摘要
以悬浮细胞作为靶标材料,在COLC诱导获得多倍性细胞后,经诱导分化和再生形成纯合的多倍体植株,是一条可行且高效的多倍体诱导途径。但该途径的前提是成熟、高效的单细胞悬浮培养和植株再生体系建立。鉴于此,在获得优良胚性愈伤组织的基础上,对半夏细胞悬浮培养、细胞分化及再生植株所涉及的关键问题及其影响因素进行研究,结果获得了以近圆形细胞为主的高质量单细胞悬浮系,细胞生长曲线呈典型的"S"型,细胞密度可达2.0×106以上,活细胞率可达86.80%,近圆形细胞率可达82.64%;明确了其细胞胚胎发生的两种可能途径,并通过诱导细胞分化,成功获得了愈伤组织增殖,最终实现了高效的植株再生,愈伤分化率可达94.46%,丛生芽诱导率可达337.42%,幼芽成苗率可达90.75%,幼苗生根率可达95.40%,移栽成活率可达98.83%,从而建立了高效的半夏悬浮细胞培养和植株再生体系,为后续的基于单细胞水平的半夏多倍体诱导研究提供技术参考和科学依据。
After suspended cells as target material had been induced into polyploidy cell via COLC,it was a viable and effective polyploid induction approach that polyploidy cell was induced to differentiate and regenerate into homozygous polyploid.But the premise of the approach was that mature and efficient suspended cell culture and plant regeneration system has been set up.Base on good embryogenesis callus obtained,the key problems involved in suspended cells culture,cell differentiation and regeneration plant and its influence factors were studied.High quality suspended cell lines had been obtained which was primarily comprise of suborbicular cell.Cell growth curve showed a typical ＂S＂ curve,cell density was 2.0×106 above,living cells rate was 86.80%,suborbicular cell rate was 82.64%.The two possible approaches of the cell embryogeny of Pinellia ternata were clear and depinite.The embryogenic callus proliferated successfully by inducing cell differentiation and the efficient plant regeneration was realized finally.The differentiation of callus was 94.46%,rate of cluster buds induced was 337.42%,planting percent of buds was 90.75%,rooting rate of seedling was 95.40%,survival rate after transplanting was 98.83%.The system of high-efficiency suspended cell culture and plant regeneration were established.It provided technical reference and scientific basis to the follow-up study on polyploid induced of Pinellia ternata based on the single-cell level.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2012年第10期39-49,共11页
China Biotechnology
基金
国家转基因生物新品种培育重大专项(2009ZX08005-013B)
甘肃省农业厅农业生物技术研究与应用开发项目(GNSW-2011-07)资助项目
