Plant tissue culture agar

Manufactured by Neogen

Plant Tissue Culture Agar is a sterile, solidified growth medium used for the in vitro cultivation and propagation of plant cells, tissues, and organs. It provides the necessary nutrients and gelling agent for the successful growth and development of plant cultures.

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Lab products found in correlation

Square dish, by SPL Life Sciences (2 mentions) Ms basal salts, by Duchefa Biochemie (1 mentions)

6 protocols using plant tissue culture agar

Automated Analysis of Rice Seedlings

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After 7 days, seedlings were analyzed. Shoots, coleoptiles, and roots were dissected, put separately on a square dish (245 × 245 × 20 mm; SPL Life Sciences) filled with 0.66% (w/v) Plant Tissue Culture Agar (Neogen), and scanned (in color at 600 dpi), whereafter shoot and coleoptile images were analyzed with the in-house developed program for the automated analysis of rice seedlings, designated Plength. For contrast purposes, a blue background was placed behind the plate and a 2-cm scale bar was included in one of the lower corners as reference. Root parameters were measured manually with ImageJ [56 , 119 ]. The Python source code of Plength can be found in Additional file 15 and a manual for the easy installation of this program’s graphical user interface in Additional file 16.

Vlaminck L., Sang-Aram C., Botterman D., Uy C.J., Harper M.K., Inzé D., Gheysen G, & Depuydt S. (2020). Development of a novel and rapid phenotype-based screening method to assess rice seedling growth. Plant Methods, 16, 139.

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Automated Coleoptile Length Analysis

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For long treatments, t‐CA or c‐CA was added immediately after the transfer of the 2‐day‐old seedlings to 50‐ml tubes, and analysis was done 1–8 days after transfer (DAT). For short treatments, the isomers were added at 3 DAT, and analysis was done 1, 3, 6, 12, or 24 h after the start of the treatment. After several hours/days of growth in 50‐ml tubes, coleoptiles were dissected from the seedlings and cleared using a protocol adjusted from Nelissen et al. (2013 (link)). Coleoptiles were treated with ethanol:acetic acid (3:1) for 2 days before being positioned on a square dish (245 × 245 mm; SPL Life Sciences) filled with 0.66% (w/v) Plant Tissue Culture Agar (Neogen) and scanned (in color at 600 dpi). The obtained coleoptile images were analyzed using Plength, an in‐house developed program for the automated analysis of rice seedling lengths (Vlaminck et al., 2020 (link)). For contrast purposes and as a reference, a black background with a 2‐cm scale bar in one of the lower corners was placed behind the plate with coleoptiles before scans were made.

Vlaminck L., De Rouck B., Desmet S., Van Gerrewey T., Goeminne G., De Smet L., Storme V., Kyndt T., Demeestere K., Gheysen G., Inzé D., Vanholme B, & Depuydt S. (2022). Opposing effects of trans‐ and cis‐cinnamic acid during rice coleoptile elongation. Plant Direct, 6(12), e465.

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Cultivation of Arabidopsis thaliana and Papaver rhoeas

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Arabidopsis thaliana plants were grown as described in^{25 (link)}. Briefly, chlorine gas sterilized seeds were sown out on LRC2 plates (2.15 g.L^-1 MS basal salts (Duchefa Biochemie), 0.1g.L^-1 MES, pH adjusted to 5.7 with KOH, 1.0 % Plant Tissue Culture Agar NEOGEN), and kept at 4 °C for three days before transfer to a growth chamber for vertical growth with continuous light emitted (white fluorescent lamps, intensity 120 μmol.m^{2 (link)}.s^-1), at 22 °C. Seedlings were transferred to Jiffy pots in soil and grown under glasshouse conditions under a 16h light/8h dark regime at 22 °C.
Papaver rhoeas plants raised from seed of known SI genotypes derived from the Suttons 'Shirley Single Mixed' cultivar were used as experimental material for obtaining mRNA for the cloning of P. rhoeas PrPGAP1 gene and for pollen used for the antisense/sense oligonucleotide experiments. P. rhoeas plants were field grown. Fresh, dry pollen was collected from newly opened flowers and stored over silica gel at -20 °C, until needed.

Lin Z., Xie F., Triviño M., Zhao T., Coppens F., Sterck L., Bosch M., Franklin-Tong V.E, & Nowack M.K. (2022). Self-incompatibility requires GPI anchor remodeling by the poppy PGAP1 orthologue HLD1. Current biology : CB, 32(9), 1909-1923.e5.

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Sterilization and Growth of Arabidopsis and Tobacco

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Arabidopsis thaliana Col-0 seeds were surface-sterilized overnight with chlorine gas in a closed container. The chlorine gas was generated by mixing 150 mL NaOCl (12 to 14%) with 8 mL 37% HCl. Sterile seeds were sown on plates containing 0.5× Murashige and Skoog (MS) agar medium (1.5 g L⁻¹ MS basal mixture powder (Duchefa), 10 g L⁻¹ sucrose, 0.5 g L⁻¹ MES monohydrate, 8 g L⁻¹ Plant Tissue Culture Agar (Neogen); pH 5.7 with KOH). After sowing, seeds were stratified for 48 h at 4 °C, whereupon plates were transferred to a growth chamber (21 °C; 16 h light/8 h dark photoperiod). N. tabacum cv. Havana 425 seeds were sown in 13-cm pots containing universal potting soil (Saniflor). Plants were grown under long-day greenhouse conditions (21 °C; 16 h light/8 h dark photoperiod).

Witvrouw K., Kim H., Vanholme R., Goeminne G., Ralph J., Boerjan W, & Vanholme B. (2023). Reassessing the claimed cytokinin-substituting activity of dehydrodiconiferyl alcohol glucoside. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2123301120.

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Arabidopsis Growth Conditions and Mutants

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Unless stated otherwise, Arabidopsis plants were grown under long-day conditions (16 h of light/8 h of darkness, Lumilux Cool White lm, 50 to 70 µmol m -2 s -1 ) at 22°C on half-strength Murashige and Skoog (MS) germination medium (Murashige and Skoog, 1962) , 10 g/L sucrose, 0.5 g/L MES, pH 5.7 and 10 g/L plant tissue culture agar (Neogen). The cka123 mutant (N67786) was acquired from the Nottingham Arabidopsis Stock Centre (NASC). The sog1-7, sog1-101, als3-1, als3-1 sog1-7, atm-2 and atr-2 have been previously described (Nezames et al., 2012; Sjogren et al., 2015) .

Pengliang W., Demulder M., David P., Eekhout T., Yoshiyama K., Nguyen L., Vercauteren I., Eeckhout D., Galle M., Jaeger G.D., Larsen P.B., Audenaert D., Desnos T., Nussaume L., Loris R., & Veylder L.D. (2020). Arabidopsis Casein Kinase 2 triggers Stem Cell Exhaustion under Al Toxicity and Phosphate Deficiency Through activation of the DNA Damage Response pathway.

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Seed Sterilization and Plant Growth

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Plants were grown as described in (22). Briefly, chlorine gas sterilized seeds were sown out on LRC2 plates (2.15g.L -1 MS basal salts Duchefa Biochemie, 0.1g.L -1 MES, pH adjusted to 5.7 with KOH, 1.0% Plant Tissue Culture Agar NEOGEN), and stored in cold room (4 o C) for three days before being moved to a growth chamber for vertical growth with continuous light emitted by white fluorescent lamps (intensity 120 µmol.m 2 .s -1 ), at 22 o C. Seedlings were transferred to Jiffy pots in soil and grown under glasshouse conditions under a 16h light/8h dark regime at 22 o C. Seeds were collected when the plants were completely dry and kept at room temperature (RT) or 4 o C for long-term storage.

Lin Z., Xie F., Triviño M., Zhao T., Coppens F., Sterck L., Bosch M., Franklin‐Tong V.E., & Nowack M.K. (2021). GPI anchor remodeling by the plant PGAP1 ortholog HLD1 is essential for Papaver self-incompatibility.

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