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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting
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An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting

机译：通过体内嫁接有效促进麻疯树的植物转化和转化植物的繁殖

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摘要

An efficient and reproducible Agrobacterium-mediated in planta transformation was developed in Jatropha curcas. The various factors affecting J. curcas in planta transformation were optimized, including decapitation, Agrobacterium strain, pin-pricking, vacuum infiltration duration and vacuum pressure. Simple vegetative in vivo cleft grafting method was adopted in the multiplication of transformants without the aid of tissue culture. Among the various parameters evaluated, decapitated plants on pin-pricking and vacuum infiltrated at 250 mmHg for 3 min with the Agrobacterium strain EHA 105 harbouring the binary vector pGA 492 was proved to be efficient in all terms with a transformation efficiency of 62.66 %. Transgene integration was evinced by the GUS histochemical analysis, and the GUS positive plants were subjected to grafting. Putatively transformed J. curcas served as "Scion" and the wild type J. curcas plant severed as "Stock". There was no occurrence of graft rejection and the plants were then confirmed by GUS histochemical analysis, polymerase chain reaction (PCR) and Southern hybridization. Genetic stability of the grafted plants was evaluated by using randomly amplified polymorphic DNA (RAPD), marker which showed 100 % genetic stability between mother and grafted plants. Thus, an efficient in planta transformation and grafting based multiplication of J. curcas was established.
机译：在麻疯树中开发了一种高效且可重现的农杆菌介导的植物转化。优化了影响植物转化中麻疯树的各种因素，包括斩首，农杆菌菌株，扎针，真空渗透持续时间和真空压力。在不借助组织培养的情况下，在转化体的繁殖中采用简单的植物体内半裂嫁接方法。在所评估的各种参数中，用针刺的去头植物并用带有二元载体pGA 492的农杆菌菌株EHA 105在250 mmHg的真空下渗入3分钟，在所有方面均证明是有效的，转化效率为62.66％。通过GUS组织化学分析证实了转基因整合，并对GUS阳性植物进行了嫁接。假定转化的麻疯树为“接穗”，野生型麻疯树为“原种”。没有发生移植排斥反应，然后通过GUS组织化学分析，聚合酶链反应（PCR）和Southern杂交确认了植株。使用随机扩增的多态性DNA（RAPD）评估嫁接植物的遗传稳定性，该标记显示出母本和嫁接植物之间100％的遗传稳定性。因此，建立了一种有效的基于植物转化和嫁接的J. curcas繁殖。

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