Murashige and skoog ms basal medium

Pisum sativum seeds were decontaminated using successive bleach and 70% ethanol treatments (10 min each, under shaking [60 rpm], 3 times) and then germinated for 4 days at room temperature on 0.8% agar plates. B. ambifaria HSJ1 and isogenic hmqA::pKnock-Cm, and hmqG::pKnock-Cm mutants, previously respectively generated by integrating a 757-bp internal fragment of hmqA and a 525-bp fragment of hmqG from HSJ1 into the suicide vector pKnock-Cm and by selecting single-crossover insertion mutants obtained following mating Escherichia coli SM10 (pKnock-Cm-hmqA or pKnock-Cm-hmqG) with B. ambifaria HSJ1 (36 (link)), were incubated at 30°C overnight in TSB with shaking at 60 rpm. Seeds were exposed by adding 10⁵ bacteria/ml in 5 ml Murashige and Skoog (MS) basal medium (Sigma) (43 (link)). Plants were grown at room temperature for 5 days. Roots were then cut, dried at 52°C overnight, and weighed. A Dunn test was used for statistical analyses. This experiment was repeated twice, with the same conclusions.

The research material was wild-type flax (Linum usitatissimum L. cv. NIKE) obtained from the Flax and Hemp Collection of the Institute of Natural Fibers (Poznań, Poland), and transgenic flax overexpressing beta-ketothiolase, as well as wild-type flax treated exogenously with the 3-HB standard. Flax plants were cultured in in vitro conditions on Murashige and Skoog (MS) basal medium (Sigma-Aldrich, St. Louis, MO, USA), supplemented with 2% sucrose (Chempur, Piekary Śląskie, Poland), pH = 5.8, solidified with 0.8% agar (Sigma-Aldrich, St. Louis, MO, USA) in the phytotron at 16 h light (22 °C), 8 h darkness (16 °C).

Pisum sativum seeds were decontaminated using successive bleach and 70% ethanol treatments (10 min each, under shaking [60 rpm], 3 times) and then germinated for 4 days at room temperature on 0.8% agar plates. B. ambifaria HSJ1 and isogenic hmqA::pKnock-Cm, and hmqG::pKnock-Cm mutants, previously respectively generated by integrating a 757-bp internal fragment of hmqA and a 525-bp fragment of hmqG from HSJ1 into the suicide vector pKnock-Cm and by selecting single-crossover insertion mutants obtained following mating Escherichia coli SM10 (pKnock-Cm-hmqA or pKnock-Cm-hmqG) with B. ambifaria HSJ1 (36 (link)), were incubated at 30°C overnight in TSB with shaking at 60 rpm. Seeds were exposed by adding 10⁵ bacteria/ml in 5 ml Murashige and Skoog (MS) basal medium (Sigma) (43 (link)). Plants were grown at room temperature for 5 days. Roots were then cut, dried at 52°C overnight, and weighed. A Dunn test was used for statistical analyses. This experiment was repeated twice, with the same conclusions.