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Atropa belladonna Species Overview

Experiment: MPGR

Sample Name HelpReplicates
Callus - None3
Flower - Buds3
Flower - Mature (Fully Expanded)3
Fruit - Immature, without seed3
Fruit - Ripe, without seed3
Leaf - Other3
Root - Lateral Roots3
Root - Primary (Tap)3
Seed - Mature3
Stem - Primary Stem, Intermediate (25-75% from soil to crown)3
Sterile Seedling - None3

Platform: Atropa LC/TOF MS Positive Ion

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Description
Seeds of A. belladonna were purchased from Horizon Herbs (http://www.horizonherbs.com/). Seeds were surface sterilized with 50 % bleach solution for 5 minutes followed by extensive washing with sterile water and were then pretreated with GA3 (0.1 % w/v) for 24 h prior to germination on moistened filter paper. Following radicle emergence, seedlings were transplanted into Jiffy Peat Pellets and grown at 22°C under 16 h / 8 h day / night cycle at 120 µmol until the emergence of the sixth true leaf after which they were transplanted into C900 pots (Nursery Supplies Incorporated) and grown in Baccto® High Porosity Professional Planting Mix (Michigan Peat Company, Houston TX) supplemented with 125 ppm 14-3-14-7Ca-1Mg fertilizer (Greencare Fertilizer Inc, Kankakee, IL) at 20°C in greenhouses at Michigan State University, East Lansing, MI. Sterile seedlings were grown for fourteen days on half strength Murashige and Skoog (MS) modified basal medium with Gamborg vitamins pH 5.7, containing 1.5% w/v sucrose and 0.6 % w/v agar at 23°C under an 18 h / 6 h day light regime using a 50 µmol fluorescent light source. Callus was induced from hypocotyls of sterile seedlings on TCIM2 media (MS modified basal medium with Gamborg vitamins pH 5.7, 1.5% w/v sucrose, 1.0 mg/L 6-benzylaminopurine, 1.0 mg/L naphthalene acetic acid and 0.6 % w/v agar). Plates were incubated as described for growth of sterile seedlings and callus was harvested after 4 weeks.
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