Advertisement

Procedure for Single-Pollen Genotyping

  • Yoshihisa Suyama
Part of the Ecological Research Monographs book series (ECOLOGICAL)

Abstract

Direct determination of multilocus genotypes and DNA sequences from single pollen grains is an efficient method and can be used to answer questions in different research areas such as pollination ecology and genomics. The essential prerequisites to perform this technique are the ability (1) to manipulate small objects in small amounts of liquid by using standard pipettes, (2) to determine optimal conditions to minimize the loss of DNA, (3) to identify the optimal conditions for polymerase chain reaction amplification using an extremely low copy template, and (4) to eliminate possible contaminants. Owing to its simple yet efficient approach, the single-pollen genotyping technique has become the technique of choice among molecular ecologists. This technique is expected to be used extensively in various disciplines in the future.

Keywords

Polymerase Chain Reaction Hybrid Zone Individual Pollen Pollination Ecology Polymerase Chain Reaction Tube 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Aziz AN, Sauve RJ (2008) Genetic mapping of Echinacea purpurea via individual pollen DNA fingerprinting. Mol Breed 21:227–232CrossRefGoogle Scholar
  2. Aziz AN, Seabrook JEA, Tai GCC (1999) Amplification of RAPD markers from single pollen grains of diploid (2N=2X=24) potato. Am J Potato Res 76:179–182CrossRefGoogle Scholar
  3. Chen PH, Pan YB, Chen RK (2008) High-throughput procedure for single pollen grain collection and polymerase chain reaction in plants. J Integr Plant Biol 50:375–383CrossRefPubMedGoogle Scholar
  4. Cozzolino S, Schiestl FP, Muller A, De Castro O, Nardella AM, Widmer A (2005) Evidence for pollinator sharing in Mediterranean nectar-mimic orchids: absence of premating barriers? Proc R Soc Lond B Biol Sci 272:1271–1278CrossRefGoogle Scholar
  5. Hasegawa Y, Suyama Y, Seiwa K (2009) Pollen donor composition during the early phases of reproduction revealed by DNA genotyping of pollen grains and seeds of Castanea crenata. New Phytol 182:994–1004CrossRefGoogle Scholar
  6. Honma H, Yokoyama T, Inoue M, Uebayashi A, Matsumoto F, Watanabe Y, Nakai Y (2007) Genetical identification of coccidia in red-crowned crane, Grus japonensis. Parasitol Res 100:637–640CrossRefPubMedGoogle Scholar
  7. Ito M, Suyama Y, Ohsawa WY (2008) Airborne-pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla. Mol Ecol 17:5092–5103CrossRefPubMedGoogle Scholar
  8. Iwamoto S, Tokumasu S, Suyama Y, Kakishima M (2002) Molecular phylogeny of four selected species of the strictly anamorphic genus Thysanophora using nuclear ribosomal DNA sequences. Mycoscience 43:169–180CrossRefGoogle Scholar
  9. Iwamoto S, Tokumasu S, Suyama Y, Kakishima M (2005) Thysanophora penicillioides includes multiple genetically diverged groups that coexist respectively in Abies mariesii forests in Japan. Mycologia 97:1238–1250CrossRefPubMedGoogle Scholar
  10. Matsuki Y, Isagi Y, Suyama Y (2007) The determination of multiple microsatellite genotypes and DNA sequences from a single pollen grain. Mol Ecol Notes 7:194–198CrossRefGoogle Scholar
  11. Matsuki Y, Tateno R, Shibata M, Isagi Y (2008) Pollination efficiencies of flower-visiting insects as determined by direct genetic analysis of pollen origin. Am J Bot 95:925–930CrossRefGoogle Scholar
  12. Matsunaga S, Schutze K, Donnison IS, Grant SR, Kuroiwa T, Kawano S (1999) Single pollen typing combined with laser-mediated manipulation. Plant J 20:371–378CrossRefPubMedGoogle Scholar
  13. Paffetti D, Vettori C, Caramelli D, Vernesi C, Lari M, Paganelli A, Paule L, Giannini R (2007) Unexpected presence of Fagus orientalis complex in Italy as inferred from 45,000-year-old DNA pollen samples from Venice lagoon. BMC Evol Biol 7:S6CrossRefPubMedGoogle Scholar
  14. Parducci L, Suyama Y, Lascoux M, Bennett KD (2005) Ancient DNA from pollen: a genetic record of plant population history. Mol Ecol 14:2873–2882CrossRefPubMedGoogle Scholar
  15. Petersen G, Johansen B, Seberg O (1996) PCR and sequencing from a single pollen grain. Plant Mol Biol 31:189–191CrossRefPubMedGoogle Scholar
  16. Sato K, Suyama Y, Saito M, Sugawara K (2005) A new primer for discrimination of arbuscular mycorrhizal fungi with polymerase chain reaction-denature gradient gel electrophoresis. Grassl Sci 51:179–181CrossRefGoogle Scholar
  17. Suyama Y, Kawamuro K, Kinoshita I, Yoshimura K, Tsumura Y, Takahara H (1996) DNA sequence from a fossil pollen of Abies spp. from Pleistocene peat. Genes Genet Syst 71:145–149CrossRefPubMedGoogle Scholar
  18. Suyama Y, Yoshimaru H, Tsumura Y (2000) Molecular phylogenetic position of Japanese Abies (Pinaceae) based on chloroplast DNA sequences. Mol Phylogenet Evol 16:271–277CrossRefPubMedGoogle Scholar
  19. Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109CrossRefPubMedGoogle Scholar
  20. Widmer A, Cozzolino S, Pellegrino G, Soliva M, Dafni A (2000) Molecular analysis of orchid pollinaria and pollinaria-remains found on insects. Mol Ecol 9:1911–1914CrossRefPubMedGoogle Scholar
  21. Zhou LJ, Pei KQ, Zhou B, Ma KP (2007) A molecular approach to species identification of Chenopodiaceae pollen grains in surface soil. Am J Bot 94:477–481CrossRefGoogle Scholar
  22. Ziegenhagen B, Schauerte M, Kormutak A, Scholz F (1996) Plastid DNA polymorphism of megagametophytes and pollen in two Abies species. Silvae Genet 45:355–358Google Scholar

Copyright information

© Springer 2011

Authors and Affiliations

  1. 1.Laboratory of Forest Ecology, Graduate School of Agricultural ScienceTohoku UniversityOsakiJapan

Personalised recommendations