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Plant regeneration from cultures of fused protoplasts in brood-leaved trees

抱叶树融合原生质体培养物的植物再生

基本信息

批准号：
07456067
负责人：
YOSHIZAWA Nobuo
金额：
$ 1.47万
依托单位：
Utsunomiya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07456067/
关键词：
Protoplasts Cell suspension cultures Callus Plant regeneraion Auxin Cytokinin Cell fusion Organogenesis オ-チシン

项目摘要

Protoplast fusion was done between 3 fast-grown trees, Alnus firma, Betula platyphylla, and Populus alba. The optimal condition of protoplast fusion was as follows : cell density, 5x10^4/ml ; fusion buffer, 0.5M mannitol + 2.5mM CaCl_2 ; Alternate current, 200v/cm ; AC frequency, 1MHz ; Direct current, 2kV/cm ; pluse width, 100 mu sec.This condition of protoplast fusion gave fusion ratios of 31% for Alnus firma, 34% for Betula platyphylla, and 36% for Populus alba.Cell suspension cultures derived from fused cells of Alnus x Betula developed into white cultures of Betula-like form, suggesting the hydrid cultures between Alnus and Betula.From protoplast cultures of Betula x Populus, some shoots regenerated in MS medium containing 1 mu M NAA and 0.1 mu M 4-PU,suggesting that these shoots obtained were derived from fused protoplasts between both species, because Betula species can not differentiate any shoots in Ms medium containing NAA.Fused cell cultures of Alnus x Populus developed into callus from which some shoots differentiated. These shoots rooted in MS medium containing 0.1 mu M NAA and 2.5 mu M IBA and developed into 5 plantlets with different characters.

对3种速生树种---桤木、白桦和银白杨进行了原生质体融合。原生质体融合的最佳条件为：细胞密度5 × 10 ~ 4/ml，融合缓冲液为0.5M甘露醇+2.5mM CaCl_2，交流电压200 V/cm，交流频率1 MHz，直流电压2kV/cm，电压200 V/cm。脉冲宽度100 μ sec，在此条件下融合的融合率，桤木为31%，白桦为34%，白桦原生质体培养在MS + NAA 1 μ M +4-PU 0.1 μ M的培养基中，可再生出部分芽，并可在MS + NAA 1 μ M +4-PU 0.1 μ M的培养基中再生出部分芽，在MS + NAA 1 μ M +4-PU 0.1 μ M的培养基中再生出部分芽。这表明，白桦和杨的原生质体在MS培养基中不能分化出任何芽，而桤木×杨的融合细胞培养物形成愈伤组织，并分化出一些芽。在MS + NAA0.1 μ M + IBA2.5 μ M的培养基上生根，形成5株不同性状的小植株。