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首页> 外文期刊>Protoplasma >Culture and fusion of pollen protoplasts of Brassica oleracea L. var. italica with haploid mesophyll protoplasts of B. rapa L. ssp. pekinensis
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Culture and fusion of pollen protoplasts of Brassica oleracea L. var. italica with haploid mesophyll protoplasts of B. rapa L. ssp. pekinensis

机译：甘蓝型油菜花粉原生质体的培养和融合italica与B. rapa L. ssp的单倍体叶肉原生质体。北京

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摘要

Hybrid callus was formed from the successful protoplast fusion between pollen protoplasts of Brassica oleracea var. italica and haploid mesophyll protoplasts of Brassica rapa. The pollen protoplast isolation frequency in broccoli was highly related to the ratio of trinucleate pollens in the male gametophyte population. Large quantities of pollen protoplasts with high vigor could be isolated, and the isolation frequency reached up to 90% in 6.0–7.0 mm long flower buds with about 94.7% trinucleate-stage pollens. Pollen protoplasts could be collected and purified by discontinuous gradient centrifugation. In 1% Na-alginate embedding culture, cell divisions were observed but no further development was found. The haploid mesophyll protoplasts were isolated from in vitro haploid plants of B. rapa. Results strongly showed the variability in culturability of mesophyll protoplasts from different haploid lines. Both pollen protoplasts and haploid mesophyll protoplasts retained a stable round shape in the designed prefusion solution with an osmotic pressure of 0.74 osmol/kg. Polyethylene glycol was used for the protoplast fusion, and 40% polyethylene glycol 4000 enabled the highest fusion frequency of about 20%. Some postfusion protoplasts showed cell divisions up to callus proliferation. Calli were screened by random amplified polymorphic DNA analysis for their hybrid character. Results revealed the existence of the hybrid calli. Some of the hybrid calli grew well with green color and shoot primordia. According to our knowledge, this is the first report about a hybrid formation between two haploid protoplasts. Potential comprehensive applications, as well as problems of this technique, are discussed.
机译：杂交愈伤组织是由甘蓝变种花粉原生质体之间成功的原生质体融合形成的。芸苔的italica和单倍体叶肉原生质体。西兰花中花粉原生质分离的频率与雄配子体种群中三核花粉的比例高度相关。可以分离出大量高活力的花粉原生质体，在6.0-7.0 mm长的花芽中，分离频率高达90％，其中三核阶段的花粉约为94.7％。花粉原生质体可以通过不连续的梯度离心来收集和纯化。在1％海藻酸钠包埋培养物中，观察到细胞分裂，但未发现进一步的发育。从B. rapa的体外单倍体植物中分离出单倍体叶肉原生质体。结果强烈显示了来自不同单倍体系的叶肉原生质体可培养性的差异。花粉原生质体和单倍体叶肉原生质体在设计的预融合溶液中均保持稳定的圆形，渗透压为0.74 osmol / kg。聚乙二醇用于原生质体融合，而40％的聚乙二醇4000可实现约20％的最高融合频率。一些融合后的原生质体显示细胞分裂直至愈伤组织增殖。通过随机扩增的多态性DNA分析筛选愈伤组织的杂种特性。结果揭示了杂种愈伤组织的存在。一些杂种愈伤组织生长良好，并带有绿色和芽原基。根据我们的知识，这是有关两个单倍体原生质体之间杂合形成的第一个报道。讨论了潜在的综合应用以及该技术的问题。

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  • 来源
    《Protoplasma》 |2007年第2期|89-97|共9页
  • 作者

    Fan Liu; U. Ryschka; F. Marthe; E. Klocke; G. Schumann; H. Zhao;

  • 作者单位

    Institute of Horticultural Crops Federal Centre for Breeding Research on Cultivated Plants Quedlinburg;

    Institute of Horticultural Crops Federal Centre for Breeding Research on Cultivated Plants Quedlinburg;

    Institute of Horticultural Crops Federal Centre for Breeding Research on Cultivated Plants Quedlinburg;

    Institute of Horticultural Crops Federal Centre for Breeding Research on Cultivated Plants Quedlinburg;

    Institute of Horticultural Crops Federal Centre for Breeding Research on Cultivated Plants Quedlinburg;

    Beijing Vegetable Research Center Beijing;

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    Keywords: Pollen protoplast; Haploid mesophyll protoplast; Fusion; Hybrid callus; Brassica oleracea; Brassica rapa.;

    机译：关键词：花粉原生质体;单倍体叶肉原生质体;融合杂交愈伤组织;甘蓝甘蓝型油菜。;

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