In vitro culture of Orobanche ramosa

Orobanche spp. (broomrapes) are holoparasites that subsist on the roots of many important crops and can considerably reduce yield. The control of Orobanche spp. includes physical, chemical and biological methods. Interactions between parasitic angiosperms and their hosts first occur at the level of parasite seed germination. The seeds of all Orobanchaceae germinate in soil under natural conditions only in response to specific chemical exudates from the host plant. This study describes the influence of different plant growth regulators and host plant root exudates on germination and development of calli from Orobanche seeds in vitro . The effect of indole-3-acetic acid, gibberellic acid and kinetin on the germination of Orobanche seeds varied with concentration. These plant growth regulators also affected the period of germination and the structure of calli and protrusions. An in vitro system for the collection of tobacco root exudates was established. Compounds released from the host roots of three different tobacco cultivars were found to provoke high levels of germination of the Orobanche seeds without any period of pre-conditioning. This study developed methods for the investigation of host–parasite interactions and the effect of germination stimulants in Orobanche spp.     

Changes in Lipid Composition After In-Vitro Selection for Glyphosate Tolerance in Tobacco

After a one-step selection procedure with glyphosate added to the callus medium, tobacco regenerants were obtained from calluses surviving on field doses of the herbicide. The lipid and sterol compositions of potted plants from the two original cultivars which underwent the selection procedure but without glyphosate, and the selected regenerants were investigated. Controls were derived from micropropagation and not from seeds. They served as appropriate controls for comparison with glyphosate regenerants, i.e. they underwent exactly the same experimental conditions except for the glyphosate treatment. Plant regeneration was strongly (at one-step selection) or fully (at stepwise selection) inhibited. Only a few glyphosate-tolerant plants (seven of Nevrokop A₂₄ and five of Zlatna arda) were obtained which were cloned and potted. At least 10 plants of one clone per genotype were used for the further biochemical analyses. The spraying of the tobacco plants from the cultivars with glyphosate led to a decrease of monogalactosyl diacylglycerol content and increase of the saturated fatty acids in all glycolipid classes. The concentration of stigmasterol increased and that of sitosterol decreased only in Zl.arda variety. In the regenerants there were different changes in lipid composition, concerning mainly a decrease of monogalactosyl diacylglycerols (MGDG) and an increase of digalactosyl diacylglycerols (DGDG) in Zl.arda-derived regenerants and increase of the amounts of neutral lipids and decrease of phospholipids (PL) in the Nevrokop A₂₄-derived regenerants. Surprisingly, after treatment of the regenerants with glyphosate, the MGDG amounts in these regenerants were higher than in the controls, which could mean a lesser ability to control ion permeability.