Pollen Biology: A Laboratory Manual

ISBN-13: 9783540551706 / Angielski / Miękka / 1992 / 119 str.

K. R. Shivanna; N. S. Rangaswamy

Opis

Instructions are given for the collection and storage of pollen, pollen culture, germination, tests for viability, incompatibility and isolation of protoplasts. Principles are explained and step-by-step protocols are complemented by personal notes and precautions.

Spis treści

1 Introduction.- 2 Pollen Collection.- 3 In Vitro Germination Methods.- Controlled Hydration/Prehydration.- Culture Medium.- Fixation of Pollen Cultures.- Scoring of Pollen Cultures.- General Precautions.- Methods.- Exercise 1 Hanging Drop Culture.- Exercise 2 Sitting Drop Culture.- Exercise 3 Suspension Culture.- Exercise 4 Surface Culture.- Exercise 5 Estimation of Pollen Tube Growth by the Alcian Blue Method.- 4 Pollen Germination and Pollen Tube Growth in Vitro.- Exercise 6 Effects of Boron and Calcium on Pollen Germination and Pollen Tube Growth.- Exercise 7 Leaching of Pollen Substances in Vitro.- Exercise 8 Regulation of pH by Pollen Leachate.- Exercise 9 Role of Transcription and Translation in Pollen Germination and Pollen Tube Growth.- 5 Tests for Pollen Viability.- Exercise 10 In Vitro Germination Test.- Exercise 11 Tetrazolium Test.- Exercise 12 Fluorochromatic Reaction (FCR) Test.- Exercise 13 Fruit Set and Seed Set Test.- Exercise 14 In Vivo Germination and Pollen Tube Growth.- 6 Pollen Storage.- Exercise 15 Effects of Temperature and RH on Viability of Stored Pollen.- Exercise 16 Storage of Pollen Grains in Organic Solvents.- 7 Pollen-Pistil Interaction.- Exercise 17 Cytochemical Localization of Esterases on Stigma Surface.- Exercise 18 Aniline Blue Fluorescence Method to Study Pollen Germination and Pollen Tube Growth in the Pistil.- Exercise 19 Multiple Staining for Localizing Pollen Tubes in the Pistil.- Exercise 20 Semi-Vivo Technique to Study Pollen Germination and Pollen Tube Growth.- Exercise 21 Use of DNA Fluorochromes for Studies on Pollen Grain and Pollen Tube Nuclei.- Exercise 22 Controlled Pollinations.- 8 Self-Incompatibility.- Exercise 23 Confirmation of Self-Incompatibility.- Exercise 24 Studies on Heteromorphic Self-Incompatibility.- Exercise 25 Bud Pollination and Delayed Pollination to Overcome Self-Incompatibility.- Exercise 26 In Vitro Placental Pollination and Fertilization.- 9 Isolation of Protoplasts.- Exercise 27 Isolation of Microspore Protoplasts.- Exercise 28 Isolation of Protoplasts from Pollen Grains.- Exercise 29 Isolation of Sperm Cells (Male Gametes).- Exercise 30 Isolation of Embryo Sacs and Protoplasts from Embryo Sacs.- 10 Appendices.- Appendix 1 Basic Requirements for Laboratory Exercises.- Appendix 2 Suitable Plant Systems.- Appendix 3 Pollen Morphology.- Appendix 4 Pollen Counts.- Appendix 5 Methods to Obtain Constant Relative Humidity.- Appendix 6 Principles of Fluorescence Microscopy.- Appendix 7 Commonly Used Fluorochromes, and Their Excitation and Fluorescence Ranges.- Appendix 8 Filter Combinations Commonly Used for Different Excitation Ranges.- Appendix 9 Basic Statistical Analyses.- References.