JPS603455B2 - Manufacturing method of wasabi-like flavoring agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a wasabi-like flavor seasoning from wasabi callus.

Currently, callus culture of various plant tissues is being carried out, but callus induction of wasabi plant tissue has not yet been achieved.

The present inventors succeeded in inducing callus from wasabi plant tissue and found that the callus of the parentheses exhibits a wasabi-like flavor.

That is, plant tissues of the genus Wasabia are heated at a low temperature of 2000 q or less, preferably 10 to 15 qo, and treated with kinetin, 2,4-
It was discovered that when cultured in a callus induction medium containing D and thiamine, callus was induced after a few months and that the callus had a wasabi-like flavor.

The present invention has been made based on this discovery. Therefore, this invention cultivates plant tissue of the Brassicaceae family belonging to the genus Wasapia in a callus-inducing medium containing kinetin, 2,4-D, and thiamine at a low temperature of 2 p.m. or less to induce and induce callus. The present invention relates to a method for producing a wasabi-like flavoring material, which comprises growing the callus, and separating and collecting the grown callus from a culture solution.

The base material used for callus transfer is a basic material consisting of sugar, inorganic salts, and myo-inositol, to which kinetin, 2,4-D, thiamine, and yeast extract are added, but only this basic material is used. In this case, it is extremely difficult to transfer callus.

The basic material is one used in normal plant tissue culture, which consists of sugars such as dulcose and sucrose, and inorganic salts as shown below. Inorganic salts used: NH4N03,
KNQ, KH2P04, HB04, K1, CaC12,
M book 04, MnS○4, CuS〇4, ZnS○4, Fe
S〇4, C and CI2, N are also Md04, The tissue of young wasabi seedlings is excellent as a wasabi tissue used for callus induction.It is sufficient to use the sterile tissue of normal wasabi seedlings, but To obtain the product at will, it is desirable to germinate and grow dormant seeds.

If this method is adopted, the first step will be as follows. 1st
Process: Dormant wasabi seeds were soaked in an aqueous solution containing gibberellin for several days to awaken them from dormancy, then washed with 95% ethanol, and washed with 95% ethanol.
．． Sterilize the seeds by soaking them in 6% antiformin.

After washing, the seeds are sown on a normal sugar and inorganic salt medium, cultured in a cool room, and germinated, and seedlings are obtained after several months. Next, the seedlings were placed on the callus transfer medium, and 1
If the culture is continued for several months at a low temperature of 5 qo, callus will begin to be induced. It is necessary to induce callus in the first step at a temperature of 2.0°C or lower, preferably at a temperature of 10 to 15°C, and callus will not be induced at 30°C.

The second callus propagation step is a step in which the induced callus obtained in the first step is aerobically liquid cultured or solid cultured in a callus growth medium to produce proliferated callus (cultured cells).

The callus growth medium used in this step may be the same as the callus induction medium described above, but since coconut milk promotes callus growth, it is preferable to add 10 to 20% coconut milk to it. .

The growth temperature of callus is preferably 10 to 15°C.At low temperatures, callus will not die even if it is frozen, but its growth will simply be delayed, but when cultured at 30°C, callus will turn black and stop growing.

Therefore, the second step, like the first step, must be performed at a temperature of 20 qo or less.

It is desirable to perform callus growth culture under aerobic conditions.
The culture method may be either solid culture method or liquid culture method, but
From an industrial standpoint, liquid culture methods are preferred.

Calli grown using this liquid culture method are called cultured cells.
It has a wasabi-like flavor, and after being separated from the culture solution and collected, it can be dried and powdered as necessary to be used as a wasabi-like flavoring agent. This will be explained below using examples. Example Commercially available seeds 1 of a wasabi plant belonging to Wasabia japonica Matsmmma and a Daruma strain.
．． Seeds were immersed in a 20' gibberellin A3 solution (concentration 10Q blood) for 4 days in a cold room at 10°C to awaken the seeds from dormancy, and the seeds were washed once with 95% ethanol.
Further, the sample was sterilized by immersing it in 50% of a 0.6% antiformin solution and gently shaking it for 3 minutes.

After sterilization, the seeds were washed three times with sterilized water, and one batch of the washed seeds were sown on the base plate (agar plate medium) shown in Table 1, and left in a dry place at 10:00 for two months. As a result 1
．． Ten seedlings of Hine were obtained. Next, these were added to kinetin, 0.2 9/so, 2,4-DI. omo/so, thiamine, HCII. 0 ta/ku, yeast extract (di o
Co., Ltd.) 4.0 and 4.0 mL were respectively layered on a test tube agar medium for callus induction, which was added to the basal medium shown in Table 1, and heated to 10°C.
After culturing for three months in a cold room, callus will begin to induce. When these were further cultured under the same conditions for 6 months, callus pieces (referred to as primary callus) each having a length of approximately 1 proboscis were obtained. Six of these callus pieces were further subcultured on the ground of the above-mentioned agar tower for callus induction to allow callus to proliferate, thereby obtaining a proliferated callus (secondary callus) weighing 150 days (wet weight).

Table 1 Basic medium NH4N〇3 1.65 Yu/MuKN
03 1.90″○aOZ2
．． 2 days 20 0.44MgS04.7 days 20
0.37KH2P04
0.17th 3B03 6.
2 Monosom KI O. 8
3″Na2Moo4.2 days 20 0.2
5000 and 2.6 days 2○ 0.025Mn
S04.4 day 20 22.30uS04.5
Day 20 0.025ZnS04.4 Day 20
8.6Na2 [EDTA
37.3FeS04.7 days 20 27.8M
yo-lnositol 100Sucros
e 30 Evening/Mu Agar
lo ″pH 6.1 (sterilization 120
(°C, 15 minutes) Approximately 2.4 days (wet weight) of nuclear proliferation callus (secondary callus) was subjected to flask shake culture in a liquid medium for callus growth (composition is the same as the medium for callus induction).

Claims (1)

[Claims] 1. Cultivate plant tissues of the Brassicaceae family belonging to the genus Wasabia at a low temperature of 20°C or lower using a callus induction medium containing kinetin, 2,4-D and thiamine to induce and proliferate callus, and to induce and proliferate callus. A method for producing a wasabi-like flavoring agent, which comprises separating and collecting it from a culture solution.