High-throughput procedure for single pollen grain collection and polymerase chain reaction in plants

Journal of Integrative Plant Biology
Ping-Hua ChenRu-Kai Chen

Abstract

Single pollen grain polymerase chain reaction (PCR) has succeeded in several species, however only limited numbers of pollen grains were involved due to difficulties in pollen isolation and lysis. This has limited its application in genetic analysis and mapping studies in plants. A high-throughput (HT) procedure for collecting and detecting genetic variation in a large number of individual pollen grains by PCR is reported. The HT procedure involved the collection of individual pollen grains by a pair of special forceps and the lysis of pollen grains in a heated alkali/detergent solution followed by neutralization with a tris-ethylenediamine tetraacetic acid (TE) buffer. These resulting template solutions yielded PCR reactions involving the 5S ribosomal RNA intergenic spacers, randomly amplified polymorphic DNA, and simple sequence repeats markers. Using this procedure, one person with experience could collect and process up to 288 single pollen grain PCR reactions per day. The method worked well on sugarcane, corn, Miscanthus spp., snap bean, sorghum, and tomato. The ability to collect and conduct PCR on individual pollen grains on a large scale offers a new approach to genetic analyses and mapping studies in an easily controll...Continue Reading

References

Jul 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·L ZhangN Arnheim
Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·H LiN Arnheim
Apr 1, 1996·Plant Molecular Biology·G PetersenO Seberg
Nov 26, 1999·The Plant Journal : for Cell and Molecular Biology·S MatsunagaS Kawano
Sep 12, 2000·Plant Physiology·G P CopenhaverD Preuss
Dec 2, 2000·Journal of Clinical Microbiology·W Abu Al-Soud, P Rådström
Apr 11, 2001·Genetica·J P Townsend, D L Hartl
Oct 10, 2002·Electrophoresis·Hanlin Li, Edward S Yeung
Apr 22, 2003·BioTechniques·Zhanguo XinJohn J Burke
Aug 16, 2003·The Journal of Heredity·N Tel-ZurY Mizrahi
Dec 25, 2004·Plant Physiology·Elodie Parre, Anja Geitmann
Jul 21, 2005·Molecular Ecology·L ParducciK D Bennett
Jul 1, 1985·Plant Physiology·F A LoewusJ J McCollum

❮ Previous
Next ❯

Citations

Jan 10, 2016·Science & Justice : Journal of the Forensic Science Society·Cassandra SchieldDavid Gangitano
Mar 13, 2014·Journal of Integrative Plant Biology·Ali Bektaş, Ignacio Chapela
Jun 9, 2016·International Journal of Molecular Sciences·Yongqiang LiuKongming Wu
Mar 9, 2017·PloS One·Rashmi Prava MohantyEstelle Levetin
Feb 14, 2018·International Journal of Molecular Sciences·Hong ChangKongming Wu
Apr 13, 2018·Proteomics·Laura MincarelliIain C Macaulay
Dec 11, 2019·The Plant Journal : for Cell and Molecular Biology·Roven Rommel FuentesSander A Peters
Jun 1, 2017·BMC Genomics·Arpita KonarJeanne Romero-Severson
Dec 12, 2018·Annals of Botany·Corneile MinnaarJeffrey D Karron

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.