JPH022343A - Method for culturing dyestuff-producing plant cell - Google Patents
Method for culturing dyestuff-producing plant cellInfo
- Publication number
- JPH022343A JPH022343A JP63141788A JP14178888A JPH022343A JP H022343 A JPH022343 A JP H022343A JP 63141788 A JP63141788 A JP 63141788A JP 14178888 A JP14178888 A JP 14178888A JP H022343 A JPH022343 A JP H022343A
- Authority
- JP
- Japan
- Prior art keywords
- culturing
- light
- dyestuff
- cells
- pigment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000012258 culturing Methods 0.000 title claims abstract description 12
- 238000000034 method Methods 0.000 title claims description 6
- 239000000975 dye Substances 0.000 title abstract 5
- 239000000049 pigment Substances 0.000 claims description 13
- 239000001056 green pigment Substances 0.000 claims description 11
- 230000001678 irradiating effect Effects 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 description 19
- 241000196324 Embryophyta Species 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 239000002609 medium Substances 0.000 description 5
- 206010020649 Hyperkeratosis Diseases 0.000 description 4
- 244000178231 Rosmarinus officinalis Species 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 239000003375 plant hormone Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 229930002875 chlorophyll Natural products 0.000 description 2
- 235000019804 chlorophyll Nutrition 0.000 description 2
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 2
- 239000006870 ms-medium Substances 0.000 description 2
- 239000005971 1-naphthylacetic acid Substances 0.000 description 1
- IIDAJRNSZSFFCB-UHFFFAOYSA-N 4-amino-5-methoxy-2-methylbenzenesulfonamide Chemical compound COC1=CC(S(N)(=O)=O)=C(C)C=C1N IIDAJRNSZSFFCB-UHFFFAOYSA-N 0.000 description 1
- NEZONWMXZKDMKF-JTQLQIEISA-N Alkannin Chemical compound C1=CC(O)=C2C(=O)C([C@@H](O)CC=C(C)C)=CC(=O)C2=C1O NEZONWMXZKDMKF-JTQLQIEISA-N 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 101000588230 Homo sapiens N-alpha-acetyltransferase 10 Proteins 0.000 description 1
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 1
- 241000207923 Lamiaceae Species 0.000 description 1
- 241001071917 Lithospermum Species 0.000 description 1
- 102100031641 N-alpha-acetyltransferase 10 Human genes 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- 235000009337 Spinacia oleracea Nutrition 0.000 description 1
- 244000172533 Viola sororia Species 0.000 description 1
- UNNKKUDWEASWDN-UHFFFAOYSA-N alkannin Natural products CC(=CCC(O)c1cc(O)c2C(=O)C=CC(=O)c2c1O)C UNNKKUDWEASWDN-UHFFFAOYSA-N 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 1
- 239000004062 cytokinin Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 1
- 229960001669 kinetin Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 210000004694 pigment cell Anatomy 0.000 description 1
- 239000001057 purple pigment Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 235000021012 strawberries Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は色素産生植物細胞を培養する方法に関し、特に
植物細胞培養による緑色色素の生産に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field] The present invention relates to a method for culturing pigment-producing plant cells, and in particular to the production of green pigments by culturing plant cells.
植物細胞培養による色素生産は、ムラサキの色素産生細
胞を用いた紫色色素（シコニン）の例など多くの報告が
ある。There are many reports on pigment production by plant cell culture, including the production of a purple pigment (shikonin) using pigment-producing cells of the purple violet.
植物の組４片（葉、茎９根など）を適当な植物ホルモン
条件下におくと、カルスと呼ばれる不定形の細胞塊が生
じること及びその時、光照射条件下では、時々緑色のカ
ルスが生じることも知られている。しかしながら、−ｆ
ｆにカルス中の色素などの二次代庫産物は、親植物のそ
れに比べて著しく低い。When a set of 4 pieces of a plant (leaves, stems, 9 roots, etc.) is placed under suitable plant hormone conditions, an amorphous cell mass called a callus will be produced, and at that time, under light irradiation conditions, green callus will sometimes occur. It is also known that However, −f
The secondary storage products such as pigments in the callus are significantly lower than those of the parent plant.
本発明は上記技術水準に鑑み植物細胞の増殖量を増すと
共に、その中に含まれる色素含有量を増加させる色素産
生植物細胞の培養方法を提供しようとするものである。In view of the above-mentioned state of the art, the present invention seeks to provide a method for culturing pigment-producing plant cells that increases the amount of proliferation of plant cells and also increases the pigment content contained therein.
本発明は誘導・選抜された緑色色素産生細胞を、第１段
階では比較的弱い光ｉ！１ｌｌｌ射条件下で、第２段階
では強い光照射条件下で培」１することを特徴とする色
素産生植物細胞の１８４方法である。The present invention uses relatively weak light i! to induce and select green pigment-producing cells in the first stage. This is a 184 method for culturing pigment-producing plant cells, which is characterized by culturing them under irradiation conditions, and in the second step under conditions of strong light irradiation.
本発明を更に具体的に説明すると、本発明は■選抜され
た色素産生細胞を、それが誘導された培地条件下で、先
ず、約３０００／レツクスの光照射下で約２〜３週間培
養し、■その後、この培養Ｍ胞を約１ａ、ｏｏｏルック
ス以上の強い光照射下で培養するものである。To explain the present invention more specifically, the present invention consists of: (1) First, the selected pigment-producing cells are cultured for about 2 to 3 weeks under light irradiation of about 3000 cells/rex under the medium conditions in which they were induced. , (2) Thereafter, this cultured M cell is cultured under strong light irradiation of about 1a, ooo lux or more.
以上のように光照射条件を２段階に分けて培養すること
により、細胞の増殖量で２〜３倍、さらに緑色色素含有
率で３〜４倍の高い値を得ることができる。By culturing under two stages of light irradiation conditions as described above, it is possible to obtain a 2 to 3 times higher cell proliferation rate and a 3 to 4 times higher green pigment content.
光照射下で、緑色色素細胞が、誘導増殖できることは、
既に公知であるが、一方これを単に強い光照射下で培養
しても、その増殖量が上がらないばかりか、時には細胞
自身が死に到ることも起こる。そこで、本発明では比較
的弱い光照射下で、細胞の増殖並びに緑色色素の代謝系
の各器官及び酵素系の機能を十分に強くしておいてから
、それを次の強い光照射下に移すことにより、細胞の増
殖並びに色素含有率を上げようとしたものである。The fact that green pigment cells can be induced to proliferate under light irradiation is
Although it is already known, simply culturing these cells under strong light irradiation not only does not increase the amount of proliferation, but also sometimes causes the cells themselves to die. Therefore, in the present invention, the functions of each organ and enzyme system of the cell proliferation and green pigment metabolic system are sufficiently strengthened under relatively weak light irradiation, and then transferred to the next strong light irradiation. This was an attempt to increase cell proliferation and pigment content.
〔実施例１〕
シソ科のローズマリーを対象に、通常の植物細胞寒天培
養培地（ムラシゲ・スコッグ培地、以下、ＭＳ培地とい
う）に植物ホルモンとして、オーキシンとして１−ナフ
タリン酢酸（以下、ＮＡＡと略す）を１０−ｓ’ｒφ、
サイトカイニンとしてカイネチン（以下、Ｋと略す）を
１０〜１０−６ル４ ｆｆｉ加した培地で、誘導・選抜
することにより、緑色の色素を産生ずるローズマリーカ
ルスを得た。これを試料に、次のような各条件でその増
殖量及び緑色色素含有量を調べた。[Example 1] Targeting rosemary of the Lamiaceae family, 1-naphthalene acetic acid (hereinafter abbreviated as NAA) was added as a plant hormone and as auxin to a normal plant cell agar culture medium (Murashige-Skogg medium, hereinafter referred to as MS medium). ) is 10-s'rφ,
Rosemary callus producing green pigment was obtained by induction and selection in a medium supplemented with 10 to 10<-6>l of kinetin (hereinafter abbreviated as K) as a cytokinin. Using this as a sample, its proliferation amount and green pigment content were examined under the following conditions.
（期間３週間毎に植継トータル６週間、温度２５°Ｃ１
光源：白色蛍光灯）
条件１：光照射 ３０００／１／ツクス条件２：光照射
１ （ＬＯ００／ｌ／ツクス条件３：光照射 ３００
０ルツクス＋光照射 ５，０００ｚｙノクス条件４：光
照身 ３０００Ｍンクス＋光照射 ＨＬＯＯＯ肩クスそ
のり果を表−１に示す。(Transplanting period every 3 weeks, total 6 weeks, temperature 25°C1
Light source: white fluorescent lamp) Condition 1: Light irradiation 3000/1/Tux Condition 2: Light irradiation 1 (LO00/l/Tux Condition 3: Light irradiation 300
0 lux + light irradiation 5,000 zynox Condition 4: light shine 3000M lux + light irradiation The results of HLOOO shoulder x are shown in Table-1.
表−１光の２段照射の効果
照射の２段階法の効果について調べた。（カッコ内は色
素産生細胞の培養ホ・シモン条件を示す）表−２にその
結果を示す。（期間：３週間毎に植継ドータ／Ｌ／６週
間、温度＝２５°Ｃ９光源：白色蛍光灯）
表−２各植物細胞への光２段照射の効果〔実施例２〕
本発明の効果を、基本培地は実施例１と同じにして、他
の６α物の緑色色素産生細胞について調べた。Table 1: Effect of two-stage irradiation of light The effects of the two-stage method of irradiation were investigated. (The numbers in parentheses indicate the culture conditions for pigment-producing cells.) Table 2 shows the results. (Period: Transplanted daughters/L/6 weeks every 3 weeks, Temperature = 25°C9 Light source: White fluorescent lamp) Table 2 Effects of two-stage light irradiation on each plant cell [Example 2] Effects of the present invention Using the same basic medium as in Example 1, other 6α green pigment producing cells were investigated.
対象として、■ローズマリー（ＮへＡ１０””モ／Ｌ’
／ Ｌ 、 Ｋ ： １０−’〜１０−６モＩＶ／Ｌ
） 、■ホウレンソウ（ＮＡ八： １ｏ １ｔ／ｌ
、に： １ｏ−’Ｍ／ｌ） 、■タバコ（Ｎ Ａ Ａ
： １０−’ Ｍ／ｌ、 Ｋ ：１０−６Ｍ／ｌ）ｌ■
イチゴ（ＮＡＡ：１０−５Ｍ／Ｌ、ベンジル・アデニン
ニ１ａ−’Ｍμ）をとシあげ、光〔実施例３〕
上記の例は固体培地（寒天使用）のケークであるが、液
体培養についても同様の効果が得られている。ローズマ
リーの色素産生細胞１゜０２を２−角程度に細分後、３
００−フラスコ中に液穢１０ ＱｍＺ（ＭＳ培地に植物
ホルモン二ＮＡＡ１０〜’ Ｍ／ｚ 、 Ｋ １０−’
Ｍ／ｌを添加）に入れ、回転培養（１００ｒｐｍ ）
を行い、その際の照射条件を上記（実施例１）と同様に
変化させた。As a target, ■Rosemary (N to A10""Mo/L'
/L, K: 10-'~10-6mo IV/L
),■Spinach (NA8: 1o 1t/l
, to: 1o-'M/l), ■Tobacco (N A A
: 10-' M/l, K: 10-6 M/l)l■
Strawberries (NAA: 10-5M/L, Benzyl adeninii 1a-'Mμ) were roasted and exposed to light [Example 3] The above example is a cake on a solid medium (using agar), but the same applies to liquid culture. effects have been obtained. After subdividing 1°02 of rosemary pigment-producing cells into approximately 2-square pieces,
00-Liquid 10QmZ in a flask (plant hormone two NAA10~' M/z, K10-' in MS medium
Add M/l) and rotate culture (100 rpm).
was carried out, and the irradiation conditions at that time were changed in the same manner as described above (Example 1).
その結果、３０００ルツクス単独ＶＣものと比べ、５０
００ ／１／ツクス＋１１１０００ルックスの２段階照
射では、増殖量で約３倍、クロロフィル含清で約五５イ
＆となった。As a result, compared to 3000 lux single VC, 50
In the two-stage irradiation of 00/1/tux + 111,000 lux, the amount of proliferation was about 3 times greater, and the amount of chlorophyll content was about 55 I&.
緑色色素産生細胞を培責する際、本発明のように光を２
段階（第１段階では、３０００ルック７−程度、第２段
階では強い光として１１０００ルックス以上）に照射す
ることにより、細胞の増殖量を約２〜３倍に上昇し、か
つその中に含まれる緑色色素（主としてクロロフイＩｖ
）の含有量も３〜４倍に上げることができる。このこと
は実際の物質生産の場で考えると、各々の効果のかけ合
せて効果が吊るだめ、通常の培養の場合に比べ色素生産
量が、約１０倍になる効果がある。When culturing green pigment producing cells, light is applied at 2 times as in the present invention.
By irradiating at a stage (1st stage, about 3000 lux 7-, and 2nd stage irradiation with strong light of 11000 lux or more), the amount of cell proliferation is increased by about 2 to 3 times, and the Green pigment (mainly chlorophyll IV)
) content can also be increased 3 to 4 times. When this is considered in the context of actual substance production, the effects of each effect are multiplied, and the amount of pigment produced is approximately 10 times greater than in the case of normal culture.
Claims (1)
較的弱い光照射条件下で、第２段階では強い光照射条件
下で培養することを特徴とする色素産生植物細胞の培養
方法。A method for culturing pigment-producing plant cells, which comprises culturing induced and selected green pigment-producing cells under relatively weak light irradiation conditions in the first step and under strong light irradiation conditions in the second step.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63141788A JPH022343A (en) | 1988-06-10 | 1988-06-10 | Method for culturing dyestuff-producing plant cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63141788A JPH022343A (en) | 1988-06-10 | 1988-06-10 | Method for culturing dyestuff-producing plant cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH022343A true JPH022343A (en) | 1990-01-08 |
Family
ID=15300169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63141788A Pending JPH022343A (en) | 1988-06-10 | 1988-06-10 | Method for culturing dyestuff-producing plant cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH022343A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007244240A (en) * | 2006-03-14 | 2007-09-27 | Hamamatsu Kagaku Gijutsu Kenkyu Shinkokai | Method for producing green callus cell |
| CN112386627A (en) * | 2019-08-14 | 2021-02-23 | 大江生医股份有限公司 | Application of rosemary callus juice and culture medium for inducing rosemary callus |
-
1988
- 1988-06-10 JP JP63141788A patent/JPH022343A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007244240A (en) * | 2006-03-14 | 2007-09-27 | Hamamatsu Kagaku Gijutsu Kenkyu Shinkokai | Method for producing green callus cell |
| CN112386627A (en) * | 2019-08-14 | 2021-02-23 | 大江生医股份有限公司 | Application of rosemary callus juice and culture medium for inducing rosemary callus |
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