Chinese Bulletin of Botany ›› 2014, Vol. 49 ›› Issue (3): 229-245.DOI: 10.3724/SP.J.1259.2014.00229

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Current Methods Used to Isolate and Purify Generative and Sperm Cells from Pollen Grains and Tubes of Flowering Plants

Yunlong Lu1, Liqin Wei2, Shaojun Dai1*, Tai Wang2*   

  1. 1Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field, Ministry of Education, Alkali Soil Natural Environmental Science Center, Northeast Forestry University, Harbin 150040, China

    2Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
  • Received:2014-01-22 Revised:2014-03-06 Online:2014-05-01 Published:2014-06-03
  • Contact: Shaojun Dai, Tai Wang

Abstract: In flowering plants, sperm cell development involves a special postmeiotic process. During the process, each microspore generated by pollen mother-cell meiosis undergoes unsymmetrical mitosis to result in a large vegetative cell and a small generative cell enclosed in the vegetative cell. The vegetative cell exits the cell cycle, and the generative cell undergoes further mitosis to generate 2 sperm cells. Functional genomics studies of generative and sperm cells are essential for understanding the molecular control of fate determination and the function of sperm cells and cell–cell recognition during double fertilization. These studies require isolated and purified generative and sperm cells. Here, we summarize the main methods used to isolate and purify generative and sperm cells from pollen grains and tubes of diverse plant species since the 1970s, discuss the advantages and disadvantages of each method, and analyze factors affecting output, purity, longevity and viability of purified generative and sperm cells.