This study focused on the identification and elimination of endophytic bacterial contaminations during in vitro propagation of European bladdernut (Staphylea pinnata). Axillary shoots were propagated on Murashige and Skoog medium with 20 mg ∙ dm⁻³ FeEDDHA, 5 μM BA and 0.5 μM NAA at 20/18°C (day/night) and a 16-h photoperiod. Clouding by endophytic bacterial colonies was observed where shoots contacted the media. Bacteria were isolated and separated by repeated streaking as two strains. Gram staining revealed that both strains were Gram-negative. The colonies were very precisely identified as Acinetobacter johnsonii, strain ATCC 17909, and Methylobacterium rhodesianum, strain DSM 5687, using VITEK^®2—a rapid bacterial identification system—and the 16S rRNA gene sequencing method. The agar disc-diffusion test proved that both bacterial strains were susceptible to 13 antibiotics (out of 25 tested), derived from the groups of fluoroquinolones, aminoglycosides and tetracyclines. Doxycycline or gentamicin (100–300 mg ∙ dm⁻³) was added to the S. pinnata shoot propagation medium to eliminate bacteria. Gentamicin 100 mg ∙ dm⁻³ showed the best effect, inhibiting the growth of endogenous bacteria (63%) when applied in the medium for 4 weeks. After the following transfer to media without antibiotics, shoots developed axillary buds and bacterial colonies were not observed.

中文翻译：

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Staphylea pinnata L体外培养过程中内生细菌的鉴定和控制。

这项研究的重点是在欧洲elimination（Staphylea pinnata）的体外繁殖过程中鉴定和消除内生细菌污染。在20/18°C（昼/夜）和16小时光照下，在20 mg∙dm ^-3 FeEDDHA，5μMBA和0.5μMNAA的Murashige和Skoog培养基上繁殖腋芽。在芽与培养基接触的地方观察到内生细菌菌落混浊。通过重复划线作为两个菌株，分离并分离细菌。革兰氏染色显示两种菌株均为革兰氏阴性。使用VITEK可以非常准确地将菌落鉴定为约氏不动杆菌（ATCC 17909）和罗得氏甲基杆菌（DSM 5687）。^®2-快速细菌鉴定系统-和16S rRNA基因测序方法。琼脂圆盘扩散试验证明，两种细菌均对13种抗生素敏感（从25种测试中），这些抗生素来自氟喹诺酮类，氨基糖苷类和四环素类。将强力霉素或庆大霉素（100–300 mg∙dm ^-3）添加到pina S. pinnata枝条繁殖培养基中以消除细菌。庆大霉素100 mg∙dm ^-3表现出最好的效果，当在培养基中施用4周时，可抑制内源细菌的生长（63％）。在随后转移至不含抗生素的培养基后，枝条发育出腋芽，未观察到细菌菌落。