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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。
It is feasible and highly efficient polyploid induction pathway to use suspension cells as a target material to induce polyploid cells to be induced by COLC, then to induce the differentiation and regeneration to form homozygous polyploid plants. However, the prerequisite for this approach is the establishment of a mature and efficient single cell suspension culture and plant regeneration system. In view of this, on the basis of obtaining excellent embryogenic callus, the key problems involved in suspension culture, cell differentiation and regenerated plantlets of Pinellia ternata and their influencing factors were studied. As a result, Mass single cell suspension system showed a typical “S” shape with a cell density of over 2.0 × 106, a viable cell ratio of 86.80% and a nearly circular cell percentage of 82.64% Two possible ways of embryogenesis, and succeeded in callus proliferation by inducing cell differentiation, finally achieving efficient plant regeneration, the callus differentiation rate can reach 94.46%, the induction rate of clustered shoots can reach 337.42% The seedling rate can reach 90.75%, the rooting rate of seedling can reach 95.40%, and the survival rate of transplanting can reach 98.83%. Thus, a highly efficient suspended cell culture and plant regeneration system was established. Polyploid induction studies provide technical reference and scientific basis.