time, and amount of pulverization can be set in advance.

The extraction means consists of a water injection device 4 and a heating device 5.

Block 6 - Analyzer. By mixing the old ATP extract obtained by the extraction means (water injection device 4 and heating device 5) with a reagent (luciferin-luciferase reagent) that exhibits an ATP-dependent fluorescent enzyme reaction, and measuring the amount of luminescence. Calculate ATP concentration.

Block 7 - Processor. The ATP 1 degree data obtained by the analyzer 6 is used to calculate graphs or values as shown in FIGS. 2, 3, Tables 1 and 2. The activity level of the seeds can be determined by comparing the slope or value of this graph with data prepared in advance to determine the relationship between germination rate and ATPffi.

In the illustrated embodiment, each component of the apparatus may be a conventionally known appropriate apparatus, so a detailed explanation thereof will be omitted here.

As explained above, in the method for measuring the activity of seeds according to the present invention, an index of the activity of the seeds is obtained by measuring the amount of increase in ATP immediately after the seeds absorb water. Compared to the conventional method, it is configured as follows.
The activity of seeds can be easily measured in 2 hours, and is extremely useful in practice.

[Brief explanation of the drawing]

Figure 1 is a block diagram showing an embodiment of the present invention, Figure 2 is the water absorption time and AT of brown rice that has been heat treated to change its germination rate.
FIG. 3 is a graph showing the relationship between the increase in ATP and the germination rate at the second hour of water absorption in FIG. 2. In FIG. 1, 1: water absorption means, 2: freezing means, 3: crushing means, 4: water injection device, 5: heating device, 6: analysis device, 7: arithmetic processing device. ==force, ″+meeting (no change in contents) Fig. 1 Fig. 2 Water absorption amount (hr) Germination rate (%) Procedural amendment (method) January 29, 1988

Claims (1)

[Claims] The seeds are made to absorb water at a constant temperature, and after a certain period of time, the seeds are frozen to stop the biological enzymatic reaction.The frozen seeds are then ground under low temperature conditions using a cryo-pulverization method to suppress the enzymatic reaction. Adenosine triphosphate is crushed and extracted from the crushed seeds, the adenosine triphosphate concentration is determined by the amount of luminescence generated from the fluorescent enzyme reaction of the extracted adenosine triphosphate, and the adenosine triphosphate concentration is determined based on the obtained adenosine triphosphate concentration. A method for measuring the activity of seeds, which comprises calculating and comparing the rate of increase or pattern of increase in triphosphoric acid to measure the activity of seeds.