Pyramiding of alien-derived Wheat streak mosaic virus (WSMV) resistance and resistance enhancing genes in wheat is a cost-effective and environmentally safe strategy for disease control. PCR-based markers and cytogenetic analysis with genomic in situ hybridisation were applied to identify alien chromatin in four genetically diverse populations of wheat (Triticum aestivum) lines incorporating chromosome segments from Thinopyrum intermedium and Secale cereale (rye). Out of 20 experimental lines, 10 carried Th. intermedium chromatin as T4DL*4Ai#2S translocations, while, unexpectedly, 7 lines were positive for alien chromatin (Th. intermedium or rye) on chromosome 1B. The newly described rye 1RS chromatin, transmitted from early in the pedigree, was associated with enhanced WSMV resistance. Under field conditions, the 1RS chromatin alone showed some resistance, while together with the Th. intermedium 4Ai#2S offered superior resistance to that demonstrated by the known resistant cultivar Mace. Most alien wheat lines carry whole chromosome arms, and it is notable that these lines showed intra-arm recombination within the 1BS arm. The translocation breakpoints between 1BS and alien chromatin fell in three categories: (i) at or near to the centromere, (ii) intercalary between markers UL-Thin5 and Xgwm1130 and (iii) towards the telomere between Xgwm0911 and Xbarc194. Labelled genomic Th. intermedium DNA hybridised to the rye 1RS chromatin under high stringency conditions, indicating the presence of shared tandem repeats among the cereals. The novel small alien fragments may explain the difficulty in developing well-adapted lines carrying Wsm1 despite improved tolerance to the virus. The results will facilitate directed chromosome engineering producing agronomically desirable WSMV-resistant germplasm.
WSMV disease scores raw data
Lines were originally scored in 2002 for response to natural infection by WSMV at Hays, KS USA and Sidney, NE USA (Divis et al., 2006). Visual subjective assessment of the degree of chlorosis and plant stunting were used (Table 1 and footnotes). Selected lines were further evaluated in the field at the University of Nebraska Agricultural Research and Development Center, Mead, NE. In 2011 individual spikes from each line were harvested and used to seed at least 20, 1.2 m replicate rows of single plant-derived lines in September 2011 as a block surrounded by early planted wheat that served as a ‘green bridge’ for the development of WCM populations. Multiple plantings of the resistant cultivar Mace and the susceptible line Tomahawk were included as a control
PCR gels of markers in TableS1 and S2_I
First of two files with images of gels used for scoring.
PCR gels of markers in TableS1 and S2_II
Second of two files of gel images used for scoring.
FISH_Experiment
Table with FISH experiments analysed for each line described
FISH_Experiment_Schedules_I
First of two files with slides and probes for FISH experiments
FISH_Experiment_Schedules_II
Second of two files with slides and probes for FISH experiments
FISH experiments of line Millenium
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
Millenium.zip
FISH experiments for line KS95H102
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
KS95H102.zip
FISH experiment for line KS96HW10-1
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
KS96HW10-1.zip
FISH experiments for line N02Y5003
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5003.zip
FISH Experiments for line N02Y5096
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5096.zip
FISH Experiments for line N02Y5018
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorscence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5018.zip
FISH experiments for line N02Y5019
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5019.zip
FISH experiments for N02Y5021
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5021.zip
FISH experiments for line N02Y5025
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorscence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5025.zip
FISH experiments for line N02Y5057
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorscence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5057.zip
FISH experiments for line N02Y5075
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorscence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5075.zip
FISH experiments for line N02Y5078
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5078.zip
FISH experiments for line N02Y5082
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5082.zip
FISH experiments for line N02Y5105
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5105.zip
FISH experiments for line N02Y5106
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5106.zip
FISH experiments for line N02Y5109
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5109.zip
FISH experiments for line N02Y5117 Mace
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5117 Mace.zip
FISH experiments for line N02Y5121
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5121.zip
FISH experiments for line N02Y5149
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5149.zip
FISH experiments for line N02Y5154
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5154.zip
FISH experiments for line N02Y5156
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y5156.zip
FISH experiments for line N02Y5163
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail
N02Y5163.zip
FISH experiments for line N02Y2016
Zip file containing individual tif files of blue (DAPI) green (FITC) and red (Alexa594) fluorescence taken on Zeiss or Nikon epifluorescence microscope. File name give Slide code and probe combination, see FISH experiment file for list and FISH schedule files for more detail.
N02Y2016.zip