Studies on the development of transformation in Phalaenopsis.

Transgenic Phalaenopsis植物曾被用一种急性颗粒枪装置捕获过。所有胆脂蛋白促动病毒（CaMV）35 S的促发者，可将尿苷和大米actin基因用β-葡萄糖酶（gus）的transient assay表达为报告器。Bialaphos抗性基因（酒吧）哪种承认对Herbicide Bialaphos的耐受性，被用作一个可选择的标记。Protocorm-like bodies （PLBs） of Phalaenopsis derived from the leaf segment culture were bombarded by gold particles coated with pMSP 38 and pWI-GUS DNA containing the bar gene and the gus gene respectively driven by 35 S promoter。Newly-formed PLBs被选定为包括bialaphos在内的中间体。最后，七种转基因植物对生物产生了抵抗力，其中一种含有并表现出了天然气基因。PCR分析证实了他们的转基因自然和西方分析以及历史化学GUS分析显示了对Bar和Gus基因反应的表达。新PLBs （T_1） ... More 该片段是由转基因PLB （T_0）分部产生的。T_1 PLBs形成T_2 PLBs在该段上的错误片段。从PCR检测到扩增DNA，所有含有棒基因的T_1和T_2基因的植物体。西方分析显示，在所有克隆物种中，将棒基因描述为良好的T_0生成，并利用Phalaenopsis中的转化、钙形成和植物再生，对Richard Shaffer 'Santa Cruz'进行了测试。《Vacin和Went中的PLB段形成的呼叫》（1949年）中的糖霜。Sucrose的最佳浓度为40克·1^<-1>。介质含有200毫升·1^<-1>椰子水对钙形成有影响。用凝胶胶制成的媒体固体是适合于Callus induction，与那些与Agar相比较。Calli形成的PLBs在没有Sucrose的情况下很容易地转移到媒体。在植物中，没有变化是通过躯体植入物再生的。“卡利轰炸了gus基因通过历史化学分析展示了gus基因的表达，并建议卡利对Phalaenopsis转化的可用性。”Less（低）

Transgenic Phalaenopsis plants have been obtained by particle bombardment with a pneumatic particle gun device. All promoters of cauliflower mosaic virus (CaMV) 35S,maize ubiquitin and rice actin genes could be expressed in Phalaenopsis using transient assay of beta-glucuronidase (gus) gene as a reporter. Bialaphos resistance gene (bar) which confers tolerance to herbicide bialaphos, was used as a selectable marker. Protocorm-like bodies (PLBs) of Phalaenopsis derived from the leaf segment culture were bombarded by gold particles coated with pMSP38 and pWI-GUS DNA containing the bar gene and the gus gene respectively driven by 35S promoter. Newly-formed PLBs were selected on the medium including bialaphos. Finally seven transgenic plants resistant to bialaphos were obtained, and one of them contained and expressed gus gene. PCR analysis confirmed their transgenic nature and western analysis and histochemical GUS assay showed the expression of bar and gus gene respectively.New PLBs (T_1 … More ) were formed from the segments obtained by the division of the transgenic PLB (T_0). The bisected segments of T_1 PLBs formed T_2 PLBs on the segments. From the detection of amplified DNA by PCR,all plantlets of the T_1 and T_2 generation contained the bar gene. The western analysis showed that the bar gene expressed in all of the clonal progenies as well T_0 generation.To use the callus of Phalaenopsis for transformation, the callus formation and plant regeneration in Phalaenopsis Richard Shaffer 'Santa Cruz' were examined. PLB segments formed calli in Vacin and Went (1949) medium with sucrose. The optimal concentration of sucrose was 40 g・1^<-1>. Media containing 200 ml・1^<-1> coconut water were effective for the callus formation. The media solidified with gellan gum was suitable for the callus induction as compared with those with agar. The calli formed PLBs easily after transferring to media without sucrose. No variation was observed in the flowering plants regenerated through somatic embryogenesis.The calli bombarded the gus gene showed the expression of the gene through the histochemical assay, suggesting the availability of calli for the transformation of Phalaenopsis. Less