JPS6260076B2 - - Google Patents
Info
- Publication number
- JPS6260076B2 JPS6260076B2 JP58194726A JP19472683A JPS6260076B2 JP S6260076 B2 JPS6260076 B2 JP S6260076B2 JP 58194726 A JP58194726 A JP 58194726A JP 19472683 A JP19472683 A JP 19472683A JP S6260076 B2 JPS6260076 B2 JP S6260076B2
- Authority
- JP
- Japan
- Prior art keywords
- fusion
- seaweed
- cell
- filaments
- culture
- 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.)
- Expired
Links
- 241001474374 Blennius Species 0.000 claims description 20
- 230000007910 cell fusion Effects 0.000 claims description 15
- 239000002202 Polyethylene glycol Substances 0.000 claims description 8
- 210000000170 cell membrane Anatomy 0.000 claims description 8
- 230000004927 fusion Effects 0.000 claims description 8
- 229920001223 polyethylene glycol Polymers 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 6
- 230000034217 membrane fusion Effects 0.000 claims description 5
- 239000000411 inducer Substances 0.000 claims description 4
- 230000001939 inductive effect Effects 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 239000013535 sea water Substances 0.000 description 10
- 210000001938 protoplast Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 210000002421 cell wall Anatomy 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 150000004804 polysaccharides Chemical class 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- 206010021033 Hypomenorrhoea Diseases 0.000 description 2
- 241000206607 Porphyra umbilicalis Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229940106157 cellulase Drugs 0.000 description 2
- 239000000819 hypertonic solution Substances 0.000 description 2
- 229940021223 hypertonic solution Drugs 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- JHJLBTNAGRQEKS-UHFFFAOYSA-M sodium bromide Chemical compound [Na+].[Br-] JHJLBTNAGRQEKS-UHFFFAOYSA-M 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 229920001221 xylan Polymers 0.000 description 2
- 150000004823 xylans Chemical class 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 229920000057 Mannan Polymers 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 101100496858 Mus musculus Colec12 gene Proteins 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 210000004423 amoeboid cell Anatomy 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- FDJOLVPMNUYSCM-UVKKECPRSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2,7, Chemical compound [Co+3].N#[C-].C1([C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)[N-]\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O FDJOLVPMNUYSCM-UVKKECPRSA-L 0.000 description 1
- 229960000633 dextran sulfate Drugs 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- WVJKHCGMRZGIJH-UHFFFAOYSA-N methanetriamine Chemical compound NC(N)N WVJKHCGMRZGIJH-UHFFFAOYSA-N 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 229960002901 sodium glycerophosphate Drugs 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- REULQIKBNNDNDX-UHFFFAOYSA-M sodium;2,3-dihydroxypropyl hydrogen phosphate Chemical compound [Na+].OCC(O)COP(O)([O-])=O REULQIKBNNDNDX-UHFFFAOYSA-M 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
- C12N5/14—Plant cells
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Cultivation Of Seaweed (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【発明の詳細な説明】
本発明は海苔の細胞融合方法、更に詳しくは、
海苔の糸状体より放出される殻胞子の細胞膜融合
させる方法に関する。[Detailed Description of the Invention] The present invention provides a method for cell fusion of seaweed, more specifically,
The present invention relates to a method for fusing cell membranes of shell spores released from seaweed filaments.
近年、遺伝子工学的手法としての細胞融合に関
する研究開発が盛んに行われるようになり、陸上
植物においては既に実用化の段階にまで成功した
例もみられている。 In recent years, research and development on cell fusion as a genetic engineering method has been actively conducted, and there have already been cases where it has reached the stage of practical application in land plants.
しかし、一方海藻類(アマノリ類も含む)にお
いては現在までのところ細胞融合についての成功
例は未だ報告されていない。 However, to date, no successful case of cell fusion has been reported in seaweeds (including laver).
このような事情は、陸上植物においては市販の
酵素剤(例えばセルラーゼ・オノヅカ、ペクトリ
アーゼ、マセロチーム)を用いて簡単にプロトプ
ラストを調製できるが海藻類、特にアマノリ類で
はそれの組織を構成している骨格成分である多糖
類が上述したような酵素剤で処理しても健全な状
態でプロトプラスト化されないことに基因してい
る。 This is because for land plants, protoplasts can be easily prepared using commercially available enzymes (e.g., cellulase, pectoliase, macerozyme), but for seaweeds, especially seaweeds, the skeleton that makes up their tissues is difficult to prepare. This is because the component polysaccharides are not converted into protoplasts in a healthy state even when treated with the enzymes mentioned above.
すなわち、アマノリ類の組織を構成している多
糖類はキシラン、マンナン及びポルフイランから
成つており、特に細胞壁は強固な繊維状に形成さ
れたキシランから成つているので上記酵素剤によ
つては加水分解されないからである。因に、現在
のところ上記3種の多糖類に対して加水分解能を
合わせ持つ酵素剤についても報告はみられない。 In other words, the polysaccharides that make up the tissues of laver are composed of xylan, mannan, and porphyrane, and the cell walls in particular are composed of xylan formed into strong fibers, so they cannot be hydrolyzed by the enzymes mentioned above. This is because it is not done. Incidentally, there are currently no reports on enzyme agents that have the ability to hydrolyze the three types of polysaccharides mentioned above.
上述したように、海苔の組織を酵素で処理する
ことにより健全なプロトプラストを調製すること
が非常に困難である現状に鑑み、本発明者は海苔
の生活史において糸状体から放出されてまもな
い、直径10〜13ミクロン程度のグニヤグニヤした
アメーバー状の細胞である殻胞子に着目し、研究
した結果、この殻胞子を融合誘導物質で処理する
と細胞膜融合が行われることの知見を得て本発明
をなすに至つた。 As mentioned above, in view of the current situation where it is extremely difficult to prepare healthy protoplasts by treating seaweed tissues with enzymes, the present inventors investigated the problem of protoplasts that are released from filaments during the life history of seaweed. As a result of our research, we focused on the shell spores, which are squishy amoeboid cells with a diameter of approximately 10 to 13 microns, and obtained the knowledge that cell membrane fusion occurs when the shell spores are treated with a fusion-inducing substance.This led us to the present invention. I arrived at the eggplant.
したがつて、本発明は上記殻胞子を用いて海苔
の細胞融合を行うための手法を提供することを目
的とする。 Therefore, an object of the present invention is to provide a method for carrying out cell fusion of seaweed using the above-mentioned conchospores.
以下本発明を詳しく説明する。 The present invention will be explained in detail below.
本発明は、海苔の糸状体より放出される殻胞子
を融合誘導物質で処理して細胞膜融合させること
を特徴とする。 The present invention is characterized in that the shell spores released from seaweed filaments are treated with a fusion-inducing substance to cause cell membrane fusion.
本発明で細胞融合に用いる殻胞子は、海苔の生
活史においてノリ網に付着して発芽して仮根を生
じ、細胞膜を形成し、分裂増殖して海苔葉体にな
るものであつて、細胞壁を形成する前の段階であ
る殻胞子は、海苔葉体から細胞壁を除去したプロ
トプラストと同様な形態にあるものと考えられ
る。 During the life history of the seaweed, the chospores used for cell fusion in the present invention attach to the seaweed web, germinate, produce rhizoids, form a cell membrane, and divide and multiply to become the seaweed thallus. The shell spore, which is the stage before the formation of a seaweed thallus, is thought to have a similar morphology to a protoplast, which is obtained by removing the cell wall from the seaweed thallus.
一般に、陸上植物における細胞融合に関する報
告によると、セルラーゼのような酵素で処理して
得られるプロトプラストは高張溶液中で処理しな
いと破裂してしまうことから、0.4〜0.7Mのマン
ニトールもしくはソルビトールを添加した高張溶
液で処理することが必要とされている。 Generally, according to reports on cell fusion in land plants, protoplasts obtained by treatment with enzymes such as cellulase burst unless treated in a hypertonic solution, so 0.4-0.7M mannitol or sorbitol was added. Treatment with hypertonic solutions is required.
しかし、このことは細胞融合させたものを再生
させる場合、培養液を高張にして培養することに
なるのでその際プロトプラストの外液の浸透圧が
高くなつてプロトプラスト表面の細胞膜が何らか
の障害を受けて正常な状態での生育を不可能にす
る要因ともなる。 However, when regenerating the fused cells, the culture medium is made hypertonic and the osmotic pressure of the external fluid of the protoplasts increases, causing some damage to the cell membrane on the surface of the protoplasts. It also becomes a factor that makes it impossible to grow under normal conditions.
ところが、本発明で細胞融合に用いる殻胞子
は、自然状態では糸状体から放出された後海水中
で浮遊してノリ網に付着することから理解される
ように、マントニールやソルビトールを添加しな
い培養液でもそれ自体破裂することがないので、
上述したような生育上の支障がみられない特性を
有する。 However, as can be understood from the fact that the chospores used for cell fusion in the present invention are released from filaments in their natural state and then float in seawater and attach to the seaweed, culture without the addition of mantonyl or sorbitol is necessary. Even liquid will not explode by itself, so
It has the characteristic that it does not cause any growth problems as described above.
海苔の糸状体から放出される殻胞子を人工的に
得るには、成熟した糸状体を人工海水中で通気撹
拌下に培養して殻胞子を放出させ(3〜4日で放
出する)、ついで殻胞子を含有する培養液を遠心
分離して残渣を採取し、この残渣に適量の滅菌人
工海水を加えて殻胞子の懸濁液を調製するとよ
い。 To artificially obtain the cholaspores released from seaweed filaments, mature filaments are cultured in artificial seawater with aeration and agitation to release the chospores (they are released in 3 to 4 days). It is preferable to centrifuge the culture solution containing the conciliar spores, collect the residue, and add an appropriate amount of sterilized artificial seawater to the residue to prepare a suspension of the conciliar spores.
次に、このようにして得られた殻胞子の懸濁液
を用いて細胞融合を行うには、該懸濁液をホール
スライドグラス上に滴下し、これに融合誘導物
質、例えばポリエチレングリコールの溶液(15〜
20%wt/wt)、高分子デキストラン硫酸、高濃度
硝酸ナトリウムもしくはHigh PH−High Ca溶
液(50mM CaCl2、PH10.5)等を室温下に40〜50
分程度放置するとよく、このポリエチレングリコ
ールのような融合誘導物質の処理により細胞融合
が起つた状態が顕微鏡下での観察で確認される。
なお、細胞融合の確認は、干田等による「植物に
おける細胞融合、“細胞”12(8)1980」に掲載され
ている細胞融合の過程を示した模式図に基づいて
行つた。因に、ポリエチレングリコールのような
融合誘導物質で処理しない場合には細胞膜融合は
実質上確認されない。 Next, in order to perform cell fusion using the suspension of conchospores obtained in this way, the suspension is dropped onto a hole slide glass, and a solution of a fusion inducer, such as polyethylene glycol, is added to the suspension. (15~
20% wt/wt), polymer dextran sulfate, high concentration sodium nitrate or High PH-High Ca solution (50mM CaCl 2 , PH10.5) etc. at room temperature for 40-50 min.
It is advisable to leave the cells for about a minute, and it is confirmed by observation under a microscope that cell fusion has occurred by treatment with a fusion-inducing substance such as polyethylene glycol.
The cell fusion was confirmed based on a schematic diagram showing the process of cell fusion published in "Cell Fusion in Plants, Cells" 12(8) 1980 by Hoshita et al. In fact, cell membrane fusion is virtually not observed unless treated with a fusion inducer such as polyethylene glycol.
上述のようにして細胞膜融合させた殻胞子を人
工海水中で培養するとその90%以上が仮根を生
じ、細胞壁を形成して分裂増殖を行つて生長し、
葉体を形成するに至る。 When the chospores with cell membrane fusion as described above are cultured in artificial seawater, more than 90% of them develop rhizoids, form cell walls, and grow by dividing and multiplying.
This leads to the formation of a thallus.
したがつて、本発明に従つて、海苔の糸状体か
ら放出される殻胞子を用いると、プロトプラスト
と同様にして細胞融合することが可能となる。 Therefore, according to the present invention, using the chelaspores released from the filaments of seaweed, it becomes possible to perform cell fusion in the same manner as protoplasts.
以下に実施例を示して本発明を更に具体的に説
明する。 EXAMPLES The present invention will be explained in more detail with reference to Examples below.
実施例
殻胞子懸濁液の調製
海苔の成熟した糸状体を、藻類の研究に一般的
に使用される下記組成の人効海水(Asp12)中で
17〜18℃の温度及び4000ルクスの照度(明期8時
間、暗期16時間)において通気撹拌下に培養し
た。Example: Preparation of schistospore suspension Mature filaments of seaweed were placed in human-strength seawater (Asp12) with the following composition, which is commonly used in algae research.
Culture was carried out at a temperature of 17-18° C. and an illuminance of 4000 lux (light period 8 hours, dark period 16 hours) with aeration and agitation.
上記培養後3〜4日で殻胞子の放出がみられた
ので培養液を遠心分離(1000r.p.m.で5分間)に
付して得られる残渣を採取し、これに滅菌した上
記人工海水の適量を加えて殻胞子懸濁液とした。 After 3 to 4 days of the above culture, the release of conchospores was observed, so the culture solution was centrifuged (1000 rpm for 5 minutes), the resulting residue was collected, and an appropriate amount of the above sterilized artificial seawater was added to this. was added to obtain a schistospore suspension.
人工海水(Asp12)の組成:
NaCl 28g
MgSO4・7H2O 7g
MgCl2・6H2O 4g
KCl 400mg
CaCl2・2H2O 1.46g
NaNO3 100mg
K2HPO4 10mg
グリセロ燐酸ナトリウム 10mg
ビタミンB12 0.02μg
ビオチン 0.1μg
チアミン 10μg
PMetal 10ml
SMetal 10ml
トリスアミノメタン 1g
蒸溜水 100ml
PH 8.0〜8.1 PMetalの組成
EDTA 1mg
H3BO3 1mg
MnCl2・4H2O 0.14mg
FeCl2・6H2O 0.05mg
ZnCl2 0.01mg
CoCl2・6H2O 4μg
CuSO4・5H2O 0.5μg
蒸溜水 1mlSMetalの組成
NaBr 1.2mg
AlCl3・6H2O 1.2mg
SrCl2・6H2O 0.6mg
NaMoO4・2H2O 0.12mg
PbCl 0.03mg
KI 1.5μg
蒸溜水 1ml
殻胞子の細胞融合
上述のようにして調製した殻胞子懸濁液の150
μlをホールスライドグラス上に滴下し、それに
下記組成のポリエチレングリコール溶液の450μ
lを融合誘導物質として徐々に加えて室温下に40
〜50分放置した。 Composition of artificial seawater (Asp12): NaCl 28g MgSO 4・7H 2 O 7g MgCl 2・6H 2 O 4g KCl 400mg CaCl 2・2H 2 O 1.46g NaNO 3 100mg K 2 HPO 4 10mg Sodium glycerophosphate 10mg Vitamin B 12 0.02 μg Biotin 0.1μg Thiamine 10μg PMetal 10ml SMetal 10ml Trisaminomethane 1g Distilled water 100ml PH 8.0-8.1 Composition of PMetal EDTA 1mg H 3 BO 3 1mg MnCl 2・4H 2 O 0.14mg FeCl 2・6H 2 O 0.05mg ZnCl 2 0 .01 mg CoCl 2・6H 2 O 4μg CuSO 4・5H 2 O 0.5μg Distilled water 1ml SMetal composition NaBr 1.2mg AlCl 3・6H 2 O 1.2mg SrCl 2・6H 2 O 0.6mg NaMoO 4・2H 2 O 0.12mg PbCl 0.03 mg KI 1.5 μg Distilled water 1 ml Cell fusion of conspores 150 mg of the conspore suspension prepared as described above
Drop 1 μl onto a hole slide glass, and add 450 μl of polyethylene glycol solution with the following composition.
Gradually add l as a fusion inducer and incubate at room temperature for 40 minutes.
Let stand for ~50 minutes.
ポリエチレングリコール溶液の組成:
ポリエチレングリコール 45g
CaCl2・2H2O 735mg
蒸溜水 100ml
上述のようにしてポリエチレングリコールで処
理したものに滅菌した上記人工海水500μlを
徐々に加え室温下に10分放置し、ついで滅菌した
人工海水を用いて緩徐に洗浄した後、上記ホール
スライドグラスを内径9cmのシヤーレに収容し、
これに滅菌した人工海水20mlを緩徐に注入し、
18℃の温度及び7000ルクスの照度(明期8時間、
暗期16時間)において培養を行つた。なお、培養
液の換水は2日毎に行つた。 Composition of polyethylene glycol solution: Polyethylene glycol 45g CaCl 2.2H 2 O 735mg Distilled water 100ml To the polyethylene glycol treated material as described above, 500 μl of the above sterilized artificial seawater was gradually added, left at room temperature for 10 minutes, and then After slowly washing with sterilized artificial seawater, the whole slide glass was placed in a shear dish with an inner diameter of 9 cm.
Slowly inject 20ml of sterilized artificial seawater into this.
Temperature of 18℃ and illuminance of 7000 lux (8 hours light period,
Culture was performed in a dark period of 16 hours). In addition, the water of the culture solution was changed every two days.
培養10日後には葉長100μ程度に生長し、培養
20日後には葉長1〜1.5mm程度に生長したことが
確認された。 After 10 days of culture, the leaves grew to a length of about 100μ, and the culture
After 20 days, it was confirmed that the leaves had grown to about 1 to 1.5 mm in length.
Claims (1)
導物質で処理して細胞膜融合させることを特徴と
する海苔の細胞融合方法。 2 融合誘導物質がポリエチレングリコールであ
る特許請求の範囲第1項記載の方法。[Scope of Claims] 1. A method for cell fusion of seaweed, which comprises treating shell spores released from filaments of seaweed with a fusion-inducing substance to cause cell membrane fusion. 2. The method according to claim 1, wherein the fusion inducer is polyethylene glycol.
Priority Applications (9)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP58194726A JPS6087791A (en) | 1983-10-18 | 1983-10-18 | Cell fusion of laver |
EP84112071A EP0141304B1 (en) | 1983-10-18 | 1984-10-09 | Method for the cell fusion of laver |
DE8484112071T DE3476558D1 (en) | 1983-10-18 | 1984-10-09 | Method for the cell fusion of laver |
AT84112071T ATE40568T1 (en) | 1983-10-18 | 1984-10-09 | METHOD OF CELL MERGING OF LETTUCE. |
NZ209829A NZ209829A (en) | 1983-10-18 | 1984-10-09 | Method for the cell fusion of seaweed |
AU34135/84A AU585486B2 (en) | 1983-10-18 | 1984-10-11 | Method for the cell fusion of laver |
CA000465577A CA1225607A (en) | 1983-10-18 | 1984-10-16 | Method for the cell fusion of laver |
KR1019840006432A KR920007397B1 (en) | 1983-10-18 | 1984-10-17 | Method for the cell fusion of laver |
ES536826A ES536826A0 (en) | 1983-10-18 | 1984-10-17 | PROCEDURE FOR THE CELL MERGER OF OVES OR EDIBLE ALGAE |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP58194726A JPS6087791A (en) | 1983-10-18 | 1983-10-18 | Cell fusion of laver |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS6087791A JPS6087791A (en) | 1985-05-17 |
JPS6260076B2 true JPS6260076B2 (en) | 1987-12-14 |
Family
ID=16329217
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP58194726A Granted JPS6087791A (en) | 1983-10-18 | 1983-10-18 | Cell fusion of laver |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS6087791A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8843057B2 (en) | 1998-09-16 | 2014-09-23 | Dialware Inc. | Physical presence digital authentication system |
US8935367B2 (en) | 1998-10-02 | 2015-01-13 | Dialware Inc. | Electronic device and method of configuring thereof |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2021036778A (en) * | 2019-08-30 | 2021-03-11 | 松田産業株式会社 | On-land aquaculture method for red alga pyropia |
-
1983
- 1983-10-18 JP JP58194726A patent/JPS6087791A/en active Granted
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8843057B2 (en) | 1998-09-16 | 2014-09-23 | Dialware Inc. | Physical presence digital authentication system |
US8935367B2 (en) | 1998-10-02 | 2015-01-13 | Dialware Inc. | Electronic device and method of configuring thereof |
Also Published As
Publication number | Publication date |
---|---|
JPS6087791A (en) | 1985-05-17 |
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