The dried buds of Lonicera hypoglauca Miq. have antipyretic, antidotal and anti-inflammatory properties and as Flos lonicerae are widely used in Chinese patent medicine. Because of its medicinal value, there is a need to establish its transformation system in order to study the biosynthesis pathway of its active substances. The phosphomannose isomerase (PMI) gene is probably the most effective selection maker for the transformation system of plants. To establish PMI selection system for transformation of L. hypoglauca, explants were cultured on the induction medium (MS + 0.1 mg/L NAA + 1.0 mg/L 6-BA + 6 g/L agar) supplemented with different ratios of mannose and sucrose, and data on callus formation were collected. After the concentration of mannose and sucrose was determined, 172 leaf explants, 61 leafstalks and 53 stem segment explants were infected with Agrobactrium containing pCAMBIA1301-PMI. The putatively transformed calluses induced from these explants were detected by PCR, RT-PCR and chlorophenol red assay (CPR). We found that callus formation was effectively suppressed on the induction medium supplemented with mannose. The appropriate concentration of mannose and sucrose in the selection medium was 8 g/L and 22 g/L, under which the transformation frequency of leaf from L. hypoglauca was 15.96%. CPR assay for callus induced from leafstalk and stem have a moderate false negative rate when RT-PCR used as criterion. Taken together, it was concluded that PMI/mannose selection system could be used in the transformation of L. hypoglauca, and CPR assay might be suited for the preliminary screening of the transformation of L. hypoglauca.

中文翻译：

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磷酸甘露糖异构酶/甘露糖选择系统在土壤杆菌介导的农杆菌介导的转化中的应用。

的干燥芽忍冬hypoglauca木麻黄。具有解热，解毒和抗炎特性，金银花被广泛应用于中成药中。由于其药用价值，需要建立其转化体系以研究其活性物质的生物合成途径。磷酸甘露糖异构酶（PMI）基因可能是植物转化系统最有效的选择工具。要建立PMI选择系统的改造L. hypoglauca，在补充有不同比例的甘露糖和蔗糖的诱导培养基（MS + 0.1 mg / L NAA + 1.0 mg / L 6-BA + 6 g / L琼脂）上培养外植体，并收集愈伤组织形成的数据。测定甘露糖和蔗糖的浓度后，用含pCAMBIA1301-PMI的土壤杆菌感染了172个叶外植体，61个叶柄和53个茎节外植体。通过PCR，RT-PCR和氯酚红测定法（CPR）检测从这些外植体诱导出的假定转化的老茧。我们发现，在添加甘露糖的诱导培养基上，愈伤组织的形成得到了有效抑制。选择培养基中甘露糖和蔗糖的适宜浓度分别为8 g / L和22 g / L，在此条件下，次要叶的转化频率为是15.96％。当以RT-PCR为标准时，从茎和茎诱导的愈伤组织的CPR测定具有中等的假阴性率。综上所述，可以得出结论，PMI /甘露糖选择系统可以用于L.hypoglauca的转化，而CPR测定法可能适合初步筛选L.glaucauca的转化。