Arabidopsis Pollen Germination Medium (Li H 1999)

Table of Contents
    Add a header to begin generating the table of contents

    Introduction

    The pollen germination medium (PGM) is used for allowing the pollen to germinate either by in vitro or semi-in vitro methods. This web page deals with the composition and preparation of PGM as mentioned in the article titled "Control of Pollen Tube Tip Growth by a Rop GTPase–Dependent Pathway That Leads to Tip-Localized Calcium Influx" published in the plant cell by Li H1999 et al.

    pH

    The pH of the medium should be adjusted to 7.0 using dilute HCl.

    Composition

    ReagentM.WWorking concentrationStock solutionAdd for 10 ml stockAdd from stock for 1X of 50ml
    Sucrose342.318%--9 g
    H2BO361.80.01%1%0.1 g 0.5 ml
    CaCl2(anhydrous)110.981mM 1M 1.1 g 50 µl
    Ca(NO3)2164.081mM 1M 1.64 g 50 µl
    MgSO4 .7H2O246.471mM1M2.46 g50 µl
    Noble Agar -0.50%--0.25 g

    Reagents

    • Sucrose
    • Boric acid
    • Calcium chloride
    • Calcium nitrate
    • Magnesium sulphate
    • Noble agar
    • Distilled water
    • HCl

    Instruments and other requirements

    • Weighing balance. paper boats.
    • Graduated cylinder and a glass beaker.
    • pH meter. Magnetic stirrer and magnetic bead/pellet.

    Procedure

    Use ddH2O only for making PGM.

    You can choose to make 1X (working concentration) or make stocks and use them to make 1X whenever you want. Making stock solutions eliminate the possibility of errors that may happen while weighing very low quantities of reagents. Making working concentration from stocks also helps you to add some other reagents without further diluting working concentration.

    1. Determine whether you want to start with stock solutions or directly making working concentration.
    2. Weigh the reagents accordingly (Refer to the table above).
    3. Dissolve salts by using a magnetic stirrer or vortex mixer.
    4. Adjust pH to 7.0 using HCl and KOH/NaOH (pH is one of the critical factors)
    5. Make the volume as planned.
    6. Sterilization of media is not required for immediate use. However, doing filter sterilization might increase its shelf life by preventing microbial growth.
    7. Aliquot to smaller volumes if needed (frequent opening and closing in open-air might result in microbial growth).
    8. Noble agar is preferable over agar as a solidifying agent.

    Storage

    Store PGM (Pollen Germination Medium) stock solutions at 4˚C for 1-2 weeks.

    References

    Control of Pollen Tube Tip Growth by a Rop GTPase–Dependent Pathway That Leads to Tip-Localized Calcium Influx

    Leave a Comment