Journal
PLANT BREEDING
Volume 135, Issue 4, Pages 439-445Publisher
WILEY-BLACKWELL
DOI: 10.1111/pbr.12382
Keywords
maize; haploid; diploid; fluorescence; sorting
Funding
- National Science Foundation Partnerships for Innovation: Building Innovation Capacity (PFI:BIC) [1237720]
- USDA's National Institute of Food and Agriculture [IOW04314, IOW01018]
- RF Baker Center for Plant Breeding
- K.J. Frey Chair in Agronomy at Iowa State University
- Div Of Industrial Innovation & Partnersh
- Directorate For Engineering [1237720] Funding Source: National Science Foundation
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A new fluorescence-based method for inbred haploid differentiation in maize kernels was developed by utilizing the R1-nj colour marker in combination with fluorescence microspectroscopy and imaging. Seven inbred lines with varying R1-nj expression were used in this study. The fluorescence response of the diploid kernels at the embryonic dye spot was shown to simultaneously exhibit lower intensity and occur at a higher wavelength than the fluorescence of the dye-lacking haploid embryos. Intensity and area thresholds were applied to fluorescence images to sort the haploids from mixed sample populations, and sorting efficiencies of greater than 80% were achieved in all seven inbred lines (with values greater than 90% for five lines). The potential for high-throughput sorting when fluorescence imaging is combined with existing technologies for seed handling as well as high sorting efficiency may make fluorescence a viable and promising alternative to current sorting methods for some inbred lines.
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