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首页> 外文会议>Annual Meeting of The Florida^State^Horticultural^Society >Yield and Transformation Ability of Citrus Protoplasts Derived from Either Cell Suspension Cultures or Embryogenic Callus
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Yield and Transformation Ability of Citrus Protoplasts Derived from Either Cell Suspension Cultures or Embryogenic Callus

机译：柑橘原生质体的产量和转化能力衍生自细胞悬浮培养或胚性愈伤组织

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Protoplasts are plant cells that have had their cell walls enzymatically removed. Protoplast transformation incorporates recombinant DNA directly into the genome to produce genetically modified plants, circumventing the traditional Agrobacterium-mediated transformation. Citrus cell suspension cultures derived from totipotent embryogenic callus cells and grown in a liquid medium are commonly utilized as source explants for the isolation of citrus protoplasts. However, these rapidly dividing cells areprone to somaclonal variation and point mutations. The objective of this study was to compare yield and transformation efficiency of citrus protoplasts obtained from embryogenic cultures that were maintained on solid medium as callus cultures or in liquid medium as suspension cultures. Microscopic comparison of cells between the two cell culture types was also conducted. Four citrus cultivars were evaluated in this study: two sweet orange (‘EVI’ and ‘Cara Cara’), one tangor (‘W Murcott’), and onegrapefruit (‘Ray Ruby’). Protoplast yield was similar for cell suspension derived protoplasts subcultured on a 14-day interval and protoplasts derived from callus cultures subcultured on a bimonthly interval. The transient transformation rate was alsonot affected by either cell type or time of harvest in any cultivar. The protoplast transformation rate was not affected by the culture type. We conclude that embryogenic callus maintained on solid medium can be a viable alternative to suspension cultures for protoplast isolation and transformation.
机译：原生质体是植物细胞，其酶促除去细胞壁。原生质体转化将重组DNA直接掺入基因组中以产生遗传修饰的植物，规避传统的农杆菌介导的转化。衍生自Totipotent胚性愈伤细胞并在液体培养基中生长的柑橘悬浮培养物通常用作分离柑橘原生质体的源外植物。然而，这些迅速分开的细胞被糖瘤变异和点突变被突出。本研究的目的是比较从胚胎培养物中获得的柑橘原生质体的产量和转化效率，该培养物在固体培养基中作为愈伤组织培养物或液体培养基作为悬浮培养物。还进行了两种细胞培养类型之间细胞的微观比较。在这项研究中评估了四种柑橘品种：两个甜橙（'evi'和'Cara Cara'），一个橘子（'W murcott'）和Onegrapefruit（'ray Ruby'）。原生质体产率类似于细胞悬浮液衍生的原生质体，其在衍生自培养的愈伤组织培养物的14天间隔和原生质体上培养的原生质体。瞬态转化率是受任何品种的细胞类型或收获时间影响的alsonot。原生质体转化率不受培养类型的影响。我们得出结论，维持在固体介质上的胚胎愈伤组织可以是用于原生质体分离和转化的悬浮培养物的可行替代品。

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