A rapid PCR protocol for marker assisted detection of heterozygotes in segregating generations involving 1BL/1RS translocation and normal wheat lines

Nadella, K. D., Peake, A. S., Bariana, H. S., Cooper, M., Godwin, I. D. and Carroll, B. J. (2002) A rapid PCR protocol for marker assisted detection of heterozygotes in segregating generations involving 1BL/1RS translocation and normal wheat lines. Australian Journal of Agricultural Research, 53 8: 931-938. doi:10.1071/AR00088


Author Nadella, K. D.
Peake, A. S.
Bariana, H. S.
Cooper, M.
Godwin, I. D.
Carroll, B. J.
Title A rapid PCR protocol for marker assisted detection of heterozygotes in segregating generations involving 1BL/1RS translocation and normal wheat lines
Journal name Australian Journal of Agricultural Research   Check publisher's open access policy
ISSN 0004-9409
Publication date 2002-01-01
Sub-type Article (original research)
DOI 10.1071/AR00088
Volume 53
Issue 8
Start page 931
End page 938
Total pages 8
Place of publication Collingwood, Vic
Publisher CSIRO Publishing
Collection year 2002
Language eng
Subject C1
300203 Plant Improvement (Selection, Breeding and Genetic Engineering)
620101 Wheat
Abstract A rapid and reliable polymerase chain reaction (PCR)-based protocol was developed for detecting zygosity of the 1BL/1RS translocation in hexaploid wheat. The protocol involved a multiplex PCR with 2 pairs of oligonucleotide primers, rye-specific Ris-1 primers, and consensus 5S intergenic spacer (IGS) primers, and digestion of the PCR products with the restriction enzyme, MseI. A small piece of alkali-treated intact leaf tissue is used as a template for the PCR, thereby eliminating the necessity for DNA extraction. The test is simple, highly sensitive, and rapid compared with the other detection systems of 1BS1RS heterozygotes in hexaploid wheat. PCR results were confirmed with AFLP analyses. Diagnostic tests for 1BL/1RS translocation based on Sec-1-specific ELISA, screening for chromosome arm 1RS controlled rust resistance locus Yr9, and the PCR test differed in their ability to detect heterozygotes. The PCR test and rust test detected more heterozygotes than the ELISA test. The PCR test is being used to facilitate S1 family recurrent selection in the Germplasm Enhancement Program of the Australian Northern Wheat Improvement Program. A combination of the PCR zygosity test with other markers currently being implemented in the breeding program makes this test economical for 1BL/1RS characterisation of S1 families.
Keyword Agriculture, Multidisciplinary
Aflp
Elisa
Pcr Zygosity Test
Stripe Rust
Recurrent Selection
Agronomic Performance
Disease Resistance
Hexaploid Wheat
Rye
1b
Organization
Stocks
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Agriculture and Food Sciences
 
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