Plant regeneration from transverse stalk sections of chive plants

An efficient system for in vitro regeneration of plants is a prerequisite for plant improvement by using advanced biotechnological techniques. In this work, regeneration of adventitious buds was induced from thin stalk sections (1-2 mm) containing apical shoot meristem of in vitro grown chive (Allium schoenoprasum) plants. Cytokinins 6-furfurylaminopurine (Kin), 6-benzylaminopurine (BA) or thidiazuron (TDZ) at 5 mu M stimulated this process, and Kin was the most efficient among them. Adventitious buds developed further on the same media for an additional 4 weeks and Kin showed the most pronounced effect on shoot and root growth. Thereafter, all acquired shoots were detached and transferred to solid Murashige and Skoog (MS) medium supplemented with 5 mu M Kin. Single shoots developed secondary adventitious shoots and formed bunches. The mean number of adventitious shoots per bunch was the highest in TDZ-induced shoots (33.6), while Kin- and BA-induced shoots formed 11.9 and 11.3 adventitious shoots per bunch, respectively. Kin-induced shoots exhibited the best shoot and root growth. Plants obtained in this study acclimatized readily at high frequency.nul

Plant regeneration from transverse stalk sections of chive plants

An efficient system for in vitro regeneration of plants is a prerequisite for plant improvement by using advanced biotechnological techniques. In this work, regeneration of adventitious buds was induced from thin stalk sections (1-2 mm) containing apical shoot meristem of in vitro grown chive (Allium schoenoprasum) plants. Cytokinins 6-furfurylaminopurine (Kin), 6-benzylaminopurine (BA) or thidiazuron (TDZ) at 5 mu M stimulated this process, and Kin was the most efficient among them. Adventitious buds developed further on the same media for an additional 4 weeks and Kin showed the most pronounced effect on shoot and root growth. Thereafter, all acquired shoots were detached and transferred to solid Murashige and Skoog (MS) medium supplemented with 5 mu M Kin. Single shoots developed secondary adventitious shoots and formed bunches. The mean number of adventitious shoots per bunch was the highest in TDZ-induced shoots (33.6), while Kin- and BA-induced shoots formed 11.9 and 11.3 adventitious shoots per bunch, respectively. Kin-induced shoots exhibited the best shoot and root growth. Plants obtained in this study acclimatized readily at high frequency.nul

Establishment of transgenic Rhazya stricta hairy roots to modulate terpenoid indole alkaloid production

Key message: Transgenic hairy roots of R. stricta were developed for investigation of alkaloid accumulations. The contents of five identified alkaloids, including serpentine as a new compound, increased compared to non-transformed roots .Abstract: Rhazya stricta Decne. is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). In order to study TIA production and enable metabolic engineering, we established hairy root cultures of R. stricta by co-cultivating cotyledon, hypocotyl, leaf, and shoot explants with wild-type Agrobacterium rhizogenes strain LBA 9402 and A. rhizogenes carrying the pK2WG7-gusA binary vector. Hairy roots initiated from the leaf explants 2 to 8 weeks. Transformation was confirmed by polymerase chain reaction and in case of GUS clones with GUS staining assay. Transformation efficiency was 74 and 83 % for wild-type and GUS hairy root clones, respectively. Alkaloid accumulation was monitored by HPLC, and identification was achieved by UPLC-MS analysis. The influence of light (16 h photoperiod versus total darkness) and media composition (modified Gamborg B5 medium versus Woody Plant Medium) on the production of TIAs were investigated. Compared to non-transformed roots, wild-type hairy roots accumulated significantly higher amounts of five alkaloids. GUS hairy roots contained higher amounts two of alkaloids compared to non-transformed roots. Light conditions had a marked effect on the accumulation of five alkaloids whereas the composition of media only affected the accumulation of two alkaloids. By successfully establishing R. stricta hairy root clones, the potential of transgenic hairy root systems in modulating TIA production was confirmed