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Successive Culture and Organogensis of haploid plants in Equisetum arvense L.

木贼单倍体植物的连续培养和器官发生。

基本信息

批准号：
60480014
负责人：
TAKEUCHI Masayuki
金额：
$ 3.01万
依托单位：
Saitama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480014/
关键词：
Equisetum Apogamy Shoot formation Antheridogenesis Archegogenesis Gametophyte Sucrose サイトカイニン

项目摘要

1. We have established cultured gametopluyte tissue originated from a spore of Equisetum arvense L. on the Murashige & Skoog's medium added 3% sucrose and 0.8% agar. It is very rare case to establish and subculture of haploid gametophyte in aseptic condition for a long time.2. Shoot formation occured by the effects of a low concentration of 6-benzil adenine (6-BA) on th gametophyte, however Kinetin, Zeatin and 6-PU have a little effects as compared with 6-BA.3. All tested phytohormones, IAA, NAA, ABA, 2,4-D and GA3, showed some inhibiting effects to shoot fromation.4. Morphological observation of the shoot formation was observed by routine method of light microscope, and transmit- and scanning electron microscope. Apogamy from a epidermal cell of cultured gametophyte was confirmed by these observation.5. In the light, differentiation of antheridia occured in the subcultured gametophyte on MS-medium added 3% sucrose. Many sperm cells were observed in the antheridia during subculture for a long period.6. Modification of sucrose concentration in the medium was required for archegonium formation on the cultured gametophyte. The arechegonia were observed after transplantation from 3% sucrose medium with 6-BA to 0.5%. It seems that the effect is specific in sucrose, because the effect was not appeared with many sugars other than sucrose.7. Protoplasts were isolated from cultured gametophyte tissue by digestive enzymes, and they were cultivated for long term. Regeneration of cell wall, cell division of the protoplasts and gametophyte regeneration were observed.

1.我们建立了培养的配子体组织起源于问荆孢子。在Murashige & Skoog培养基上加入3%蔗糖和0.8%琼脂。在无菌条件下长时间建立和继代培养单倍体配子体的情况非常罕见.低浓度的6-苄基腺嘌呤（6-BA）对配子体的分化有促进作用，而激动素、玉米素和6-PU对配子体分化的影响较小. IAA、NAA、阿坝、2，4-D和GA 3对不定芽的形成均有一定的抑制作用.用常规的光学显微镜、透射电镜和扫描电镜对不定芽发生进行了形态学观察。这些观察证实了配子体培养表皮细胞的无配子生殖.在光照条件下，继代培养的配子体在MS +3%蔗糖培养基上可分化出精子器。在长期的继代培养过程中，精子器中可见大量的精细胞.在培养的配子体上颈卵器的形成需要改变培养基中的蔗糖浓度。在3%蔗糖+0.5% 6-BA的培养基上移栽后，可观察到不定芽的形成。这一效应似乎是蔗糖特有的，因为除蔗糖外，其他许多糖都没有出现这种效应.用酶消化法从培养的配子体组织中分离出原生质体，并进行长期培养。观察了细胞壁再生、原生质体分裂和配子体再生。